Information
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Patent Application
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20040030110
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Publication Number
20040030110
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Date Filed
April 02, 200222 years ago
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Date Published
February 12, 200420 years ago
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CPC
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US Classifications
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International Classifications
Abstract
The present invention provides novel isolated polynucleotides and small molecule target polypeptides encoded by the polynucleotides. Antibodies that immunospecifically bind to a novel small molecule target polypeptide or any derivative, variant, mutant or fragment of that polypeptide, polynucleotide or antibody are disclosed, as are methods in which the small molecule target polypeptide, polynucleotide and antibody are utilized in the detection and treatment of a broad range of pathological states. More specifically, the present invention discloses methods of using recombinantly expressed and/or endogenously expressed proteins in various screening procedures for the purpose of identifying therapeutic antibodies and therapeutic small molecules associated with diseases.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to novel polypeptides that are targets of small molecule drugs and that have properties related to stimulation of biochemical or physiological responses in a cell, a tissue, an organ or an organism. More particularly, the novel polypeptides are gene products of novel genes, or are specified biologically active fragments or derivatives thereof. Methods of use encompass diagnostic and prognostic assay procedures as well as methods of treating diverse pathological conditions.
BACKGROUND
[0002] Eukaryotic cells are characterized by biochemical and physiological processes which under normal conditions are exquisitely balanced to achieve the preservation and propagation of the cells. When such cells are components of multicellular organisms such as vertebrates, or more particularly organisms such as mammals, the regulation of the biochemical and physiological processes involves intricate signaling pathways. Frequently, such signaling pathways are constituted of extracellular signaling proteins, cellular receptors that bind the signaling proteins and signal transducing components located within the cells.
[0003] Signaling proteins may be classified as endocrine effectors, paracrine effectors or autocrine effectors. Endocrine effectors are signaling molecules secreted by a given organ into the circulatory system, which are then transported to a distant target organ or tissue. The target cells include the receptors for the endocrine effector, and when the endocrine effector binds, a signaling cascade is induced. Paracrine effectors involve secreting cells and receptor cells in close proximity to each other, for example two different classes of cells in the same tissue or organ. One class of cells secretes the paracrine effector, which then reaches the second class of cells, for example by diffusion through the extracellular fluid. The second class of cells contains the receptors for the paracrine effector; binding of the effector results in induction of the signaling cascade that elicits the corresponding biochemical or physiological effect. Autocrine effectors are highly analogous to paracrine effectors, except that the same cell type that secretes the autocrine effector also contains the receptor. Thus the autocrine effector binds to receptors on the same cell, or on identical neighboring cells. The binding process then elicits the characteristic biochemical or physiological effect.
[0004] Signaling processes may elicit a variety of effects on cells and tissues including by way of nonlimiting example induction of cell or tissue proliferation, suppression of growth or proliferation, induction of differentiation or maturation of a cell or tissue, and suppression of differentiation or maturation of a cell or tissue.
[0005] Many pathological conditions involve dysregulation of expression of important effector proteins. In certain classes of pathologies the dysregulation is manifested as diminished or suppressed level of synthesis and secretion protein effectors. In a clinical setting a subject may be suspected of suffering from a condition brought on by diminished or suppressed levels of a protein effector of interest. Therefore there is a need to be able to assay for the level of the protein effector of interest in a biological sample from such a subject, and to compare the level with that characteristic of a nonpathological condition. There further is a need to provide the protein effector as a product of manufacture. Administration of the effector to a subject in need thereof is useful in treatment of the pathological condition, or the protein effector deficiency or suppression may be favorably acted upon by the administration of another small molecule drug product. Accordingly, there is a need for a method of treatment of a pathological condition brought on by a diminished or suppressed levels of the protein effector of interest.
[0006] Small molecule targets have been implicated in various disease states or pathologies. These targets may be proteins, and particularly enzymatic proteins, which are acted upon by small molecule drugs for the purpose of altering target function and achieving a desired result. Cellular, animal and clinical studies can be performed to elucidate the genetic contribution to the etiology and pathogenesis of conditions in which small molecule targets are implicated in a variety of physiologic, pharmacologic or native states. These studies utilize the core technologies at CuraGen Corporation to look at differential gene expression, protein-protein interactions, large-scale sequencing of expressed genes and the association of genetic variations such as, but not limited to, single nucleotide polymorphisms (SNPs) or splice variants in and between biological samples from experimental and control groups. The goal of such studies is to identify potential avenues for therapeutic intervention in order to prevent, treat the consequences or cure the conditions.
[0007] In order to treat diseases, pathologies and other abnormal states or conditions in which a mammalian organism has been diagnosed as being, or as being at risk for becoming, other than in a normal state or condition, it is important to identify new therapeutic agents. Such a procedure includes at least the steps of identifying a target component within an affected tissue or organ, and identifying a candidate therapeutic agent that modulates the functional attributes of the target. The target component may be any biological macromolecule implicated in the disease or pathology. Commonly the target is a polypeptide or protein with specific functional attributes. Other classes of macromolecule may be a nucleic acid, a polysaccharide, a lipid such as a complex lipid or a glycolipid; in addition a target may be a sub-cellular structure or extra-cellular structure that is comprised of more than one of these classes of macromolecule. Once such a target has been identified, it may be employed in a screening assay in order to identify favorable candidate therapeutic agents from among a large population of substances or compounds.
[0008] In many cases the objective of such screening assays is to identify small molecule candidates; this is commonly approached by the use of combinatorial methodologies to develop the population of substances to be tested. The implementation of high throughput screening methodologies is advantageous when working with large, combinatorial libraries of compounds.
SUMMARY OF THE INVENTION
[0009] The invention is based in part upon the discovery of nucleic acid sequences encoding novel polypeptides. These nucleic acids and polypeptides, as well as derivatives, homologs, analogs and fragments thereof, will hereinafter be collectively designated as “NOVX” nucleic acid, which represents the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or polypeptide sequences, which represents the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106.
[0010] In one aspect, the invention provides an isolated polypeptide comprising a mature form of a NOVX amino acid. One example is a variant of a mature form of a NOVX amino acid sequence, wherein any amino acid in the mature form is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence of the mature form are so changed. The amino acid can be, for example, a NOVX amino acid sequence or a variant of a NOVX amino acid sequence, wherein any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence are so changed. The invention also includes fragments of any of these. In another aspect, the invention also includes an isolated nucleic acid that encodes a NOVX polypeptide, or a fragment, homolog, analog or derivative thereof.
[0011] Also included in the invention is a NOVX polypeptide that is a naturally occurring allelic variant of a NOVX sequence. In one embodiment, the allelic variant includes an amino acid sequence that is the translation of a nucleic acid sequence differing by a single nucleotide from a NOVX nucleic acid sequence. In another embodiment, the NOVX polypeptide is a variant polypeptide described therein, wherein any amino acid specified in the chosen sequence is changed to provide a conservative substitution. In one embodiment, the invention discloses a method for determining the presence or amount of the NOVX polypeptide in a sample. The method involves the steps of: providing a sample; introducing the sample to an antibody that binds immunospecifically to the polypeptide; and determining the presence or amount of antibody bound to the NOVX polypeptide, thereby determining the presence or amount of the NOVX polypeptide in the sample. In another embodiment, the invention provides a method for determining the presence of or predisposition to a disease associated with altered levels of a NOVX polypeptide in a mammalian subject. This method involves the steps of: measuring the level of expression of the polypeptide in a sample from the first mammalian subject; and comparing the amount of the polypeptide in the sample of the first step to the amount of the polypeptide present in a control sample from a second mammalian subject known not to have, or not to be predisposed to, the disease, wherein an alteration in the expression level of the polypeptide in the first subject as compared to the control sample indicates the presence of or predisposition to the disease.
[0012] In a further embodiment, the invention includes a method of identifying an agent that binds to a NOVX polypeptide. This method involves the steps of: introducing the polypeptide to the agent; and determining whether the agent binds to the polypeptide. In various embodiments, the agent is a cellular receptor or a downstream effector.
[0013] In another aspect, the invention provides a method for identifying a potential therapeutic agent for use in treatment of a pathology, wherein the pathology is related to aberrant expression or aberrant physiological interactions of a NOVX polypeptide. The method involves the steps of: providing a cell expressing the NOVX polypeptide and having a property or function ascribable to the polypeptide; contacting the cell with a composition comprising a candidate substance; and determining whether the substance alters the property or function ascribable to the polypeptide; whereby, if an alteration observed in the presence of the substance is not observed when the cell is contacted with a composition devoid of the substance, the substance is identified as a potential therapeutic agent. In another aspect, the invention describes a method for screening for a modulator of activity or of latency or predisposition to a pathology associated with the NOVX polypeptide. This method involves the following steps: administering a test compound to a test animal at increased risk for a pathology associated with the NOVX polypeptide, wherein the test animal recombinantly expresses the NOVX polypeptide. This method involves the steps of measuring the activity of the NOVX polypeptide in the test animal after administering the compound of step; and comparing the activity of the protein in the test animal with the activity of the NOVX polypeptide in a control animal not administered the polypeptide, wherein a change in the activity of the NOVX polypeptide in the test animal relative to the control animal indicates the test compound is a modulator of latency of, or predisposition to, a pathology associated with the NOVX polypeptide. In one embodiment, the test animal is a recombinant test animal that expresses a test protein transgene or expresses the transgene under the control of a promoter at an increased level relative to a wild-type test animal, and wherein the promoter is not the native gene promoter of the transgene. In another aspect, the invention includes a method for modulating the activity of the NOVX polypeptide, the method comprising introducing a cell sample expressing the NOVX polypeptide with a compound that binds to the polypeptide in an amount sufficient to modulate the activity of the polypeptide.
[0014] The invention also includes an isolated nucleic acid that encodes a NOVX polypeptide, or a fragment, homolog, analog or derivative thereof. In a preferred embodiment, the nucleic acid molecule comprises the nucleotide sequence of a naturally occurring allelic nucleic acid variant. In another embodiment, the nucleic acid encodes a variant polypeptide, wherein the variant polypeptide has the polypeptide sequence of a naturally occurring polypeptide variant. In another embodiment, the nucleic acid molecule differs by a single nucleotide from a NOVX nucleic acid sequence. In one embodiment, the NOVX nucleic acid molecule hybridizes under stringent conditions to the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or a complement of the nucleotide sequence. In another aspect, the invention provides a vector or a cell expressing a NOVX nucleotide sequence.
[0015] In one embodiment, the invention discloses a method for modulating the activity of a NOVX polypeptide. The method includes the steps of: introducing a cell sample expressing the NOVX polypeptide with a compound that binds to the polypeptide in an amount sufficient to modulate the activity of the polypeptide. In another embodiment, the invention includes an isolated NOVX nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising a NOVX amino acid sequence or a variant of a mature form of the NOVX amino acid sequence, wherein any amino acid in the mature form of the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence of the mature form are so changed. In another embodiment, the invention includes an amino acid sequence that is a variant of the NOVX amino acid sequence, in which any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence are so changed.
[0016] In one embodiment, the invention discloses a NOVX nucleic acid fragment encoding at least a portion of a NOVX polypeptide or any variant of the polypeptide, wherein any amino acid of the chosen sequence is changed to a different amino acid, provided that no more than 10% of the amino acid residues in the sequence are so changed. In another embodiment, the invention includes the complement of any of the NOVX nucleic acid molecules or a naturally occurring allelic nucleic acid variant. In another embodiment, the invention discloses a NOVX nucleic acid molecule that encodes a variant polypeptide, wherein the variant polypeptide has the polypeptide sequence of a naturally occurring polypeptide variant. In another embodiment, the invention discloses a NOVX nucleic acid, wherein the nucleic acid molecule differs by a single nucleotide from a NOVX nucleic acid sequence.
[0017] In another aspect, the invention includes a NOVX nucleic acid, wherein one or more nucleotides in the NOVX nucleotide sequence is changed to a different nucleotide provided that no more than 15% of the nucleotides are so changed. In one embodiment, the invention discloses a nucleic acid fragment of the NOVX nucleotide sequence and a nucleic acid fragment wherein one or more nucleotides in the NOVX nucleotide sequence is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% of the nucleotides are so changed. In another embodiment, the invention includes a nucleic acid molecule wherein the nucleic acid molecule hybridizes under stringent conditions to a NOVX nucleotide sequence or a complement of the NOVX nucleotide sequence. In one embodiment, the invention includes a nucleic acid molecule, wherein the sequence is changed such that no more than 15% of the nucleotides in the coding sequence differ from the NOVX nucleotide sequence or a fragment thereof.
[0018] In a further aspect, the invention includes a method for determining the presence or amount of the NOVX nucleic acid in a sample. The method involves the steps of: providing the sample; introducing the sample to a probe that binds to the nucleic acid molecule; and determining the presence or amount of the probe bound to the NOVX nucleic acid molecule, thereby determining the presence or amount of the NOVX nucleic acid molecule in the sample. In one embodiment, the presence or amount of the nucleic acid molecule is used as a marker for cell or tissue type.
[0019] In another aspect, the invention discloses a method for determining the presence of or predisposition to a disease associated with altered levels of the NOVX nucleic acid molecule of in a first mammalian subject. The method involves the steps of: measuring the amount of NOVX nucleic acid in a sample from the first mammalian subject; and comparing the amount of the nucleic acid in the sample of step (a) to the amount of NOVX nucleic acid present in a control sample from a second mammalian subject known not to have or not be predisposed to, the disease; wherein an alteration in the level of the nucleic acid in the first subject as compared to the control sample indicates the presence of or predisposition to the disease.
[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.
[0021] Other features and advantages of the invention will be apparent from the following detailed description and claims.
DETAILED DESCRIPTION OF THE INVENTION
[0022] The present invention provides novel nucleotides and polypeptides encoded thereby. Included in the invention are the novel nucleic acid sequences, their encoded polypeptides, antibodies, and other related compounds. The sequences are collectively referred to herein as “NOVX nucleic acids” or “NOVX polynucleotides” and the corresponding encoded polypeptides are referred to as “NOVX polypeptides” or “NOVX proteins.” Unless indicated otherwise, “NOVX” is meant to refer to any of the novel sequences disclosed herein. Table 1 provides a summary of the NOVX nucleic acids and their encoded polypeptides.
1TABLE 1
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Sequences and Corresponding SEQ ID Numbers
SEQ ID
NO
NOVXInternal(nucleic)SEQ ID NO
AssignmentIdentificationacid)(polypeptide)Homology
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1aCG55912-0112CACNG4
2aCG55918-0134Zinc Transporter
2bCG55918_0256Zinc Transporter
3aCG56641-0278Thrombospondin
4aCG56832-0l910Guanylate Kinase
4bCG56832-021112Guanylate Kinase
4cCG56832-031314Guanylate Kinase
5aCG58618-011516Catechol O-Methyltransferase
5bCG58618-021718Catechol O-Methyltransferase
6aCG59580-011920GPCR
7aCG59611-012122GPCR
8aCG59617-012324GPCR
9aCG59826-012526Transformation Sensitive Protein
IEF SSP 3521
10aCG59839-012728Cation Transporting ATPase
11aCG59847-012930Intracellular Protein
12aCG59905-013132Sushi Containing Membrane
Protein
13aCG59930-013334Leucine Rich Repeat
13bCG59930-023536Leucine Rich Repeat
14aCG59934-013738Leucine Rich Repeat
14bCG59934-023940Leucine Rich Repeat
15aCG88565-014142UDP N-Acetylglucosamine
Transporter
16aCG88623-014344Potassium Channel Alpha
Subunit
17aCG88645-014546Cardiac Potassium Channel
Subunit(KV6.2)
18aCG88738-014748Synaptotagmin Interacting
Protein STIP1
19aCG88902-014950UDP Glucuronosyltransferase
19bCG88902-025152UDP Glucuronosyltransferase
20aCG89048_015354Kelch
20bCG89048_025556Kelch
21aCG89098-015758Cytoplasmic G-box Protein
22aCG89126-015960Cytochrome P-450
23aCG89367_016162db1/cdc24rhoGEF
24aCG89645-016364Mitochondrial Protein
25aCG89677-016566Arylsulfatase B
26aCG90697_016768RIS
26bCG89697_017970RIS
27aCG90001-017172Peptidylprolyl Isomerase A
28aCG90011-017374ATP Specific Succinyl Coa
Synthetase Beta Subunit
Precursor
29aCG90204-017576Semaphorin Cytoplasmic
Domain
30aCG90385-017778Mitogen Activated Kinase
30bCG90385-027980Mitogen Activated Kinase
31aCG90635-018182Nuclear Body-Associated Kinase
2B
32aCG90729-018384Proline Rich Inositol
Polyphosphate 5 Phophotase
33aCG90760-018586Transcription Factor
34aCG90770-0187889530058B02R1K Lysosomal
35aCG91002-018990Steroid Dehydrogenase
35bCG91002_029192Steroid Dehydrogenase
36aCG91298-019394Phosphatidylglycerophosphate
Synthase
37aCG91383-019596Aldehyde Dehydrogenase
38aCG91403-019798Proline Rich Synapse Associated
Protein 2
39aCG91434-0199100Aldehyde Dehydrogenase
40aCG91484-01101102GPCR
41aCG91514-01103104Telokin
42aCG91587-01105106Tripartite Motif Protein TRIM4
Isoform Alpha
43aCG91631_01107108NAG5
44aCG91643_01109110Protein Kinase-ERK1
45aCG91911-01111112Mitogen-Activated Protein
Kinase Kinase Kinase 8
46aCG91931-01113114Rab-like Small GTPases
47aCG91941-01115116Serine/Threonine-Protein Kinase
48aCG91951-01117118Serine/Threonine-Protein Kinase
49aCG92025-01119120Gamma-Glutamyltransferase
49bCG92025-02121122Gamma-Glutamyltransferase
50aCG92078-01123124Yolk Sac Permease-Like YSPL-
1 Form
51aCG92088-01125126UDP-galactose: beta-d-
galactosyl-1,4-glucosylceramide
alpha-1,3-galactosyltransferase
(iGb(3) synthase)
52aCG92142-01127128Glycerol-3-Phosphate
Acyltransferase
53aCG92152_01129130Plasminogen Activator SPA
54aCG92228-01131132Transmembrane Tryptase
54bCG92228-02133134Transmembrane Tryptase
55aCG92425-01135136Retinol Dehydrogenase
55bCG92425-02137138Retinol Dehydrogenase
56aCG92477_01139140Secretin
57aCG92499-01141142Seven Transmembrane Domain
Protein
57bCG92499-02143144Seven Transmembrane Domain
Protein
58aCG92541-01145146Munc13-4
59aCG92662-01147148Prostaglandin-E2 9-Reductase
60aCG92683-01149150C2PA
60bCG92683-02151152C2PA
61aCG92694-01153154Long Chain 2-Ketoacyl-CoA
Thiolase
62aCG92896-01155156Phosphatidylinositol 5-
Phosphate 4-Kinase
62bCG92896-02157158Phosphatidylinositol 5-
Phosphate 4-Kinase
63aCG92987_01159160Annexin VI
64aCG93042_01161162GPCR
65aCG93265-01163164L-Serine Dehydratase
65bCG93265-02165166L-Serine Dehydratase
66aCG93464_01167168Phosphatidylinositol 3-Kinase
66bCG93464_02169170Phosphatidylinositol 3-Kinase
66cCG93464_03171172Phosphatidylinositol 3-Kinase
67aCG93495-01173174MAP Kinase-Activating Death
Domain Protein
68aCG93529-01175176Macrophage ABC Transporter
69aCG93594-01177178Phosphatidylinositol-Specific
Phospholipase C
70aCG93669-01179180Serine/Threonine Kinase NEK3
70bCG93669-02181182Serine/Threonine Kinase NEK3
70cCG93669-03183184Serine/Threonine Kinase NEK3
71aCG93896-01185186NEK
71bCG93896-02187188NEK
72aCG93939-01189190Sodium-And Chloride-
Dependent Transporter NTT4
73aCG94245-01191192Hepatocyte Nuclear Factor 4
74aCG94302-01193194MUNC13-1 (KIAA1032)
75aCG94356-01195196Carboxylesterase
76aCG94421_01197198Kelch-BTB
77aCG94465-01199200Protein Kinase
77bCG94465-02201202Protein Kinase
77cCG94465-03203204Protein Kinase
78aCG94511-01205206Pyruvate Dehydrogenase
79aCG94551-01207208MUNC13-3
80aCG94682_02209210Renal Organic Anion
Transporter 1
81aCG90214_01211212Beta 1,4N-
Acetylgalactosaminyltransferase
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[0023] Table 1 indicates homology of NOVX nucleic acids to known protein families. Thus, the nucleic acids and polypeptides, antibodies and related compounds according to the invention corresponding to a NOVX as identified in column 1 of Table 1 will be useful in therapeutic and diagnostic applications implicated in, for example, pathologies and disorders associated with the known protein families identified in column 5 of Table 1.
[0024] NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and contexts. The various NOVX nucleic acids and polypeptides according to the invention are useful as novel members of the protein families according to the presence of domains and sequence relatedness to previously described proteins. Additionally, NOVX nucleic acids and polypeptides can also be used to identify proteins that are members of the family to which the NOVX polypeptides belong.
[0025] Consistent with other known members of the family of proteins, identified in column 5 of Table 1, the NOVX polypeptides of the present invention show homology to, and contain domains that are characteristic of, other members of such protein families. Details of the sequence relatedness and domain analysis for each NOVX are presented in Example A.
[0026] The NOVX nucleic acids and polypeptides can also be used to screen for molecules, which inhibit or enhance NOVX activity or function. Specifically, the nucleic acids and polypeptides according to the invention may be used as targets for the identification of small molecules that modulate or inhibit diseases associated with the protein families listed in Table 1.
[0027] The NOVX nucleic acids and polypeptides are also useful for detecting specific cell types. Details of the expression analysis for each NOVX are presented in Example C. Accordingly, the NOVX nucleic acids, polypeptides, antibodies and related compounds according to the invention will have diagnostic and therapeutic applications in the detection of a variety of diseases with differential expression in normal vs. diseased tissues, e.g. a variety of cancers.
[0028] Additional utilities for NOVX nucleic acids and polypeptides according to the invention are disclosed herein.
[0029] The present invention is based on the identification of biological macromolecules differentially modulated in a pathologic state, disease, or an abnormal condition or state. Among the pathologies or diseases of present interest include metabolic diseases including those related to endocrinologic disorders, cancers, various tumors and neoplasias, inflammatory disorders, central nervous system disorders, and similar abnormal conditions or states. In very significant embodiments of the present invention, the biological macromolecules implicated in the pathologies and conditions are proteins and polypeptides, and in such cases the present invention is related as well to the nucleic acids that encode them. Methods that may be employed to identify relevant biological macromolecules include any procedures that detect differential expression of nucleic acids encoding proteins and polypeptides associated with the disorder, as well as procedures that detect the respective proteins and polypeptides themselves. Significant methods that have been employed by the present inventors, include GeneCalling® technology and SeqCalling TM technology, disclosed respectively, in U.S. Pat. No. 5,871,697, and in U.S. Ser. No. 09/417,386, filed Oct. 13, 1999, each of which is incorporated herein by reference in its entirety. GeneCalling® is also described in Shimkets, et al., “Gene expression analysis by transcript profiling coupled to a gene database query” Nature Biotechnology 17:198-803 (1999).
[0030] The invention provides polypeptides and nucleotides encoded thereby that have been identified as having novel associations with a disease or pathology, or an abnormal state or condition, in a mammal. The present invention further identifies a set of proteins and polypeptides, including naturally occurring polypeptides, precursor forms or proproteins, or mature forms of the polypeptides or proteins, which are implicated as targets for therapeutic agents in the treatment of various diseases, pathologies, abnormal states and conditions. A target may be employed in any of a variety of screening methodologies in order to identify candidate therapeutic agents which interact with the target and in so doing exert a desired or favorable effect. The candidate therapeutic agent is identified by screening a large collection of substances or compounds in an important embodiment of the invention. Such a collection may comprise a combinatorial library of substances or compounds in which, in at least one subset of substances or compounds, the individual members are related to each other by simple structural variations based on a particular canonical or basic chemical structure. The variations may include, by way of nonlimiting example, changes in length or identity of a basic framework of bonded atoms; changes in number, composition and disposition of ringed structures, bridge structures, alicyclic rings, and aromatic rings; and changes in pendent or substituents atoms or groups that are bonded at particular positions to the basic framework of bonded atoms or to the ringed structures, the bridge structures, the alicyclic structures, or the aromatic structures.
[0031] A polypeptide or protein described herein, and that serves as a target in the screening procedure, includes the product of a naturally occurring polypeptide or precursor form or proprotein. The naturally occurring polypeptide, precursor or proprotein includes, e.g., the full-length gene product, encoded by the corresponding gene. The naturally occurring polypeptide also includes the polypeptide, precursor or proprotein encoded by an open reading frame described herein. A “mature” form of a polypeptide or protein arises as a result of one or more naturally occurring processing steps as they may occur within the cell, including a host cell. The processing steps occur as the gene product arises, e.g., via cleavage of the amino-terminal methionine residue encoded by the initiation codon of an open reading frame, or the proteolytic cleavage of a signal peptide or leader sequence. Thus, a mature form arising from a precursor polypeptide or protein that has residues 1 to N, where residue 1 is the N-terminal methionine, would have residues 2 through N remaining. Alternatively, a mature form arising from a precursor polypeptide or protein having residues 1 to N, in which an amino-terminal signal sequence from residue 1 to residue M is cleaved, includes the residues from residue M+1 to residue N remaining. A “mature” form of a polypeptide or protein may also arise from non-proteolytic post-translational modification. Such non-proteolytic processes include, e.g., glycosylation, myristylation or phosphorylation. In general, a mature polypeptide or protein may result from the operation of only one of these processes, or the combination of any of them.
[0032] As used herein, “identical” residues correspond to those residues in a comparison between two sequences where the equivalent nucleotide base or amino acid residue in an alignment of two sequences is the same residue. Residues are alternatively described as “similar” or “positive” when the comparisons between two sequences in an alignment show that residues in an equivalent position in a comparison are either the same amino acid or a conserved amino acid as defined below.
[0033] As used herein, a “chemical composition” relates to a composition including at least one compound that is either synthesized or extracted from a natural source. A chemical compound may be the product of a defined synthetic procedure. Such a synthesized compound is understood herein to have defined properties in terms of molecular formula, molecular structure relating the association of bonded atoms to each other, physical properties such as chromatographic or spectroscopic characterizations, and the like. A compound extracted from a natural source is advantageously analyzed by chemical and physical methods in order to provide a representation of its defined properties, including its molecular formula, molecular structure relating the association of bonded atoms to each other, physical properties such as chromatographic or spectroscopic characterizations, and the like.
[0034] As used herein, a “candidate therapeutic agent” is a chemical compound that includes at least one substance shown to bind to a target biopolymer. In important embodiments of the invention, the target biopolymer is a protein or polypeptide, a nucleic acid, a polysaccharide or proteoglycan, or a lipid such as a complex lipid. The method of identifying compounds that bind to the target effectively eliminates compounds with little or no binding affinity, thereby increasing the potential that the identified chemical compound may have beneficial therapeutic applications. In cases where the “candidate therapeutic agent” is a mixture of more than one chemical compound, subsequent screening procedures may be carried out to identify the particular substance in the mixture that is the binding compound, and that is to be identified as a candidate therapeutic agent.
[0035] As used herein, a “pharmaceutical agent” is provided by screening a candidate therapeutic agent using models for a disease state or pathology in order to identify a candidate exerting a desired or beneficial therapeutic effect with relation to the disease or pathology. Such a candidate that successfully provides such an effect is termed a pharmaceutical agent herein. Nonlimiting examples of model systems that may be used in such screens include particular cell lines, cultured cells, tissue preparations, whole tissues, organ preparations, intact organs, and nonhuman mammals. Screens employing at least one system, and preferably more than one system, may be employed in order to identify a pharmaceutical agent. Any pharmaceutical agent so identified may be pursued in further investigation using human subjects.
[0036] NOVX Nucleic Acids and Polypeptides
[0037] NOVX Clones
[0038] NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and contexts. The various NOVX nucleic acids and polypeptides according to the invention are useful as novel members of the protein families according to the presence of domains and sequence relatedness to previously described proteins. Additionally, NOVX nucleic acids and polypeptides can also be used to identify proteins that are members of the family to which the NOVX polypeptides belong.
[0039] The NOVX genes and their corresponding encoded proteins are useful for preventing, treating or ameliorating medical conditions, e.g., by protein or gene therapy. Pathological conditions can be diagnosed by determining the amount of the new protein in a sample or by determining the presence of mutations in the new genes. Specific uses are described for each of the NOVX genes, based on the tissues in which they are most highly expressed. Uses include developing products for the diagnosis or treatment of a variety of diseases and disorders.
[0040] The NOVX nucleic acids and proteins of the invention are useful in potential diagnostic and therapeutic applications and as a research tool. These include serving as a specific or selective nucleic acid or protein diagnostic and/or prognostic marker, wherein the presence or amount of the nucleic acid or the protein are to be assessed, as well as potential therapeutic applications such as the following: (i) a protein therapeutic, (ii) a small molecule drug target, (iii) an antibody target (therapeutic, diagnostic, drug targeting/cytotoxic antibody), (iv) a nucleic acid useful in gene therapy (gene delivery/gene ablation), and (v) a composition promoting tissue regeneration in vitro and in vivo (vi) biological defense weapon.
[0041] In one specific embodiment, the invention includes an isolated polypeptide comprising an amino acid sequence selected from the group consisting of: (a) a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106; (b) a variant of a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106, wherein any amino acid in the mature form is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence of the mature form are so changed; (c) an amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106; (d) a variant of the amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 106 wherein any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence are so changed; and (e) a fragment of any of (a) through (d).
[0042] In another specific embodiment, the invention includes an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: (a) a mature form of the amino acid sequence given SEQ ID NO: 2n, wherein n is an integer between 1 and 106; (b) a variant of a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106 wherein any amino acid in the mature form of the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence of the mature form are so changed; (c) the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106; (d) a variant of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106, in which any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence are so changed; (e) a nucleic acid fragment encoding at least a portion of a polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106 or any variant of said polypeptide wherein any amino acid of the chosen sequence is changed to a different amino acid, provided that no more than 10% of the amino acid residues in the sequence are so changed; and (f) the complement of any of said nucleic acid molecules.
[0043] In yet another specific embodiment, the invention includes an isolated nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of: (a) the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106; (b) a nucleotide sequence wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% of the nucleotides are so changed; (c) a nucleic acid fragment of the sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106; and (d) a nucleic acid fragment wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% of the nucleotides are so changed.
[0044] One aspect of the invention pertains to isolated nucleic acid molecules that encode NOVX polypeptides or biologically active portions thereof. Also included in the invention are nucleic acid fragments sufficient for use as hybridization probes to identify NOVX-encoding nucleic acids (e.g., NOVX mRNAs) and fragments for use as PCR primers for the amplification and/or mutation of NOVX nucleic acid molecules. As used herein, the term “nucleic acid molecule” is intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), analogs of the DNA or RNA generated using nucleotide analogs, and derivatives, fragments and homologs thereof. The nucleic acid molecule may be single-stranded or double-stranded, but preferably is comprised double-stranded DNA.
[0045] An NOVX nucleic acid can encode a mature NOVX polypeptide. As used herein, a “mature” form of a polypeptide or protein disclosed in the present invention is the product of a naturally occurring polypeptide or precursor form or proprotein. The naturally occurring polypeptide, precursor or proprotein includes, by way of nonlimiting example, the full-length gene product, encoded by the corresponding gene. Alternatively, it may be defined as the polypeptide, precursor or proprotein encoded by an ORF described herein. The product “mature” form arises, again by way of nonlimiting example, as a result of one or more naturally occurring processing steps as they may take place within the cell, or host cell, in which the gene product arises. Examples of such processing steps leading to a “mature” form of a polypeptide or protein include the cleavage of the N-terminal methionine residue encoded by the initiation codon of an ORF, or the proteolytic cleavage of a signal peptide or leader sequence. Thus a mature form arising from a precursor polypeptide or protein that has residues 1 to N, where residue 1 is the N-terminal methionine, would have residues 2 through N remaining after removal of the N-terminal methionine. Alternatively, a mature form arising from a precursor polypeptide or protein having residues 1 to N, in which an N-terminal signal sequence from residue 1 to residue M is cleaved, would have the residues from residue M+1 to residue N remaining. Further as used herein, a “mature” form of a polypeptide or protein may arise from a step of post-translational modification other than a proteolytic cleavage event. Such additional processes include, by way of non-limiting example, glycosylation, myristoylation or phosphorylation. In general, a mature polypeptide or protein may result from the operation of only one of these processes, or a combination of any of them.
[0046] The term “probes”, as utilized herein, refers to nucleic acid sequences of variable length, preferably between at least about 10 nucleotides (nt), 100 nt, or as many as approximately, e.g., 6,000 nt, depending upon the specific use. Probes are used in the detection of identical, similar, or complementary nucleic acid sequences. Longer length probes are generally obtained from a natural or recombinant source, are highly specific, and much slower to hybridize than shorter-length oligomer probes. Probes may be single- or double-stranded and designed to have specificity in PCR, membrane-based hybridization technologies, or ELISA-like technologies.
[0047] The term “isolated” nucleic acid molecule, as utilized herein, is one, which is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid. Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′- and 3′-termini of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated NOVX nucleic acid molecules can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell/tissue from which the nucleic acid is derived (e.g., brain, heart, liver, spleen, etc.). Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or of chemical precursors or other chemicals when chemically synthesized.
[0048] A nucleic acid molecule of the invention, e.g., a nucleic acid molecule having the nucleotide sequence SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or a complement of this aforementioned nucleotide sequence, can be isolated using standard molecular biology techniques and the sequence information provided herein. Using all or a portion of the nucleic acid sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 as a hybridization probe, NOVX molecules can be isolated using standard hybridization and cloning techniques (e.g., as described in Sambrook, et al., (eds.), MOLECULAR CLONING: A LABORATORY MANUAL 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; and Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, N.Y., 1993.) A nucleic acid of the invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to NOVX nucleotide sequences can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.
[0049] As used herein, the term “oligonucleotide” refers to a series of linked nucleotide residues, which oligonucleotide has a sufficient number of nucleotide bases to be used in a PCR reaction. A short oligonucleotide sequence may be based on, or designed from, a genomic or cDNA sequence and is used to amplify, confirm, or reveal the presence of an identical, similar or complementary DNA or RNA in a particular cell or tissue. Oligonucleotides comprise portions of a nucleic acid sequence having about 10 nt, 50 nt, or 100 nt in length, preferably about 15 nt to 30 nt in length. In one embodiment of the invention, an oligonucleotide comprising a nucleic acid molecule less than 100 nt in length would further comprise at least 6 contiguous nucleotides SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or a complement thereof. Oligonucleotides may be chemically synthesized and may also be used as probes.
[0050] In another embodiment, an isolated nucleic acid molecule of the invention comprises a nucleic acid molecule that is a complement of the nucleotide from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or a portion of this nucleotide sequence (e.g., a fragment that can be used as a probe or primer or a fragment encoding a biologically-active portion of an NOVX polypeptide). A nucleic acid molecule that is complementary to the nucleotide sequence from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 is one that is sufficiently complementary to the nucleotide sequence from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 that it can hydrogen bond with little or no mismatches to the nucleotide sequence from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, thereby forming a stable duplex.
[0051] As used herein, the term “complementary” refers to Watson-Crick or Hoogsteen base pairing between nucleotides units of a nucleic acid molecule, and the term “binding” means the physical or chemical interaction between two polypeptides or compounds or associated polypeptides or compounds or combinations thereof. Binding includes ionic, non-ionic, van der Waals, hydrophobic interactions, and the like. A physical interaction can be either direct or indirect. Indirect interactions may be through or due to the effects of another polypeptide or compound. Direct binding refers to interactions that do not take place through, or due to, the effect of another polypeptide or compound, but instead are without other substantial chemical intermediates.
[0052] Fragments provided herein are defined as sequences of at least 6 (contiguous) nucleic acids or at least 4 (contiguous) amino acids, a length sufficient to allow for specific hybridization in the case of nucleic acids or for specific recognition of an epitope in the case of amino acids, respectively, and are at most some portion less than a full length sequence. Fragments may be derived from any contiguous portion of a nucleic acid or amino acid sequence of choice. Derivatives are nucleic acid sequences or amino acid sequences formed from the native compounds either directly or by modification or partial substitution. Analogs are nucleic acid sequences or amino acid sequences that have a structure similar to, but not identical to, the native compound but differs from it in respect to certain components or side chains. Analogs may be synthetic or from a different evolutionary origin and may have a similar or opposite metabolic activity compared to wild type. Homologs are nucleic acid sequences or amino acid sequences of a particular gene that are derived from different species.
[0053] A full-length NOVX clone is identified as containing an ATG translation start codon and an in-frame stop codon. Any disclosed NOVX nucleotide sequence lacking an ATG start codon therefore encodes a truncated C-terminal fragment of the respective NOVX polypeptide, and requires that the corresponding full-length cDNA extend in the 5′ direction of the disclosed sequence. Any disclosed NOVX nucleotide sequence lacking an in-frame stop codon similarly encodes a truncated N-terminal fragment of the respective NOVX polypeptide, and requires that the corresponding full-length cDNA extend in the 3′ direction of the disclosed sequence.
[0054] Derivatives and analogs may be full length or other than full length, if the derivative or analog contains a modified nucleic acid or amino acid, as described below. Derivatives or analogs of the nucleic acids or proteins of the invention include, but are not limited to, molecules comprising regions that are substantially homologous to the nucleic acids or proteins of the invention, in various embodiments, by at least about 70%, 80%, or 95% identity (with a preferred identity of 80-95%) over a nucleic acid or amino acid sequence of identical size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art, or whose encoding nucleic acid is capable of hybridizing to the complement of a sequence encoding the aforementioned proteins under stringent, moderately stringent, or low stringent conditions. See e.g. Ausubel, et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, N.Y., 1993, and below.
[0055] A “homologous nucleic acid sequence” or “homologous amino acid sequence,” or variations thereof, refer to sequences characterized by a homology at the nucleotide level or amino acid level as discussed above. Homologous nucleotide sequences encode those sequences coding for isoforms of NOVX polypeptides. Isoforms can be expressed in different tissues of the same organism as a result of, for example, alternative splicing of RNA. Alternatively, isoforms can be encoded by different genes. In the invention, homologous nucleotide sequences include nucleotide sequences encoding for an NOVX polypeptide of species other than humans, including, but not limited to: vertebrates, and thus can include, e.g., frog, mouse, rat, rabbit, dog, cat cow, horse, and other organisms. Homologous nucleotide sequences also include, but are not limited to, naturally occurring allelic variations and mutations of the nucleotide sequences set forth herein. A homologous nucleotide sequence does not, however, include the exact nucleotide sequence encoding human NOVX protein. Homologous nucleic acid sequences include those nucleic acid sequences that encode conservative amino acid substitutions (see below) in SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, as well as a polypeptide possessing NOVX biological activity. Various biological activities of the NOVX proteins are described below.
[0056] An NOVX polypeptide is encoded by the open reading frame (“ORF”) of an NOVX nucleic acid. An ORF corresponds to a nucleotide sequence that could potentially be translated into a polypeptide. A stretch of nucleic acids comprising an ORF is uninterrupted by a stop codon. An ORF that represents the coding sequence for a full protein begins with an ATG “start” codon and terminates with one of the three “stop” codons, namely, TAA, TAG, or TGA. For the purposes of this invention, an ORF may be any part of a coding sequence, with or without a start codon, a stop codon, or both. For an ORF to be considered as a good candidate for coding for a bona fide cellular protein, a minimum size requirement is often set, e.g., a stretch of DNA that would encode a protein of 50 amino acids or more.
[0057] The nucleotide sequences determined from the cloning of the human NOVX genes allows for the generation of probes and primers designed for use in identifying and/or cloning NOVX homologues in other cell types, e.g. from other tissues, as well as NOVX homologues from other vertebrates. The probe/primer typically comprises substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 25, 50, 100, 150, 200, 250, 300, 350 or 400 consecutive sense strand nucleotide sequence SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106; or an anti-sense strand nucleotide sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106.
[0058] Probes based on the human NOVX nucleotide sequences can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In various embodiments, the probe further comprises a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a part of a diagnostic test kit for identifying cells or tissues which mis-express an NOVX protein, such as by measuring a level of an NOVX-encoding nucleic acid in a sample of cells from a subject e.g., detecting NOVX mRNA levels or determining whether a genomic NOVX gene has been mutated or deleted.
[0059] “A polypeptide having a biologically-active portion of an NOVX polypeptide” refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of a polypeptide of the invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. A nucleic acid fragment encoding a “biologically-active portion of NOVX” can be prepared by isolating a portion SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, that encodes a polypeptide having an NOVX biological activity (the biological activities of the NOVX proteins are described below), expressing the encoded portion of NOVX protein (e.g., by recombinant expression in vitro) and assessing the activity of the encoded portion of NOVX.
[0060] NOVX Nucleic Acid and Polypeptide Variants
[0061] The invention further encompasses nucleic acid molecules that differ from the nucleotide sequences shown in SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 due to degeneracy of the genetic code and thus encode the same NOVX proteins as that encoded by the nucleotide sequences shown in SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106. In another embodiment, an isolated nucleic acid molecule of the invention has a nucleotide sequence encoding a protein having an amino acid sequence shown in SEQ ID NO: 2n, wherein n is an integer between 1 and 106.
[0062] In addition to the human NOVX nucleotide sequences shown in SEQ ID NO: 2n−1, wherein n is an integer between 1 and 106, it will be appreciated by those skilled in the art that DNA sequence polymorphisms that lead to changes in the amino acid sequences of the NOVX polypeptides may exist within a population (e.g., the human population). Such genetic polymorphism in the NOVX genes may exist among individuals within a population due to natural allelic variation. As used herein, the terms “gene” and “recombinant gene” refer to nucleic acid molecules comprising an open reading frame (ORF) encoding an NOVX protein, preferably a vertebrate NOVX protein. Such natural allelic variations can typically result in 1-5% variance in the nucleotide sequence of the NOVX genes. Any and all such nucleotide variations and resulting amino acid polymorphisms in the NOVX polypeptides, which are the result of natural allelic variation and that do not alter the functional activity of the NOVX polypeptides, are intended to be within the scope of the invention.
[0063] Moreover, nucleic acid molecules encoding NOVX proteins from other species, and thus that have a nucleotide sequence that differs from the human SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 are intended to be within the scope of the invention. Nucleic acid molecules corresponding to natural allelic variants and homologues of the NOVX cDNAs of the invention can be isolated based on their homology to the human NOVX nucleic acids disclosed herein using the human cDNAs, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions.
[0064] Accordingly, in another embodiment, an isolated nucleic acid molecule of the invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 750, 1000, 1500, or 2000 or more nucleotides in length. In yet another embodiment, an isolated nucleic acid molecule of the invention hybridizes to the coding region. As used herein, the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% homologous to each other typically remain hybridized to each other.
[0065] Homologs (i.e., nucleic acids encoding NOVX proteins derived from species other than human) or other related sequences (e.g., paralogs) can be obtained by low, moderate or high stringency hybridization with all or a portion of the particular human sequence as a probe using methods well known in the art for nucleic acid hybridization and cloning.
[0066] As used herein, the phrase “stringent hybridization conditions” refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target sequence hybridize to the target sequence at equilibrium. Since the target sequences are generally present at excess, at Tm, 50% of the probes are occupied at equilibrium. Typically, stringent conditions will be those in which the salt concentration is less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 60° C. for longer probes, primers and oligonucleotides. Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide.
[0067] Stringent conditions are known to those skilled in the art and can be found in Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Preferably, the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other. A non-limiting example of stringent hybridization conditions are hybridization in a high salt buffer comprising 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 65° C., followed by one or more washes in 0.2×SSC, 0.01% BSA at 50° C. An isolated nucleic acid molecule of the invention that hybridizes under stringent conditions to the sequences SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, corresponds to a naturally-occurring nucleic acid molecule. As used herein, a “naturally-occurring” nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).
[0068] In a second embodiment, a nucleic acid sequence that is hybridizable to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or fragments, analogs or derivatives thereof, under conditions of moderate stringency is provided. A non-limiting example of moderate stringency hybridization conditions are hybridization in 6×SSC, 5× Denhardt's solution, 0.5% SDS and 100 mg/ml denatured salmon sperm DNA at 55° C., followed by one or more washes in 1×SSC, 0.1% SDS at 37° C. Other conditions of moderate stringency that may be used are well-known within the art. See, e.g., Ausubel, et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990; GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY.
[0069] In a third embodiment, a nucleic acid that is hybridizable to the nucleic acid molecule comprising the nucleotide sequences SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or fragments, analogs or derivatives thereof, under conditions of low stringency, is provided. A non-limiting example of low stringency hybridization conditions are hybridization in 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 mg/ml denatured salmon sperm DNA, 10% (wt/vol) dextran sulfate at 40° C., followed by one or more washes in 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS at 50C. Other conditions of low stringency that may be used are well known in the art (e.g., as employed for cross-species hybridizations). See, e.g., Ausubel, et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY; Shilo and Weinberg, 1981. Proc Natl Acad Sci USA 78: 6789-6792.
[0070] Conservative Mutations
[0071] In addition to naturally-occurring allelic variants of NOVX sequences that may exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation into the nucleotide sequences SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, thereby leading to changes in the amino acid sequences of the encoded NOVX proteins, without altering the functional ability of said NOVX proteins. For example, nucleotide substitutions leading to amino acid substitutions at “non-essential” amino acid residues can be made in the sequence SEQ ID NO: 2n, wherein n is an integer between 1 and 106. A “non-essential” amino acid residue is a residue that can be altered from the wild-type sequences of the NOVX proteins without altering their biological activity, whereas an “essential” amino acid residue is required for such biological activity. For example, amino acid residues that are conserved among the NOVX proteins of the invention are predicted to be particularly non-amenable to alteration. Amino acids for which conservative substitutions can be made are well-known within the art.
[0072] Another aspect of the invention pertains to nucleic acid molecules encoding NOVX proteins that contain changes in amino acid residues that are not essential for activity. Such NOVX proteins differ in amino acid sequence from SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 yet retain biological activity. In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein comprises an amino acid sequence at least about 45% homologous to the amino acid sequences SEQ ID NO: 2n, wherein n is an integer between 1 and 106. Preferably, the protein encoded by the nucleic acid molecule is at least about 60% homologous to SEQ ID NO: 2n, wherein n is an integer between 1 and 106; more preferably at least about 70% homologous SEQ ID NO: 2n, wherein n is an integer between 1 and 106; still more preferably at least about 80% homologous to SEQ ID NO: 2n, wherein n is an integer between 1 and 106; even more preferably at least about 90% homologous to SEQ ID NO: 2n, wherein n is an integer between 1 and 106; and most preferably at least about 95% homologous to SEQ ID NO: 2n, wherein n is an integer between 1 and 106.
[0073] An isolated nucleic acid molecule encoding an NOVX protein homologous to the protein of SEQ ID NO: 2n, wherein n is an integer between 1 and 106 can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein.
[0074] Mutations can be introduced into SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted, non-essential amino acid residues. A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined within the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted non-essential amino acid residue in the NOVX protein is replaced with another amino acid residue from the same side chain family. Alternatively, in another embodiment, mutations can be introduced randomly along all or part of an NOVX coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for NOVX biological activity to identify mutants that retain activity. Following mutagenesis SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, the encoded protein can be expressed by any recombinant technology known in the art and the activity of the protein can be determined.
[0075] The relatedness of amino acid families may also be determined based on side chain interactions. Substituted amino acids may be fully conserved “strong” residues or fully conserved “weak” residues. The “strong” group of conserved amino acid residues may be any one of the following groups: STA, NEQK, NHQK, NDEQ, QHRK, MILV, MILF, HY, FYW, wherein the single letter amino acid codes are grouped by those amino acids that may be substituted for each other. Likewise, the “weak” group of conserved residues may be any one of the following: CSA, ATV, SAG, STNK, STPA, SGND, SNDEQK, NDEQHK, NEQHRK, HFY, wherein the letters within each group represent the single letter amino acid code.
[0076] In one embodiment, a mutant NOVX protein can be assayed for (i) the ability to form protein:protein interactions with other NOVX proteins, other cell-surface proteins, or biologically-active portions thereof, (ii) complex formation between a mutant NOVX protein and an NOVX ligand; or (iii) the ability of a mutant NOVX protein to bind to an intracellular target protein or biologically-active portion thereof; (e.g. avidin proteins).
[0077] In yet another embodiment, a mutant NOVX protein can be assayed for the ability to regulate a specific biological function (e.g., regulation of insulin release).
[0078] Antisense Nucleic Acids
[0079] Another aspect of the invention pertains to isolated antisense nucleic acid molecules that are hybridizable to or complementary to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or fragments, analogs or derivatives thereof. An “antisense” nucleic acid comprises a nucleotide sequence that is complementary to a “sense” nucleic acid encoding a protein (e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence). In specific aspects, antisense nucleic acid molecules are provided that comprise a sequence complementary to at least about 10, 25, 50, 100, 250 or 500 nucleotides or an entire NOVX coding strand, or to only a portion thereof. Nucleic acid molecules encoding fragments, homologs, derivatives and analogs of an NOVX protein of SEQ ID NO: 2n, wherein n is an integer between 1 and 106, or antisense nucleic acids complementary to an NOVX nucleic acid sequence of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, are additionally provided.
[0080] In one embodiment, an antisense nucleic acid molecule is antisense to a “coding region” of the coding strand of a nucleotide sequence encoding an NOVX protein. The term “coding region” refers to the region of the nucleotide sequence comprising codons which are translated into amino acid residues. In another embodiment, the antisense nucleic acid molecule is antisense to a “noncoding region” of the coding strand of a nucleotide sequence encoding the NOVX protein. The term “noncoding region” refers to 5′ and 3′ sequences which flank the coding region that are not translated into amino acids (i.e., also referred to as 5′ and 3′ untranslated regions).
[0081] Given the coding strand sequences encoding the NOVX protein disclosed herein, antisense nucleic acids of the invention can be designed according to the rules of Watson and Crick or Hoogsteen base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of NOVX mRNA, but more preferably is an oligonucleotide that is antisense to only a portion of the coding or noncoding region of NOVX mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of NOVX mRNA. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides in length. An antisense nucleic acid of the invention can be constructed using chemical synthesis or enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally-occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids (e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used).
[0082] Examples of modified nucleotides that can be used to generate the antisense nucleic acid include: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N-6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described farther in the following subsection).
[0083] The antisense nucleic acid molecules of the invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an NOVX protein to thereby inhibit expression of the protein (e.g., by inhibiting transcription and/or translation). The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. An example of a route of administration of antisense nucleic acid molecules of the invention includes direct injection at a tissue site. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface (e.g., by linking the antisense nucleic acid molecules to peptides or antibodies that bind to cell surface receptors or antigens). The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient nucleic acid molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol III promoter are preferred.
[0084] In yet another embodiment, the antisense nucleic acid molecule of the invention is an α-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other. See, e.g., Gaultier, et al., 1987. Nucl. Acids Res. 15: 6625-6641. The antisense nucleic acid molecule can also comprise a 2′-o-methylribonucleotide (See, e.g., Inoue, et al. 1987. Nucl. Acids Res. 15: 6131-6148) or a chimeric RNA-DNA analogue (See, e.g., Inoue, et al., 1987. FEBS Lett. 215: 327-330.
[0085] Ribozymes and PNA Moieties
[0086] Nucleic acid modifications include, by way of non-limiting example, modified bases, and nucleic acids whose sugar phosphate backbones are modified or derivatized. These modifications are carried out at least in part to enhance the chemical stability of the modified nucleic acid, such that they may be used, for example, as antisense binding nucleic acids in therapeutic applications in a subject.
[0087] In one embodiment, an antisense nucleic acid of the invention is a ribozyme. Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes as described in Haselhoff and Gerlach 1988. Nature 334: 585-591) can be used to catalytically cleave NOVX mRNA transcripts to thereby inhibit translation of NOVX mRNA. A ribozyme having specificity for an NOVX-encoding nucleic acid can be designed based upon the nucleotide sequence of an NOVX cDNA disclosed herein (i.e., SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106). For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an NOVX-encoding mRNA. See, e.g., U.S. Pat. No. 4,987,071 to Cech, et al. and U.S. Pat. No. 5,116,742 to Cech, et al. NOVX mRNA can also be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., (1993) Science 261:1411-1418.
[0088] Alternatively, NOVX gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the NOVX nucleic acid (e.g., the NOVX promoter and/or enhancers) to form triple helical structures that prevent transcription of the NOVX gene in target cells. See, e.g., Helene, 1991. Anticancer Drug Des. 6: 569-84; Helene, et al. 1992. Ann. N.Y. Acad. Sci. 660:27-36; Maher, 1992. Bioassays 14: 807-15.
[0089] In various embodiments, the NOVX nucleic acids can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids. See, e.g., Hyrup, et al., 1996. Bioorg Med Chem 4: 5-23. As used herein, the terms “peptide nucleic acids” or “PNAs” refer to nucleic acid mimics (e.g., DNA mimics) in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup, et al., 1996. supra; Perry-O'Keefe, et al., 1996. Proc. Natl. Acad. Sci. USA 93: 14670-14675.
[0090] PNAs of NOVX can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication. PNAs of NOVX can also be used, for example, in the analysis of single base pair mutations in a gene (e.g., PNA directed PCR clamping; as artificial restriction enzymes when used in combination with other enzymes, e.g., S1 nucleases (See, Hyrup, et al., 1996.supra); or as probes or primers for DNA sequence and hybridization (See, Hyrup, et al., 1996, supra; Perry-O'Keefe, et al., 1996. supra).
[0091] In another embodiment, PNAs of NOVX can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras of NOVX can be generated that may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes (e.g., RNase H and DNA polymerases) to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation (see, Hyrup, et al., 1996. supra). The synthesis of PNA-DNA chimeras can be performed as described in Hyrup, et al., 1996. supra and Finn, et al., 1996. Nucl Acids Res 24: 3357-3363. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry, and modified nucleoside analogs, e.g., 5′-(4-methoxytrityl)amino-5′-deoxy-thymidine phosphoramidite, can be used between the PNA and the 5′ end of DNA. See, e.g., Mag, et al., 1989. Nucl Acid Res 17: 5973-5988. PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5′ PNA segment and a 3′ DNA segment. See, e.g., Finn, et al., 1996. supra. Alternatively, chimeric molecules can be synthesized with a 5′ DNA segment and a 3′ PNA segment. See, e.g., Petersen, et al., 1975. Bioorg. Med. Chem. Lett. 5: 1119-11124.
[0092] In other embodiments, the oligonucleotide may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger, et al., 1989. Proc. Natl. Acad. Sci. U.S.A. 86: 6553-6556; Lemaitre, et al., 1987. Proc. Natl. Acad. Sci. 84: 648-652; PCT Publication No. WO88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO 89/10134). In addition, oligonucleotides can be modified with hybridization triggered cleavage agents (see, e.g., Krol, et al., 1988. BioTechniques 6:958-976) or intercalating agents (see, e.g., Zon, 1988. Pharm. Res. 5: 539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, a hybridization triggered cross-linking agent, a transport agent, a hybridization-triggered cleavage agent, and the like.
[0093] NOVX Polypeptides
[0094] A polypeptide according to the invention includes a polypeptide including the amino acid sequence of NOVX polypeptides whose sequences are provided in SEQ ID NO: 2n, wherein n is an integer between 1 and 106. The invention also includes a mutant or variant protein any of whose residues may be changed from the corresponding residues shown in SEQ ID NO: 2n, wherein n is an integer between 1 and 106 while still encoding a protein that maintains its NOVX activities and physiological functions, or a functional fragment thereof.
[0095] In general, an NOVX variant that preserves NOVX-like function includes any variant in which residues at a particular position in the sequence have been substituted by other amino acids, and further include the possibility of inserting an additional residue or residues between two residues of the parent protein as well as the possibility of deleting one or more residues from the parent sequence. Any amino acid substitution, insertion, or deletion is encompassed by the invention. In favorable circumstances, the substitution is a conservative substitution as defined above.
[0096] One aspect of the invention pertains to isolated NOVX proteins, and biologically-active portions thereof, or derivatives, fragments, analogs or homologs thereof. Also provided are polypeptide fragments suitable for use as immunogens to raise anti-NOVX antibodies. In one embodiment, native NOVX proteins can be isolated from cells or tissue sources by an appropriate purification scheme using standard protein purification techniques. In another embodiment, NOVX proteins are produced by recombinant DNA techniques. Alternative to recombinant expression, an NOVX protein or polypeptide can be synthesized chemically using standard peptide synthesis techniques.
[0097] An “isolated” or “purified” polypeptide or protein or biologically-active portion thereof is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the NOVX protein is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. The language “substantially free of cellular material” includes preparations of NOVX proteins in which the protein is separated from cellular components of the cells from which it is isolated or recombinantly-produced. In one embodiment, the language “substantially free of cellular material” includes preparations of NOVX proteins having less than about 30% (by dry weight) of non-NOVX proteins (also referred to herein as a “contaminating protein”), more preferably less than about 20% of non-NOVX proteins, still more preferably less than about 10% of non-NOVX proteins, and most preferably less than about 5% of non-NOVX proteins. When the NOVX protein or biologically-active portion thereof is recombinantly-produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% of the volume of the NOVX protein preparation.
[0098] The language “substantially free of chemical precursors or other chemicals” includes preparations of NOVX proteins in which the protein is separated from chemical precursors or other chemicals that are involved in the synthesis of the protein. In one embodiment, the language “substantially free of chemical precursors or other chemicals” includes preparations of NOVX proteins having less than about 30% (by dry weight) of chemical precursors or non-NOVX chemicals, more preferably less than about 20% chemical precursors or non-NOVX chemicals, still more preferably less than about 10% chemical precursors or non-NOVX chemicals, and most preferably less than about 5% chemical precursors or non-NOVX chemicals.
[0099] Biologically-active portions of NOVX proteins include peptides comprising amino acid sequences sufficiently homologous to or derived from the amino acid sequences of the NOVX proteins (e.g., the amino acid sequence shown in SEQ ID NO: 2n, wherein n is an integer between 1 and 106) that include fewer amino acids than the full-length NOVX proteins, and exhibit at least one activity of an NOVX protein. Typically, biologically-active portions comprise a domain or motif with at least one activity of the NOVX protein. A biologically-active portion of an NOVX protein can be a polypeptide which is, for example, 10, 25, 50, 100 or more amino acid residues in length. Moreover, other biologically-active portions, in which other regions of the protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the functional activities of a native NOVX protein.
[0100] In an embodiment, the NOVX protein has an amino acid sequence shown SEQ ID NO: 2n, wherein n is an integer between 1 and 106. In other embodiments, the NOVX protein is substantially homologous to SEQ ID NO: 2n, wherein n is an integer between 1 and 106, and retains the functional activity of the protein of SEQ ID NO: 2n, wherein n is an integer between 1 and 106, yet differs in amino acid sequence due to natural allelic variation or mutagenesis, as described in detail, below. Accordingly, in another embodiment, the NOVX protein is a protein that comprises an amino acid sequence at least about 45% homologous to the amino acid sequence SEQ ID NO: 2n, wherein n is an integer between 1 and 106, and retains the functional activity of the NOVX proteins of SEQ ID NO: 2n, wherein n is an integer between 1 and 106.
[0101] Determining Homology Between Two or More Sequences
[0102] To determine the percent homology of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison purposes (e.g. gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are homologous at that position (i.e., as used herein amino acid or nucleic acid “homology” is equivalent to amino acid or nucleic acid “identity”).
[0103] The nucleic acid sequence homology may be determined as the degree of identity between two sequences. The homology may be determined using computer programs known in the art, such as GAP software provided in the GCG program package. See, Needleman and Wunsch, 1970. J Mol Biol 48: 443-453. Using GCG GAP software with the following settings for nucleic acid sequence comparison: GAP creation penalty of 5.0 and GAP extension penalty of 0.3, the coding region of the analogous nucleic acid sequences referred to above exhibits a degree of identity preferably of at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%, with the CDS (encoding) part of the DNA from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106.
[0104] The term “sequence identity” refers to the degree to which two polynucleotide or polypeptide sequences are identical on a residue-by-residue basis over a particular region of comparison. The term “percentage of sequence identity” is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I, in the case of nucleic acids) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The term “substantial identity” as used herein denotes a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 80 percent sequence identity, preferably at least 85 percent identity and often 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison region.
[0105] Chimeric and Fusion Proteins
[0106] The invention also provides NOVX chimeric or fusion proteins. As used herein, an NOVX “chimeric protein” or “fusion protein” comprises an NOVX polypeptide operatively-linked to a non-NOVX polypeptide. An “NOVX polypeptide” refers to a polypeptide having an amino acid sequence corresponding to an NOVX protein SEQ ID NO: 2n, wherein n is an integer between 1 and 106, whereas a “non-NOVX polypeptide” refers to a polypeptide having an amino acid sequence corresponding to a protein that is not substantially homologous to the NOVX protein, e.g., a protein that is different from the NOVX protein and that is derived from the same or a different organism. Within an NOVX fusion protein the NOVX polypeptide can correspond to all or a portion of an NOVX protein. In one embodiment, an NOVX fusion protein comprises at least one biologically-active portion of an NOVX protein. In another embodiment, an NOVX fusion protein comprises at least two biologically-active portions of an NOVX protein. In yet another embodiment, an NOVX fusion protein comprises at least three biologically-active portions of an NOVX protein. Within the fusion protein, the term “operatively-linked” is intended to indicate that the NOVX polypeptide and the non-NOVX polypeptide are fused in-frame with one another. The non-NOVX polypeptide can be fused to the N-terminus or C-terminus of the NOVX polypeptide.
[0107] In one embodiment, the fusion protein is a GST-NOVX fusion protein in which the NOVX sequences are fused to the C-terminus of the GST (glutathione S-transferase) sequences. Such fusion proteins can facilitate the purification of recombinant NOVX polypeptides.
[0108] In another embodiment, the fusion protein is an NOVX protein containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g. mammalian host cells), expression and/or secretion of NOVX can be increased through use of a heterologous signal sequence.
[0109] In yet another embodiment, the fusion protein is an NOVX-immunoglobulin fusion protein in which the NOVX sequences are fused to sequences derived from a member of the immunoglobulin protein family. The NOVX-immunoglobulin fusion proteins of the invention can be incorporated into pharmaceutical compositions and administered to a subject to inhibit an interaction between an NOVX ligand and an NOVX protein on the surface of a cell, to thereby suppress NOVX-mediated signal transduction in vivo. The NOVX-immunoglobulin fusion proteins can be used to affect the bioavailability of an NOVX cognate ligand. Inhibition of the NOVX ligand/NOVX interaction may be useful therapeutically for both the treatment of proliferative and differentiative disorders, as well as modulating (e.g. promoting or inhibiting) cell survival. Moreover, the NOVX-immunoglobulin fusion proteins of the invention can be used as immunogens to produce anti-NOVX antibodies in a subject, to purify NOVX ligands, and in screening assays to identify molecules that inhibit the interaction of NOVX with an NOVX ligand.
[0110] An NOVX chimeric or fusion protein of the invention can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, e.g., by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, e.g., Ausubel, et al. (eds.) CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). An NOVX-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the NOVX protein.
[0111] NOVX Agonists and Antagonists
[0112] The invention also pertains to variants of the NOVX proteins that function as either NOVX agonists (i.e., mimetics) or as NOVX antagonists. Variants of the NOVX protein can be generated by mutagenesis (e.g., discrete point mutation or truncation of the NOVX protein). An agonist of the NOVX protein can retain substantially the same, or a subset of, the biological activities of the naturally occurring form of the NOVX protein. An antagonist of the NOVX protein can inhibit one or more of the activities of the naturally occurring form of the NOVX protein by, for example, competitively binding to a downstream or upstream member of a cellular signaling cascade which includes the NOVX protein. Thus, specific biological effects can be elicited by treatment with a variant of limited function. In one embodiment, treatment of a subject with a variant having a subset of the biological activities of the naturally occurring form of the protein has fewer side effects in a subject relative to treatment with the naturally occurring form of the NOVX proteins.
[0113] Variants of the NOVX proteins that function as either NOVX agonists (i.e., mimetics) or as NOVX antagonists can be identified by screening combinatorial libraries of mutants (e.g., truncation mutants) of the NOVX proteins for NOVX protein agonist or antagonist activity. In one embodiment, a variegated library of NOVX variants is generated by combinatorial mutagenesis at the nucleic acid level and is encoded by a variegated gene library. A variegated library of NOVX variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential NOVX sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g., for phage display) containing the set of NOVX sequences therein. There are a variety of methods which can be used to produce libraries of potential NOVX variants from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene then ligated into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all of the sequences encoding the desired set of potential NOVX sequences. Methods for synthesizing degenerate oligonucleotides are well-known within the art. See, e.g., Narang, 1983. Tetrahedron 39: 3; Itakura, et al., 1984. Annu. Rev. Biochem. 53: 323; Itakura, et al., 1984. Science 198: 1056; Ike, et al., 1983. Nuci. Acids Res. 11: 477.
[0114] Polypeptide Libraries
[0115] In addition, libraries of fragments of the NOVX protein coding sequences can be used to generate a variegated population of NOVX fragments for screening and subsequent selection of variants of an NOVX protein. In one embodiment, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of an NOVX coding sequence with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, renaturing the DNA to form double-stranded DNA that can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with S1 nuclease, and ligating the resulting fragment library into an expression vector. By this method, expression libraries can be derived which encodes N-terminal and internal fragments of various sizes of the NOVX proteins.
[0116] Various techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. Such techniques are adaptable for rapid screening of the gene libraries generated by the combinatorial mutagenesis of NOVX proteins. The most widely used techniques, which are amenable to high throughput analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation of the vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a new technique that enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify NOVX variants. See, e.g., Arkin and Yourvan, 1992. Proc. Natl. Acad. Sci. USA 89: 7811-7815; Delgrave, et al., 1993. Protein Engineering 6:327-331.
[0117] NOVX Antibodies
[0118] The term “antibody” as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, i.e., molecules that contain an antigen binding site that specifically binds (immunoreacts with) an antigen. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, Fab, Fab′ and F(ab′)2 fragments, and an Fab expression library. In general, antibody molecules obtained from humans relates to any of the classes IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature of the heavy chain present in the molecule. Certain classes have subclasses as well, such as IgG1, IgG2, and others. Furthermore, in humans, the light chain may be a kappa chain or a lambda chain. Reference herein to antibodies includes a reference to all such classes, subclasses and types of human antibody species.
[0119] An isolated protein of the invention intended to serve as an antigen, or a portion or fragment thereof, can be used as an immunogen to generate antibodies that immunospecifically bind the antigen, using standard techniques for polyclonal and monoclonal antibody preparation. The full-length protein can be used or, alternatively, the invention provides antigenic peptide fragments of the antigen for use as immunogens. An antigenic peptide fragment comprises at least 6 amino acid residues of the amino acid sequence of the full length protein, such as an amino acid sequence shown in SEQ ID NO: 2n, wherein n is an integer between 1 and 106, and encompasses an epitope thereof such that an antibody raised against the peptide forms a specific immune complex with the full length protein or with any fragment that contains the epitope. Preferably, the antigenic peptide comprises at least 10 amino acid residues, or at least 15 amino acid residues, or at least 20 amino acid residues, or at least 30 amino acid residues. Preferred epitopes encompassed by the antigenic peptide are regions of the protein that are located on its surface; commonly these are hydrophilic regions.
[0120] In certain embodiments of the invention, at least one epitope encompassed by the antigenic peptide is a region of NOVX that is located on the surface of the protein, e.g., a hydrophilic region. A hydrophobicity analysis of the human NOVX protein sequence will indicate which regions of a NOVX polypeptide are particularly hydrophilic and, therefore, are likely to encode surface residues useful for targeting antibody production. As a means for targeting antibody production, hydropathy plots showing regions of hydrophilicity and hydrophobicity may be generated by any method well known in the art, including, for example, the Kyte Doolittle or the Hopp Woods methods, either with or without Fourier transformation. See, e.g., Hopp and Woods, 1981, Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte and Doolittle 1982, J. Mol. Biol. 157: 105-142, each incorporated herein by reference in their entirety. Antibodies that are specific for one or more domains within an antigenic protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.
[0121] A protein of the invention, or a derivative, fragment, analog, homolog or ortholog thereof, may be utilized as an immunogen in the generation of antibodies that immunospecifically bind these protein components.
[0122] Various procedures known within the art may be used for the production of polyclonal or monoclonal antibodies directed against a protein of the invention, or against derivatives, fragments, analogs homologs or orthologs thereof (see, for example, Antibodies: A Laboratory Manual, Harlow E, and Lane D, 1988, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., incorporated herein by reference). Some of these antibodies are discussed below.
[0123] Polyclonal Antibodies
[0124] For the production of polyclonal antibodies, various suitable host animals (e.g., rabbit, goat, mouse or other mammal) may be immunized by one or more injections with the native protein, a synthetic variant thereof, or a derivative of the foregoing. An appropriate immunogenic preparation can contain, for example, the naturally occurring immunogenic protein, a chemically synthesized polypeptide representing the immunogenic protein, or a recombinantly expressed immunogenic protein. Furthermore, the protein may be conjugated to a second protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. The preparation can further include an adjuvant. Various adjuvants used to increase the immunological response include, but are not limited to, Freund's (complete and incomplete), mineral gels (e.g., aluminum hydroxide), surface active substances (e.g., lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, dinitrophenol, etc.), adjuvants usable in humans such as Bacille Calmette-Guerin and Corynebacterium parvum, or similar immunostimulatory agents. Additional examples of adjuvants which can be employed include MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).
[0125] The polyclonal antibody molecules directed against the immunogenic protein can be isolated from the mammal (e.g., from the blood) and further purified by well known techniques, such as affinity chromatography using protein A or protein G, which provide primarily the IgG fraction of immune serum. Subsequently, or alternatively, the specific antigen which is the target of the immunoglobulin sought, or an epitope thereof, may be immobilized on a column to purify the immune specific antibody by immunoaffinity chromatography. Purification of immunoglobulins is discussed, for example, by D. Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia Pa., Vol. 14, No. 8 (Apr. 17, 2000), pp. 25-28).
[0126] Monoclonal Antibodies
[0127] The term “monoclonal antibody” (MAb) or “monoclonal antibody composition”, as used herein, refers to a population of antibody molecules that contain only one molecular species of antibody molecule consisting of a unique light chain gene product and a unique heavy chain gene product. In particular, the complementarity determining regions (CDRs) of the monoclonal antibody are identical in all the molecules of the population. MAbs thus contain an antigen binding site capable of immunoreacting with a particular epitope of the antigen characterized by a unique binding affinity for it.
[0128] Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975). In a hybridoma method, a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes can be immunized in vitro.
[0129] The immunizing agent will typically include the protein antigen, a fragment thereof or a fusion protein thereof. Generally, either peripheral blood lymphocytes are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell [Goding, Monoclonal Antibodies: Principles and Practice, Academic Press, (1986) pp. 59-103]. Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells can be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (“HAT medium”), which substances prevent the growth of HGPRT-deficient cells.
[0130] Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, Calif. and the American Type Culture Collection, Manassas, Va. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies [Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, Marcel Dekker, Inc., New York, (1987) pp. 51-63].
[0131] The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980). It is an objective, especially important in therapeutic applications of monoclonal antibodies, to identify antibodies having a high degree of specificity and a high binding affinity for the target antigen.
[0132] After the desired hybridoma cells are identified, the clones can be subcloned by limiting dilution procedures and grown by standard methods (Goding,1986). Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.
[0133] The monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
[0134] The monoclonal antibodies can also be made by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also can be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, Nature 368, 812-13 (1994)) or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody of the invention to create a chimeric bivalent antibody.
[0135] Humanized Antibodies
[0136] The antibodies directed against the protein antigens of the invention can further comprise humanized antibodies or human antibodies. These antibodies are suitable for administration to humans without engendering an immune response by the human against the administered immunoglobulin. Humanized forms of antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)2 or other antigen-binding subsequences of antibodies) that are principally comprised of the sequence of a human immunoglobulin, and contain minimal sequence derived from a non-human immunoglobulin. Humanization can be performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science. 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. (See also U.S. Pat. No. 5,225,539.) In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., 1986; Riechmann et al., 1988; and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992)).
[0137] Human Antibodies
[0138] Fully human antibodies essentially relate to antibody molecules in which the entire sequence of both the light chain and the heavy chain, including the CDRs, arise from human genes. Such antibodies are termed “human antibodies”, or “fully human antibodies” herein. Human monoclonal antibodies can be prepared by the trioma technique; the human B-cell hybridoma technique (see Kozbor, et al., 1983 Immunol Today 4: 72) and the EBV hybridoma technique to produce human monoclonal antibodies (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). Human monoclonal antibodies may be utilized in the practice of the present invention and may be produced by using human hybridomas (see Cote, et al., 1983. Proc Natl Acad Sci USA 80: 2026-2030) or by transforming human B-cells with Epstein Barr Virus in vitro (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96).
[0139] In addition, human antibodies can also be produced using additional techniques, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol. 222:581 (1991)). Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in Marks et al. (Bio/Technology 10, 779-783 (1992)); Lonberg et al. (Nature 368 856-859 (1994)); Morrison (Nature 368 812-13 (1994)); Fishwild et al,(Nature Biotechnology 14, 845-51 (1996)); Neuberger (Nature Biotechnology 14, 826 (1996)); and Lonberg and Huszar (Intern. Rev. Immunol. 13 65-93 (1995)).
[0140] Human antibodies may additionally be produced using transgenic nonhuman animals which are modified so as to produce fully human antibodies rather than the animal's endogenous antibodies in response to challenge by an antigen. (See PCT publication WO94/02602). The endogenous genes encoding the heavy and light immunoglobulin chains in the nonhuman host have been incapacitated, and active loci encoding human heavy and light chain immunoglobulins are inserted into the host's genome. The human genes are incorporated, for example, using yeast artificial chromosomes containing the requisite human DNA segments. An animal which provides all the desired modifications is then obtained as progeny by crossbreeding intermediate transgenic animals containing fewer than the full complement of the modifications. The preferred embodiment of such a nonhuman animal is a mouse, and is termed the XenomouseTM as disclosed in PCT publications WO 96/33735 and WO 96/34096. This animal produces B cells which secrete fully human immunoglobulins. The antibodies can be obtained directly from the animal after immunization with an immunogen of interest, as, for example, a preparation of a polyclonal antibody, or alternatively from immortalized B cells derived from the animal, such as hybridomas producing monoclonal antibodies. Additionally, the genes encoding the immunoglobulins with human variable regions can be recovered and expressed to obtain the antibodies directly, or can be further modified to obtain analogs of antibodies such as, for example, single chain Fv molecules.
[0141] An example of a method of producing a nonhuman host, exemplified as a mouse, lacking expression of an endogenous immunoglobulin heavy chain is disclosed in U.S. Pat. No. 5,939,598. It can be obtained by a method including deleting the J segment genes from at least one endogenous heavy chain locus in an embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector containing a gene encoding a selectable marker; and producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker.
[0142] A method for producing an antibody of interest, such as a human antibody, is disclosed in U.S. Pat. No. 5,916,771. It includes introducing an expression vector that contains a nucleotide sequence encoding a heavy chain into one mammalian host cell in culture, introducing an expression vector containing a nucleotide sequence encoding a light chain into another mammalian host cell, and fusing the two cells to form a hybrid cell. The hybrid cell expresses an antibody containing the heavy chain and the light chain.
[0143] In a further improvement on this procedure, a method for identifying a clinically relevant epitope on an immunogen, and a correlative method for selecting an antibody that binds immunospecifically to the relevant epitope with high affinity, are disclosed in PCT publication WO 99/53049.
[0144] Fab Fragments and Single Chain Antibodies
[0145] According to the invention, techniques can be adapted for the production of single-chain antibodies specific to an antigenic protein of the invention (see e.g., U.S. Pat. No. 4,946,778). In addition, methods can be adapted for the construction of Fab expression libraries (see e.g., Huse, et al., 1989 Science 246: 1275-1281) to allow rapid and effective identification of monoclonal Fab fragments with the desired specificity for a protein or derivatives, fragments, analogs or homologs thereof. Antibody fragments that contain the idiotypes to a protein antigen may be produced by techniques known in the art including, but not limited to: (i) an F(ab′)2 fragment produced by pepsin digestion of an antibody molecule; (ii) an Fab fragment generated by reducing the disulfide bridges of an F(ab′)2 fragment; (iii) an Fab fragment generated by the treatment of the antibody molecule with papain and a reducing agent and (iv) Fv fragments.
[0146] Bispecific Antibodies
[0147] Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens. In the present case, one of the binding specificities is for an antigenic protein of the invention. The second binding target is any other antigen, and advantageously is a cell-surface protein or receptor or receptor subunit. Methods for making bispecific antibodies are known in the art. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 305:537-539 (1983)). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of ten different antibody molecules, of which only one has the correct bispecific structure. The purification of the correct molecule is usually accomplished by affinity chromatography steps. Similar procedures are disclosed in WO 93/08829, published May 13, 1993, and in Traunecker et al., EMBO J., 10:3655-3659 (1991).
[0148] Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy-chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light-chain binding present in at least one of the fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. For further details of generating bispecific antibodies see, for example, Suresh et al., Methods in Enzymology, 121:210 (1986).
[0149] According to another approach described in WO 96/27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture. The preferred interface comprises at least a part of the CH3 region of an antibody constant domain. In this method, one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.
[0150] Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′)2 bispecific antibodies). Techniques for generating bispecific antibodies from antibody fragments have been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. Brennan et al., Science 229:81 (1985) describe a procedure wherein intact antibodies are proteolytically cleaved to generate F(ab′)2 fragments. These fragments are reduced in the presence of the dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The Fab′ fragments generated are then converted to thionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives is then reconverted to the Fab′-thiol by reduction with mercaptoethylamine and is mixed with an equimolar amount of the other Fab′-TNB derivative to form the bispecific antibody. The bispecific antibodies produced can be used as agents for the selective immobilization of enzymes.
[0151] Additionally, Fab′ fragments can be directly recovered from E. coli and chemically coupled to form bispecific antibodies. Shalaby et al., J. Exp. Med. 175:217-225 (1992) describe the production of a fully humanized bispecific antibody F(ab′)2 molecule. Each Fab′ fragment was separately secreted from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody. The bispecific antibody thus formed was able to bind to cells overexpressing the ErbB2 receptor and normal human T cells, as well as trigger the lytic activity of human cytotoxic lymphocytes against human breast tumor targets.
[0152] Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et al., J. Immunol. 148(5): 1547-1553 (1992). The leucine zipper peptides from the Fos and Jun proteins were linked to the Fab′ portions of two different antibodies by gene fusion. The antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The “diabody” technology described by Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism for making bispecific antibody fragments. The fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the VH and VL domains of one fragment are forced to pair with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (sFv) dimers has also been reported. See, Gruber et al., J. Immunol. 152:5368 (1994). Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al., J. Immunol. 147:60 (1991).
[0153] Exemplary bispecific antibodies can bind to two different epitopes, at least one of which originates in the protein antigen of the invention. Alternatively, an anti-antigenic arm of an immunoglobulin molecule can be combined with an arm which binds to a triggering molecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2, CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD16) so as to focus cellular defense mechanisms to the cell expressing the particular antigen. Bispecific antibodies can also be used to direct cytotoxic agents to cells which express a particular antigen. These antibodies possess an antigen-binding arm and an arm which binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Another bispecific antibody of interest binds the protein antigen described herein and further binds tissue factor (TF).
[0154] Heteroconjugate Antibodies
[0155] Heteroconjugate antibodies are also within the scope of the present invention. Heteroconjugate antibodies are composed of two covalently joined antibodies. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360; WO 92/200373; EP 03089). It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Pat. No. 4,676,980.
[0156] Effector Function Engineering
[0157] It can be desirable to modify the antibody of the invention with respect to effector function, so as to enhance, e.g., the effectiveness of the antibody in treating cancer. For example, cysteine residue(s) can be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated can have improved internalization capability and/or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195 (1992) and Shopes, J. Immunol., 148: 2918-2922 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al. Cancer Research, 53: 2560-2565 (1993). Alternatively, an antibody can be engineered that has dual Fc regions and can thereby have enhanced complement lysis and ADCC capabilities. See Stevenson et al., Anti-Cancer Drug Design, 3: 219-230 (1989).
[0158] Immunoconjugates
[0159] The invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate).
[0160] Chemotherapeutic agents useful in the generation of such immunoconjugates have been described above. Enzymatically active toxins and fragments thereof that can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. A variety of radionuclides are available for the production of radioconjugated antibodies. Examples incude 212Bi 131I, 131In, 90Y, and 186Re.
[0161] Conjugates of the antibody and cytotoxic agent are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science, 238: 1098 (1987). Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See WO94/11026.
[0162] In another embodiment, the antibody can be conjugated to a “receptor” (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a “ligand” (e.g., avidin) that is in turn conjugated to a cytotoxic agent.
[0163] Immunoliposomes
[0164] The antibodies disclosed herein can also be formulated as inmuunoliposomes. Liposomes containing the antibody are prepared by methods known in the art, such as described in Epstein et al., Proc. Natl. Acad. Sci. USA, 82: 3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA, 77: 4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556.
[0165] Particularly useful liposomes can be generated by the reverse-phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter. Fab′ fragments of the antibody of the present invention can be conjugated to the liposomes as described in Martin et al., J. Biol. Chem., 257: 286-288 (1982) via a disulfide-interchange reaction. A chemotherapeutic agent (such as Doxorubicin) is optionally contained within the liposome. See Gabizon et al., J. National Cancer Inst., 81(19): 1484 (1989).
[0166] Diagnostic Applications of Antibodies Directed Against the Proteins of the Invention
[0167] Antibodies directed against a protein of the invention may be used in methods known within the art relating to the localization and/or quantitation of the protein (e.g., for use in measuring levels of the protein within appropriate physiological samples, for use in diagnostic methods, for use in imaging the protein, and the like). In a given embodiment, antibodies against the proteins, or derivatives, fragments, analogs or homologs thereof, that contain the antigen binding domain, are utilized as pharmacologically-active compounds (see below).
[0168] An antibody specific for a protein of the invention can be used to isolate the protein by standard techniques, such as immunoaffinity chromatography or immunoprecipitation. Such an antibody can facilitate the purification of the natural protein antigen from cells and of recombinantly produced antigen expressed in host cells. Moreover, such an antibody can be used to detect the antigenic protein (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression of the antigenic protein. Antibodies directed against the protein can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 125I, 131I, 35S or 3H.
[0169] Antibody Therapeutics
[0170] Antibodies of the invention, including polyclonal, monoclonal, humanized and fully human antibodies, may used as therapeutic agents. Such agents will generally be employed to treat or prevent a disease or pathology in a subject. An antibody preparation, preferably one having high specificity and high affinity for its target antigen, is administered to the subject and will generally have an effect due to its binding with the target. Such an effect may be one of two kinds, depending on the specific nature of the interaction between the given antibody molecule and the target antigen in question. In the first instance, administration of the antibody may abrogate or inhibit the binding of the target with an endogenous ligand to which it naturally binds. In this case, the antibody binds to the target and masks a binding site of the naturally occurring ligand, wherein the ligand serves as an effector molecule. Thus the receptor mediates a signal transduction pathway for which ligand is responsible.
[0171] Alternatively, the effect may be one in which the antibody elicits a physiological result by virtue of binding to an effector binding site on the target molecule. In this case the target, a receptor having an endogenous ligand which may be absent or defective in the disease or pathology, binds the antibody as a surrogate effector ligand, initiating a receptor-based signal transduction event by the receptor.
[0172] A therapeutically effective amount of an antibody of the invention relates generally to the amount needed to achieve a therapeutic objective. As noted above, this may be a binding interaction between the antibody and its target antigen that, in certain cases, interferes with the functioning of the target, and in other cases, promotes a physiological response. The amount required to be administered will furthermore depend on the binding affinity of the antibody for its specific antigen, and will also depend on the rate at which an administered antibody is depleted from the free volume other subject to which it is administered. Common ranges for therapeutically effective dosing of an antibody or antibody fragment of the invention may be, by way of nonlimiting example, from about 0.1 mg/kg body weight to about 50 mg/kg body weight. Common dosing frequencies may range, for example, from twice daily to once a week.
[0173] Pharmaceutical Compositions of Antibodies
[0174] Antibodies specifically binding a protein of the invention, as well as other molecules identified by the screening assays disclosed herein, can be administered for the treatment of various disorders in the form of pharmaceutical compositions. Principles and considerations involved in preparing such compositions, as well as guidance in the choice of components are provided, for example, in Remington: The Science And Practice Of Pharmacy 19th ed. (Alfonso R. Gennaro, et al., editors) Mack Pub. Co., Easton, Pa.:11995; Drug Absorption Enhancement: Concepts, Possibilities, Limitations, And Trends, Harwood Academnic Publishers, Langhorne, Pa., 1994; and Peptide And Protein Drug Delivery (Advances In Parenteral Sciences, Vol. 4), 1991, M. Dekker, New York.
[0175] If the anti genic protein is intracellular and whole antibodies are used as inhibitors, internalizing antibodies are preferred. However, liposomes can also be used to deliver the antibody, or an antibody fragment, into cells. Where antibody fragments are used, the smallest inhibitory fragment that specifically binds to the binding domain of the target protein is preferred. For example, based upon the variable-region sequences of an antibody, peptide molecules can be designed that retain the ability to bind the target protein sequence. Such peptides can be synthesized chemically and/or produced by recombinant DNA technology. See, e.g., Marasco et al., Proc. Natl. Acad. Sci. USA, 90: 7889-7893 (1993). The formulation herein can also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Alternatively, or in addition, the composition can comprise an agent that enhances its function, such as, for example, a cytotoxic agent, cytokine, chemotherapeutic agent, or growth-inhibitory agent. Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
[0176] The active ingredients can also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacrylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles, and nanocapsules) or in macroemulsions.
[0177] The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
[0178] Sustained-release preparations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.
[0179] ELISA Assay
[0180] An agent for detecting an analyte protein is an antibody capable of binding to an analyte protein, preferably an antibody with a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab)2) can be used. The term “labeled”, with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently-labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently-labeled streptavidin. The term “biological sample” is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. Included within the usage of the term “biological sample”, therefore, is blood and a fraction or component of blood including blood serum, blood plasma, or lymph. That is, the detection method of the invention can be used to detect an analyte mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of an analyte mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of an analyte protein include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence. In vitro techniques for detection of an analyte genomic DNA include Southern hybridizations. Procedures for conducting immunoassays are described, for example in “ELISA: Theory and Practice: Methods in Molecular Biology”, Vol. 42, J. R. Crowther (Ed.) Human Press, Totowa, N.J., 1995; “Immunoassay”, E. Diamandis and T. Christopoulus, Academic Press, Inc., San Diego, Calif., 1996; and “Practice and Thory of Enzyme Immunoassays”, P. Tijssen, Elsevier Science Publishers, Amsterdam, 1985. Furthermore, in vivo techniques for detection of an analyte protein include introducing into a subject a labeled anti-an analyte protein antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.
[0181] NOVX Recombinant Expression Vectors and Host Cells
[0182] Another aspect of the invention pertains to vectors, preferably expression vectors, containing a nucleic acid encoding an NOVX protein, or derivatives, fragments, analogs or homologs thereof. As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively-linked. Such vectors are referred to herein as “expression vectors”. In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, “plasmid” and “vector” can be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.
[0183] The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, that is operatively-linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, “operably-linked” is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner that allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell).
[0184] The term “regulatory sequence” is intended to includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or peptides, including fusion proteins or peptides, encoded by nucleic acids as described herein (e.g., NOVX proteins, mutant forms of NOVX proteins, fusion proteins, etc.).
[0185] The recombinant expression vectors of the invention can be designed for expression of NOVX proteins in prokaryotic or eukaryotic cells. For example, NOVX proteins can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors) yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase. Expression of proteins in prokaryotes is most often carried out in Escherichia coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: (i) to increase expression of recombinant protein; (ii) to increase the solubility of the recombinant protein; and (iii) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988. Gene 67: 31-40), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) that fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein. Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amrann et al., (1988) Gene 69:301-315) and pET 11d (Studier et al., GENE ExPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 60-89).
[0186] One strategy to maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein. See, e.g., Gottesman, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli (see, e.g., Wada, et al., 1992. Nucl. Acids Res. 20: 2111-2118). Such alteration of nucleic acid sequences of the invention can be carried out by standard DNA synthesis techniques.
[0187] In another embodiment, the NOVX expression vector is a yeast expression vector. Examples of vectors for expression in yeast Saccharomyces cerivisae include pYepSec1 (Baldari, et al., 1987. EMBO J. 6: 229-234), pMFa (Kuijan and Herskowitz, 1982. Cell 30: 933-943), pJRY88 (Schultz et al., 1987. Gene 54: 113-123), pYES2 (Invitrogen Corporation, San Diego, Calif.), and picZ (InVitrogen Corp, San Diego, Calif.).
[0188] Alternatively, NOVX can be expressed in insect cells using baculovirms expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., SF9 cells) include the pAc series (Smith, et al., 1983. Mol. Cell. Biol. 3: 2156-2165) and the pVL series (Lucklow and Summers, 1989. Virology 170: 31-39).
[0189] In yet another embodiment, a nucleic acid of the invention is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 (Seed, 1987. Nature 329: 840) and pMT2PC (Kaufman, et al., 1987. EMBO J. 6: 187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2, cytomegalovirus, and simian virus 40. For other suitable expression systems for both prokaryotic and eukaryotic cells see, e.g., Chapters 16 and 17 of Sambrook, et al., MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.
[0190] In another embodiment, the recombinant mammalian expression vector is capable of directing expression of the nucleic acid preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Tissue-specific regulatory elements are known in the art. Non-limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert, et al., 1987. Genes Dev. 1: 268-277), lymphoid-specific promoters (Calame and Eaton, 1988. Adv. Immunol. 43: 235-275), in particular promoters of T cell receptors (Winoto and Baltimore, 1989. EMBO J. 8: 729-733) and immunoglobulins (Baneri, et al., 1983. Cell 33: 729-740; Queen and Baltimore, 1983. Cell 33: 741-748), neuron-specific promoters (e.g., the neurofilament promoter; Byrne and Ruddle, 1989. Proc. Nat. Acad. Sci. USA 86: 5473-5477), pancreas-specific promoters (Edlund, et al., 1985. Science 230: 912-916), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are also encompassed, e.g., the murine hox promoters (Kessel and Gruss, 1990. Science 249: 374-379) and the c-fetoprotein promoter (Campes and Tilghman, 1989. Genes Dev. 3: 537-546).
[0191] The invention further provides a recombinant expression vector comprising a DNA molecule of the invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively-linked to a regulatory sequence in a manner that allows for expression (by transcription of the DNA molecule) of an RNA molecule that is antisense to NOVX mRNA. Regulatory sequences operatively linked to a nucleic acid cloned in the antisense orientation can be chosen that direct the continuous expression of the antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen that direct constitutive, tissue specific or cell type specific expression of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced. For a discussion of the regulation of gene expression using antisense genes see, e.g., Weintraub, et al., “Antisense RNA as a molecular tool for genetic analysis,” Reviews-Trends in Genetics, Vol. 1(1) 1986.
[0192] Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms “host cell” and “recombinant host cell” are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.
[0193] A host cell can be any prokaryotic or eukaryotic cell. For example, NOVX protein can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are known to those skilled in the art.
[0194] Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection, or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals.
[0195] For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Various selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding NOVX or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die).
[0196] A host cell of the invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) NOVX protein. Accordingly, the invention further provides methods for producing NOVX protein using the host cells of the invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding NOVX protein has been introduced) in a suitable medium such that NOVX protein is produced. In another embodiment, the method further comprises isolating NOVX protein from the medium or the host cell.
[0197] Transgenic NOVX Animals
[0198] The host cells of the invention can also be used to produce non-human transgenic animals. For example, in one embodiment, a host cell of the invention is a fertilized oocyte or an embryonic stem cell into which NOVX protein-coding sequences have been introduced. Such host cells can then be used to create non-human transgenic animals in which exogenous NOVX sequences have been introduced into their genome or homologous recombinant animals in which endogenous NOVX sequences have been altered. Such animals are useful for studying the function and/or activity of NOVX protein and for identifying and/or evaluating modulators of NOVX protein activity. As used herein, a “transgenic animal” is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more of the cells of the animal includes a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, etc. A transgene is exogenous DNA that is integrated into the genome of a cell from which a transgenic animal develops and that remains in the genome of the mature animal, thereby directing the expression of an encoded gene product in one or more cell types or tissues of the transgenic animal. As used herein, a “homologous recombinant animal” is a non-human animal, preferably a mammal, more preferably a mouse, in which an endogenous NOVX gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell of the animal, e.g., an embryonic cell of the animal, prior to development of the animal.
[0199] A transgenic animal of the invention can be created by introducing NOVX-encoding nucleic acid into the male pronuclei of a fertilized oocyte (e.g., by microinjection, retroviral infection) and allowing the oocyte to develop in a pseudopregnant female foster animal. The human NOVX cDNA sequences SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 can be introduced as a transgene into the genome of a non-human animal. Alternatively, a non-human homologue of the human NOVX gene, such as a mouse NOVX gene, can be isolated based on hybridization to the human NOVX cDNA (described further supra) and used as a transgene. Intronic sequences and polyadenylation signals can also be included in the transgene to increase the efficiency of expression of the transgene. A tissue-specific regulatory sequence(s) can be operably-linked to the NOVX transgene to direct expression of NOVX protein to particular cells. Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Pat. Nos. 4,736,866; 4,870,009; and 4,873,191; and Hogan, 1986. In: MANIPULATING THE MOUSE EMBRYO, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Similar methods are used for production of other transgenic animals. A transgenic founder animal can be identified based upon the presence of the NOVX transgene in its genome and/or expression of NOVX mRNA in tissues or cells of the animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene-encoding NOVX protein can further be bred to other transgenic animals carrying other transgenes.
[0200] To create a homologous recombinant animal, a vector is prepared which contains at least a portion of an NOVX gene into which a deletion, addition or substitution has been introduced to thereby alter, e.g., functionally disrupt, the NOVX gene. The NOVX gene can be a human gene (e.g., the cDNA of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106), but more preferably, is a non-human homologue of a human NOVX gene. For example, a mouse homologue of human NOVX gene of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 can be used to construct a homologous recombination vector suitable for altering an endogenous NOVX gene in the mouse genome. In one embodiment, the vector is designed such that, upon homologous recombination, the endogenous NOVX gene is functionally disrupted (i.e., no longer encodes a functional protein; also referred to as a “knock out” vector).
[0201] Alternatively, the vector can be designed such that, upon homologous recombination, the endogenous NOVX gene is mutated or otherwise altered but still encodes functional protein (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous NOVX protein). In the homologous recombination vector, the altered portion of the NOVX gene is flanked at its 5′- and 3′-termini by additional nucleic acid of the NOVX gene to allow for homologous recombination to occur between the exogenous NOVX gene carried by the vector and an endogenous NOVX gene in an embryonic stem cell. The additional flanking NOVX nucleic acid is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several kilobases of flanking DNA (both at the 5′- and 3′-termini) are included in the vector. See, e.g., Thomas, et al., 1987. Cell 51: 503 for a description of homologous recombination vectors. The vector is ten introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced NOVX gene has homologously-recombined with the endogenous NOVX gene are selected. See, e.g., Li, et al., 1992. Cell 69: 915.
[0202] The selected cells are then injected into a blastocyst of an animal (e.g., a mouse) to form aggregation chimeras. See, e.g., Bradley, 1987. In: TERATOCARCINOMAS AND EMBRYONIC STEM CELLS: A PRACTICAL APPROACH, Robertson, ed. IRL, Oxford, pp. 113-152. A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal and the embryo brought to term. Progeny harboring the homologously-recombined DNA in their germ cells can be used to breed animals in which all cells of the animal contain the homologously-recombined DNA by germline transmission of the transgene. Methods for constructing homologous recombination vectors and homologous recombinant animals are described further in Bradley, 1991. Curr. Opin. Biotechnol. 2: 823-829; PCT International Publication Nos.: WO 90/11354; WO 91/01140; WO 92/0968; and WO 93/04169.
[0203] In another embodiment, transgenic non-humans animals can be produced that contain selected systems that allow for regulated expression of the transgene. One example of such a system is the cre/loxP recombinase system of bacteriophage P1. For a description of the cre/loxP recombinase system, See, e.g., Lakso, et al., 1992. Proc. Natl. Acad. Sci. USA 89: 6232-6236. Another example of a recombinase system is the FLP recombinase system of Saccharomyces cerevisiae. See, O'Gorman, et al., 1991. Science 251:1351-1355. If a cre/loxP recombinase system is used to regulate expression of the transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein are required. Such animals can be provided through the construction of “double” transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.
[0204] Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, et al., 1997. Nature 385: 810-813. In brief, a cell (e.g., a somatic cell) from the transgenic animal can be isolated and induced to exit the growth cycle and enter Go phase. The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal of the same species from which the quiescent cell is isolated. The reconstructed oocyte is then cultured such that it develops to morula or blastocyte and then transferred to pseudopregnant female foster animal. The offspring borne of this female foster animal will be a clone of the animal from which the cell (e.g., the somatic cell) is isolated.
[0205] Pharmaceutical Compositions
[0206] The NOVX nucleic acid molecules, NOVX proteins, and anti-NOVX antibodies (also referred to herein as “active compounds”) of the invention, and derivatives, fragments, analogs and homologs thereof, can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically comprise the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, finger's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.
[0207] A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
[0208] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EC™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.
[0209] Sterile injectable solutions can be prepared by incorporating the active compound (e.g., an NOVX protein or anti-NOVX antibody) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
[0210] Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
[0211] For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.
[0212] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.
[0213] The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.
[0214] In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.
[0215] It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.
[0216] The nucleic acid molecules of the invention can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see, e.g., U.S. Pat. No. 5,328,470) or by stereotactic injection (see, e.g., Chen, et al., 1994. Proc. Natl. Acad. Sci. USA 91: 3054-3057). The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells that produce the gene delivery system.
[0217] The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.
[0218] Screening and Detection Methods
[0219] The isolated nucleic acid molecules of the invention can be used to express NOVX protein (e.g., via a recombinant expression vector in a host cell in gene therapy applications), to detect NOVX mRNA (e.g., in a biological sample) or a genetic lesion in an NOVX gene, and to modulate NOVX activity, as described further, below. In addition, the NOVX proteins can be used to screen drugs or compounds that modulate the NOVX protein activity or expression as well as to treat disorders characterized by insufficient or excessive production of NOVX protein or production of NOVX protein forms that have decreased or aberrant activity compared to NOVX wild-type protein (e.g.; diabetes (regulates insulin release); obesity (binds and transport lipids); metabolic disturbances associated with obesity, the metabolic syndrome X as well as anorexia and wasting disorders associated with chronic diseases and various cancers, and infectious disease (possesses anti-microbial activity) and the various dyslipidemias. In addition, the anti-NOVX antibodies of the invention can be used to detect and isolate NOVX proteins and modulate NOVX activity. In yet a further aspect, the invention can be used in methods to influence appetite, absorption of nutrients and the disposition of metabolic substrates in both a positive and negative fashion.
[0220] The invention further pertains to novel agents identified by the screening assays described herein and uses thereof for treatments as described, supra.
[0221] Screening Assays
[0222] The invention provides a method (also referred to herein as a “screening assay”) for identifying modulators, i.e., candidate or test compounds or agents (e.g., peptides, peptidomimetics, small molecules or other drugs) that bind to NOVX proteins or have a stimulatory or inhibitory effect on, e.g., NOVX protein expression or NOVX protein activity. The invention also includes compounds identified in the screening assays described herein. In one embodiment, the invention provides assays for screening candidate or test compounds which bind to or modulate the activity of the membrane-bound form of an NOVX protein or polypeptide or biologically-active portion thereof. The test compounds of the invention can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the “one-bead one-compound” library method; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds. See, e.g., Lam, 1997. Anticancer Drug Design 12: 145.
[0223] A “small molecule” as used herein, is meant to refer to a composition that has a molecular weight of less than about 5 kD and most preferably less than about 4 kD. Small molecules can be, e.g., nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, lipids or other organic or inorganic molecules. Libraries of chemical and/or biological mixtures, such as fungal, bacterial, or algal extracts, are known in the art and can be screened with any of the assays of the invention.
[0224] Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt, et al., 1993. Proc. Natl. Acad. Sci. U.S.A. 90: 6909; Erb, et al., 1994. Proc. Natl. Acad. Sci. U.S.A. 91: 11422; Zuckermann, et al., 1994. J. Med. Chem. 37: 2678; Cho, et al., 1993. Science 261: 1303; Carrell, et al., 1994. Angew. Chem. Int. Ed. Engl. 33: 2059; Carell, et al., 1994. Angew. Chem. Int. Ed. Engl. 33: 2061; and Gallop, et al., 1994. J. Med. Chem. 37:1233.
[0225] Libraries of compounds may be presented in solution (e.g., Houghten, 1992. Biotechniques 13: 412-421), or on beads (Lam, 1991. Nature 354: 82-84), on chips (Fodor, 1993. Nature 364: 555-556), bacteria (Ladner, U.S. Pat. No. 5,223,409), spores (Ladner, U.S. Pat. No. 5,233,409), plasmids (Cull, et al., 1992. Proc. Natl. Acad. Sci. USA 89: 1865-1869) or on phage (Scott and Smith, 1990. Science 249: 386-390; Devlin, 1990. Science 249: 404-406; Cwirla, et al., 1990. Proc. Natl. Acad. Sci. U.S.A. 87: 6378-6382; Felici, 1991. J. Mol. Biol. 222: 301-310; Ladner, U.S. Pat. No. 5,233,409.).
[0226] In one embodiment, an assay is a cell-based assay in which a cell which expresses a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface is contacted with a test compound and the ability of the test compound to bind to an NOVX protein determined. The cell, for example, can of mammalian origin or a yeast cell. Determining the ability of the test compound to bind to the NOVX protein can be accomplished, for example, by coupling the test compound with a radioisotope or enzymatic label such that binding of the test compound to the NOVX protein or biologically-active portion thereof can be determined by detecting the labeled compound in a complex. For .example, test compounds can be labeled with 125I, 35S, 14C, or 3H, either directly or indirectly, and the radioisotope detected by direct counting of radioemission or by scintillation counting. Alternatively, test compounds can be enzymatically-labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product. In one embodiment, the assay comprises contacting a cell which expresses a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface with a known compound which binds NOVX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an NOVX protein, wherein determining the ability of the test compound to interact with an NOVX protein comprises determining the ability of the test compound to preferentially bind to NOVX protein or a biologically-active portion thereof as compared to the known compound.
[0227] In another embodiment, an assay is a cell-based assay comprising contacting a cell expressing a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface with a test compound and determining the ability of the test compound to modulate (e.g., stimulate or inhibit) the activity of the NOVX protein or biologically-active portion thereof. Determining the ability of the test compound to modulate the activity of NOVX or a biologically-active portion thereof can be accomplished, for example, by determining the ability of the NOVX protein to bind to or interact with an NOVX target molecule. As used herein, a “target molecule” is a molecule with which an NOVX protein binds or interacts in nature, for example, a molecule on the surface of a cell which expresses an NOVX interacting protein, a molecule on the surface of a second cell, a molecule in the extracellular milieu, a molecule associated with the internal surface of a cell membrane or a cytoplasmic molecule. An NOVX target molecule can be a non-NOVX molecule or an NOVX protein or polypeptide of the invention. In one embodiment, an NOVX target molecule is a component of a signal transduction pathway that facilitates transduction of an extracellular signal (e.g. a signal generated by binding of a compound to a membrane-bound NOVX molecule) through the cell membrane and into the cell. The target, for example, can be a second intercellular protein that has catalytic activity or a protein that facilitates the association of downstream signaling molecules with NOVX.
[0228] Determining the ability of the NOVX protein to bind to or interact with an NOVX target molecule can be accomplished by one of the methods described above for determining direct binding. In one embodiment, determining the ability of the NOVX protein to bind to or interact with an NOVX target molecule can be accomplished by determining the activity of the target molecule. For example, the activity of the target molecule can be determined by detecting induction of a cellular second messenger of the target (i.e. intracellular Ca2+, diacylglycerol, IP3, etc.), detecting catalytic/enzymatic activity of the target an appropriate substrate, detecting the induction of a reporter gene (comprising an NOVX-responsive regulatory element operatively linked to a nucleic acid encoding a detectable marker, e.g., luciferase), or detecting a cellular response, for example, cell survival, cellular differentiation, or cell proliferation.
[0229] In yet another embodiment, an assay of the invention is a cell-free assay comprising contacting an NOVX protein or biologically-active portion thereof with a test compound and determining the ability of the test compound to bind to the NOVX protein or biologically-active portion thereof. Binding of the test compound to the NOVX protein can be determined either directly or indirectly as described above. In one such embodiment, the assay comprises contacting the NOVX protein or biologically-active portion thereof with a known compound which binds NOVX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an NOVX protein, wherein determining the ability of the test compound to interact with an NOVX protein comprises determining the ability of the test compound to preferentially bind to NOVX or biologically-active portion thereof as compared to the known compound.
[0230] In still another embodiment, an assay is a cell-free assay comprising contacting NOVX protein or biologically-active portion thereof with a test compound and determining the ability of the test compound to modulate (e.g. stimulate or inhibit) the activity of the NOVX protein or biologically-active portion thereof. Determining the ability of the test compound to modulate the activity of NOVX can be accomplished, for example, by determining the ability of the NOVX protein to bind to an NOVX target molecule by one of the methods described above for determining direct binding. In an alternative embodiment, determining the ability of the test compound to modulate the activity of NOVX protein can be accomplished by determining the ability of the NOVX protein further modulate an NOVX target molecule. For Example, the catalytic/enzymatic activity of the target molecule on an appropriate substrate can be determined as described, supra.
[0231] In yet another embodiment, the cell-free assay comprises contacting the NOVX protein or biologically-active portion thereof with a known compound which binds NOVX protein to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an NOVX protein, wherein determining the ability of the test compound to interact with an NOVX protein comprises determining the ability of the NOVX protein to preferentially bind to or modulate the activity of an NOVX target molecule.
[0232] The cell-free assays of the invention are amenable to use of both the soluble form or the membrane-bound form of NOVX protein. In the case of cell-free assays comprising the membrane-bound form of NOVX protein, it may be desirable to utilize a solubilizing agent such that the membrane-bound form of NOVX protein is maintained in solution. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton® X-100, Triton® X-114, Thesit®, Isotridecypoly(ethylene glycol ether)n, N-dodecyl—N,N-dimethyl-3-ammonio-1-propane sulfonate, 3-(3-cholamidopropyl) dimethylamminiol-1-propane sulfonate (CHAPS), or 3-(3-cholamidopropyl)dimethylamminiol-2-hydroxy-1-propane sulfonate (CHAPSO).
[0233] In more than one embodiment of the above assay methods of the invention, it may be desirable to immobilize either NOVX protein or its target molecule to facilitate separation of complexed from uncomplexed forms of one or both of the proteins, as well as to accommodate automation of the assay. Binding of a test compound to NOVX protein, or interaction of NOVX protein with a target molecule in the presence and absence of a candidate compound, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein can be provided that adds a domain that allows one or both of the proteins to be bound to a matrix. For example, GST-NOVX fusion proteins or GST-target fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtiter plates, that are then combined with the test compound or the test compound and either the non-adsorbed target protein or NOVX protein, and the mixture is incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads or microtiter plate wells are washed to remove any unbound components, the matrix immobilized in the case of beads, complex determined either directly or indirectly, for example, as described, supra. Alternatively, the complexes can be dissociated from the matrix, and the level of NOVX protein binding or activity determined using standard techniques.
[0234] Other techniques for immobilizing proteins on matrices can also be used in the screening assays of the invention. For example, either the NOVX protein or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin. Biotinylated NOVX protein or target molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well-known within the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with NOVX protein or target molecules, but which do not interfere with binding of the NOVX protein to its target molecule, can be derivatized to the wells of the plate, and unbound target or NOVX protein trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the NOVX protein or target molecule, as well as enzyme-linked assays that rely on detecting an enzymatic activity associated with the NOVX protein or target molecule.
[0235] In another embodiment, modulators of NOVX protein expression are identified in a method wherein a cell is contacted with a candidate compound and the expression of NOVX mRNA or protein in the cell is determined. The level of expression of NOVX mRNA or protein in the presence of the candidate compound is compared to the level of expression of NOVX mRNA or protein in the absence of the candidate compound. The candidate compound can then be identified as a modulator of NOVX mRNA or protein expression based upon this comparison. For example, when expression of NOVX mRNA or protein is greater (i.e., statistically significantly greater) in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of NOVX mRNA or protein expression. Alternatively, when expression of NOVX mRNA or protein is less (statistically significantly less) in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of NOVX mRNA or protein expression. The level of NOVX mRNA or protein expression in the cells can be determined by methods described herein for detecting NOVX mRNA or protein.
[0236] In yet another aspect of the invention, the NOVX proteins can be used as “bait proteins” in a two-hybrid assay or three hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos, et al., 1993. Cell 72: 223-232; Madura, et al., 1993. J. Biol. Chem. 268: 12046-12054; Bartel, et al., 1993. Biotechniques 14: 920-924; Iwabuchi, et al., 1993. Oncogene 8: 1693-1696; and Brent WO 94/10300), to identify other proteins that bind to or interact with NOVX (“NOVX-binding proteins” or “NOVX-bp”) and modulate NOVX activity. Such NOVX-binding proteins are also likely to be involved in the propagation of signals by the NOVX proteins as, for example, upstream or downstream elements of the NOVX pathway.
[0237] The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene that codes for NOVX is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4). In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor. If the “bait” and the “prey” proteins are able to interact, in vivo, forming an NOVX-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., LacZ) that is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene that encodes the protein which interacts with NOVX.
[0238] The invention further pertains to novel agents identified by the aforementioned screening assays and uses thereof for treatments as described herein.
[0239] Detection Assays
[0240] Portions or fragments of the cDNA sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. By way of example, and not of limitation, these sequences can be used to: (i) map their respective genes on a chromosome; and, thus, locate gene regions associated with genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (iii) aid in forensic identification of a biological sample. Some of these applications are described in the subsections, below.
[0241] Chromosome Mapping
[0242] Once the sequence (or a portion of the sequence) of a gene has been isolated, this sequence can be used to map the location of the gene on a chromosome. This process is called chromosome mapping. Accordingly, portions or fragments of the NOVX sequences, SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or fragments or derivatives thereof, can be used to map the location of the NOVX genes, respectively, on a chromosome. The mapping of the NOVX sequences to chromosomes is an important first step in correlating these sequences with genes associated with disease.
[0243] Briefly, NOVX genes can be mapped to chromosomes by preparing PCR primers (preferably 15-25 bp in length) from the NOVX sequences. Computer analysis of the NOVX, sequences can be used to rapidly select primers that do not span more than one exon in the genomic DNA, thus complicating the amplification process. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to the NOVX sequences will yield an amplified fragment.
[0244] Somatic cell hybrids are prepared by fusing somatic cells from different mammals (e.g., human and mouse cells). As hybrids of human and mouse cells grow and divide, they gradually lose human chromosomes in random order, but retain the mouse chromosomes. By using media in which mouse cells cannot grow, because they lack a particular enzyme, but in which human cells can, the one human chromosome that contains the gene encoding the needed enzyme will be retained. By using various media, panels of hybrid cell lines can be established. Each cell line in a panel contains either a single human chromosome or a small number of human chromosomes, and a full set of mouse chromosomes, allowing easy mapping of individual genes to specific human chromosomes. See, e.g., D'Eustachio, et al., 1983. Science 220: 919-924. Somatic cell hybrids containing only fragments of human chromosomes can also be produced by using human chromosomes with translocations and deletions.
[0245] PCR mapping of somatic cell hybrids is a rapid procedure for assigning a particular sequence to a particular chromosome. Three or more sequences can be assigned per day using a single thermal cycler. Using the NOVX sequences to design oligonucleotide primers, sub-localization can be achieved with panels of fragments from specific chromosomes. Fluorescence in situ hybridization (FISH) of a DNA sequence to a metaphase chromosomal spread can further be used to provide a precise chromosomal location in one step. Chromosome spreads can be made using cells whose division has been blocked in metaphase by a chemical like colcemid that disrupts the mitotic spindle. The chromosomes can be treated briefly with trypsin, and then stained with Giemsa. A pattern of light and dark bands develops on each chromosome, so that the chromosomes can be identified individually. The FISH technique can be used with a DNA sequence as short as 500 or 600 bases. However, clones larger than 1,000 bases have a higher likelihood of binding to a unique chromosomal location with sufficient signal intensity for simple detection. Preferably 1,000 bases, and more preferably 2,000 bases, will suffice to get good results at a reasonable amount of time. For a review of this technique, see, Verma, et al., HUMAN CHROMOSOMES: A MANUAL OF BASIC TECHNIQUES (Pergamon Press, New York 1988).
[0246] Reagents for chromosome mapping can be used individually to mark a single chromosome or a single site on that chromosome, or panels of reagents can be used for marking multiple sites and/or multiple chromosomes. Reagents corresponding to noncoding regions of the genes actually are preferred for mapping purposes. Coding sequences are more likely to be conserved within gene families, thus increasing the chance of cross hybridizations during chromosomal mapping.
[0247] Once a sequence has been mapped to a precise chromosomal location, the physical position of the sequence on the chromosome can be correlated with genetic map data. Such data are found, e.g., in McKusick, MENDELIAN INHERITANCE IN MAN, available on-line through Johns Hopkins University Welch Medical Library). The relationship between genes and disease, mapped to the same chromosomal region, can then be identified through linkage analysis (co-inheritance of physically adjacent genes), described in, e.g., Egeland, et al., 1987. Nature, 325: 783-787.
[0248] Moreover, differences in the DNA sequences between individuals affected and unaffected with a disease associated with the NOVX gene, can be determined. If a mutation is observed in some or all of the affected individuals but not in any unaffected individuals, then the mutation is likely to be the causative agent of the particular disease. Comparison of affected and unaffected individuals generally involves first looking for structural alterations in the chromosomes, such as deletions or translocations that are visible from chromosome spreads or detectable using PCR based on that DNA sequence. Ultimately, complete sequencing of genes from several individuals can be performed to confirm the presence of a mutation and to distinguish mutations from polymorphisms.
[0249] Tissue Typing
[0250] The NOVX sequences of the invention can also be used to identify individuals from minute biological samples. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, and probed on a Southern blot to yield unique bands for identification. The sequences of the invention are useful as additional DNA markers for RFLP (“restriction fragment length polymorphisms,” described in U.S. Pat. No. 5,272,057).
[0251] Furthermore, the sequences of the invention can be used to provide an alternative technique that determines the actual base-by-base DNA sequence of selected portions of an individual's genome. Thus, the NOVX sequences described herein can be used to prepare two PCR primers from the 5′- and 3′-termini of the sequences. These primers can then be used to amplify an individual's DNA and subsequently sequence it.
[0252] Panels of corresponding DNA sequences from individuals, prepared in this manner, can provide unique individual identifications, as each individual will have a unique set of such DNA sequences due to allelic differences. The sequences of the invention can be used to obtain such identification sequences from individuals and from tissue. The NOVX sequences of the invention uniquely represent portions of the human genome. Allelic variation occurs to some degree in the coding regions of these sequences, and to a greater degree in the noncoding regions. It is estimated that allelic variation between individual humans occurs with a frequency of about once per each 500 bases. Much of the allelic variation is due to single nucleotide polymorphisms (SNPs), which include restriction fragment length polymorphisms (RFLPs).
[0253] Each of the sequences described herein can, to some degree, be used as a standard against which DNA from an individual can be compared for identification purposes. Because greater numbers of polymorphisms occur in the noncoding regions, fewer sequences are necessary to differentiate individuals. The noncoding sequences can comfortably provide positive individual identification with a panel of perhaps 10 to 1,000 primers that each yield a noncoding amplified sequence of 100 bases. If predicted coding sequences, such as those in SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 are used, a more appropriate number of primers for positive individual identification would be 500-2,000.
[0254] Predictive Medicine
[0255] The invention also pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, pharmacogenomics, and monitoring clinical trials are used for prognostic (predictive) purposes to thereby treat an individual prophylactically. Accordingly, one aspect of the invention relates to diagnostic assays for determining NOVX protein and/or nucleic acid expression as well as NOVX activity, in the context of a biological sample (e.g., blood, serum, cells, tissue) to thereby determine whether an individual is afflicted with a disease or disorder, or is at risk of developing a disorder, associated with aberrant NOVX expression or activity. The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers. The invention also provides for prognostic (or predictive) assays for determining whether an individual is at risk of developing a disorder associated with NOVX protein, nucleic acid expression or activity. For example, mutations in an NOVX gene can be assayed in a biological sample. Such assays can be used for prognostic or predictive purpose to thereby prophylactically treat an individual prior to the onset of a disorder characterized by or associated with NOVX protein, nucleic acid expression, or biological activity.
[0256] Another aspect of the invention provides methods for determining NOVX protein, nucleic acid expression or activity in an individual to thereby select appropriate therapeutic or prophylactic agents for that individual (referred to herein as “pharmacogenomics”). Pharmacogenomics allows for the selection of agents (e.g., drugs) for therapeutic or prophylactic treatment of an individual based on the genotype of the individual (e.g., the genotype of the individual examined to determine the ability of the individual to respond to a particular agent.)
[0257] Yet another aspect of the invention pertains to monitoring the influence of agents (e.g., drugs, compounds) on the expression or activity of NOVX in clinical trials.
[0258] These and other agents are described in further detail in the following sections.
[0259] Diagnostic Assays
[0260] An exemplary method for detecting the presence or absence of NOVX in a biological sample involves obtaining a biological sample from a test subject and contacting the biological sample with a compound or an agent capable of detecting NOVX protein or nucleic acid (e.g., mRNA, genomic DNA) that encodes NOVX protein such that the presence of NOVX is detected in the biological sample. An agent for detecting NOVX mRNA or genomic DNA is a labeled nucleic acid probe capable of hybridizing to NOVX mRNA or genomic DNA. The nucleic acid probe can be, for example, a full-length NOVX nucleic acid, such as the nucleic acid of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or a portion thereof, such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to NOVX mRNA or genomic DNA. Other suitable probes for use in the diagnostic assays of the invention are described herein.
[0261] An agent for detecting NOVX protein is an antibody capable of binding to NOVX protein, preferably an antibody with a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab′)2) can be used. The term “labeled”, with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently-labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently-labeled streptavidin. The term “biological sample” is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection method of the invention can be used to detect NOVX mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of NOVX mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of NOVX protein include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence. In vitro techniques for detection of NOVX genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of NOVX protein include introducing into a subject a labeled anti-NOVX antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.
[0262] In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject.
[0263] In another embodiment, the methods further involve obtaining a control biological sample from a control subject, contacting the control sample with a compound or agent capable of detecting NOVX protein, mRNA, or genomic DNA, such that the presence of NOVX protein, mRNA or genomic DNA is detected in the biological sample, and comparing the presence of NOVX protein, mRNA or genomic DNA in the control sample with the presence of NOVX protein, mRNA or genomic DNA in the test sample. The invention also encompasses kits for detecting the presence of NOVX in a biological sample. For example, the kit can comprise: a labeled compound or agent capable of detecting NOVX protein or mRNA in a biological sample; means for determining the amount of NOVX in the sample; and means for comparing the amount of NOVX in the sample with a standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instructions for using the kit to detect NOVX protein or nucleic acid.
[0264] Prognostic Assays
[0265] The diagnostic methods described herein can furthermore be utilized to identify subjects having or at risk of developing a disease or disorder associated with aberrant NOVX expression or activity. For example, the assays described herein, such as the preceding diagnostic assays or the following assays, can be utilized to identify a subject having or at risk of developing a disorder associated with NOVX protein, nucleic acid expression or activity. Alternatively, the prognostic assays can be utilized to identify a subject having or at risk for developing a disease or disorder. Thus, the invention provides a method for identifying a disease or disorder associated with aberrant NOVX expression or activity in which a test sample is obtained from a subject and NOVX protein or nucleic acid (e.g., mRNA, genomic DNA) is detected, wherein the presence of NOVX protein or nucleic acid is diagnostic for a subject having or at risk of developing a disease or disorder associated with aberrant NOVX expression or activity. As used herein, a “test sample” refers to a biological sample obtained from a subject of interest. For example, a test sample can be a biological fluid (e.g., serum), cell sample, or tissue.
[0266] Furthermore, the prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate) to treat a disease or disorder associated with aberrant NOVX expression or activity. For example, such methods can be used to determine whether a subject can be effectively treated with an agent for a disorder. Thus, the invention provides methods for determining whether a subject can be effectively treated with an agent for a disorder associated with aberrant NOVX expression or activity in which a test sample is obtained and NOVX protein or nucleic acid is detected (e.g., wherein the presence of NOVX protein or nucleic acid is diagnostic for a subject that can be administered the agent to treat a disorder associated with aberrant NOVX expression or activity).
[0267] The methods of the invention can also be used to detect genetic lesions in an NOVX gene, thereby determining if a subject with the lesioned gene is at risk for a disorder characterized by aberrant cell proliferation and/or differentiation. In various embodiments, the methods include detecting, in a sample of cells from the subject, the presence or absence of a genetic lesion characterized by at least one of an alteration affecting the integrity of a gene encoding an NOVX-protein, or the misexpression of the NOVX gene. For example, such genetic lesions can be detected by ascertaining the existence of at least one of: (i) a deletion of one or more nucleotides from an NOVX gene; (ii) an addition of one or more nucleotides to an NOVX gene; (iii) a substitution of one or more nucleotides of an NOVX gene, (iv) a chromosomal rearrangement of an NOVX gene; (v) an alteration in the level of a messenger RNA transcript of an NOVX gene, (vi) aberrant modification of an NOVX gene, such as of the methylation pattern of the genomic DNA, (vii) the presence of a non-wild-type splicing pattern of a messenger RNA transcript of an NOVX gene, (viii) a non-wild-type level of an NOVX protein, (ix) allelic loss of an NOVX gene, and (x) inappropriate post-translational modification of an NOVX protein. As described herein, there are a large number of assay techniques known in the art which can be used for detecting lesions in an NOVX gene. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.
[0268] In certain embodiments, detection of the lesion involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran, et al., 1988. Science 241: 1077-1080; and Nakazawa, et al., 1994. Proc. Natl. Acad. Sci. USA 91: 360-364), the latter of which can be particularly useful for detecting point mutations in the NOVX-gene (see, Abravaya, et al., 1995. Nucl. Acids Res. 23: 675-682). This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells of the sample, contacting the nucleic acid sample with one or more primers that specifically hybridize to an NOVX gene under conditions such that hybridization and amplification of the NOVX gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample. It is anticipated that PCR and/or LCR may be desirable to use as a preliminary amplification step in conjunction with any of the techniques used for detecting mutations described herein.
[0269] Alternative amplification methods include: self sustained sequence replication (see, Guatelli, et al., 1990. Proc. Natl. Acad. Sci. USA 87: 1874-1878), transcriptional amplification system (see, Kwoh, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 1173-1177); QP Replicase (see, Lizardi, et al, 1988. BioTechnology 6: 1197), or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers.
[0270] In an alternative embodiment, mutations in an NOVX gene from a sample cell can be identified by alterations in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, amplified (optionally), digested with one or more restriction endonucleases, and fragment length sizes are determined by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DNA indicates mutations in the sample DNA. Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Pat. No. 5,493,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.
[0271] In other embodiments, genetic mutations in NOVX can be identified by hybridizing a sample and control nucleic acids, e.g., DNA or RNA, to high-density arrays containing hundreds or thousands of oligonucleotides probes. See, e.g., Cronin, et al., 1996. Human Mutation 7: 244-255; Kozal, et al., 1996. Nat. Med. 2: 753-759. For example, genetic mutations in NOVX can be identified in two dimensional arrays containing light-generated DNA probes as described in Cronin, et al., supra. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild-type gene and the other complementary to the mutant gene.
[0272] In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the NOVX gene and detect mutations by comparing the sequence of the sample NOVX with the corresponding wild-type (control) sequence. Examples of sequencing reactions include those based on techniques developed by Maxim and Gilbert, 1977. Proc. Natl. Acad. Sci. USA 74: 560 or Sanger, 1977. Proc. Natl. Acad. Sci. USA 74: 5463. It is also contemplated that any of a variety of automated sequencing procedures can be utilized when performing the diagnostic assays (see, e.g., Naeve, et al., 1995. Biotechniques 19: 448), including sequencing by mass spectrometry (see, e.g., PCT International Publication No. WO 94/16101; Cohen, et al., 1996. Adv. Chromatography 36: 127-162; and Griffin, et al., 1993. Appl. Biochem. Biotechnol. 38: 147-159).
[0273] Other methods for detecting mutations in the NOVX gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes. See, e.g., Myers, et al., 1985. Science 230: 1242. In general, the art technique of “mismatch cleavage” starts by providing heteroduplexes of formed by hybridizing (labeled) RNA or DNA containing the wild-type NOVX sequence with potentially mutant RNA or DNA obtained from a tissue sample. The double-stranded duplexes are treated with an agent that cleaves single-stranded regions of the duplex such as which will exist due to basepair mismatches between the control and sample strands. For instance, RNA/DNA duplexes can be treated with RNase and DNA/DNA hybrids treated with S1 nuclease to enzymatically digesting the mismatched regions. In other embodiments, either DNA/DNA or RNA/DNA duplexes can be treated with hydroxylamine or osmium tetroxide and with piperidine in order to digest mismatched regions. After digestion of the mismatched regions, the resulting material is then separated by size on denaturing polyacrylamide gels to determine the site of mutation. See, e.g., Cotton, et al., 1988. Proc. Natl. Acad. Sci. USA 85: 4397; Saleeba, et al., 1992. Methods Enzymol. 217: 286-295. In an embodiment, the control DNA or RNA can be labeled for detection.
[0274] In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called “DNA mismatch repair” enzymes) in defined systems for detecting and mapping point mutations in NOVX cDNAs obtained from samples of cells. For example, the mutY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches. See, e.g., Hsu, et al., 1994. Carcinogenesis 15: 1657-1662. According to an exemplary embodiment, a probe based on an NOVX sequence, e.g. a wild-type NOVX sequence, is hybridized to a cDNA or other DNA product from a test cell(s). The duplex is treated with a DNA mismatch repair enzyme, and the cleavage products, if any, can be detected from electrophoresis protocols or the like. See, e.g., U.S. Pat. No. 5,459,039.
[0275] In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in NOVX genes. For example, single strand conformation polymorphism (SSCP) may be used to detect differences in electrophoretic mobility between mutant and wild type nucleic acids. See, e.g., Orita, et al., 1989. Proc. Natl. Acad. Sci. USA: 86: 2766; Cotton, 1993. Mutat. Res. 285: 125-144; Hayashi, 1992. Genet. Anal. Tech. Appl. 9: 73-79. Single-stranded DNA fragments of sample and control NOVX nucleic acids will be denatured and allowed to renature. The secondary structure of single-stranded nucleic acids varies according to sequence, the resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments may be labeled or detected with labeled probes. The sensitivity of the assay may be enhanced by using RNA (rather than DNA), in which the secondary structure is more sensitive to a change in sequence. In one embodiment, the subject method utilizes heteroduplex analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility. See, e.g., Keen, et al., 1991. Trends Genet. 7: 5.
[0276] In yet another embodiment, the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGE). See, e.g., Myers, et al., 1985. Nature 313: 495. When DGGE is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA. See, e.g., Rosenbaum and Reissner, 1987. Biophys. Chem. 265: 12753.
[0277] Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension. For example, oligonucleotide primers may be prepared in which the known mutation is placed centrally and then hybridized to target DNA under conditions that permit hybridization only if a perfect match is found. See, e.g., Saiki, et al., 1986. Nature 324: 163; Saiki, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 6230. Such allele specific oligonucleotides are hybridized to PCR amplified target DNA or a number of different mutations when the oligonucleotides are attached to the hybridizing membrane and hybridized with labeled target DNA.
[0278] Alternatively, allele specific amplification technology that depends on selective PCR amplification may be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification may carry the mutation of interest in the center of the molecule (so that amplification depends on differential hybridization; see, e.g., Gibbs, et al., 1989. Nucl. Acids Res. 17: 2437-2448) or at the extreme 3′-terminus of one primer where, under appropriate conditions, mismatch can prevent, or reduce polymerase extension (see, e.g., Prossner, 1993. Tibtech. 11: 238). In addition it may be desirable to introduce a novel restriction site in the region of the mutation to create cleavage-based detection. See, e.g., Gasparini, et al., 1992. Mol. Cell Probes 6: 1. It is anticipated that in certain embodiments amplification may also be performed using Taq ligase for amplification. See, e.g. Barany, 1991. Proc. Natl. Acad. Sci. USA 88: 189. In such cases, ligation will occur only if there is a perfect match at the 3′-terminus of the 5′ sequence, making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification.
[0279] The methods described herein may be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one probe nucleic acid or antibody reagent described herein, which may be conveniently used, e.g., in clinical settings to diagnose patients exhibiting symptoms or family history of a disease or illness involving an NOVX gene.
[0280] Furthermore, any cell type or tissue, preferably peripheral blood leukocytes, in which NOVX is expressed may be utilized in the prognostic assays described herein. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.
[0281] Pharmacogenomics
[0282] Agents, or modulators that have a stimulatory or inhibitory effect on NOVX activity (e.g., NOVX gene expression), as identified by a screening assay described herein can be administered to individuals to treat (prophylactically or therapeutically) disorders (The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.) In conjunction with such treatment, the pharmacogenomics (i.e., the study of the relationship between an individual's genotype and that individual's response to a foreign compound or drug) of the individual may be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Thus, the pharmacogenomics of the individual permits the selection of effective agents (e.g., drugs) for prophylactic or therapeutic treatments based on a consideration of the individual's genotype. Such pharmacogenomics can further be used to determine appropriate dosages and therapeutic regimens. Accordingly, the activity of NOVX protein, expression of NOVX nucleic acid, or mutation content of NOVX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual.
[0283] Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See e.g., Eichelbaum, 1996. Clin. Exp. Pharmacol. Physiol., 23: 983-985; Linder, 1997. Clin. Chem., 43: 254-266. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare defects or as polymorphisms. For example, glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited enzymopathy in which the main clinical complication is hemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.
[0284] As an illustrative embodiment, the activity of drug metabolizing enzymes is a major determinant of both the intensity and duration of drug action. The discovery of genetic polymorphisms of drug metabolizing enzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome PREGNANCY ZONE PROTEIN PRECURSOR enzymes CYP2D6 and CYP2C 19) has provided an explanation as to why some patients do not obtain the expected drug effects or show exaggerated drug response and serious toxicity after taking the standard and safe dose of a drug. These polymorphisms are expressed in two phenotypes in the population, the extensive metabolizer (EM) and poor metabolizer (PM). The prevalence of PM is different among different populations. For example, the gene coding for CYP2D6 is highly polymorphic and several mutations have been identified in PM, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2Cl9 quite frequently experience exaggerated drug response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, PM show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite morphine. At the other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.
[0285] Thus, the activity of NOVX protein, expression of NOVX nucleic acid, or mutation content of NOVX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual. In addition, pharmacogenetic studies can be used to apply genotyping of polymorphic alleles encoding drug-metabolizing enzymes to the identification of an individual's drug responsiveness phenotype. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with an NOVX modulator, such as a modulator identified by one of the exemplary screening assays described herein.
[0286] Monitoring of Effects During Clinical Trials
[0287] Monitoring the influence of agents (e.g., drugs, compounds) on the expression or activity of NOVX (e.g., the ability to modulate aberrant cell proliferation and/or differentiation) can be applied not only in basic drug screening, but also in clinical trials. For example, the effectiveness of an agent determined by a screening assay as described herein to increase NOVX gene expression, protein levels, or upregulate NOVX activity, can be monitored in clinical trails of subjects exhibiting decreased NOVX gene expression, protein levels, or downregulated NOVX activity. Alternatively, the effectiveness of an agent determined by a screening assay to decrease NOVX gene expression, protein levels, or downregulate NOVX activity, can be monitored in clinical trails of subjects exhibiting increased NOVX gene expression, protein levels, or upregulated NOVX activity. In such clinical trials, the expression or activity of NOVX and, preferably, other genes that have been implicated in, for example, a cellular proliferation or immune disorder can be used as a “read out” or markers of the immune responsiveness of a particular cell.
[0288] By way of example, and not of limitation, genes, including NOVX, that are modulated in cells by treatment with an agent (e.g., compound, drug or small molecule) that modulates NOVX activity (e.g., identified in a screening assay as described herein) can be identified. Thus, to study the effect of agents on cellular proliferation disorders, for example, in a clinical trial, cells can be isolated and RNA prepared and analyzed for the levels of expression of NOVX and other genes implicated in the disorder. The levels of gene expression (i.e., a gene expression pattern) can be quantified by Northern blot analysis or RT-PCR, as described herein, or alternatively by measuring the amount of protein produced, by one of the methods as described herein, or by measuring the levels of activity of NOVX or other genes. In this manner, the gene expression pattern can serve as a marker, indicative of the physiological response of the cells to the agent. Accordingly, this response state may be determined before, and at various points during, treatment of the individual with the agent.
[0289] In one embodiment, the invention provides a method for monitoring the effectiveness of treatment of a subject with an agent (e.g., an agonist, antagonist, protein, peptide, peptidomimetic, nucleic acid, small molecule, or other drug candidate identified by the screening assays described herein) comprising the steps of (i) obtaining a pre-administration sample from a subject prior to administration of the agent; (ii) detecting the level of expression of an NOVX protein, mRNA, or genomic DNA in the preadministration sample; (iii) obtaining one or more post-administration samples from the subject; (iv) detecting the level of expression or activity of the NOVX protein, mRNA, or genomic DNA in the post-administration samples; (v) comparing the level of expression or activity of the NOVX protein, mRNA, or genomic DNA in the pre-administration sample with the NOVX protein, mRNA, or genomic DNA in the post administration sample or samples; and (vi) altering the administration of the agent to the subject accordingly. For example, increased administration of the agent may be desirable to increase the expression or activity of NOVX to higher levels than detected, ie., to increase the effectiveness of the agent. Alternatively, decreased administration of the agent may be desirable to decrease expression or activity of NOVX to lower levels than detected, i.e., to decrease the effectiveness of the agent.
[0290] Methods of Treatment
[0291] The invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disorder or having a disorder associated with aberrant NOVX expression or activity. The disorders include cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, adrenoleukodystrophy, congenital adrenal hyperplasia, prostate cancer, neoplasm; adenocarcinoma, lymphoma, uterus cancer, fertility, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, immunodeficiencies, graft versus host disease, AIDS, bronchial asthma, Crohn's disease; multiple sclerosis, treatment of Albright Hereditary Ostoeodystrophy, and other diseases, disorders and conditions of the like.
[0292] These methods of treatment will be discussed more fully, below.
[0293] Disease and Disorders
[0294] Diseases and disorders that are characterized by increased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that antagonize (i.e., reduce or inhibit) activity. Therapeutics that antagonize activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to: (i) an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof, (ii) antibodies to an aforementioned peptide; (iii) nucleic acids encoding an aforementioned peptide; (iv) administration of antisense nucleic acid and nucleic acids that are “dysfunctional” (i.e., due to a heterologous insertion within the coding sequences of coding sequences to an aforementioned peptide) that are utilized to “knockout” endogenous function of an aforementioned peptide by homologous recombination (see, e.g., Capecchi, 1989. Science 244: 1288-1292); or (v) modulators (i.e., inhibitors, agonists and antagonists, including additional peptide mimetic of the invention or antibodies specific to a peptide of the invention) that alter the interaction between an aforementioned peptide and its binding partner.
[0295] Diseases and disorders that are characterized by decreased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that increase (i.e., are agonists to) activity. Therapeutics that upregulate activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to, an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof; or an agonist that increases bioavailability.
[0296] Increased or decreased levels can be readily detected by quantifying peptide and/or RNA, by obtaining a patient tissue sample (e.g. from biopsy tissue) and assaying it in vitro for RNA or peptide levels, structure and/or activity of the expressed peptides (or mRNAs of an aforementioned peptide). Methods that are well-known within the art include, but are not limited to, immunoassays (e.g., by Western blot analysis, immunoprecipitation followed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, immunocytochemistry, etc.) and/or hybridization assays to detect expression of mRNAs (e.g., Northern assays, dot blots, in situ hybridization, and the like).
[0297] Prophylactic Methods
[0298] In one aspect, the invention provides a method for preventing, in a subject, a disease or condition associated with an aberrant NOVX expression or activity, by administering to the subject an agent that modulates NOVX expression or at least one NOVX activity. Subjects at risk for a disease that is caused or contributed to by aberrant NOVX expression or activity can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of the NOVX aberrancy, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending upon the type of NOVX aberrancy, for example, an NOVX agonist or NOVX antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein. The prophylactic methods of the invention are further discussed in the following subsections.
[0299] Therapeutic Methods
[0300] Another aspect of the invention pertains to methods of modulating NOVX expression or activity for therapeutic purposes. The modulatory method of the invention involves contacting a cell with an agent that modulates one or more of the activities of NOVX protein activity associated with the cell. An agent that modulates NOVX protein activity can be an agent as described herein, such as a nucleic acid or a protein, a naturally-occurring cognate ligand of an NOVX protein, a peptide, an NOVX peptidomimetic, or other small molecule. In one embodiment, the agent stimulates one or more NOVX protein activity. Examples of such stimulatory agents include active NOVX protein and a nucleic acid molecule encoding NOVX that has been introduced into the cell. In another embodiment, the agent inhibits one or more NOVX protein activity. Examples of such inhibitory agents include antisense NOVX nucleic acid molecules and anti-NOVX antibodies. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, the invention provides methods of treating an individual afflicted with a disease or disorder characterized by aberrant expression or activity of an NOVX protein or nucleic acid molecule. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., up-regulates or down-regulates) NOVX expression or activity. In another embodiment, the method involves administering an NOVX protein or nucleic acid molecule as therapy to compensate for reduced or aberrant NOVX expression or activity.
[0301] Stimulation of NOVX activity is desirable in situations in which NOVX is abnormally downregulated and/or in which increased NOVX activity is likely to have a beneficial effect. One example of such a situation is where a subject has a disorder characterized by aberrant cell proliferation and/or differentiation (e.g., cancer or immune associated disorders). Another Example of such a situation is where the subject has a gestational disease (e.g., preclampsia).
[0302] Determination of the Biological Effect of the Therapeutic
[0303] In various embodiments of the invention, suitable in vitro or in vivo assays are performed to determine the effect of a specific Therapeutic and whether its administration is indicated for treatment of the affected tissue.
[0304] In various specific embodiments, in vitro assays may be performed with representative cells of the type(s) involved in the patient's disorder, to determine if a given Therapeutic exerts the desired effect upon the cell type(s). Compounds for use in therapy may be tested in suitable animal model systems including, but not limited to rats, mice, chicken, cows, monkeys, rabbits, and the like, prior to testing in human subjects. Similarly, for in vivo testing, any of the animal model system known in the art may be used prior to administration to human subjects.
[0305] Prophylactic and Therapeutic Uses of the Compositions of the Invention
[0306] The NOVX nucleic acids and proteins of the invention are useful in potential prophylactic and therapeutic applications implicated in a variety of disorders including, but not limited to: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.
[0307] As an example, a cDNA encoding the NOVX protein of the invention may be useful in gene therapy, and the protein may be useful when administered to a subject in need thereof. By way of non-limiting example, the compositions of the invention will have efficacy for treatment of patients suffering from: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyslipidemias.
[0308] Both the novel nucleic acid encoding the NOVX protein, and the NOVX protein of the invention, or fragments thereof, may also be useful in diagnostic applications, wherein the presence or amount of the nucleic acid or the protein are to be assessed. A further use could be as an anti-bacterial molecule (i.e., some peptides have been found to possess anti-bacterial properties). These materials are further useful in the generation of antibodies, which immunospecifically-bind to the novel substances of the invention for use in therapeutic or diagnostic methods.
[0309] Sequence Analyses
[0310] The sequence of NOVX was derived by laboratory cloning of cDNA fragments, by in silico prediction of the sequence. cDNA fragments covering either the full length of the DNA sequence, or part of the sequence, or both, were cloned. In silico prediction was based on sequences available in CuraGen's proprietary sequence databases or in the public human sequence databases, and provided either the full length DNA sequence, or some portion thereof.
[0311] The laboratory cloning was performed using one or more of the methods summarized below:
[0312] SeqCalling™ Technology: cDNA was derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then sequenced using CuraGen Corporation's SeqCalling technology which is disclosed in fuill in U.S. Ser. No. 09/417,386 filed Oct. 13, 1999, and Ser. No. 09/614,505 filed Jul. 11, 2000. Sequence traces were evaluated manually and edited for corrections if appropriate. cDNA sequences from all samples were assembled together, sometimes including public human sequences, using bioinformatics programs to produce a consensus sequence for each assembly. Each assembly is included in CuraGen Corporation's database. Sequences were included as components for assembly when the extent of identity with another component was at least 95% over 50 bp. Each assembly represents a gene or portion thereof and includes information on variants, such as splice forms single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations.
[0313] Variant sequences are also included in this application. A variant sequence can include a single nucleotide polymorphism (SNP). A SNP can, in some instances, be referred to as a “cSNP” to denote that the nucleotide sequence containing the SNP originates as a cDNA. A SNP can arise in several ways. For example, a SNP may be due to a substitution of one nucleotide for another at the polymorphic site. Such a substitution can be either a transition or a transversion. A SNP can also arise from a deletion of a nucleotide or an insertion of a nucleotide, relative to a reference allele. In this case, the polymorphic site is a site at which one allele bears a gap with respect to a particular nucleotide in another allele. SNPs occurring within genes may result in an alteration of the amino acid encoded by the gene at the position of the SNP. Intragenic SNPs may also be silent, when a codon including a SNP encodes the same amino acid as a result of the redundancy of the genetic code. SNPs occurring outside the region of a gene, or in an intron within a gene, do not result in changes in any amino acid sequence of a protein but may result in altered regulation of the expression pattern. Examples include alteration in temporal expression, physiological response regulation, cell type expression regulation, intensity of expression, and stability of transcribed message.
[0314] Presented information includes that associated with genomic clones, public genes and ESTs sharing sequence identity with the disclosed sequence and CuraGen Corporation's Electronic Northern bioinformatic tool.
Example 1
[0315] The NOV1 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 1A.
2TABLE 1A
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NOV1 Sequence Analysis
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SEQ ID NO:11173 bp
NOV1a,AATATAGCCGGTCCTGTCCCCGTGTTAACTGGAGTGCTGAAGCGCTGGAACGAAATGC
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CG55912-01 DNA SequenceGGGGCCTCTGGTGCGAGAAGGGGGTGCAGGTGCTGCTGACGACGGTGGGCGCCTTCGC
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CGCCTTCGGCCTCATGACCATCGCCATCAGCACTGACTACTGGCTCTACACGGGGCAA
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CAAGAGCGAAAATCTGTCTCAAAAAATAAAAGAAGTAAGAAGGACCCCGGCGGCCTCA
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CGCACTCGGCCCTCTGGAGGATCTGCTGCCTGGAAGGGTTGAAAAGAGGCGTCTGCGT
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GAAGATCAATCATTTCCCGGAGGACACGGACTACGACCACGACAGCGCGGAGTATCTA
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CTCCGTACGGTCCGGGCCTCCAGCATCTTCCCCATCCTTAGCGCCATCCTGCTGCTGC
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TCGGGGGTGTGTGCGTGGCGGCCTCCCGCGTCTACAAGTCCAAGAGGAACATCATTCT
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GGGCGCAGGGATCCTGTTCGTGGCAGCAGGTCTGAGCAACATCATCGGCGTGATCGTG
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TACATCTCCGCCAACGCGGGCGAGCCGGGCCGAAGCAGAGCCAAGAAAAACCACTACT
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CGTACGGCTGGTCCTTCTACTTCGGCGGGCTGTCGTTCATCCTGGCCGAGGTGATAGG
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CGTGCTGGCCGTCAACATCTACATCGAGCCAGCCGAGGCGCACTGCCAGTCTCGGAGC
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GGGACCGCGGGGGGTCGTCCGGCTTCCTCACGCTGCACAACGCCTTCCCCAAGGAGGC
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GGGCGGCGGCGTCACGGTCACGTTCACCCGGCCGCCCGCCCCGCCCGCCCCACGCCAC
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CCCGGCCAACACCAACTCCACGGACATCTCCATGTACACGCTCAGCCGCGACCCCTCC
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AAGGGCAGCCCCCATTCCAATGCCACCACCCCCACCCCCACTAGCCTCAAGGATAGGA
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AAAACTCATTTGTGTCCATAAAAATAAAGGTAAAAAAGAAAAAAAGAAATATATATAT
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ATATATATATACGCTCAACAGGAAAACCACGCCTGTGTAGGGGCGCGGCGGGGGAGCC
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GAGGGGCGTGTCCGCGGCGCGTGCGGGCGCGCGTGCATCGAGGCTGCCOGGGTCGGGG
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GCGCCCCCGCTTTCCCCCGTGAGCGCGCTGGAGACTGCTGGGCCCGCCCCACGCCCAC
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CCTCCCCGCCCCC
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ORF Start: ATG at 55ORF Stop: TAG at 1024
SEQ ID NO:2323 aa MW at 35342.3 kD
NOV1a,MRGLWCEKGVQVLLTTVGAFAAFGLMTIAISTDYWLYTCQQERKSVSKNKRSKKDPGG
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CG55912-01 Protein SequenceLTHSGLWRICCLEGLKRGVCVKINHFPEDTDYDHDSAEYLLRTVRASSIFPILSAILL
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LLGGVCVAASRVYKSKRNIILGAGILPVAAGLSNIIGVIVYTSANAGEPGRSRAKKNH
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YSYGWSFYFGGLSFILAEVIGVLAVNIYIEPAEAHCQSRSGTAGGRPASSRCTTPSPR
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RRAAASRSRSPGRPPRPREATPANTNSTDISMYTLSRDPSKCSPHSNATTPTPTSLKD
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RKNSFVSIKIKVKKKRNIYIYIYTLNRKTTPV
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[0316] Further analysis of the NOV1 a protein yielded the following properties shown in Table 1B.
3TABLE 1B
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Protein Sequence Properties NOV1a
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PSort0.6400 probability located in plasma membrane;
analysis:0.4600 probability located in Golgi body;
0.3700 probability located in endoplasmic
reticulum (membrane);
0.1000 probability located in endoplasmic reticulum (lumen)
SignalPCleavage site between residues 22 and 23
analysis:
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[0317] A search of the NOV la protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 1C.
4TABLE 1C
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Geneseq Results for NOV1a
NOV1aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAY70462Human membrane channel protein-127 . . . 323183/327 (55%)5e−93
(MECHP-12)-Homo sapiens, 323 aa.5 . . . 323233/327 (70%)
[WO20001271l-A2, 09-MAR-2000]
ABB11805Human voltage gated Ca channel7 . . . 323183/327 (55%)le−92
subunit homologue, SEQ ID NO: 2175-7 . . . 325233/327 (70%)
Homo sapiens, 325 aa.
[WO200157188-A2, 09-AUG-2001]
AAY84376A human voltage-gated calcium7 . . . 323183/327 (55%)le−92
channel designated CACNGLIKE1-5 . . . 323233/327 (70%)
Homo sapiens, 323 aa.
[WO200014223-A1, 16-MAR-2000]
AAY84374A human a neuronal voltage-gated6 . . . 323173/324 (53%)9e−86
calcium chanel polypeptide-Homo4 . . . 315217/324 (66%)
sapiens, 315 aa. [WO200014225-A1,
16-MAR-2000]
AAB43007Human ORFX ORF2771 polypeptide6 . . . 323173/324 (53%)9e−86
sequence SEQ ID NO: 5542-Homo4 . . . 315217/324 (66%)
sapiens, 315 aa. [WO200058473-A2,
05-OCT-2000]
|
[0318] In a BLAST search of public sequence datbases, the NOV1a protein was found to have homology to the proteins shown in the BLASTP data in Table 1D.
5TABLE 1D
|
|
Public BLASTP Results for NOV1a
NOV1aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
AAL50041VOLTAGE-DEPENDENT 2 . . . 275226/309 (73%) e−113
CALCIUM CHANNEL GAMMA-811 . . . 317234/309 (75%)
SUBUNIT-Rattus norvegicus (Rat),
421 aa.
AAL50045VOLTAGE-DEPENDENT 2 . . . 275225/309 (72%) e−113
CALCIUM CHANNEL GAMMA-811 . . . 317234/309 (74%)
SUBUNIT-Mus musculus (Mouse).
423 aa.
AAL50049VOLTAGE-DEPENDENT 2 . . . 275227/309 (73%) e−112
CALCIUM CHANNEL GAMMA-811 . . . 318234/309 (75%)
SUBUNIT-Homo sapiens (Human),
426 aa.
Q9BXT0CALCIUM CHANNEL GAMMA 3 . . . 275228/309 (73%) e−112
SUBUNIT 8-Homo sapiens (Human), 1 . . . 306235/309 (75%)
414 aa (fragment).
Q9Y698Voltage-dependent calcium channel 7 . . . 323183/327 (55%)3e−92
gamma-2 subunit (Neuronal voltage- 5 . . . 323233/327 (70%)
gated calcium channel gamma-2
subunit)-Homo sapiens (Human), 323
aa.
|
[0319] PFam analysis predicts that the NOV1a protein contains the domains shown in the
6TABLE 1E
|
|
Domain Analysis of NOV1a
Identities/
Similarities
NOV1a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
PMP22_Claudin: domain 18 . . . 198 51/198 (26%)7.6e−53
of 1161/198 (81%)
|
Example 2
[0320] The NOV2 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 2A.
7TABLE 2A
|
|
NOV2 Sequence Analysis
|
|
SEQ ID NO:31116 bp
NOV2a,AAATGGGAGCCAGGTCATACACGGGATCTCTGTGGCAGGAACGGGCTGGCTGGATTCC
|
CG55918-01 DNA SequenceTCTGCCCCTGGACTTGCAGGCCATTGAGCTGGCTGCCCAGAGCAACCATCACTGCCAT
|
GCTCAGAAGGGTCCTGACAGTCACTGTGACCCCAAGAAGGGGAAGGCCCAGCGCCAGC
|
TGTATGTAGCCTCTGCCATCTGCCTGTTGTTCATGATCGGAGAAGTCGTTGGTGGGTA
|
CCTGGCACACAGCTTGGCTGTCATGACTGACGCAGCACACCTGCTCACTGACTTTGCC
|
AGCATGCTCATCAGCCTCTTCTCCCTCTGGATGTCCTCCCGGCCAGCCACCAAGACCA
|
TGAACTTTGGCTGGCAGAGAGCTGAGATCTTGGGAGCCCTGGTCTCTGTACTGTCCAT
|
CTGGGTCGTGACGGGGGTACTGGTGTACCTGGCTGTGGAGCGGCTGATCTCTGGGGAC
|
TATGAAATTGACGGGGGGACCATGCTGATCACGTCGGGCTGCGCTGTGGCTGTGAACA
|
TCATGATGGGGTTGACCCTTCACCAGTCTGGCCATGGGCACAGCCACCGCACCACCAA
|
CCAGCAGGAGGAGAACCCCAGCGTCCGAGCTGCCTTCATCCATGTGATCGGCGACTTT
|
ATGCAGAGCATGGGTGTCCTAGTGGCAGCCTATATTTTATACTTCAAGCCAGAATACA
|
AGTATGTAGACCCCATCTGCACCTTCGTCTTCTCCATCCTGGTCCTGGGGACAACCTT
|
GACCATCCTGAGAGATGTGATCCTGGTGTTGATGGAAGGTACCCCCAAGGGCGTTGAC
|
TTCACAGCTGTTCGTCATCTGCTGCTGTCGGTGGAGGGGGTAGAAGCCCTGCACAGCC
|
TGCATATCTGGGCACTGACGGTGGCCCAGCCTGTTCTGTCTGTCCACATCGCCATTGC
|
TCAGAATACAGACGCCCAGGCTGTGCTGAAGACAGCCAGCAGCCGCCTCCAAGGGAAG
|
TTCCACTTCCACACCGTGACCATCCAGATCGAGGACTACTCGGAGGACATGAAGGACT
|
GTCAGGCATGCCAGGGCCCCTCAGACTGACTGCTCAGCCAGCCACCAACTGGGGCATG
|
AACAGGACCTGCAG
|
ORF Start: ATG at 3ORF Stop: TGA at 1071
SEQ ID NO:4356 aa MW at 38698.4 kD
NOV2a,MGARSYTGSLWQEGAGWIPLPLDLQAIELAAQSNHHCHAQKGPDSHCDPKKGKAQRQL
|
CG55918-01 Protein SequenceYVASAICLLFMIGEVVGGYLAHSLAVMTDAAHLLTDFASMLISLFSLWMSSRPATKTM
|
NFGWQRAEILGALVSVLSIWVVTGVLVYLAVERLISGDYEIDGGTMLITSGCAVAVNI
|
MMGLTLHQSGHGHSHGTTNQQEENPSVRAAFIHVIGDFMQSMGVLVAAYILYFKPEYK
|
YVDPICTFVFSILVLGTTLTILRDVILVLMEGTPKGVDFTAVRDLLLSVEGVEALHSL
|
HIWALTVAQPVLSVHIAIAQNTDAQAVLKTASSRLQGKFHFHTVTIQIEDYSEDMKDC
|
QACQGPSD
|
SEQ ID NO:511154 bp
NOV2b,AAATGGGAGCCAGGTCATACACGGGATCTCTGTGGCAGGAAGGGGCTGGCTGGATTCC
|
CG55918-02 DNA SequenceTCTGCCCCGACCTGGCCTGGACTTGCAGGCCATTGAGCTGGCTGCCCAGAGCAACCAT
|
CACTGCCATGCTCAGAAGGGTCCTGACAGTCACTGTGACCCCAAGAAGGGGAAGGCCC
|
AGCGCCAGCTGTATGTAGCCTCTGCCATCTGCCTGTTGTTCATGATCGGAGAAGTCGT
|
TGATGATGAAACTGAGGCACGTTCAGGTGGGTACCTGGCACACAGCTTGGCTGTCATG
|
ACTGACGCAGCACACCTGCTCACTGACTTTGCCAGCATGCTCATCAGCCTCTTCTCCC
|
TCTGGATGTCCTCCCGGCCAGCCACCAAGACCATGAACTTTGGCTGGCAGAGAGCTGA
|
GATCTTGGGAGCCCTGGTCTCTGTACTGTCCATCTGGGTCGTGACGGGGGTACTGGTG
|
TACCTGGCTGTGGAGCGGCTGATCTCTGGGGACTATGAAATTGACGGGGGGACCATGC
|
TGATCACGTCGGGCTGCGCTGTGGCTGTGAACATCATAATGCGGTTGACCCTTCACCA
|
GTCTGGCCATGGGCACAGCCACGGCACCACCAACCAGCAGGAGGAGAACCCCAGCGTC
|
CGAGCTGCCTTCATCCATGTGATCGGCGACTTTATGCAGAGCATGGGTGTCCTAGTGG
|
CAGCCTATATTTTATACTTCAAGCCAGAATACAAGTATGTAGACCCCATCTGCACCTT
|
CGTCTTCTCCATCCTGGTCCTGGGGACAACCTTGACCATCCTGAGAGATGTGATCCTG
|
GTGTTGATGGAAGGGACCCCCAAGGGCGTTGACTTCACAGCTGTTCGTGATCTGCTGC
|
TGTCGGTGGAGGGGGTAGAAGCCCTGCACAGCCTGCATATCTGGGCACTGACGGTGGC
|
CCAGCCTGTTCTGTCTGTCCACATCGCCATTGGCCCCCAGCTCAGAATACAGACGCCC
|
AGGCTGTGCTGAAGACAGCCAGCAGCCGCCTCCAAGGGAAGTTCCACTTCCACACCGT
|
GACCATCCAGATCGAGGACTACTCGGAGGACATGAAGGACTGTCAGGCATGCCAGGGC
|
CCCTCAGACTGACTGCTCAGCCAGGCACCAACTGGGGCATGAACAGGACCTG
|
ORF Start: ATG at 3ORF Stop: at 9721
SEQ ID NO:6323 aa MW at 35057.5 kD
NOV2b,MGARSYTGSLWQEGAGWIPLPRPGLDLQAIELAAQSNHHCHAQKGPDSHCDPKKGKAQ
|
CG55918-02 Protien SequenceRQLYVASAICLLFMIGEVVGGYLAHSLAVMTDAAHLLTDFASMLISLFSLWMSSRPAT
|
KTMNFGWQRAEILGALVSVLSIWVVTGVLVYLAVERLISGDYEIDGGTMLITSGCAVA
|
VNIIMGLTLHQSCHGHSHGTTNQQEENPSVRAAFIHVIGDFMQSMGVLVAAYILYFKP
|
EYKYVDPICTFVFSILVLGTTLTILRDVILVLMEGTPKGVDFTAVRDLLLSVECVEAL
|
HSLHIWALTVAQPVLSVHIAIGPQLRIQTPRLC
|
[0321] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 2B.
8TABLE 2B
|
|
Comparison of NOV2a against NOV2b.
Identities/
NOV2a Residues/Similarities for
Protein SequenceMatch Residuesthe Matched Region
|
NOV2b1 . . . 308288/311 (92%)
1 . . . 311289/311 (92%)
|
[0322] Further analysis of the NOV2a protein yielded the following properties shown in Table 2C.
9TABLE 2C
|
|
Protein Sequence Properties NOV2a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body;
0.3000 probability located in endoplasmic
reticulum (membrane); 0.3000 probability
located in microbody (peroxisome)
SignalPCleavage site between residues 40 and 41
analysis:
|
[0323] A search of the NOV2a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 2D.
10TABLE 2D
|
|
Geneseq Results for NOV2a
NOV2aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAB60094Human transport protein TPPT-14-35 . . . 356171/322 (53%)6e−97
Homo sapiens, 320 aa. 2 . . . 320232/322 (71%)
[WO200078953-A2, 28-DEC-2000]
AAG31823Arabidopsis thaliana protein fragment46 . . . 339105/331 (31%)1e−46
SEQ ID NO: 38281-Arabidopsis27 . . . 357180/331 (53%)
thaliana, 359 aa. [EP1033405-A2, 06-
SEP-2000]
AAG31822Arabidopsis thaliana protein fragment46 . . . 339105/331 (31%)1e−46
SEQ ID NO: 38280-Arabidopsis43 . . . 373180/331 (53%)
thaliana, 375 aa. [EP1033405-A2, 06-
SEP-2000]
AAG31821Arabidopsis thaliana protein fragment46 . . . 339105/331 (31%)1e−46
SEQ ID NO: 38279-Arabidopsis53 . . . 383180/331 (53%)
thaliana, 385 aa. [EP1033405-A2, 06-
SEP-2000]
ABB53142Human 0RF48 protein-Homo35 . . . 176 84/142 (59%)7e−42
sapiens , 144 aa. [WO200177155-A2, 1 . . . 142105/142 (73%)
18-OCT-2001]
|
[0324] In a BLAST search of public sequence datbases, the NOV2a protein was found to have homology to the proteins shown in the BLASTP data in Table 2E.
11TABLE 2E
|
|
Public BLASTP Results for NOV2a
NOV2aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q62941Zinc transporter 2 (ZnT-2)-Rattus 1 . . . 356290/360 (80%) e−167
norvegicus (Rat), 359 aa. 1 . . . 359327/360 (90%)
Q9BRI3SIMILAR TO ZINK 4 . . . 356303/356 (85%) e−166
TRANSPORTER 2-Homo17 . . . 323304/356 (85%)
sapiens (Human), 323 aa.
P97441Zinc transporter 3 (ZnT-3)-Mus19 . . . 355193/350 (55%) e−107
musculus (Mouse), 388 aa.40 . . . 386242/350 (69%)
Q99726Zinc transporter 3 (ZnT-3)-Homo19 . . . 354189/349 (54%) e−105
sapiens (Human), 388 aa.40 . . . 385239/349 (68%)
CAD28545HYPOTHETICAL 35.1 KDA35 . . . 356170/322 (52%)2e−95
PROTEIN-Homo sapiens 2 . . . 320230/322 (70%)
(Human), 320 aa.
|
[0325] PFam analysis predicts that the NOV2a protein contains the domains shown in the Table 2F.
12TABLE 2F
|
|
Domain Analysis of NOV2a
Identities/
NOV2aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Cation_efflux: domain 158 . . . 345105/322 (33%)4.1e−109
of 1257/322 (80%)
|
Example 3
[0326] The NOV3 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 3A.
13TABLE 3A
|
|
NOV3 Sequence Analysis
|
|
SEQ ID NO:71814 bp
NOVa3,TCACCTCTGACCCGCAGCACTTCGTGGCCCTGGGGGCCGGTCCATCATCAATGGGAAC
|
CG56641-01 DNA SequenceTGGGCTGTGGATCCCCCTGGGTCCTACAGGGCCGGCGGGACCGTCTTTCGATATAACC
|
GTCCTCCCAGGCAGGAGGGCAAAGGGGAGAGTCTGTCGGCTGAAGGCCCCACCACCCA
|
GCCTGTGGATGTCTATGTGATGATCTTTCAGGAGCAAAACCCAGGCGTTTTTTATCAG
|
TATGTCATCTCTTCACCTCCTCCAATCCTTGAGAACCCCACCCCAGAGCCCCCTGTCC
|
CCCAGCTTCACCCGGGTAACACTCTGACCCCTGCACTTGGAAGGAGGAGGGAGAGGCT
|
GCAGGGCTGGCTCGGGGCAGTGGGGTGGCATCTGATTCGCCTGCTCCCCTGCACAGAG
|
ATTCTGACGGTGGAGCCCCCACTTGCTCCGGCACCCCGCCCAGCCCGGACCCCAGCCA
|
CCCTCCAGCGTCAGGTGCGGATCCCCCAGATGCCCGCCCCGCCCCATCCCAGGACACC
|
CCTGGGGTCTCCAGCTGCGTACTGGAAACGAGTGCGACACTCTGCATGCTCAGCGTCC
|
TGCGCGAAAGGTTCCTCTGAGGCCCCCACGTCCAGTGTCTCTTCCATCTCTCCTGCCA
|
CGGCAGCCCCACACATCTCATCTATGTCTCCGCCTCTTCCCTGCCCTGCTGGTGCCTG
|
CAGCTGGGAGGCTGGCGAGTGGACATCCTCCAGCCGCTCCTGTGGCCCCGGCACCCAG
|
CACCGCCAGCTGCAGTCCCGGCAGGAATTTGGGGCGGGTGGCTCCTCGGTGCCCCCGG
|
AGCGCTGTGGACATCTCCCCCCGCCCAACATCACCCAGTCTTGCCAGCTGCGCCTCTG
|
TGGCCATTGGGAAGTTGGCTCTCCTTGGAGCCAGTGCTCCGTGCGGTGCGCCCGGGGC
|
CAGAGAAGCCGGCAGGTTCGCTGTGTTGGGAACAATGGTGATGAAGTGAGCGAGCAGG
|
AGTGTGCGTCAGGCCCCCCGCAGCCCCCCAGCAGAGAGGCCTGTCACATGGGGCCCTG
|
TACTACTGCCTGGTTCCACAGCGACTCGAGCTCCAAGGTGTCAGCCGAGTGTGGGACG
|
GGAATCCAGCGGCGCTCTGTGGTCTGCCTTGGGAGTGGGGCAGCCCTCGGGCCAGGCC
|
AGGGGGAAGCAGGAGCAGGAACTGGGCAGAGCTGTCCAACAGGAAGCCGGCCCCCTGA
|
CATGCGCGCCTGCAGCCTGGGGCCCTGTGAGAGAACTTGGCGCTGGTACACAGGGCCC
|
TGGGGTGAGTCCTCCTCCCAATGTGCCTCTGGCACACACCGTAGAGACATCATCTGTG
|
TATCCAAACTGGGGACGGAGTTCAACGTGACTTCTCCGAGCAACTGTTCTCACCTCCC
|
CAGGCCCCCTGCCCTGCAGCCCTGTCAAGGGCAGGCCTGCCAGGACCGATGGTTTTCC
|
ACGCCCTGGAGCCCATGTTCTCGCTCCTGCCAAGGGGGAACGCAGACACGGGAGGTCC
|
AGTGCCTGAGCACCAACCAGACCCTCACCACCCGATGCCCTCCTCAACTGCGGCCCTC
|
CAGGAAGCGCCCCTGTAACAGCCAACCCTGCGATGATCAATGCAAGGACAGCTCTCCA
|
CATTGCCCCCTGGTGGTACAGGCCCGGCTCTGCGTCTACCCCTACTACACAGCCACCT
|
GTTGCCGCTCTTGCGCACATGTCCTGGAGCGGTCTCCCCAGGATCCCTCCTGAAAGGG
|
GTCCGGGGCACCTTCACGGTTTTCTGTGCCACCATCGGTCACCCATTGATCGGCCCAC
|
TCTGAACCCCCTGCCT
|
ORF Start: ATG at 194ORF Stop: TGA at 1733
SEQ ID NO:8513 aa MW at 55199.7 kD
NOV3a,MIFQEEWPGVFYQYVISSPPPILENPTPEPPVPQLQPCKTLTPALGRRRSRLQCWLGA
|
CG56641-01 Protein SequenceVCWHLIRLLPCTEILRVEPPLAPAPRPARTPGTLQRQVRIPQMPAPPHPRTPLGSPAA
|
YWKRVCHSACSASCGKGSSEAPTSSVSSISAATAAPHISSMSPPLPCPAGACSWEAGE
|
WTSCSRSCGPGTQHRQLQCRQEFCGGGSSVPPERCGHLPRPNITQSCQLRLCGHWEVG
|
SPWSQCSVRCGRGQRSRQVRCVGNNGDEVSEQECASGPPQPPSREACDMGPCTTAWFH
|
SDWSSKVSAECGTGIQRRSVVCLGSGAALGPGQGEACAGTGQSCPTGSRPPDMPACSL
|
GPCERTWRWYTGPWGECSSECGSGTQRRDIICVSRLGTEFNVTSPSNCSHLPRPPALQ
|
PCQGQACQDRWFSTPWSPCSRSCQGCTQTREVQCLSTNQTLSTRCPPQLRPSRKRPCN
|
SQPCDDQCKDSSPHCPLVVQARLCVYPYYTATCCRSCAHVLERSPQDPS
|
[0327] Further analysis of the NOV3a protein yielded the following properties shown in Table 3B.
14TABLE 3B
|
|
Protein Sequence Properties NOV3a
|
|
PSort0.3000 probability located in microbody (peroxisome);
analysis:0.3000 probability located in nucleus;
0.1000 probability located in mitochondrial matrix space;
0.1000 probability located in lysosome (lumen)
SignalPCleavage site between residues 26 and 27
analysis:
|
[0328] A search of the NOV3a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 3C.
15TABLE 3C
|
|
Geneseq Results for NOV3a
NOV3aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAB50936ADAM protein #2-Homo sapiens, 1 . . . 513454/528 (85%)0.0
491 aa. [WO200073323-A2, 07-DEC- 1 . . . 491455/528 (85%)
2000]
AAE09698Human gene 9 encoding novel protein160 . . . 513347/360 (96%)0.0
HUCMO06, SEQ ID NO: 45-Homo 13 . . . 372347/360 (96%)
sapiens, 372 aa. [WO200155202-A1,
02-AUG-2001]
AAE09713Human gene 9 encoding novel protein160 . . . 403234/246 (95%)e−147
HUCMO06, SEQ ID NO: 60-Homo 13 . . . 258235/246 (95%)
sapiens, 260 aa. [WO200155202-A1,
02-AUG-2001]
AAU32725Novel human secreted protein #3216-229 . . . 500224/285 (78%)e−130
Homo sapiens, 416 aa. 9 . . . 293226/285 (78%)
[WO200179449-A2, 25-OCT-2001]
AAB94727Human protein sequence SEQ ID 1 . . . 507218/528 (41%)e−112
NO: 15753-Homo sapiens, 538 aa. 32 . . . 530287/528 (54%)
[EP1074617-A2, 07-FEB-2001]
|
[0329] In a BLAST search of public sequence datbases, the NOV3a protein was found to have homology to the proteins shown in the BLASTP data in Table 3D.
16TABLE 3D
|
|
Public BLASTP Results for NOV3a
NOV3aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9HBS6HYPOTHETICAL 25.7 KDA281 . . . 513232/237 (97%) e−146
PROTEIN-Homo sapiens 1 . . . 237232/237 (97%)
(Human), 237 aa.
Q91YP4SIMILAR TO HYPOTHETICAL281 . . . 513200/237 (84%) e−122
PROTEIN FLJ13710-Mus 1 . . . 235210/237 (88%)
musculus (Mouse), 235 aa.
Q9H8E4PLACE2000373 PROTEIN-Homo 1 . . . 507218/528 (41%) e-111
sapiens (Human), 538 aa. 32 . . . 530287/528 (54%)
Q96H81HYPOTHETICAL 51.1 KDA 1 . . . 423172/440 (39%)2e−81
PROTEIN-Homo sapiens 32 . . . 442232/440 (52%)
(Human), 454 aa.
Q9UFG7HYPOTHETICAL 12.5 KDA407 . . . 513107/111 (96%)2e−62
PROTEIN-Homo sapiens 1 . . . 111107/111 (96%)
(Human), 111 aa (fragment).
|
[0330] PFam analysis predicts that the NOV3a protein contains the domains shown in the Table 3E.
17TABLE 3E
|
|
Domain Analysis of NOV3a
Identities/
NOV3aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
tsp_1: domain 1 of 4170 . . . 22615/62 (24%)0.18
39/62 (63%)
tsp_1: domain 2 of 4230 . . . 28418/62 (29%)0.00028
41/62 (66%)
tsp_1: domain 3 of 4357 . . . 41310/62 (16%)1.4
35/62 (56%)
tsp_1: domain 4 of 4419 . . . 46814/55 (25%)0.0035
34/55 (62%)
|
Example 4
[0331] The NOV4 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 4A.
18TABLE 4A
|
|
NOV4 Sequence Analysis
|
|
SEQ ID NO:91396 bp
NOV4a,TTCTCCCTCCACTGCCAGACAATATCCCTGAGAGTGAGGAAGCAATGAGGATTGTTTG
|
CG56832-01 DNA SequenceTTTAGTGAAAAACCAACAGCCCCTGGGAGCCACCATCAAGCGCCACGAGATGACAGGG
|
GACATCTTGGTGGCCAGGATCATCCACGGTGGGCTGGCGGAGAGAAGTGGGTTGCTAT
|
ACAAGTGATCCATATTCTGGCCATGTCTCGAGGCACAATCATGTTCAAGGTGGTTCCA
|
GTCTCTGACCCTCCTGTGAATAGCCAGCAGATGGTGTACGTCCGTGCCATGACTGAGT
|
ACTGGCCCCAGGAGGATCCCGACATCCCCTGCATGGACGCTGGATTGCCTTTCCAGAA
|
GGGGCACATCCTCCAGATTGTGGACCAGAATGATGCCCTCTGGTGGCAGGCCCGAAAA
|
ATCTCAGACCCTGCTACCTGCGCTGGGCTTGTCCCTTCTAACCACCTTCTGAAGAGGA
|
AGCAACGGGAATTCTGGTGGTCTCAGCCGTACCAGCCTCACACCTGCCTCAAGTCAAC
|
CCTATACAAGGAGGAGTTTGTTGGCTACGGTCAGAAGTTCTTTATAGGTAGCCCGCTG
|
CATGCCAGTGTGTGCTGCACCGGCAGCTGCTACAGTGCAGTGGGTGCCCCTTACGAGG
|
AGGTGGTGAGGTACCAGCGACGCCCTTCAGACAAGTACCGCCTCATAGTGCTCATGGG
|
ACCCTCTGGTGTTGGAGTAAATGACCTCAGAAGACAACTTATTGAATTTAATCCCAGC
|
CATTTTCAAAGTGCTGTGCCACACACTACTCGTACTAAAAAGAGTTACGAAATGAATG
|
GGCGTGAGTATCACTATGTGTCCAAGGAAACATTTGAAAACCTCATATATAGTCACAG
|
GATGCTGGAGTATGGTGAGTACAAAGGCCACCTGTATGGCACTAGTGTGGATGCTGTT
|
CAAACAGTCCTTGTCGAAGGAAAGATCTGTGTCATGGACCTAGAGCCTCAGGATATTC
|
AAGGGGTTCGAACCCATGAACTGAAGCCCTATGTCATATTTATAAAGCCATCGAATAT
|
GAGGTGTATGAAACAATCTCGGAAAAATGCCAAGGTTATTACTGACTACTATGTGGAC
|
ATGAAGTTCAAGGATGAAGACCTACAAGAGATGGAAAATTTAGCCCAAAGAATGGAAA
|
CTCAGTTTGGCCAATTTTTTGATCATGTGATTGTGAATGACAGCTTGCACGATGCATG
|
TGCCCAGTTGTTGTCTGCCATACAGAAGGCTCAGGAGGAGCCTCAGTGGGTACCAGCA
|
ACATGGATTTCCTCAGATACTGAGTCTCAATGAGACTTCTTGTTTAATGCTGGAGTTT
|
TAAC
|
ORF Start: ATG at 45ORF Stop: TGA at 1365
SEQ ID NO:10440 aa MW at 50266.1 kD
NOV4a,MRIVCLVKNQQPLGATIKRHEMTGDILVARIIHGGLAERSGLLYAGDKLVEVNGVSVE
|
CG56832-01 Protein SequenceGLDPEQVIHILAMSRGTIMFKVVPVSDPPVNSQQMVYVRAMTEYWPQEDPDIPCMDAG
|
LPFQKGDILQIVDQNDALWWQARKISDPATCAGLVPSNHLLKRKQREFWWSQPYQPHT
|
CLKSTLYKEEFVGYGQKFFIGSPLHASVCCTGSCYSAVGAPYEEVVRYQRRPSDKYRL
|
IVLMGPSGVGVNELRRQLIEFNPSHFQSAVPHTTRTKKSYEMNGREYHYVSKETFENL
|
IYSHRMLEYGEYKGHLYGTSVDAVQTVLVEGKICVMDLEPQDIQGVRTHELKPYVIFI
|
KPSNMRCMKQSRKNAKVITDYYVDMKFKDEDLQEMENLAQRMETQFGQFFDHVIVNDS
|
LHDACAQLLSAIQKAQEEPQWVPATWISSDTESQ
|
SEQ ID NO:111371 bp
NOV4b,CAATGAGGATTGTTTGTTTAGTGAAAAACCAACAGCCCCTGGGAGCCACCATCAAGCG
|
CG56832-02 DNA SequenceCCACGAGATGACAGGGGACATCTTGGTGGCCAGGATCATCCACGGTGGGCTGGCGGAG
|
AGAAGTGGGTTGCTATATGCTGGAGACAAGCTGGTAGAAGTGAATGGAGTTTCAGTTG
|
AGGGACTGGACCCTGAACAAGTGATCCATATTCTCGCCATGTCTCGAGGCACAATCAT
|
GTTCAAGGTGGTTCCAGTCTCTGACCCTCCTGTGAATAGCCAGCAGATGGTGTACGTC
|
CGTGCCATGACTGAGTACTGGCCCCAGGAGGATCCCGACATCCCCTGCATGGACGCTG
|
GATTGCCTTTCCAGAAGGGGGACATCCTCCAGATTGTGGACCAGAATGATGCCCTCTG
|
GTGGCAGGCCCGAAAAATCTCAGACCCTGCTACCTGCGCTGGGCTTGTCCCTTCTAAC
|
CACCTTCTGAAGAGGAAGCAACGGGAATTCTGGTGGTCTCAGCCGTACCAGCCTCACA
|
CCTGCCTCAAGTCAACCCTATACAAGGAGGAGTTTGTTGGCTACGGTCAGAAGTTCTT
|
TATAGCTGGCTTCCGCCGCAGCATGCGCCTTTGTCGCAGGAAGTCTCACCTCAGCCCG
|
CTGCATGCCAGTGTGTGCTGCACCGGCAGCTGCTACAGTGCAGTGGGTGCCCCTTACG
|
AGGAGGTGGTGAGGTACCAGCGACGCCCTTCAGACAAGTACCGCCTCATAGTGCTCAT
|
GGGACCCTCTGGTGTTGGAGTAAATGAGCTCAGAAGACAACTTATTGAATTTAATCCC
|
AGCCATTTTCAAAGTGCTGTGCCACACACTACTCGTACTAAAAAGAGTTACGAAATGA
|
ATGGGCGTGAGTATCACTATGTGTCCAAGGAAACATTTGAAAACCTCATATATAGTCA
|
CAGGATGCTGGAGTATGGTGAGTACAAAGGCCACCTGTATGGCACTAGTGTGGATGCT
|
GTTCAAACAGTCCTTGTCGAAGGAAAGATCTGTGTCATGGACCTAGAGCCTCAGGATA
|
TTCAAGGGGTTCGAACCCATGAACTGAAGCCCTATGTCATATTTATAAAGCCATCGAA
|
TATGAGGTGTATGAAACAATCTCGGAAAAATGCCAAGGTTATTACTGACTACTATGTG
|
GACATGAAGTTCAAGGATGAAGACCTACAAGAGATGGAAAATTTAGCCCAAAGAATGG
|
AAACTCAGTTTGGCCAATTTTTTGATCATGTGATTGTGAATGACAGCTTGCACGATGC
|
ATGTGCCCAGTTGTTGTCTGCCATACAGAAGGCTCAGCAGGAGCCTCAGTGGGTACCA
|
GCAACATGGATTTCCTCAGATACTGAGTCTCAATGAG
|
ORF Start: ATG at 3ORF Stop: TGA at 1368
SEQ ID NO:12455 aa MW at 52165.4 kD
NOV4b,MRIVCLVKNQQPLGATIKRHEMTGDILVARIIHGGLAERSGLLYAGDKLVEVNGVSVE
|
CG56832-02 Protien SequenceGLDPEQVIHILAMSRGTIMFKVVPVSDPPVNSQQMVYVRAMTEYWPQEDPDIPCMDAG
|
LPFQKGDILQIVDQNDALWWQARKISDPATCAGLVPSNHLLKRKQREFWWSQPYQPHT
|
CLKSTLYKEEFVGYGQKFFIAGFRRSMRLCRRKSHLSPLHASVCCTGSCYSAVGAPYE
|
EVVRYQRRPSDKYRLIVLMGPSGVGVNELRRQLIEFNPSHFQSAVPHTTRTKKSYEMN
|
GREYHYVSKETFENLIYSHRMLEYGEYKGHLYGTSVDAVQTVLVEGKICVMDLEPQDI
|
QGVRTHELKPYVIFIKPSNMRCMKQSRKNAKVITDYYVDMKFKDEDLQEMENLAQRME
|
TQFGQFFDHVIVNDSLHDACAQLLSAIQKAQEEPQWVPATWISSDTESQ
|
SEQ ID NO:131935 bp
NOV4c,TCAGAAGCTCCGGCAGGGAGGATGATACAGTCAGACAAAGGAGCAGATCCACCAGACA
|
CG56832-03 DNA SequenceAGAAGGACATGAAGCTTTCTACAGCCACCAATCCACAGAATGGTCTCTCCCAGATCCT
|
GAGGCTTGTGCTGCAAGAGCTGAGTCTGTTCTACGGCACAGATGTGAATGGAGTGTGT
|
CTCTTGTACGATCTCCTCCACTCGCCGTGGCTTCAGGCTCTGCTAAAGATTTATGACT
|
GCCTCCAGGAATTTAAAGAAAAGAAACTAGTTCCTGCCACACCACATGCACAGGTGTT
|
ATCCTATGAGGTAGTGGAGTTATTACGTGAAACCCCTACTTCCCCTGAGATCCAAGAG
|
CTGAGACAAATGCTCCAGGCTCCACACTTCAAGGCATTGCTCAGTGCCCATGACACGA
|
TAGCTCAGAAAGATTTTGAACCCCTTCTCCCTCCACTGCCAGACAATATCCCTGACAG
|
TGAGGAAGCAATGAGGATTGTTTGTTTAGTGAAAAACCAACAGCCCCTGGGAGCCACC
|
ATCAAGCCCCACGAGATGACAGGGGACATCTTGGTGGCCAGGATCATCCACGGTGGGC
|
TGGCGGAGAGAAGTGGGTTGCTATATGCTGGAGACAAACTGGTAGAAGTGAATGGAGT
|
TTCAGTTGAGGGACTGGACCCTGAACAAGTGATCCATATTCTGGCCATGTCTCGAGGC
|
ACAATCATGTTCAAGGTGGTTCCAGTCTCTGACCCTCCTGTGAATAGCCAGCAGATGG
|
TGTACGTCCGTGCCATGACTGAGTACTGGCCCCAGGAGGATCCCGACATCCCCTGCAT
|
GGACGCTGGATTGCCTTTCCAGAAGGGGGACATCCTCCAGATTGTGGACCAGAATGAT
|
GCCCTCTGGTGGCAGGCCCGAAAAATCTCAGACCCTGCTACCTGCGCTGGGCTTGTCC
|
CTTCTAACCACCTTCTGAAGAGGAAGCAACGGGAATTCTGGTGGTCTCAGCCCTACCA
|
GCCTCACACCTGCCTCAAGTCAACCCTAGAAGATGACATGAAGATTGATGAGAAATGT
|
GTGGAAGCAGACAAGGAGGAGTTTGTTGGCTACGGTCAGAAGTTCTTTATAGCTGGCT
|
TCCGCCGCAGCATGCGCCTTTGTCGCAGGAAGTCTCACCTCAGCCCGCTGCATGCCAG
|
AGGTACCAGCGACGCCCTTCAGACAAGTACCGCCTCATAGTGCTCATGGGTCCCTCTG
|
GTGTTGGAGTAAATGAGCTCAGAAGACAACTTATTGAATTTAATCCCAGCCATTTTCA
|
AAGTGCTGTGCCAACTACTCGTACTAAAAAGAGTTACGAAATGAATGGGCGTGAGTAT
|
CACTATGTGTCCAAGGAAACATTTGAAAACCTCATATATAGTCACAGGATGCTGGAGT
|
ATGGTGAGTACAAAGGCCACCTGTATGGCACTAGTGTGGATGCTGTTCAAACAGTCCT
|
TGTCGAAGGAAAGATCTGTGTCATGGACCTAGAGCCTCAGGATATTCAAGGGGTTCGA
|
ACCCATGAACTGAAGCCCTATGTCATATTTATAAAGCCATCGAATATGAGCTGTATGA
|
AACAATCTCGGAAAAATGCCAAGGTTATTACTGACTACTATGTGGACATGAAGTTCAA
|
GGATGAAGACCTACAAGAGATGGAAAATTTAGCCCAAAGAATGGAAACTCAGTTTGGC
|
CAATTTTTTGATCATGTGATTGTGAATGACAGCTTGCACGATGCATGTGCCCAGTTGT
|
TGTCTGCCATACAGAAGGCTCAGGAGGAGCCTCAGTCGGTACCAGCAACATGGATTTC
|
CTCAGATACTGAGTCTCAATGAGACTTCTTGTTTAATGCTGGAGTTTTAACACTGIAC
|
CCTTGATACAGCGATCCATAG
|
ORF Start: ATG at 22ORF Stop: TGA at 1876
SEQ ID NO:14618 aa MW at 70539.2 kD
NOV4c,MIQSDKGADPPDKKDMKLSTATNPQNGLSQILRLVLQELSLFYGRDVNGVCLLYDLLH
|
CG56832-03 Protein SequenceSPWLQALLKIYDCLQEFKEKKLVPATPHAQVLSYEVVELLRETPTSPEIQELRQMLQA
|
PHFKALLSAHDTIAQKDFEPLLPPLPDNIPESEEAMRIVCLVKMQQPLGATIKRHEMT
|
GDILVARIIHGGLAERSGLLYAGDKLVEVNGVSVEGLDPEQVIHILAMSRGTIMFKVV
|
PVSDPPVNSQQMVYVRAMTEYWPQEDPDIPCMDAGLPFQKGDILQIVDQNDALWWQAR
|
KISDPATCAGLVPSNHLLKRKQREFWWSQPYQPHTCLKSTLEDDMKIDEKCVFADKEE
|
FVGYGQKFFIAGFRRSMRLCRRKSHLSPLHASVCCTGSCYSAVGAPYEEVVRYQRRPS
|
DKYRLIVLMGPSGVGVNELRRQLIEFNPSHFQSAVPTTRTKKSYEMNGREYHYVSKET
|
FENLIYSHRMLEYGEYKGHLYGTSVDAVQTVLVEGKICVMDLEPQDIQGVRTHELKPY
|
VIFIKPSNMRCMKQSRKNAKVITDYYVDMKFKDEDLQEMENLAQRMETQFGQFFDHVI
|
VNDSLHDACAQLLSAIQKAQEEPQWVPATWISSDTESQ
|
[0332] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 4B.
19TABLE 4B
|
|
Comparison of NOV4a against NOV4b and NOV4c.
Identities/
ProteinNOV4a Residues/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV4b 1 . . . 440439/455 (96%)
1 . . . 455439/455 (96%)
NOV4c 1 . . . 440437/468 (93%)
152 . . . 618437/468 (93%)
|
[0333] Further analysis of the NOV4a protein yielded the following properties shown in Table 4C.
20TABLE 4C
|
|
Protein Sequence Properties NOV4a
|
|
PSort0.6500 probability located in cytoplasm;
analysis:0.1000 probability located in mitochondrial
matrix space; 0.1000 probability located in
lysosome (lumen); 0.0587 probability located in
microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0334] A search of the NOV4a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 4D.
21TABLE 4D
|
|
Geneseq Results for NOV4a
NOV4aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAE11774Human kinase (PKIN)-8 protein- 1 . . . 440437/442 (98%)0.0
Homo sapiens, 442 aa. 1 . . . 442438/442 (98%)
[WO200181555-A2, 01-Nov-2001]
AAU07123Human novel human protein, NHP 1 . . . 435195/443 (44%) e−105
#23-Homo sapiens, 576 aa.137 . . . 576292/443 (65%)
[WO200161016-A2, 23-AUG-2001]
AAU07119Human novel human protein, NHP 1 . . . 407186/415 (44%)2e−99
#19-Homo sapiens, 560 aa.137 . . . 548278/415 (66%)
[WO200161016-A2, 23-AUG-2001]
AAU07115Human novel human protein, NHP 1 . . . 367167/372 (44%)5e−90
#15-Homo sapiens, 520 aa.137 . . . 505254/372 (67%)
[WO200161016-A2, 23-AUG-2001]
AAU07111Human novel human protein, NHP 1 . . . 330149/335 (44%)3e−78
#11-Homo sapiens, 473 aa.137 . . . 468225/335 (66%)
[WO200161016-A2, 23-AUG-2001]
|
[0335] In a BLAST search of public sequence datbases, the NOV4a protein was found to have homology to the proteins shown in the BLASTP data in Table 4E.
22TABLE 4E
|
|
Public BLASTP Results for NOV4a
NOV4aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96JB8MEMBRANE PROTEIN 1 . . . 440438/486 (90%)0.0
PALMITOYLATED 4-Homo152 . . . 637438/486 (90%)
sapiens (Human), 637 aa.
Q96Q44ALS2CR5 PROTEIN-Homo 1 . . . 440426/455 (93%)0.0
sapiens (Human), 593 aa.152 . . . 593426/455 (93%)
Q9QYH1DLG6 ALPHA-Rattus norvegicus 1 . . . 439363/441 (82%)0.0
(Rat), 441 aa. 1 . . . 441395/441 (89%)
Q920P7MDLG6B-Mus musculus (Mouse), 1 . . . 439369/479 (77%)0.0
479 aa. 1 . . . 479399/479 (83%)
Q9QYG9DLG6 GAMMA-Rattus 1 . . . 439348/441 (78%)0.0
norvegicus (Rat), 423 aa. 1 . . . 423380/441 (85%)
|
[0336] PFam analysis predicts that the NOV4a protein contains the domains shown in the Table 4F.
23TABLE 4F
|
|
Domain Analysis of NOV4a
Identities/
Similarities
NOV4a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
PDZ: domain 1 of 1 3 . . . 8325/84 (30%)4.3e−09
59/84 (70%)
SH3: domain 1 of 1 94 . . . 15919/68 (28%)0.002
47/68 (69%)
Guanylate_kin: domain 1266 . . . 37040/108 (37%) 1e−32
of 179/108 (73%)
I_LWEQ: domain 1 of 1403 . . . 420 6/18 (33%)6.3
15/18 (83%)
|
Example 5
[0337] The NOV5 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 5A.
24TABLE 5A
|
|
NOV5 Sequence Analysis
|
|
SEQ ID NO:15894 bp
NOV5a,ACACAAATAGGCAAATTGTGGGTATGGGATTCCCTCCCTACCTCCCTCCACCCCAGGG
|
CG58618-01 DNA SequenceCCCAGGTAGCGACCATGTCCCCTGCCATTGCATTGGCCTTCCTGCCACTGGTGGTAAC
|
ATTGCTGGTCCGGTACCCGCACTACTTCCCATTOCTGGTGCGCACGGTCTTGCTGCGA
|
AGCCTCCGAGACTGCCTGTCAGGGCTGCGGATCGAGGACCCGGCCTTCAGCPACGTGC
|
TCACCCATGCCCTGCCCGGTGACCCTGGTCACATCCTCACCACCCTGGACCACTGGAG
|
CAGCCGCTGCCAGTACTTGAGCCACATCGGGCCTGTCAAAGGTCAGATCCTGATGCGG
|
CTGGTGGAGGAGAAGGCCCCTGCTTGTGTGCTGGAATTGGGAACCTACTCTGCATACT
|
CTACCCTGCTTATTGCCCGAGCCCTGCCCCCTGCGCGTCGCCTTCTTACTGTGGAGCG
|
GGACCCACGCACGGCAGCAGTGGCTGAAAAACTCATCCGCCTGGCCGGCTTTGATGAG
|
CACCACGTGGAGCTCATCCTGGGCAGCTCACAGGACGTCATCCCGTGCCTACGCACCC
|
AGTATCAGCTCAGTCCCCCAGACCTCCTCCTCCTGGCACACCGGCCACGATGTTACCT
|
GAGGGACCTGCAGCTGCTGGAGGCCCATGCCCTACTGCCAGCACGTGCCACCGTGCTC
|
GCTGACCATGTGCTCTTCCCTGGTGCACCCCGCTTCTTGCAGTATGCTAAGAGCTGTG
|
GCCGCTACCGCTGCCGCCTCCACCACACTGQCCTTCCAGACTTCCCTGCCATCAACGA
|
TGGAATAGCTCAGCTCACCTATCCTGGACCAGGCTGAGGTCCACGCCCAGCGGTACTT
|
ACTGATGCCCACCCCCACCCCCAC
|
ORF Staff: ATG at 73ORF Stop: TGA at 847
SEQ ID NO:16258 aa MW at 28694.2 kD
NOVA5a,MSPAIALAFLPLVVTLLVRYRHYFRLLVRTVLLRSLRDCLSGLRIEERAFSYVLTHAL
|
CG58618-01 Protein SequencePGDPGHILTTLDHWSSRCEYLSHMGPVKGQILMRLVEEKAPACVLELGTYCGYSTLLI
|
ARALPPGGRLLTVERDPRTAAVAEKLIRLAGFDEHQVELIVGSSEDVIPCLRTQYQLS
|
RADLVLLAHRPRCYLRDLQLLEAHALLPAGATVLADHVLFPGAPRFLQYAKSCGRYRC
|
RLHHTGLPDFPAIKDGIAQLTYAGPG
|
SEQ ID NO:17 897 bp
NOV5b,AAATAGGCAAATTGTGGGTATGGGATTCCCTCCCTACCTCCCTCCACCCCAGGGCCCA
|
CG5818-02 DNA SequenceGGTAGGGACCATGTCCCCTGCCATTGCATTGGCCTTCCTGCCACTGGTGGTAACATTG
|
CTGGTGCGGTACCGGCACTACTTCCGATTGCTGGTGCGCACGGTCTTGCTGCGAAGCC
|
TCCGAGACTGCCTGTCAGGGCTGCGGATCGAGGACCGGGCCTTCAGCTACGTGCTCAC
|
CCATGCCCTGCCCGGTGACCCTGGTCACATCCTCACCACCCTGGACCACTGGAGCAGC
|
CGCTGCGAGTACTTGAGCCACATGGGGCCTGTCAAAGGTCAGATCCTGATGCGGCTGC
|
TGGAGGAGAAGGCCCCTCCTTGTGTGCTGGAATTGGGAACCTACTCTCGATACTCTAC
|
CCTGCTTATTGCCCGAGCCCTGCCCCCTGGGGGTCGCCTTCTTACTGTGGAGCGGGAC
|
CCACGCACGCCAGCAGTCCCTGAAAAACTCATCCGCCTCCCCGGCTTTGATGACCACA
|
TGGTGGAGCTCATCGTGGGCAGCTCAGAGGACGTGATCCCGTGCCTACGCACCCAGTA
|
TCAGCTGAGTCGGGCAGACCTGGTGCTCCTGGCACACCGGCCACGATGTTACCTGACC
|
GACCTGCAGCTGCTGGAGGCCCATGCCCTACTGCCAGCACGTGCCACCGTGCTGGCTG
|
ACCATGTGCTCTTCCCTGGTGCACCCCGCTTCTTGCAGTATGCTAAGAGCTGTGGCCG
|
CTACCGCTGCCGCCTCCACCACACTGGCCTTCCAGACTTCCCCGCCATCAAGGATGGA
|
ATAGCTCAGCTCACCTATGCTGGACCAGGCTGAGGTCCAGGCCCAGGGGTACTTACTG
|
ATGCCCACCCCCACAATTTGCCTATTT
|
ORF Staff: ATG at 69ORF Stop: TGA at 843
SEQ ID NO:18258 aa MW at 28697.3 kD
NOV5b,MSPAIALAFLPLVVTLLVRYRHYFRLLVRTVLLRSLEDCLSGLRIEERAFSYVLTHAL
|
CG58618-02 Protein SequencePGDPGHILTTLDHWSSRCEYLSHMGPVKGQILMRLVEEKAPACVLELGTYCGYSTLLI
|
ARALPPGGRLLTVERDPRTAAVAEKLIRLAGFDEHMVELIVGSSEDVIPCLRTQYQLS
|
RADLVLLAHRPRCYLRDLQLLEAHALLPAGATVLADHVLFPGAPRFLQYAKSCGRYRC
|
RLHHTGLPDFPAIKDGIAQLTYAGPG
|
[0338] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 5B.
25TABLE 5B
|
|
Comparison of NOV5a against NOV5b.
Identities/
ProteinNOV5a Residues/Similarities for
SequenceMatch Residuesthe Matched Region
|
NOV5b1 . . . 258244/258 (94%)
1 . . . 258244/258 (94%)
|
[0339] Further analysis of the NOV5a protein yielded the following properties shown in Table 5C.
26TABLE 5C
|
|
Protein Sequence Properties NOV5a
|
|
PSort0.8200 probability located in endoplasmic reticulum
analysis:(membrane); 0.1900 probability located in plasma membrane;
0.1000 probability located in endoplasmic reticulum (lumen);
0.1000 probability located in outside
SignalPCleavage site between residues 42 and 43
analysis:
|
[0340] A search of the NOV5a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 5D.
27TABLE 5D
|
|
Geneseq Results for NOV5a
NOV5aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAE07183Catechol-O-methyltransferase-like 1 . . . 258257/258 (99%) e−148
human enzyme #1-Homo sapiens, 1 . . . 258257/258 (99%)
258 aa. [WO200157220-A2, 09-
AUG-2001]
AAE07184Catechol-O-methyltransferase-like91 . . . 258167/168 (99%)6e−94
human enzyme #2-Homo sapiens, 1 . . . 168167/168 (99%)
168 aa. [WO200157220-A2, 09-
AUG-2001]
AAW70989Murine catechol-O-methyltransferase 4 . . . 258 93/257 (36%)5e−43
(COMT)-Mus sp, 265 aa. 5 . . . 259149/257 (57%)
[WO9832878-A1, 30-JUL-1998]
AAB43663Human cancer associated protein 4 . . . 258 94/259 (36%)7e−42
sequence SEQ ID NO: 1108-Homo38 . . . 294151/259 (58%)
sapiens, 299 aa. [WO200055350-A1,
21-SEP-2000]
AAW70988Human catechol-O-methyltransferase 4 . . . 258 94/259 (36%)7e−42
(COMT)-Homo sapiens, 271 aa.10 . . . 266151/259 (58%)
[WO9832878-A1, 30-JUL-1998]
|
[0341] In a BLAST search of public sequence datbases, the NOV5a protein was found to have homology to the proteins shown in the BLASTP data in Table 5E.
28TABLE 5E
|
|
Public BLASTP Results for NOV5a
NOV5aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q91XH4CATECHOL-O- 4 . . . 258 94/257 (36%)3e−43
METHYLTRANSFERASE-Mus 5 . . . 259150/257 (57%)
musculus(Mouse), 265 aa.
O88587Catechol O-methyltransferase, 4 . . . 258 93/257 (36%)1e−42
membrane-bound form (EC 2.1.1.6) 5 . . . 259149/257 (57%)
(MB-COMT) [Contains: Catechol O-
methyltransferase, soluble form (S-
COMT)]-Mus musculus(Mouse), 265 aa.
P21964Catechol O-methyltransferase, 4 . . . 258 94/259 (36%)2e−41
membrane-bound form (EC 2.1.1.6)10 . . . 266151/259 (58%)
(MB-COMT) [Contains: Catechol O-
methyltransferase, soluble form (S-
COMT)]-Homo sapiens(Human), 271 aa.
P22734Catechol O-methyltransferase, 4 . . . 257 91/256 (35%)2e−41
membrane-bound form (EC 2.1.1.6) 5 . . . 258151/256 (58%)
(MB-COMT) [Contains: Catechol O-
methyltransferase, soluble form (S-
COMT)]-Rattus norvegicus(Rat), 264 aa.
Q99028Catechol-O-methyltransferase, soluble77 . . . 258 77/182 (42%)2e−36
form (EC 2.1.1.6) (S-COMT)-Sus 2 . . . 181116/182 (63%)
scrofa(Pig), 186 aa (fragment).
|
[0342] PFam analysis predicts that the NOV5a protein contains the domains shown in the Table 5F.
29TABLE 5F
|
|
Domain Analysis of NOV5a
Identities/
Similarities
NOV5a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Methyltransf_3: domain 155 . . . 252 49/218 (22%)8.9e−05
of 1109/218 (50%)
|
Example 6
[0343] The NOV6 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 6A.
30TABLE 6A
|
|
NOV6 Sequence Analysis
|
|
SEQ ID NO:191115 bp
NOV6a,TCTTGACTGTAGAATATAAGTGAGAATATTTCTGATATGTTTGTAATGCATGCTTGTG
|
CG59580-01 DNA SequenceTCATGTTCTCTTTCTCTCTCTTTTGGAAGTCACATCAACAAGATGAAACCAGGGAATG
|
AGACACAAATTTCACAATTCCTTCTCCTGGGACTTTCAGAGGAACCAGAATTGCAGCC
|
CTTCCTCTTTGGGCTATTTCTGTCCATGTACCTGGTCACCGTGCTCGGGAACCTGCTC
|
ATCATCCTGGCCACAATCTCAGACTCCCACCTCCACACCCCCATGTACTTCTTCCTCT
|
CCAACCTGTCCTTTGCAGACATCTGTTTTGTGTCTACCACTGTCCCAAAGATGCTGGT
|
GAACATCCAGACAGAGAGCAGAGTCATCACCTATGCAGACTGCATCACCCAGATGTGC
|
TTTTTTATACTCTTTGTAGTGTTGGACAGCTTACTCCTGACTGTGATGGCCTATGACC
|
GGTTTGTGGCCATCTGTCACCCCCTGCACTACACAGTCATTATGAACTCCTGGCTCTG
|
TGGACTGCTGGTTCTGGTGTCCTGGATCGTGAGCATCCTATATTCTCTGTTACAAAGC
|
ATAATGGCATTGCAGCTGTCCTTCTGTACAGAATTGAAAATCCCTCATTTTTTCTGTG
|
AACTTAATCAGGTCATCCACCTTGCCTGTTCCGACACTTTTATTAATGACATGATCAT
|
GAATTTTACAAGTGTGCTGCTGGGTGGGGGATGCCTCGCTGGAATATTTACTTACTTT
|
AAGATACTTTGTTGCATATGTTCGATCTCATCAGCTCAGGGGATGAATAAAGCACTTT
|
CCACCTGTGCATCTCACCTCTCAGTTGTCTCCTTATTTTATTGTACAGGCGTAGGTGT
|
GTACCTTAGTTCTGCTGCAACCCATAACTCACTCTCAAATGCTGCAGCCTCGGTGATG
|
TACACTGTGGTCACCTCCATGCTCAACCCCTTCATCTACAGCCTGAGGAATAAAGACA
|
TAAACAGAGCTCTGAATCGATTCTTCAGAGAGCAGAAACAGGAGGGCCATTTTCCAGA
|
AGTTCTTGAGATTTCAGAGCTCTAAATCCCCTAGCCAGAAATTATGATTCATGGATCA
|
GATTGTGAAAGTA
|
ORF Start: ATG at 50ORF Stop: TAA at 1067
SEQ ID NO:20339 aa MW at 37821.1 kD
NOV6a,MLVSCSLSLSFGSHINKMKPGNETQISQFLLLGLSEEPELQPFLFGLFLSMYLVTVLG
|
CG59580-01 Protein SequenceNLLIILATISDSHLHTPMYFFLSNLSFADICFVSTTVPKMLVNIQTQSRVITYADCIT
|
QMCFFILFVVLDSLLLTVMAYDRFVAICHPLHYTVIMNSWLCCLLVLVSWIVSILYSL
|
LQSIMALQLSFCTELKIPHFFCELNQVIHLACSDTFINDMMMNFTSVLLGGGCLAGIF
|
TYFKILCCICSISSAQCMNKALSTCASELSVVSLFYCTGVCVYLSSAATHNSLSNAAA
|
SVMYTVVTSMLNPFIYSLRNKDINRALNRFFREOKQEGHFPEVLEISEL
|
[0344] Further analysis of the NOV6a protein yielded the following properties shown in Table 6B.
31TABLE 6B
|
|
Protein Sequence Properties NOV6a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body;
0.3000 probability located in endoplasmic
reticulum (membrane); 0.3000 probability located
in microbody (peroxisome)
SignalPCleavage site between residues 69 and 70
analysis:
|
[0345] A search of the NOV6a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 6C.
32TABLE 6C
|
|
Geneseq Results for NOV6a
NOV6aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAG72100Human olfactory receptor polypeptide, 18 . . . 330312/314 (99%)e−179
SEQ ID NO: 1781-Homo sapiens, 1 . . . 314313/314 (99%)
314 aa. [WO200127158-A2, 19-APR-
2001]
AAG72195Human olfactory receptor polypeptide, 18 . . . 322244/306 (79%)e−136
SEQ ID NO: 1876-Homo sapiens, 1 . . . 306267/306 (86%)
309 aa. [WO200127158-A2, 19-APR-
2001]
ABG29229Novel human diagnostic protein 16 . . . 322239/308 (77%)e−133
#29220-Homo sapiens, 788 aa.478 . . . 785264/308 (85%)
[WO200175067-A2, 11-OCT-2001]
ABG26552Novel human diagnostic protein 16 . . . 322239/308 (77%)e−133
#26543-Homo sapiens, 379 aa. 69 . . . 376264/308 (85%)
[WO200175067-A2, 11-OCT-2001]
ABG29229Novel human diagnostic protein 16 . . . 322239/308 (77%)e−133
#29220-Homo sapiens. 788 aa.478 . . . 785264/308 (85%)
[WO200175067-A2, 11-OCT-2001]
|
[0346] In a BLAST search of public sequence datbases, the NOV6a protein was found to have homology to the proteins shown in the BLASTP data in Table 6D.
33TABLE 6D
|
|
Public BLASTP Results for NOV6a
NOV6aIdentities/
ProteinResidues/Similarities for
AccessionProtein/Organism/Matchthe MatchedExpect
NumberLengthResiduesPortionValue
|
O14581Olfactory receptor18 . . . 322244/306 (79%)e−136
7A17-Homo sapiens 1 . . . 306267/306 (86%)
(Human), 309 aa.
O76100Olfactory receptor18 . . . 322238/306 (77%)e−132
7A10 (OST027)- 1 . . . 306262/306 (84%)
Homo
sapiens(Human),
309 aa.
Q15622Olfactory receptor18 . . . 330232/314 (73%)e−127
7A5 (Olfactory 1 . . . 314260/314 (81%)
receptor TPCR92)-
Homo sapiens
(Human), 319 aa.
Q95157Olfactory18..322216/306 (70%)e−121
receptor-like 1 . . . 306254/306 (82%)
protein OLF4-Canis
familiaris(Dog),
309 aa.
Q9JHB2M12 ODORANT18 . . . 324220/308 (71%)e−121
RECEPTOR-Mus 1 . . . 308258/308 (83%)
musculus(Mouse),
309 aa.
|
[0347] PFam analysis predicts that the NOV6a protein contains the domains shown in the Table 6E.
34TABLE 6E
|
|
Domain Analysis of NOV6a
Identities/
Similarities
NOV6a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Bac_export_3: domain 1138 . . . 221 18/87 (21%)8.4
of 1 52/87 (60%)
7tm_1:domain 1 of 1 58 . . . 306 55/269 (20%)5.9e−38
182/269 (68%)
|
Example 7
[0348] The NOV7 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 7A.
35TABLE 7A
|
|
NOV7 Sequence Analysis
|
|
SEQ ID NO:21982 bp
NOV7a,ATGAATGGAGGGGTTTAACTATTCCAGAGTATCTGAATTCATGTTACTTGGACTTACT
|
CG59611-01 DNA SequenceGATTCTCCTGAACTCCAGATATTCTTTTTTGTGGTGTTTTCTGTCTTCTATTTAATGA
|
CCATGTTGGGCAACTGCCTGATTTTACTCACTGTCCTATCCACCTCACACCTTCACTC
|
TCGCACGTACTTCCTGCTCAGCAACCTGTCTCATATTGACATGTGCCTGTCCTCCTTT
|
GCCACACCAAAGATGATTATGGACTTTTTTGCTCTGCGTAAGACCATCTCTTTTGAAG
|
GCTGCATTTCTCAGATCTTTTTTTTACACCTCTTCAATGGGACTGAGATTGTGCTGCT
|
GATCTCCATGTCTTTTGACAGGTATATTGCCATATGTAAACCTCTCCGCTATTCAACA
|
ATTATGAGCCAAAGAGTGTGTGTTGAGCTTGTGGCAGTTTCTTGTTGGACAGTGGGCT
|
TTCTACATACAATGAGCCAATTAAGTTTTTCCCTCTATTTGCCCTTCTGTGTTCCCAA
|
TGTTGTAGACAGTTTTTTCTGTGATCTTCCTTTGGTCATCCAGTTAGCTTGTATAGAT
|
ATTTATGTTCTTGGGACCTCCATGATTTCAACCAGTGGTGTGACTGCTCTTACAAGTT
|
TTCTGCTTTTGCTCACCTCCTACATCATTGTTCTTAATACTATCAGGGACTACTCCTC
|
CACAGGATCCTCCAAGGCTCTTTCTACCTGTACAGCACATTTTATTGTTCTGTTAATG
|
TTCTTTGGGCCCTGTATTTTCATTTATGTGTGGCCTTCCACAAACTTCCTGGTAGACA
|
AAATTCTCTCTGTTTTCTATACCATCTTCACTCCCTTTCTGAATCCACTTATCTATAC
|
TTTGAGAAACCAGGAAGTGAAGACAGCAATGAAGAAGAAACTGAATATTCAGTATTTC
|
AGTCTTGGGAAAACTGCTCCGTGATACTTCATGCAATGAATAGCGATCTCCTTT
|
ORF Start: ATG at 5ORF Stop: TGA at 950
SEQ ID NO:22315 aa MW at 35742.0 kD
NOV7a,MEGFNYSRVDEFMLLGLTDSPELQIFFFVVFSVFYLMTMLGNCLILLTVLSTSHLHSR
|
CG59611-01 Protein SequenceTYFLLSNLSHIDMCLSSFATPKMIMDFFALRKTISFEGCISQIFFLHLFNGTEIVLLI
|
SMSFDRYIAICKPLRYSTIMSQRVCVELVAVSCWTVQFLHTMSQLSFSLYLPFCVPNV
|
VDSFFCDLPLVIQLACIDIYVLGTSMISTSGVTALTSFLLLLTSYIIVLNTIRDYSST
|
GSSKALSTCTAHFIVVLMFFGPCIFIYVWPSTNFLVDKILSVFYTIFTPFLNPLIYTL
|
RNQEVKTAMKKKLNIQYFSLGKTAP
|
[0349] Further analysis of the NOV7a protein yielded the following properties shown in Table 7B.
36TABLE 7B
|
|
Protein Sequence Properties NOV7a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body;
0.3000 probability located in endoplasmic
reticulum (membrane); 0.3000 probability
located in microbody (peroxisome)
SignalPCleavage site between residues 52 and 53
analysis:
|
[0350] A search of the NOV7a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 7C.
37TABLE 7C
|
|
Geneseq Results for NOV7a
NOV7aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAG71880Human olfactory receptor polypeptide,1 . . . 315312/315 (99%)e−178
SEQ ID NO: 1561-Homo sapiens,1 . . . 314313/315 (99%)
314 aa. [WO200127158-A2, 19-APR-
2001]
AAG72471Human OR-like polypeptide query1 . . . 315304/315 (96%)e−173
sequence, SEQ ID NO: 2152-Homo1 . . . 314307/315 (96%)
sapiens, 314 aa. [WO200127158-A2,
19-APR-2001]
AAG71767Human olfactory receptor polypeptide,1 . . . 315304/315 (96%)e−173
SEQ ID NO: 1448-Homo sapiens.1 . . . 314307/315 (96%)
314 aa. [WO200127158-A2, 19-APR-
2001]
AAG71495Human olfactory receptor polypeptide,1 . . . 315294/315 (93%)e−164
SEQ ID NO: 1176-Homo sapiens,1 . . . 313299/315 (94%)
313 aa. [WO200127158-A2, 19-APR-
2001]
AAG71508Human olfactory receptor polypeptide,1 . . . 292280/292 (95%)e−159
SEQ ID NO: 1189-Homo sapiens,1 . . . 291283/292 (96%)
291 aa. [WO200127158-A2, 19-APR-
2001]
|
[0351] In a BLAST search of public sequence datbases, the NOV7a protein was found to have homology to the proteins shown in the BLASTP data in Table 7D.
38TABLE 7D
|
|
Public BLASTP Results for NOV7a
NOV7aIdentities/
ProteinResidues/Similarities for
AccessionProtein/Organism/Matchthe MatchedExpect
NumberLengthResiduesPortionValue
|
AAL61393OLFACTORY1 . . . 315262/315 (83%)e−147
RECEPTOR1 . . . 314278/315 (88%)
MOR246-5-
Mus musculus
(Mouse), 326 aa.
AAL61267OLFACTORY5 . . . 301205/297 (69%)e−118
RECEPTOR6 . . . 301252/297 (84%)
MOR247-1-
Mus musculus
(Mouse), 318 aa.
AAL61437OLFACTORY1 . . . 314193/314 (61%)e−109
RECEPTOR1 . . . 312247/314 (78%)
MOR246-2-
Mus musculus
(Mouse), 324 aa.
AAL61440OLFACTORY5 . . . 314188/310 (60%)e−107
RECEPTOR5 . . . 312239/310 (76%)
MOR246-4-
Mus musculus
(Mouse), 321 aa.
AAL61478OLFACTORY1 . . . 301180/301 (59%)e−105
RECEPTOR1 . . . 300234/301 (76%)
MOR246-6-
Mus musculus
(Mouse), 323 aa.
|
[0352] PFam analysis predicts that the NOV7a protein contains the domains shown in the Table 7E.
39TABLE 7E
|
|
Domain Analysis of NOV7a
NOV7aIdentities/
MatchSimilaritiesExpect
Pfam DomainRegionfor the Matched RegionValue
|
7tm_1: domain 1 of 141 . . . 288 51/276 (18%)3.3e−19
174/276 (63%)
|
Example 8
[0353] The NOV8 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 8A.
40TABLE 8A
|
|
NOV8 Sequence Analysis
|
|
SEQ ID NO:23963 bp
NOV8a,CTGCAAAAAGACATGGGAAAGACCAAAAACACATCGCTGGACACTGTGGTGAGAGATT
|
CG59617-01 DNA SequenceTCATTCTTCTGGGTTTGTCTCACCCCCCGAATATAAGAAGCCTCCTCTTCCTGGTCTT
|
CTTCGTCATTTACATCCTCACTCAGCTGGGGAACCTGCTCATTCTGCTCACCGTGTGG
|
GCTGACCCGAAGCTCCGTGCTCGCCCCATGTACATTCTTCTGGGAGTGCTCTCATTCC
|
TGGACATGTGGCTCTCCTCAGTCATCGTTCCTGGAATTATTTTAAACTTCACTCCTGC
|
CAACAAGGCTATCCCGTTTGGTGGCTGTGTGGCTCAACTGTATTTCTTTCACTTCCTG
|
GGCAGCACCCAGTGCTTCCTCTACACCTTGATGGCCTATGACAGGTACCTGGCAATAT
|
GTCAGCCCCTGCGCTACCCAGTGCTCATGAATGGGAGGTTATGCACAGTCCTTGTGGC
|
TGGAGCTTGGGTCGCCGGCTCCATGCATGGGTCTATCCAGGCCACCCTGACCTTCCGC
|
CTGCCCTACTGTGGGCCCAATCAGGTAGATTACTTTATCTGTGACATCCCCGCAGTAT
|
TGAGACTGGCCTGTGCTGACACAACTGTCAATGAGCTTGTGACCTTTGTGGACATCGG
|
GGTAGTGGCCGCCAGTTGCTTCATGTTAATTCTGCTCTCGTATGCCAACATAGTAAAT
|
GCCATCCTGAAGATACGCACCACTGATGGGAGGCGCCGGGCCTTCTCCACCTGTGGCT
|
CCCACCTAATCGTGGTCACAGTCTACTATGTCCCCTGTATTTTCATCTACCTTAGGGC
|
TGGCTCCAAAGGCCCCCTGGATGGGGCAGCGGCTGTGTTTTACACTGTTGTCACTCCA
|
TTACTGAACCCCCTCATCTATACACTGAGGAACCAGGAAGTGAAGTCTGCCCTGAAGA
|
GGATAACAGCAGGTCAAGGGACTGAATGAAAATAA
|
ORF Start: ATG at 13ORF Stop: at 853
SEQ ID NO:24280 aa MW at 30929.4 kD
NOV8a,MGKTKNTSLDTVVRDFILLGLSHPPNIRSLLFLVFFVIYILTQLGNLLILLTVWADPK
|
CG59617-01 Protein SequenceLRARPMYILLGVLSFLDMWLSSVIVPGIILNFTPANKAIPFGGCVAQLYFFHFLGSTQ
|
CFLYTLMAYDRYLAICQPLRYPVLMNGRLCTVLVAGAWVAGSMHGSIQATLTFRLPYC
|
GPNQVDYFICDIPAVLRLACADTTVNELVTFVDIGVVAASCFMLILLSYANIVNAILK
|
IRTTDGRRRAFSTCGSHLIVVTVYYVPCIFlYLRAGSKGPLDGAAAVF
|
[0354] Further analysis of the NOV8a protein yielded the following properties shown in Table 8B.
41TABLE 8B
|
|
Protein Sequence Properties NOV8a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body;
0.3000 probability located in endoplasmic reticulum
(membrane); 0.3000 probability located in microbody
(peroxisome)
SignalPCleavage site between residues 56 and 57
analysis:
|
[0355] A search of the NOV8a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 8C.
42TABLE 8C
|
|
Geneseq Results for NOV8a
NOV8aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAG72301Human olfactory receptor polypeptide.1 . . . 280279/280 (99%)e−161
SEQ ID NO: 1982-Homo sapiens,
318 aa. [WO200127158-A2, 19-APR-
2001]
AAG72302Human olfactory receptor polypeptide,1 . . . 280261/280 (93%)e−151
SEQ ID NO: 1983-Homo sapiens,1 . . . 280269/280 (95%)
310 aa. [WO200127158-A2, 19-APR-
2001]
AAY90874Human G protein-coupled receptor1 . . . 280261/280 (93%)e−151
GTAR14-5 SEQ ID NO: 6-Homo1 . . . 280269/280 (95%)
sapiens, 310 aa. [WO200021999-A1,
20-APR-2000]
AAG72708Murine OR-like polypeptide query1 . . . 280252/280 (90%)e−149
sequence, SEQ ID NO: 2390-Mus7 . . . 286268/280 (95%)
musculus, 324 aa. [WO200127158-
A2, 19-APR-2001]
AAG73014Olfactory receptor-like polypeptide,1 . . . 280249/280 (88%)e−146
SEQ ID NO: 2696-Unidentified, 3181 . . . 280263/280 (93%)
aa. [WO200127158-A2, 19-APR-
2001]
|
[0356] In a BLAST search of public sequence datbases, the NOV8a protein was found to have homology to the proteins shown in the BLASTP data in Table 8D.
43TABLE 8D
|
|
Public BLASTP Results for NOV8a
NOV8aIdentities/
ProteinProtein/Residues/Similarities for
AccessionOrganism/Matchthe MatchedExpect
NumberLengthResiduesPortionValue
|
AAL61330OLFACTORY 1 . . . 280252/280 (90%)e−149
RECEPTOR 1 . . . 280268/280 (95%)
MOR223-6-
Mus musculus
(Mouse),
318 aa.
Q62944TASTE BUD 1 . . . 280249/280 (88%)e−146
RECEPTOR 1 . . . 280263/280 (93%)
PROTEIN
TB 641-
Rattus
norvegicus
(Rat), 318 aa.
AAL61485OLFACTORY 1 . . . 280248/280 (88%)e−146
RECEPTOR 1 . . . 280263/280 (93%)
MOR223-9-
Mus musculus
(Mouse),
318 aa.
AAL61329OLFACTORY11 . . . 280202/270 (74%)e−118
RECEPTOR 7 . . . 276232/270 (85%)
MOR223-5-
Mus musculus
(Mouse),
313 aa.
AAL61470OLFACTORY11 . . . 280169/270 (62%)3e−94
RECEPTOR 7 . . . 275199/270 (73%)
MOR223-8-
Mus musculus
(Mouse),
310 aa.
|
[0357] PFam analysis predicts that the NOV8a protein contains the domains shown in the Table 8E.
44TABLE 8E
|
|
Domain Analysis of NOV8a
NOV8aIdentities/
MatchSimilaritiesExpect
Pfam DomainRegionfor the Matched RegionValue
|
7tm_1: domain 1 of 145 . . . 253 42/215 (20%)1.5e−23
149/215 (69%)
|
Example 9
[0358] The NOV9 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 9A.
45TABLE 9A
|
|
NOV9 Sequence Analysis
|
|
SEQ ID NO:251704 bp
NOV9a,GCCGCGCTGTGCTATGAAGCAGGTCAATGAGCTGAAAGAGAACGCCAACAAGGCCCTC
|
CG59826-01 DNA SequenceGGCGTGGAAAACATCGACAATGCCTTACAGTGCTACTCTGAAGCCATTAAGCTGGATC
|
CCCACAACCACGTGCTCTACACCAACTGTTCTGCCACCTATGCCAAGAAAGGAGACTA
|
CCAGAAGGCTTACAAGGACCGCTGCAAGACCGTCGACCTAAAATTTGACTGGGCCAAG
|
GGATATTCACGAAAAGCAGCAGCTCTAGAGTTCTTAAACCGATTTGAAGAAGCCAAGG
|
GAACCTACGAGGAGAGCTTAAAACACGAGGCAGATAACCCTCAACTGAAAGAGGGTTT
|
ACAGAATATGGAGGCCAGGTTGGCAGAGAGAAAATGTATGAATGCTTTCAACATGCCC
|
AATCTGAAACAGAAGTTGGAGAGTGATCCCAGGACAAGGACACTGCTCAGTGATCCTA
|
CCTACCGGGAGCTGATAGAGCAACTAAAAAACAAGCCTTCTGATCTGGGCACAAAACT
|
ACGAGATCCCCGGATCCTGACCACTCTCAGTGTCCTCCTTGAGGTCGATCGGGGCAGT
|
ATGGATGAGGAGGAACAGGTTGCAACACCTTCACCACCACCCCTTCCCAAAAAGGAGA
|
CCAAGCCAGAGCCAGTGGAAGAAGATCTTCCAGAGAATAAGAAGCAGGCACTGAAAGA
|
AAAAGAGCTGAGGAACGATGCCTACAAGAAGAAAGACAGACTGACAGCCTTGAAGCAT
|
TACGACAAAGCCAAGGAGCTGGACCCCAGCAACATGACTTACATTACCAATCAAGCAG
|
CGGTATACTTTGAAAAGGGCGACTACAATAAGTGTTGGGAGCTTTGTGAGAAGTCCAT
|
TGAAGTGGGGAGAGAAAACAGAGAAGACTATCTACAGATTGCCAAAGTGTGTGCTCGA
|
ATTGGCAACTCCTACTTCAAAGAAGAAAAGTACAAGGATGCCATCCATTTCTATAACA
|
AGTCTCTGGCAGAGCACCGAACTCCAGATGTGCTCAAGAAATGCCAGCAGGCAGAGAA
|
AATCCTGAAGGAGCAAGAACGGCTGGCCTACATAAACCCCGACCTGGCTTTAGAAGAG
|
AAGAACAAAGGCAACGAGTGTTTTCAGAAAGGGGACTATCCCCACGCCATGAAGCATT
|
ATACGGAAGCCATCAAAAGGAACCCGAAAGATGCCAAATTATACAGCAATCGAGCTGC
|
CTGTTACACCAAACTCCTGGATTTCCAGCTGGCACTCAAGGACTGTGAAGAATGTATC
|
CAGCTAGAACCGACCTTCATCAAGGGTTATACACGGAAAGCCGTTGCCCTGGAAGCGA
|
TGAAGGACTACACCAAAGTCATGGATGTGTACCGGAAGGCGCTAGACCCGGACTCCAG
|
CTGTAAGGAGGCGGCAGACGGCTACCAGCGCTGTATGATGGCGCAGTACAACCGGCAC
|
CACAGCCCCGAAGATGTGAAGCGACGAGCCATGGCCGACCCCGAGGTGCAGCAGATCA
|
TGAGTGACATAGCCATGCACCTTATCCTGGAGCAGATGCAGAAGGACCCCCAGGCACT
|
CAGCGAACACTTAAAGAATCCTGTAATCGCAAAGAAGATCCAGAAGCTGATGGATGTG
|
GGTCTGATTGCAATTCGACAATGACTTGTCCATCCCCGCTTCCCTTCGCCCTCATCCG
|
GAAAGAGGAGCGGGGACCGCGG
|
ORF Start: ATG at 14ORF Stop:TGA at 1646
SEQ ID NO:26544 aa MW at 62861.1 kD
NOV9a,MKQVNELKEKGNKALGVENIDNALQCYSEAIKLDPHNHVLYSNCSATYAKKGDYQKAY
|
CG59826-01 Protein SequenceKDGCKTVDLKFDWCKGYSRKAAALEFLNRFEEAKGTYEESLKHEADNPQLKEGLQNME
|
ARLAERKCMNAFNMPNLKQKLESDPRTRTLLSDPTYRELIEQLKNKPSDLGTKLRDPR
|
ILTTLSVLLEVDRGSMDEEEQVATPSPPPLPKKETKPEPVEEDLPENKKQALKEKBLR
|
NDAYKKKDRLTALKHYDKAKELDPSNMTYITNQAAVYFEKGDYNKCWELCEKSIEVGR
|
ENREDYLQIAKVCARIGNSYFKEEKYKDAIHFYNKSLAEHRTPDVLKKCQQAEKILKE
|
QERLAYINPDLALEEKNKGNECFQKGDYPHAMKHYTEAIKRNPKDAKLYSNRAACYTK
|
LLDFQLALKDCEECIQLEPTFIKGYTRKAVALEAMKDYTKVMDVYRKALDPDSSCKEA
|
ADGYQRCMMAQYNRHHSPEDVKRRAMADPEVQQIMSDIAMHLILEQMQKDPQALSEHL
|
KNPVIAKKIQKLMDVGLIAIRQ
|
[0359] Further analysis of the NOV9a protein yielded the following properties shown in Table 9B.
46TABLE 9B
|
|
Protein Sequence Properties NOV9a
|
|
PSort0.4500 probability located in cytoplasm;
analysis:0.3000 probability located in microbody
(peroxisome); 0.1000 probability located/
in mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0360] A search of the NOV9a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 9C.
47TABLE 9C
|
|
Geneseq Results for NOV9a
NOV9aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAY07058Renal cancer associated antigen 1 . . . 543500/543 (92%)0.0
precursor sequence-Homo sapiens, 1 . . . 543515/543 (94%)
543 aa. [WO9904265-A2, 28-JAN-
1999]
ABB57064Mouse ischaemic condition related 1 . . . 543488/543 (89%)0.0
protein sequence SEQ ID NO: 127- 1 . . . 543509/543 (92%)
Mus musculus, 543 aa.
[WO200188188-A2, 22-NOV-2001]
ABG18854 Novel human diagnostic protein 1 . . . 538472/540 (87%)0.0
#18845-Homo sapiens, 629 aa.33 . . . 572499/540 (92%)
[WO200175067-A2, 11-OCT-2001]
ABG18854Novel human diagnostic protein 1 . . . 538472/540 (87%)0.0
#18845-Homo sapiens, 629 aa.33 . . . 572499/540 (92%)
[WO200175067-A2, 11-OCT-2001]
ABG18853Novel human diagnostic protein26 . . . 348279/323 (86%)e−160
#18844-Homo sapiens, 319 aa. 1 . . . 313290/323 (89%)
[WO200175067-A2, 11-OCT-2001]
|
[0361] In a BLAST search of public sequence datbases, the NOV9a protein was found to have homology to the proteins shown in the BLASTP data in Table 9D.
48TABLE 9D
|
|
Public BLASTP Results for NOV9a
NOV9aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P31948Stress-induced-phosphoprotein 1 (STI1)1 . . . 543500/543 (92%)0.0
(Hsp70/Hsp90-organizing protein)1 . . . 543515/543 (94%)
(Transformation-sensitive protein IEF
SSP 3521)-Homo sapiens (Human),
543 aa.
O35814P60 PROTEIN-Rattus norvegicus1 . . . 543489/543 (90%)0.0
(Rat), 543 aa.1 . . . 543509/543 (93%)
Q99L66STRESS-INDUCED1 . . . 543488/543 (89%)0.0
PHOSPHOPROTEIN 1-Mus1 . . . 543509/543 (92%)
musculus (Mouse), 543 aa.
Q60864MSTI1-Mus musculus (Mouse), 5431 . . . 543488/543 (89%)0.0
aa.1 . . . 543509/543 (92%)
O54981HSP70/HSP90 ORGANIZING1 . . . 543485/543 (89%)0.0
PROTEIN- Cricetulus griseus (Chinese1 . . . 543508/543 (93%)
hamster), 543 aa.
|
[0362] PFam analysis predicts that the NOV9a protein contains the domains shown in the Table 9E.
49TABLE 9E
|
|
Domain Analysis of NOV9a
Identities/
NOV9aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
TPR: domain 1 of 8 4 . . . 3711/34 (32%)0.0001
25/34 (74%)
TPR: domain 2 of 8 72 . . . 105 9/34 (26%)0.07
25/34 (74%)
TPR: domain 3 of 8225 . . . 25813/34 (38%)18
23/34 (68%)
TPR: domain 4 of 8259 . . . 292 9/34 (26%)4.5
24/34 (71%)
TPR: domain 5 of 8300 . . . 33312/34 (35%) 7e−05
26/34 (76%)
TPR: domain 6 of 8360 . . . 39310/34 (29%)7.4e−06
26/34 (76%)
TPR: domain 7 of 8394 . . . 42711/34 (32%)7.5e−08
29/34 (85%)
TPR: domain 8 of 8428 . . . 461 8/34 (24%)0.89
26/34 (76%)
|
Example 10
[0363] The NOV10 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 10A.
50TABLE 10A
|
|
NOV10 Sequence Analysis
|
|
SEQ ID NO: 272649 bp
NOV10a,CAACTGATTTTAAACTCTACAGAGATGCCTACTTGTTTCTACTATGTCTTGTGGCAGT
CG59839-01 DNA Sequence
TGCTGGCATTGGGTTTATCTACACTATTATTAATAGCATTTTAAATGAGGTACAAGTT
|
GGGGTCATAATTATCGAGTCTCTTGATATTATCACAATTACTGTGCCCCCTGCACTTC
|
CTGCTGCAATGACTGCTGGTATTGTGTATGCTCAGAGAAGACTGAAAAAAATCGGTAT
|
TTTCTGTATCAGTCCTCAAAGAATAAATATTTGTGGACAGCTCAATCTTGTTTGCTTT
|
GACAAGACTGGAACTCTAACTGAAGATGGTTTAGATCTTTGGGGGATTCAACGAGTGG
|
AAAATGCACGATTTCTTTCACCAGAAGAAAATGTGTGCAATGAGATGTTGGTAAAATC
|
CCAGTTTGTTGCTTGTATGGCTACTTGTCATTCACTTACAAAAATTGAAGGAGTGCTC
|
TCTGGTGATCCACTTGATCTGAAAATGTTTGAGGCTATTGGATGGATTCTGGAAGAAG
|
CAACTGAAGAAGAAACAGCACTTCATAATCGAATTATGCCCACAGTGGTTCGTCCTCC
|
CAAACAACTGCTTCCTGAATCTACCCCTGCAGGAAACCAAGAAATGGAGCTGTTTGAA
|
CTTCCAGCTACTTATGAGATAGGAATTGTTCGCCAGTTCCCATTTTCTTCTGCTTTGC
|
AACGTATGAGTGTGGTTGCCAGGGTGCTGGGGGATAGGAAAATGGACGCCTACATGAA
|
AGGAGCGCCCGAGGCCATTGCCGGTCTCTGTAAACCTGAAACAGTTCCTGTCGATTTT
|
CAAAACGTTTTGGAAGACTTCACTAAACAGGGCTTCCGTGTGATTGCTCTTGCACACA
|
GAAAATTGGAGTCAAAACTGACATGGCATAAAGTACAGAATATTAGCAGAGATGCAAT
|
TGAGAACAACATGGATTTTATGGGATTAATTATAATGCAGAACAAATTAAAGCAAGAA
|
ACCCCTGCAGTACTTGAAGATTTGCATAAAGCCAACATTCGCACCGTCATGGTCACAG
|
GTGACAGTATGTTGACTGCTGTCTCTGTGGCCAGAGATTGTGGAATGATTCTACCTCA
|
GGATAAAGTGATTATTGCTGAAGCATTACCTCCAAAGGATGGGAAAGTTGCCAAAATA
|
AATTGGCATTATGCAGACTCCCTCACGCAGTGCAGTCATCCATCAGCAATTGACCCAG
|
AGGCTATTCCGGTTAAATTGGTCCATGATAGCTTAGAGGATCTTCAAATGACTCGTTA
|
TCATTTTGCAATGAATGGAAAATCATTCTCAGTGATACTGGAGCATTTTCAAGACCTT
|
GTTCCTAAGTTGATGTTGCATGGCACCGTGTTTGCCCGTATGGCACCTGATCAGAAGA
|
CACAGTTGATAGAAGCATTGCAAAATGTTGATTATTTTGTTGGGATGTGTGGTGATGG
|
CGCAAATGATTGTGGTGCTTTGAAGAGGGCACACGGAGGCATTTCCTTATCGGAGCTC
|
GAAGCTTCAGTGGCATCTCCCTTTACCTCTAAGACTCCTAGTATTTCCTGTGTGCCAA
|
ACCTTATCAGGGAAGGCCGTGCTGCTTTAATAACTTCCTTCTGTGTGTTTAAATTCAT
|
GGCATTGTACAGCATTATCCAGTACTTCAGTGTTACTCTGCTGTATTCTATCTTAAGT
|
AACCTAGGAGACTTCCAGTTTCTCTTCATTGATCTGGCAATCATTTTGGTAGTGGTAT
|
TTACAATGAGTTTAAATCCTGCCTGGAAAGAACTTGTGGCACAAAGACCACCTTCGGG
|
TCTTATATCTGGGGCCCTTCTCTTCTCCGTTTTGTCTCAGATTATCATCTGCATTGGA
|
TTTCAATCTTTGGGTTTTTTTTGGGTCAAACAGCAACCTTGGTATGAAGTGTGGCATC
|
CAAAATCAGATGCTTGTAATACAACAGGAAGCGGGTTTTGGAATTCTTCACACGTAGA
|
CAATGAAACCGAACTTGATGAACTAATATACAAAATTATGAAAATACCACAGTGTTTT
|
TTATTTCCAGTTTTCAGTACCTCATAGTGGCAATTGCCTTTTCAAAAGGAAAACCCTT
|
CAGGCAACCTTGCTACAAAAATTATTTTTTTGTTTTTTCTGTGATTTTTTTATATATT
|
TTTATATTATTCATCATGTTGTATCCAGTTGCCTCTGTTGACCAGGTTCTTCAGATAG
|
TGTGTGTACCATATCAGTGGCGTGTAACTATGCTCATCATTGTTCTTGTCAATGCCTT
|
TGTGTCTATCACAGTGGAGGAGTCAGTGGATCGGTGGGGAAAATGCTGCTTACCCTGG
|
GCCCTGGGCTGTAGAAAGAAGACACCAAAGGCAAAGTACATGTATCTGGCGCAGGAGC
|
TCTTGGTTGATCCAGAATGGCCACCAAAACCTCAGACAACCACAGAAGCTAAAGCTTT
|
AGTTAAGGAGAATGGATCATGTCAAATCATCACCATAACATAGCAGTGAATCAGTCTC
|
AGTGGTATTGCTGATAGCAGTATTCAGGAATATGTGATTTTAGGAGTTTCTGATCCTG
|
TGTGTCAGAATGGCACTAGTTCAGTTTATGTCCCTTCTGATATGTAGCTTATTTGAC
|
AGCTTTGCTCTTCCTTAAAATAAAAAAAAAAAAAAAAAA
|
ORF Start: ATG at 183ORF Stop: TAG at 2055
SEQ ID NO: 28624 aaMW at 69590.1 kD
NOV10a,MTAGIVYAQRRLKKIGIFCISPQRINICGQLNLVCFDKTGTLTEDGLDLWGIQRVENA
CG59839-01 Protein Sequence
RFLSPEENVCNEMLVKSQFVACMATCHSLTKIEGVLSGDPLDLKMFEAIGWILEEATE
|
EETALHNRIMPTVVRPPKQLLPESTPAGNQEMELFELPATYEIGIVRQFPFSSALQRM
|
SVVARVLGDRKMDAYMKGAPEAIAGLCKPETVPVDFQNVLEDFTKQGFRVIALAHRKL
|
ESKLTWHKVQNISRDAIENNMDFMGLIIMQNKLKQETPAVLEDLHKANIRTVMVTGDS
|
MLTAVSVARDCGMILPQDKVIIAEALPPKDGKVAKINWHYADSLTQCSHPSAIDPEAI
|
PVKLVHDSLEDLQMTRYHFAMNGKSFSVILEHFQDLVPKLMLHGTVFARMAPDQKTQL
|
IEALQNVDYFVGMCGDGANDCGALKRAHGGISLSELEASVASPFTSKTPSISCVPNLI
|
REGRAALITSFCVFKFMALYSIIQYFSVTLLYSILSNLGDFQFLFIDLAIILVVVFTM
|
SLNPAWKELVAQRPPSGLISGALLFSVLSQIIICIGFQSLGFFWVKQQPWYEVWHPKS
|
DACNTTGSGFWNSSHVDNETELDELIYKIMKIPQCFLFPVFSTS
|
[0364] Further analysis of the NOV10a protein yielded the following properties shown in Table 10B.
51TABLE 10B
|
|
Protein Sequence Properties NOV10a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4617 probability located in mitochondrial
inner membrane; 0.4000 probability located
in Golgi body; 0.3000 probability located
in endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0365] A search of the NOV10a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 10C.
52TABLE 10C
|
|
Geneseq Results for NOV10a
NOV10aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAB40996Human ORFX ORF760 polypeptide 48 . . . 604557/557 (100%)0.0
sequence SEQ ID NO:1520-Homo 2 . . . 558557/557 (100%)
sapiens, 692 aa. [WO200058473-A2,
05-OCT-2000]
AAM93525Human polypeptide, SEQ ID NO:261 . . . 604341/344 (99%)0.0
3259-Homo sapiens, 505 aa. 1 . . . 344341/344 (99%)
[EP1130094-A2, 05-SEP-2001]
AAU23078Novel human enzyme polypeptide297 . . . 604308/308 (100%)0.0
#164-Homo sapiens, 476 aa. 8 . . . 315308/308 (100%)
[WO200155301-A2, 02-AUG-2001]
AAM93906Human polypeptide, SEQ ID NO: 1 . . . 573263/575 (45%)e−138
4053-Homo sapiens, 842 aa.139 . . . 691370/575 (63%)
[EP1130094-A2, 05-SEP-2001]
AAM79751Human protein SEQ ID NO 3397- 39 . . . 573241/537 (44%)e−124
Homo sapiens, 666 aa. 1 . . . 515342/537 (62%)
[WO200157190-A2, 09-AUG-2001]
|
[0366] In a BLAST search of public sequence datbases, the NOV10a protein was found to have homology to the proteins shown in the BLASTP data in Table 10D.
53TABLE 10D
|
|
Public BLASTP Results for NOV10a
NOV10aIdentities/
ProteinProtein/Residues/Similarities for
AccessionOrganism/Matchthe MatchedExpect
NumberLengthResiduePortionValue
|
Q9H7F0Probable 1 . . . 624 624/624 (100%)0.0
cation- 61 . . . 684 624/624 (100%)
transporting
ATPase 3
(EC 3.6.3.-)-
Homo sapiens
(Human),
684 aa
(fragment).
Q96KS1HYPO- 1 . . . 499482/499 (96%)0.0
THETICAL185 . . . 680486/499 (96%)
77.3 KDA
PROTEIN-
Homo
sapiens
(Human),
701 aa.
Q9NQ11Probable 1 . . . 573264/575 (45%)e−138
cation- 477 . . . 1029371/575 (63%)
transporting
ATPase 1
(EC 3.6.1.-)-
Homo sapiens
(Human),
1180 aa.
O74431Probable 1 . . . 588 250/609 (41%)e−124
cation- 583 . . . 1181367/609 (60%)
transporting
ATPase
C1672.11C
(EC 3.6.3.-)-
Schizo-
saccharomyces
pombe (Fission
yeast), 1315 aa.
Q21286Probable 1 . . . 573248/587 (42%)e−122
cation- 467 . . . 1034351/587 (59%)
transporting
ATPase
K07E3.7 in
chromosome
X (EC 3.6.3.-)-
Caenorhabditis
elegans,
1152 aa.
|
[0367] PFam analysis predicts that the NOV Oa protein contains the domains shown in the Table 10E.
54TABLE 10E
|
|
Domain Analysis of NOV10a
Identities/
NOV10aSimilarities for
Matchthe MatchedExpect
Pfam DomainRegionRegionValue
|
Hydrolase: domain 1 of 1 31 . . . 443 40/423 (9%)0.009
247/423 (58%)
Hemagglutinin: domain 1 of 1555 . . . 561 4/7 (57%)8.9
7/7 (100%)
|
Example 11
[0368] The NOV11 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 11A.
55TABLE 11A
|
|
NOV11 Sequence Analysis
|
|
SEQ ID NO: 293880 bp
NOV 11a,CTAACATCATCATGACAGGATCAAATTCACACATAACAATATTAACTTTAAATGTAAA
CG59847-01 DNA Sequence
TGGACTAAATGCTCCAATTAAAAGACACAGACTGGCAAATTGGATAAAGAGTCAAGAC
|
CCATCAGTGTGCTGTATTCAGGAAACCCATCTCATGTGCAGAGACACACATAGGCTCA
|
AAATAAAAGGATGGAGGAAGATCTATCAAAAACAAAAAAAGGCAGGGGTTGCAACCCT
|
AGTCTCTGATAAAACAGACAAACCAATAAAGATCAAAAGAGACAAAGAAGGCCATTAC
|
ATAATGGTAAAGGGATCAATTCAACAAGAAGAGCTAACTATCCTAAATATATATGCAC
|
CCAATACAGCAGCACCCACATTCATAAAGCAAGTCCTGAGTGACCTACAAAGAGACTT
|
AGACTCCCACACATTAATAATGGGAGACTTTAACACCCCACTGTCAACATTAGACAGA
|
TCAACGAGACAGAAAGTCAACAAGGATACCCAGGAATTGAACTCAGCTCTGCACCAAG
|
TGGACCTAATAGACATCTATACAACTCTCCACCCCAAATCAACAGAATATACATTTTT
|
TTCAGCACCACACCCCACCTATTCCAAAATTGACCACATACTGGGAAGTAAAGCTCTC
|
CTCAGCAAATGTAAAAGAACAGAAATTATAACAAACTATCTCTCAGACCACAGTGCAA
|
TCAAACTAGAACTCAGGATTCAGAATCTCACTCAAAACTGCTCAACTACATGGAAACT
|
GAACAACCTGCTCCTGAATGACTACTGGGTACATAACGAAATGAAGGCAGAAATAAAG
|
ATGTTCTTTGAAACCAACAAGAACAAAGACACAACATACCAGAATCTCTGGGACGCAT
|
TCAAAGGAGTGTGTAGAGGGAAATTTATAGCACTAAATGCCCACAAGAGAAAGCAGGA
|
AAGATCCAAAATTGACATCCTAACATCACAATTAAAAGAACTAGAAAAGCAAGAGCAA
|
ACACACTCAAAAGGTAGCAGAAGGCAAGAAATAACTAAAATCAGAGCAGAACTGAAGG
|
AAATAGAGACACGAAAAACCCTTCAAAAAATTAATGAATCCAGGAGCATCAACAAAAC
|
AGATAGACCACTAGCAAGACTAATAAAGAAAAAAAGAGAGAAGAATCAAATAGACGCA
|
ATAAAAAATGATAAAGGGGATATCACCACCGATCCCACAGAAATACAAACTACCATCA
|
GAGAATACTACAAACACCTCTACGCAAATAAACTAGAAAATCTAGAAGAAATGGATAA
|
ATTCCTCGACACATACACTCTCCCAAGACTAAACCAGGAAGAAGTTGAATCTCTGAAT
|
AGACCAATAACAGGATCTGAAACTGTGGCAATAATCAATAGCTTACCAACCAAAAAGA
|
GTCCAGGACCAGATGGATTCACAGCTGAATTCTACCAGAGGTACAAGGAGGAACTGGT
|
ACCATTCCCTCTGAAACTATTCCAATCAATAGAAAAAGAGGGAATCCTCCCTAACTCA
|
TTTTATGAGGCCAGCATCATTCTGATACCAAAGCCTGGCAGAGACACAACCAAAAAAG
|
AGAATTTTAGACCAATATCCTTGATGAACATTGATGCAAAAATCCTCAATAAAATACT
|
GGCAAAACGAATCCAGCAGCACAGCAAAAAGCTTATCCACCATGATCAAGTGGGCTTC
|
ATCCCTGGGATGCAAGGCTGGTTCAATATATGCAAATCAATAAATGTAATCCAGCATA
|
TAAACAGAGCCAAAGACAAAAACCACATGATTATCTCAATAGATGCAGAAAAGGCCTT
|
TGACAAAATTCAACAACGCTTCATGCTAAAAACTCTCAATAAATCAGGTATTGATGGG
|
ACGTATTTCAAAATAATAAGAGCTATCTATGACAAACCCACAGCCAATATCATACTGA
|
ATGGACAAAAACTGGAAGCATTCCCTTTGAAAACCGGCACAAGACAGGGATACCCTCT
|
CTCACCACTCCTATTCAACATAGTGTTGGAAGTTCTGGCCAGGGCAATTAGGCAGGAG
|
AAGGAAATAAAGTGTATTCAATTAGGAAAAGAGGAAGTCAAATTGTCCCTGTTTGCAG
|
ACGACATGATTGTATATCTAGAAAACCCCATTGTCTCAGCCCAAAATCTCCTTAAGCT
|
GATAAGCAACTTCAGCAAAGTCTCAGGATACAAAATCAATGTACAAAAATCACAAGCA
|
TTCTTCTACACCAACTACAGACAAACAGAGAGCCAAATCATGAGTGAACTCCCATTCA
|
CAATTGCTTCAAAGAGAATAAAATACCTAGGAATCCAACTTACAAGGGATGTGAAGGA
|
CCTCTTCAAGGAGAACTACAAACCACTGCTCAAGGAAATAAAAGAGGATACAAACAAA
|
TGGAAGAACATTCCATGCTCATGGGTAGGAAGAATCAATATCGTGAAAATGGCCATAC
|
TGCCCAAGGTAATTTACAGATTCAATGCCATCCCCATCAAGCTACCAATGCCTTTCTT
|
CACAGAATTGGAAAAAACTACTTTAAAGTTCATATGGAACCAAAAAAGAGCCTGCATC
|
GCCAAGTCAATCCTAAGCCAAAAGGACAAAGCTGGAGGCATCACATTACCTGACTTCA
|
CACTACACTACAAGGCTACAGTAACCAAAACAGCATGGTACTGGTACCAAAACAGAGA
|
TATAGATCAATGGAACAGAACAGAGCCCTCAGAAATAATGCCGCATATCTACAACTAT
|
CGGATCTTTGACAAACCTGAGAAAAACAAGCAATGGGGAAAGGATTCCCTATTTAATA
|
AATGGTGCTGGGAAACCTGCCTAGCCATATGTAGAAAGCTGAAACTGGATCCCTTCCT
|
TACACCTTATACAAAAATCAATTCAAGATGGATTAAAGACTTAAACGTTAGACCTAAA
|
ACCATAAAAACTCTAGAAGAAAACCTAGGCATTACCATTCAGGACGTAGCTATGGGCA
|
AGGACTTCATGTCTAAAACACCAAAAGCAATGGCAACAAAAGCCAAAATTGACAAATG
|
GGATCTAATTAAACTAAAGAGCTTCTGCACAGCAAAAGAAACTACCATCAGAGTGAAC
|
AGGCAACCTACAAAATGGGAGAAAATTTTTGCAGCCTACTCATCTGACAAAGGGCTAA
|
TATCCAGAATCTACAATGAACTCAAACAAATTTACAAGAAAAAAACAAACAACCCCAT
|
CAAAAAGTGGGCAAAGGACATGAACAGACACTTCTCAAAAGAAGACATTTATGCAGCC
|
AAAAAACACATGAAAAAATGCTCATCATCACTGGCCATCAGAGAAATGCAAATCAAAA
|
CCACAATGAGATACCATCTCACACCAGTTAGAATGGCAATCATTAAAAAGTCAGGAAA
|
CAACAGGTGCTGGAGAGGATGTGGAGAAATAGGAACACTTTTACACTGTTGGTGGGAC
|
TGTAAACTAGTTCAACCCTTGTGGAAGTCAGTGTGGCGATTCCTCAGGGATCTAGACC
|
TAGAAATACCATTTGACCCAGCCATCCCATTACTGGGTATATACCCAAAGGACTATAA
|
ATCATGCTGCTATAAAGATACATGCACACATATGTTTATTGCAGCACTATTCACAATA
|
GAAAGACTTGGAACCAACCCAAAGACTTGGAACCAACCCAAATGTCCAACAATGATAG
|
ACTGGATTAAGAAAATGTGGCACATATACACCATGGAATACTATGCAGCCATAAAAAA
|
TGAAGAGTTCATGTCCTTTGTAGGGACATGGATGAAATTGGAAATCATCATTCTCAGT
|
AAACTATCGCAAGAACAAAAAACCAAACACCGCATATTCTCACTCATAGGTGGGAATT
|
GAACAATGAGATCACATGGACACAGGAAGGGGAACATCACACTCTGGGGACT
|
ORF Start: ATG at 12ORF Stop: TGA at 3828
SEQ ID NO: 301272 aaMW at 148630.6 kD
NOV11a,MTGSNSHITILTLNVNGLNAPIKRHRLANWIKSQDPSVCCIQETHLMCRDTHRLKIKG
CG59847-01 Protein Sequence
WRKIYQKQKKAGVATLVSDKTDKPIKIKRDKEGHYIMVKGSIQQEELTILNIYAPNTG
|
APTFIKQVLSDLQRDLDSHTLIMGDFNTPLSTLDRSTRQKVNKDTQELNSALHQVDLI
|
DIYRTLHPKSTEYTFFSAPHPTYSKIDHILGSKALLSKCKRTEIITNYLSDHSAIKLE
|
LRIQNLTQNCSTTWKLNNLLLNDYWVHNEMKAEIKMFFETNKNKDTTYQNLWDAFKGV
|
CRGKFIALNAHKRKQERSKIDILTSQLKELEKQEQTHSKGSRRQEITKIRAELKEIET
|
RKTLQKINESRSINKTDRPLARLIKKKREKNQIDAIKNDKGDITTDPTEIQTTIREYY
|
KHLYANKLENLEEMDKFLDTYTLPRLNQEEVESLNRPITGSETVAIINSLPTKKSPGP
|
DGFTAEFYQRYKEELVPFPLKLFQSIEKEGILPNSFYEASIILIPKPGRDTTKKENFR
|
PISLMNIDAKILNKILAKRIQQHSKKLIHHDQVGFIPGMQGWFNICKSINVIQHINRA
|
KDKNHMIISIDAEKAFDKIQQRFMLKTLNKSGIDGTYFKIIRAIYDKPTANIILNGQK
|
LEAFPLKTGTRQGYPLSPLLFNIVLEVLARAIRQEKEIKCIQLGKEEVKLSLFADDMI
|
VYLENPIVSAQNLLKLISNFSKVSGYKINVQKSQAFFYTNYRQTESQIMSELPFTIAS
|
KRIKYLGIQLTRDVKDLFKENYKPLLKEIKEDTNKWKNIPCSWVGRINIVKMAILPKV
|
IYRFNAIPIKLPMPFFTELEKTTLKFIWNQKRACIAKSILSQKDKAGGITLPDFTLHY
|
KATVTKTAWYWYQNRDIDQWNRTEPSEIMPHIYNYRIFDKPEKNKQWGKDSLFNKWCW
|
ETWLAICRKLKLDPFLTPYTKINSRWIKDLNVRPKTIKTLEENLGITIQDVAMGKDFM
|
SKTPKAMATKAKIDKWDLIKLKSFCTAKETTIRVNRQPTKWEKIFAAYSSDKGLISRI
|
YNELKQIYKKKTNNPIKKWAKDMNRHFSKEDIYAAKKHMKKCSSSLAIREMQIKTTMR
|
YHLTPVRMAIIKKSGNNRCWRGCGEIGTLLHCWWDCKLVQPLWKSVWRFLRDLDLEIP
|
FDPAIPLLGIYPKDYKSCCYKDTCTHMFIAALFTIERLGTNPKTWNQPKCPTMIDWIK
|
KMWHIYTMEYYAAIKNEEFMSFVGTWMKLEIIILSKLSQEQKTKHRIFSLIGGN
|
[0369] Further analysis of the NOV11 a protein yielded the following properties shown in Table 11B.
56TABLE 11B
|
|
Protein Sequence Properties NOV11a
|
|
PSort0.5500 probability located in endoplasmic reticulum
analysis:(membrane); 0.1900 probability located in lysosome
(lumen); 0.1363 probability located in microbody
(peroxisome); 0.1000 probability located in endoplasmic
reticulum (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0370] A search of the NOV11a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 11C.
57TABLE 11C
|
|
Geneseq Results for NOV11a
NOV11aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB64943Human secreted protein sequence1..12721205/1281 (94%)0.0
encoded by gene 7 SEQ ID NO:121 -6..12801213/1281 (94%)
Homo sapiens, 1280 aa.
[WO200076530-A1, 21 Dec. 2000]
ABG14889Novel human diagnostic protein 1..12721196/1281 (93%)0.0
#14880 - Homo sapiens, 1284 aa.10..12841218/1281 (94%)
[WO200175067-A2, 11 Oct. 2001]
ABG10795Novel human diagnostic protein 1..12721196/1281 (93%)0.0
#10786 - Homo sapiens, 1284 aa.10..12841218/1281 (94%)
[WO200175067-A2, 11 Oct. 2001]
ABG09636Novel human diagnostic protein 1..12721196/1281 (93%)0.0
#9627 - Homo sapiens, 1284 aa.10..12841218/1281 (94%)
[WO200175067-A2, 11 Oct. 2001]10..12841218/1281
ABG06053Novel human diagnostic protein 1..12721196/1281 (93%)0.0
#6044 - Homo sapiens, 1284 aa.
[WO200175067-A2, 11-OCT-2001]
|
[0371] In a BLAST search of public sequence datbases, the NOV11a protein was found to have homology to the proteins shown in the BLASTP data in Table 11D.
58TABLE 11D
|
|
Public BLASTP Results for NOV11a
NOV11aIdentities/
ProteinResidues/Similarities
AccessionMatchfor theExpect
NumberProtein/Organism/LengthResiduesMatched PortionValue
|
O00360PUTATIVE P150 - Homo sapiens1..12721209/1281 (94%)0.0
(Human), 1275 aa.1..12751224/1281 (95%)
Q9YSK0HYPOTHETICAL 149.0 KDA1..12721208/1281 (94%)0.0
PROTEIN - Homo sapiens1..12751226/1281 (95%)
(Human), 1275 aa.
Q9UN80HYPOTHETICAL 149.0 KDA1..12721207/1281 (94%)0.0
PROTEIN - Homo sapiens1..12751226/1281 (95%)
(Human), 1275 aa.
Q15604ORF2 CONTAINS A REVERSE1..12721205/1281 (94%)0.0
TRANSCRIPTASE DOMAIN -1..12751224/1281 (95%)
Homo sapiens (Human), 1275 aa.
AAL50637HYPOTHETICAL 149.0 KDA1..12721206/1281 (94%)0.0
PROTEIN - Homo sapiens1..12751223/1281 (95%)
(Human), 1275 aa.
|
[0372] PFam analysis predicts that the NOV1 la protein contains the domains shown in the Table 11E.
59TABLE 11E
|
|
Domain Analysis of NOV11a
Identities/
NOV11aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
AP_endonucleas1: 8..234 79/297 (27%)1e-83
domain 1 of 1202/297 (68%)
rvt: domain 1 of 1494..764 84/278 (30%)1.1e-71
218/278 (78%)
|
Example 12
[0373] The NOV12 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 12A.
60TABLE 12A
|
|
NOV12 Sequence Analysis
|
|
SEQ ID NO: 313145 bp
NOV12a,ATTTTACTTGCTGGATCTTCAGCCTTGACCTGTATGGCAAATGGCTTATGGGACCGAT
CG59905-01 DNA Sequence
CCCTGCCCAAGTGTTTGGCTATATCCTGTGGACACCCAGGGGTCCCTGCCAACGCCGT
|
CCTCACTGGAGAGCTGTTTACCTATGGCGCCGTCGTGCACTACTCCTGCAGAGGGAGC
|
GAGAGCCTCATAGGCAACGACACGAGAGTGTGCCAGGAAGACAGTCACTCGAGCGGGG
|
CACTGCCCCACTGCACAGGAAATAATCCTGGATTCTGTGGTGATCCGGGGACCCCAGC
|
ACATGGGTCTCGGCTTGGTGATGACTTTAAGACAAAGAGTCTTCTCCGCTTCTCCTGT
|
GAAATGGGGCACCAGCTGAGGGGCTCCCCTGAACGCACGTGTTTGCTCAATGGGTCAT
|
GGTCAGGACTGCAGCCGGTGTGTGAGGCCGTGTCCTGTGGCAACCCTGGCACACCCAC
|
CAACGGAATGATTGTCAGTAGTGATGGCATTCTGTTCTCCAGCTCGGTCATCTATGCC
|
TGCTGGGAAGGCTACAAGACCTCAGGGCTCATGACACGGCATTGCACAGCCAATGGGA
|
CCTGGACAGGCACTGCTCCCGACTGCACAATTATAAGTTGTGGGGATCCAGGCACACT
|
AGCAAATGGCATCCAGTTTGGGACCGACTTCACCTTCAACAAGACTGTGAGCTATCAG
|
TGTAACCCAGGCTATGTCATGGAAGCAGTCACATCCGCCACTATTCGCTGTACCAAAG
|
ACGGCAGGTGGAATCCGAGCAAACCTGTCTGCAAAGCCGTGCTGTGTCCTCAGCCGCC
|
GCCGGTGCAGAATGGAACAGTGGAGGGAAGTGATTTCCGCTGGGGCTCCAGCATAAGT
|
TACAGCTGCATGGACGGTTACCAGCTCTCTCACTCCGCCATCCTCTCCTGTGAAGGTC
|
GCGGGGTGTGGAAAGGAGAGATCCCCAGTGTCTGCCTGTGTTCTGCGGAGACCCTGGC
|
ATCCCCGCAGAAGGGCGACTTAGTGGGAAAAGTTTCACCTATAAGTCCGAAGTCTTCT
|
TCCAGTGCAAATCTCCATTTATACTCGTGGGATCCTCCAGAAGAGTCTGCCAAGCTGA
|
CGGCACGTGGAGCGGCATACAACCCACCTGCATTGGTAATAATTATCATACAGCTCTG
|
GGGATACCTGGGAGTATTTGGAGATGAGGACGCTTCATTCCGAAATTGGGTCATTTGT
|
GATTACATAGAAAGTGTTTCCATAGACAGTTTTCTGTACAAGGTTGGAAGCACGGTTT
|
TTTTCAGGTGCAGAAAAGGCTACCATATTCAAGGTTCCACGACTCGCACCTGCCTTGC
|
CAATTTAACATGGAGTGGGATACAGACCGAATGTATACCTCATGCCTGCAGACAGCCA
|
GAAACCCCGGCACACGCGGATGTGAGAGCCATCGATCTTCCTACTTTCGGCTACACCT
|
TAGTGTACACCTGCCATCCAGGCTTTTTCCTCGCAGGGGGATCTCAGCACAGAACATG
|
TAAAGCAGACATGAAATGGACAGGAAAGTCGCCTGTGTGTAAAATTCCTTCAGATGTC
|
TTTTTCGTCAATTCACTGTGGAAGGGGTATTATGAATATTTAGGGAAAAGACAACCCG
|
CCACTCTAACTGTTGACTGGTTCAATGCAACAAGCAGTAAGGTGAATGCCACCTTCAG
|
CGAAGCCTCGCCAGTGGAGCTGAAGTTGACAGGCATTTACAAGAAGGAGGAGGCCCAC
|
TTACTCCTGAAAGCTTTTCAAATTAAAGGCCAGGCAGATATTTTTGTAAGCAAGTTCG
|
AAAATGACAACTGGGGACTAGATGGTTATGTGTCATCTGGACTTGAAAGAGGAGGATT
|
TACTTTTCAAGGTGACATTCATGGAAAAGACTTTGGAAAATTTAAGCTAGAAAGGCAA
|
GGATGGGTCACAATATTCTTGAGCCTATTTCTTCATCTTAAATCTCAGTATAGAAGTT
|
CCCAAGGTTGTTACGAGATTGAGAGGCCACATCCTTTAAACCCAGATCAAGACTCTTC
|
CAGTCATTACCACGGCACCAGCAGTGGCTCTGTGGCGGCTGCCATTCTGGTTCCTTTC
|
TTTGCTCTAATTTTATCAGGGTTTGCATTTTACCTCTACAAACACAGAACGAGACCAA
|
AAGTTCAATACAATGGCTATGCTGGGCATGAAAACAGCAATGGACAAGCATCGTTTGA
|
AAACCCCATGTATGATACAAACTTAAAACCCACAGAAGCCAAGGCTAAAACCACACAC
|
GGGCTGCTCACGTCTCTGGACTCCAAGAGAAAGATGGTTATAACCCTGGCAGAGTCTT
|
TACCCGCCCTGCGAACCTGCCTGTTGATCTGCTTCCCACTCCTCTTTAGACTTGCTCG
|
TGAATCTTCAGCACAGCCATTTGTTTTGAGTATTCAAACACATAAAAAGATATCTGGC
|
AACTTCCCACTTCTTATGACTTCTCAGACACCTTGTAACGGAGCTTCCCTTGGAGGTG
|
GACAAACTTCTCACTCACAAGAGAGACGGAGAGCAACAGAGAGAGATGGGCAGGGATT
|
GATTCGAGGATTGGCTGACTCGATTATGGAGGGTGAGAAGTCCCAGGACAGGCCGTCT
|
GCAAGGCGTAAACCCAGGGAAGCTGCTGGTATGGCTCGGTTCAAGTCCAAAGGCCTCA
|
GCACCAAGGAAACCAAGGTGATAACTCTCAGTTCGGGTCAAAGTCCTGGGAGTCTGCA
|
AGGCCTCTCATTGAGTAAGTCTCACAAGATCCGATGGTTTTATAAAGGGCAGTTCCCC
|
TGCACAAGCTCTCTTCTCTGCCACCACGCGTTTGCTGTTCATTCACCTTTCACCATGA
|
TTGTGAGGCCTCCCAGCCATGTGGAACTGTGCAAAGAGGATGTTCAAAGGGCATCTTG
|
GACCAGGAGGAGGAAGCCCATTGTGAGGATGGCAAAGAACAATAAATGGAAGGCCCGT
|
GGAACCCTGAGCGGCTGCCCCATCACTGTCCTCTCCATCCTTTCCATGCTTCCCATGG
|
TGCATGGCTCTCAGGCTCTCCAAGCACCATGAGCTCCTGGCGCGTATGGCCTGGCCCC
|
TGAGTGTGGAGTCTGTGGACTTTAAATGTGCCTTTACTCAGCCAGCATCTCCCTGGCT
|
GGGTGCGATGTTG
|
ORF Start: ATG at 34ORF Stop: TGA at 3046
SEQ ID NO: 321004 aaMW at 109752.7 kD
NOV12a,MANGLWIDRSLPKCLAISCGHPGVPANAVLTGELFTYGAVVHYSCRGSESLIGNDTRVC
CG59905-01 Protein Sequence
QEDSHWSGALPHCTGNNPGFCGDPGTPAHGSRLGDDFKTKSLLRFSCEMGHQLRGSPE
|
RTCLLNGSWSGLQPVCEAVSCGNPGTPTNGMIVSSDGILFSSSVIYACWEGYKTSGLM
|
TRHCTANGTWTGTAPDCTIISCGDPGTLANGIQFGTDFTFNKTVSYQCNPGYVMEAVT
|
SATIRCTKDGRWNPSKPVCKAVLCPQPPPVQNGTVEGSDFRWGSSISYSCMDGYQLSH
|
SAILSCEGRGVWKCEIPSVCLCSAETLASPQKGDLVGKVSPISPKSSSSANLHLYSWD
|
PPEESAKLTARGAAYNPPALVIIIIQLWGYLGVFGDEDASFRNWVICDYIESVSTDSF
|
LYKVGSTVFFRCRKGYHIQGSTTRTCLANLTWSGIQTECIPHACRQPETPAHADVRAI
|
DLPTFGYTLVYTCHPGFFLAGGSEHRTCKADMKWTGKSPVCKIPSDVFFVNSLWKGYY
|
EYLGKRQPATLTVDWFNATSSKVNATFSEASPVELKLTGIYKKEEAHLLLKAFQIKGQ
|
ADIFVSKFENDNWGLDGYVSSGLERGGFTFQGDIHGKDFGKFKLERQGWVTIFLSLFL
|
HLKSQYRSSQGCYEIERPHPLNPDQDSSSNYNGTSSGSVAAAILVPFFALILSGFAFY
|
LYKHRTRPKVQYNGYAGHENSNGQASFENPMYDTNLKPTEAKAKTTHGLLTSLDSKRK
|
MVITLAESLPALRTCLLICFPLLFRLARESSAQPFVLSIQTHKKISGNFPLIMTSQTP
|
CNGASLGGGQTSHSQERRRATERDGQGLIRGLADSIMEGEKSQDRPSARRKPREAAGM
|
ARFKSKGLSTKETKVITLSSGQSPGSLQGLSLSKSHKIRWFYKGQFPCTSSLLCHHAF
|
AVHSPFTMIVRPPSHVELCKEDVQRASWTRRRKPIVRMAKNNKWKARGTLSGCPITVL
|
SILSMLPMVHGSQALQAP
|
[0374] Further analysis of the NOV12a protein yielded the following properties shown in Table 12B.
61TABLE 12B
|
|
Protein Sequence Properties NOV12a
|
|
PSort0.6000 probability located in nucleus; 0.6000 probability
analysis:located in plasma membrane; 0.4000 probability located
in Golgi body; 0.3000 probability located in
in endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0375] A search of the NOV12a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 12C.
62TABLE 12C
|
|
Geneseq Results for NOV12a
NOV12aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Indentifier[Patent #, Date]ResiduesRegionValue
|
AAU00816Human Immunoglobulin superfamily, 3..204118/202 (58%)4e-75
IgSF, protein #2 - Homo sapiens, 613375..576152/202 (74%)
aa. [WO200118176-A1,
15 Mar. 2001]
AAG68264Human POLY11 protein sequence 3..498147/507 (28%)2e-64
SEQ ID NO:22 - Homo sapiens, 2050 775..1242223/507 (42%)
aa. [WO200179294-A2,
25 Oct. 2001]
AAU19902Novel human calcium-binding protein 3..498147/507 (28%)2e-64
#11 - Homo sapiens, 1139 aa.270..737223/507 (42%)
[WO200155304-A2, 02 Aug. 2001]
AAG66398Receptor 222 - Unidentified, 979 aa. 3..498147/507 (28%)2e-64
[CN1296952-A, 30 May 2001]110..577223/507 (42%)
AAU16963Human novel secreted protein, SEQ 3..498147/507 (28%)2e-64
ID 204 - Homo sapiens, 1139 aa.270..737223/507 (42%)
[WO200155441-A2, 02 Aug. 2001]
|
[0376] In a BLAST search of public sequence datbases, the NOV12a protein was found to have homology to the proteins shown in the BLASTP data in Table 12D.
63TABLE 12D
|
|
Public BLASTP Results for NOV12a
NOV12aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96Q03KIAA1884 PROTEIN - Homo 6..739375/757 (49%)0.0
sapiens (Human), 946 aa (fragment).221..932480/757 (62%)
Q96RM4CUB AND SUSHI MULTIPLE 1..328312/328 (95%)0.0
DOMAINS 1 PROTEIN - Homo2786..3113313/328(95%)
sapiens (Human), 3508 aa.
Q923L3CSMD1 - Mus musculus (Mouse), 1..328279/328 (85%)e-176
3564 aa.2842..3169297/328 (90%)
Q96QU9CUB AND SUSHI MULTIPLE408..739300/347 (86%)e-174
DOMAINS PROTEIN 1 SHORT3058..3375302/347 (86%)
FORM - Homo sapiens (Human),
3389 aa.
Q96PZ3KIAA1894 PROTEIN - Homo218..739239/544 (43%)e-125
sapiens (Human), 514 aa (fragment). 2..500310/544 (56%)
|
[0377] PFam analysis predicts that the NOV12a protein contains the domains shown in the Table 12E.
64TABLE 12E
|
|
Domain Analysis of NOV12a
Identities/
NOV12aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
sushi: domain 1 of 718..7118/63 (29%)2.9e-11
43/63 (68%)
sushi: domain 2 of7 79..13215/63 (24%) 4e-11
41/63 (65%)
sushi: domain 3 of 7137..19120/64 (31%)1.4e-08
41/64 (64%)
sushi: domain 4 of 7196..25121/65 (32%)5.2e-14
46/65 (71%)
sushi: domain 5 of 7256..31020/64 (31%) 4e-09
45/64 (70%)
myosin_head:398..417 8/22 (36%)8.1
domain 1 of 116/22 (73%)
sushi: domain 6 of 7395..44518/62 (29%)3.2e-07
39/62 (63%)
sushi: domain 7 of 7450..50518/64 (28%)1.4e-09
41/64 (64%)
|
Example 13
[0378] The NOV13 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 13A.
65TABLE 13A
|
|
NOV13 Sequence Analysis
|
|
SEQ ID NO: 33 1463 bp
NOV13a,TGGCACGAGGATTACATTTCACTTGGTTACAGCTGACTGGGGATTTTAATCATTTGGA
CG59930-01 DNA Sequence
GCCACACTGAAAAAAAATTTCCCCCTGCAATCTACTCTTAACTGATAGTCGCTAACT
|
TTGGTTATGTGTGTCTATTCAGAATATTACATTAATTGATTTTCTCAAACAAAGGTCCT
|
TTCTGAAATGGTATCTATGATTCAGCTATTCAAAACCTAATGAAGTTGGTGACTATGA
|
CAATGTGGAGAAATCATGACAGAAATGTGGTTTGTACTGGGGCTGTCAATGCTGTAA
|
AGGAAGTTTGGGAAAAAAGAATAAAGAAACTCAATGAAGACCTGAAGCGAGAGAAGGA
|
ATTTCAACACAAGCTAGTGCGGATCTGGGAAGAACGAGTAAGCTTAACCAAGCTAAGA
|
GAAAAGGTCACCAGGGAAGATGGAAGAGTCATTTTGAAGATAGAAAAAGAGGAATGGA
|
AGACCTCCTTCTTCTCTGCTGAAACTGAATCAACTACAGGAATGGCAACTTCATAG
|
AACTGGTTTGCTGAAAATTCCTGAATTCATTGGAAGATTCCAGAACCTCATTGTGTTA
|
GATTTATCTCGAAACACAATTTCAGAGATACCACCAGGGATTGGACTGCTTACTAGAC
|
TTCAGGAACTGATTCTCAGCTACAACAAAATCAAGACTGTCCCCAAGGAACTAAGTAA
|
TTGTGCCAGCTTGGAGAAACTAGAACTGGCTGTTAACAGAGATATATGTGATCTTCCA
|
CAAGAGCTCAGCAATCTGCTAAAACTTACTCACCTTGATCTGAGTATGAACGATTTTA
|
CTACAATCCCTCTTGCTGTGTTGAACATGCCTGCCCTTGAGTGGCTGGACATGGGAAG
|
CAACAAACTTGAACAACTTCCTGATACTATAGAAAGAATGCAAAATCTACATACGTTA
|
TGGCTGCAACGAAATGAAATAACATGCTTGCCTCAAACAATCAGCAATATGAAAAATC
|
TGGGTACTCTTGTTCTCAGCAACAATAAACTGCAAGATATTCCAGTATGCATGGAAGA
|
AATGGCAAATCTGAGGTTTGTCAACTTCAGAGACAACCCACTGAAATTGAAAGTATCA
|
CTTCCTCCCAGTGAAGGCACAGATGAAGAAGAGGAACGGGAATTATTTGGCCTTCAGT
|
TTATGCACACATACATACAAGAGTCACGGAGAAGAGCAGATCACCAAGTCAACGGTTC
|
AACTACTTTACCAATCTCCATAAATACGGATGGATAATATAATTCAAGATGCCCTTCT
|
AAAGAGGATTACTTTGGTGAATTCTCTAATGTTTGGATTTCTGAAGTAAAAAGAAAAG
|
CTATTACCAAAGTTTAATGAGGCCACAGCATTTTTTAAAGTTCATATTATTTGCTATT
|
TAAAGTATATATTTTTTTGATATAAAAATATAATTAAAAATTTTTTGGTGCAAAAAAA
|
AAAAAAAAAAAAA
|
ORF Start: ATG at 248 ORF Stop: TAA at 1253
SEQ ID NO: 34 335 aa MW at 38792.5kD
NOV13a,MTENVVCTGAVNAVKEVWEKRIKKLNEDLKREKEFQHKLVRIWEERVSLTKLREKVTR
CG59930-01 Protein Sequence
EDGRVILKIEKEEWKTLPSSLLKLNQLQEWQLHRTGLLKIPEFIGRFQNLIVLDLSRN
|
TISEIPPGIGLLTRLQELILSYNKIKTVPKELSNCASLEKLELAVNRDICDLPQELSN
|
LLKLTHLDLSMNDFTTIPLAVLNMPALEWLDMGSNKLEQLPDTIERMQNLHTLWLQRN
|
EITCLPQTISNMKNLGTLVLSNNKLQDIPVCMEEMANLRFVNFRDNPLKLKVSLPPSE
|
GTDEEEERELFGLQFMHTYIQESRRRADHQVNGSTTLPISINTDG
|
SEQ ID NO: 35 1276 bp
NOV13b,CTAGAATTCAGCGGCCGCTGAATTCTAGCTGAAAAAAAAATTTCCCCCTGCAATCTAC
CG59930-02 DNA Sequence
TCTTAACTGATAGTCGCTAACTTTGGTTATGTGTGGTAAGTATTTGACAATGTCTATT
|
CAGAATATTACATTAATTGATTTCTCAAACAAAGGTCCTTTCTGAAATGGTATCTATG
|
ATTCAGCTATTCAAAACCTAATGAAGTTGGTGACTATGACAATGTGGAGAAATCATGA
|
CAGAAAATGTGGTTTGTACTGGGGCTGTCAATGCTGTAAAGGAAGTTTGGGAAAAAAG
|
AATAAAGAAACTCAATGAAGACCTGAAGCGAGAGAAGGAATTTCAACACAAGCTAGTG
|
CGGATCTGGGAAGAACGAGTAAGCTTAACCAAGCTAAGAGAAAGGTCACCAGGGAAG
|
ATGGAAGAGTCATTTTGAAGATAGAAAAAGAGGAATGGAAGACCCTCCCTTCTTCTCT
|
GCTGAAACTGAATCAACTACAGGAATGGCAACTTCATAGAACTGGTTTGCTGAAAATT
|
CCTGAATTCATTGGAAGATTCCAGAACCTCATTGTGTTAGATTTATCTCGAAACACAA
|
TTTCAGAGATACCACCAGGGATTGGACTGCTTACTAGACTTCAGGAACTGATTCTCAG
|
CTACAACAAAATCAAGACTGTCCCCAAGGAACTAAGTAATTGTGCCAGCTTGGAGAAA
|
CTAGAACTGGCTGTTAACAGAGATATATGTGATCTTCCACAAGAGCTCAGCAATCTGC
|
TAAAACTTACTCACCTTGATCTGAGTATGAACGATTTTACTACAATCCCTCTTGCTGT
|
GTTGAACATGCCTGCCCTTGAGTGGCTGGACATGGGAAGCAACAAACTTGAACAACTT
|
CCTGATACTATAGAAAGAATGCAAAATCTACATACGTTATGGCTGCAACGAAATGAAA
|
TAACATGCTTGCCTCAAACAATCAGCAATATGAAAAATCTGGGTACTCTTGTTCTCAG
|
CAACAATAAACTGCAAGATATTCCAGTATGCATGGAAGAAATGGCAAATCTGAGGTTT
|
GTCAACTTCAGAGACAACCCACTGAAATTGAAAGTATCACTTCCTCCCAGTGAAGGCA
|
CAGATGAAGAAGAGGAACGGGAATTATTTGGCCTTCAGTTTATGCACACATACATACA
|
AGAGTCACGGAGAAGAGCAGATCACCAAGTCAACGGTTCAACTACTTTACCAATCTCC
|
ATAAATACGGATGGATAATATAATTCAAGATGCCCTTTTAAAGAGGATTACTTTGGTG
|
ORF Start: ATG at 229 ORF Stop: TAA at 1234
SEQ ID NO: 36 335 aa MW at 38792.5kD
NOV13bMTENVVCTGAVNAVKEVWEKRIKKLNEDLKREKEFQHKLVRIWEERVSLTKLREKVTR
CG59930-02 Protein Sequence
EDGRVILKIEKEEWKTLPSSLLKLNQLQEWQLHRTGLLKIPEFIGRFQNLIVLDLSRN
|
TISEIPPGIGLLTRLQELILSYNIKTVPKELSNCASLEKLELAVNRDICDLPQELSN
|
LLKLTHLDLSMNDFTTIPLAVLNMPALEWLDMGSNKLEQLPDTIERMQNLHTLWLQRN
|
EITCLPQTISNMKNLGTLVLSNNKLQDIPVCMEEMANLRFVNFRDNPLKLKVSLPPSE
|
GTDEEEERELFGLQFMHTYIQESRRRADHQVNGSTTLPISINTDG
|
[0379] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 13B.
66TABLE 13B
|
|
Comparison of NOV13a against NOV13b.
ProteinNOV13a Residues/Identities/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV13b1..335335/335 (100%)
1..335335/335 (100%)
|
[0380] Further analysis of the NOV13a protein yielded the following properties shown in Table 13C.
67TABLE 13C
|
|
Protein Sequence Properties NOV13a
|
|
PSort0.6500 probability located in cytoplasm; 0.1000
analysis:probability located in mitochondrial matrix space;
0.1000 probability located in lysosome (lumen);
0.0000 probability located in endoplasmic
reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0381] A search of the NOV13a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 13D.
68TABLE 13D
|
|
Geneseq Results for NOV13a
NOV13aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAU70173Rat secreted factor protein encoded by1..140116/141 (82%)2e−59
DNA clone P0269_H08 - Rattus1..141127/141 (89%)
norvegicus, 142 aa.
[WO200174901-A2, 11 Oct. 2001]
AAU21299Human novel foetal antigen, SEQ ID1..120110/120 (91%)7e−58
NO 1543 - Homo sapiens, 133 aa.6..125114/120 (94%)
[WO200155312-A2, 02 Aug. 2001]
AAB31563Amino acid sequence ofa human10..316 108/318 (33%)1e−48
SGT4-1 polypeptide - Homo sapiens,39..355 186/318 (57%)
371 aa. [WO200078959-A1,
28 Dec. 2000]
AAB31564Amino acid sequence of a human110..316 78/208 (37%)2e−38
SGT4-2 polypeptide - Homo sapiens,3..210132/208 (62%)
226 aa. [WO200078959-A1,
28 Dec. 2000]
ABG04747Novel human diagnostic protein #4738 -64..281 68/218 (31%)5e−25
Homo sapiens, 1271 aa.65..281 117/218 (53%)
[WO200175067-A2, 11 Oct. 2001]
|
[0382] In a BLAST search of public sequence datbases, the NOV13a protein was found to have homology to the proteins shown in the BLASTP data in Table 13E.
69TABLE 13E
|
|
Public BLASTP Results for NOV13a
NOV13aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organim/LengthResiduesPortionValue
|
Q96DD0SIMILAR TO RIKEN CDNA 1..335335/335 (100%)0.0
2010005E21 GENE - Homo 1..335335/335 (100%)
sapiens (Human), 335 aa.
Q9D8D72010005E21RIK PROTEIN - Mus 1..175156/175 (89%)1e−84
musculus (Mouse), 175 aa. 1..175167/175 (95%)
CAD20990LEUCINE-RICH REPEAT-11..316117/346 (33%)4e−51
CONTAINING 2 PROTEIN - Mus10..355188/346 (53%)
musculus (Mouse), 371 aa.
Q9CX042400002D05RIK PROTEIN - Mus11..316117/346 (33%)4e−51
musculus (Mouse), 371 aa.10..355188/346 (53%)
Q9BYS8LEUCINE-RICH REPEAT-10..316108/318 (33%)2e−48
CONTAINING 2 PROTEIN -39..355186/318 (57%)
Homo sapiens (Human), 371 aa.
|
[0383] PFam analysis predicts that the NOV13a protein contains the domains shown in the Table 13F.
70TABLE 13F
|
|
Domain Analysis of NOV13a
NOV13aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
LRR: domain 1 of 7107..12912/25 (48%)0.00049
20/25 (80%)
LRR: domain 2 of 7130..152 9/25 (36%)0.0094
20/25 (80%)
LRR: domain 3 of 7153..17610/26 (38%)75
17/26 (65%)
LRR: domain 4 of 7177..199 9/25 (36%)0.12
18/25 (72%)
LRR: domain 5 of 7200..222 8/25 (32%)0.54
20/25 (80%)
LRR: domain 6 of 7223..245 8/25 (32%)0.021
19/25 (76%)
LRR: domain 7 of 7246..268 9/25 (36%)10
16/25 (64%)
|
Example 14
[0384] The NOV14 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 14A.
71TABLE 14A
|
|
NOV14 Sequence Analysis
|
|
SEQ ID NO: 371933 bp
NOV14a,CTCCTTTCCACTTCTTTCCCCAGTCTGAGAAGAATACAGAAAGCCCCAAAGCCCTGGG
CG59934-01 DNA Sequence
TCTTGGTCCTGCCGCATGTTGGAGCTCGCTGTCCAGTGCTGGGGTCACTATGCCCTGG
|
TCACTGTGCCCTGGTCACTGTGCCATGCTGTGTGATGATCAAGTTTCTCTTCCTGTTC
|
TCTCTCCCTCCTCGAATCTAGAAGATGCTCCCTGGAGCCTGGCTGCTCTGGACCTCCC
|
TCCTGCTCCTGGCCAGGCCTGCCCAGCCCTGTCCCATGGGTTGTGACTGCTTCGTCCA
|
GGAGGTGTTCTGCTCAGATGAGGAGCTTGCCACCGTCCCGCTGGACATCCCGCCATAT
|
ACGAAAAACATCATCTTTGTGGAGACCTCGTTCACCACATTGGAAACCAGAGCTTTTG
|
GCAGTAACCCCAACTTGACCAAGGTGGTCTTCCTCAACACTCAGCTCTGCCAGTTTAG
|
GCCGGATGCCTTTGGGGGGCTGCCCAGGCTGGAGGACCTGGAGGTCACAGGCAGTAGC
|
TTCTTGAACCTCAGCACCAACATCTTCTCCAACCTGACCTCGCTGGGCAAGCTCACCC
|
TCAACTTCAACATGCTGGAGGCTCTGCCCGAGGGTCTTTTCCAGCACCTGGCTGCCCT
|
GGAGTCCCTCCACCTGCAGGGGAACCAGCTCCAGGCCCTGCCCAGGAGGCTCTTCCAG
|
CCTCTGACCCATCTGAAGACACTCAACCTGGCCCAGAACCTCCTGGCCCAGCTCCCGG
|
AGGAGCTGTTCCACCCACTCACCAGCCTGCAGACCCTGAAGCTGAGCAACAACGCGCT
|
CTCTGGTCTCCCCCAGGGTGTGTTTGGCAAACTGGGCAGCCTGCAGGAGCTCTTCCTG
|
GACAGCAACAACATCTCGGAGCTGCCCCCTCAGGTGTTCTCCCAGCTCTTCTGCCTAG
|
AGAGGCTGTGGCTGCAACGCAACGCCATCACGCACCTGCCGCTCTCCATCTTTGCCTC
|
CCTGGGTAATCTGACCTTTCTGAGCTTGCAGTGGAACATGCTTCGGGTCCTGCCTGCC
|
GGCCTCTTTGCCCACACCCCATGCCTGGTTGGCCTGTCTCTGACCCATAACCAGCTGG
|
AGACTGTCGCTGAGGGCACCTTTGCCCACCTGTCCAACCTGCGTTCCCTCATGCTCTC
|
ATACAATGCCATTACCCACCTCCCAGCTGGCATCTTCAGAGACCTGGAGGAGTTGGTC
|
AAACTCTACCTGGGCAGCAACAACCTTACGGCGCTGCACCCAGCCCTCTTCCAGAACC
|
TGTCCAAGCTGGAGCTGCTCAGCCTCTCCAAGAACCAGCTGACCACACTTCCGGAGGG
|
CATCTTCGACACCAACTACAACCTGTTCAACCTGGCCCTGCACGGTAACCCCTGGCAG
|
TGCGACTGCCACCTGGCCTACCTCTTCAACTGGCTGCAGCAGTACACCGATCGGCTCC
|
TGAACATCCAGACCTACTGCGCTGGCCCTGCCTACCTCAAAGGCCAGGTGGTGCCCGC
|
CTTGAATGAGAAGCAGCTGGTGTGTCCCGTCACCCGGGACCACTTGGGCTTCCAGGTC
|
ACGTGGCCGGACGAAAGCAAGGCAGGGGGCAGCTGGGATCTGGCTGTGCAGGAAAGGG
|
CAGCCCGGAGCCAGTGCACCTACAGCAACCCCGAGGGCACCGTGGTGCTCGCCTGTGA
|
CCAGGCCCAGTGTCGCTGGCTGAACGTCCAGCTCTCTCCTTGGCAGGGCTCCCTGGGA
|
CTGCAGTACAATGCTAGTCAGGAGTGGGACCTGAGGTCGAGCTGCGGTTCTCTGCGGC
|
TCACCGTGTCTATCGAGGCTCGGGCAGCAGGGCCCTAGTAGCAGCGCATACAGGAGCT
|
GGGGAAGGGGGCTTTGGGGCCTGCCCACGCGACAGGTAGGGGCGGAGGGGAGCTGAGT
|
CTCCGAAGCTTGGCTTTAC
|
ORF Start: ATG at 199ORF Stop: TAG at 1834
SEQ ID NO: 38545 aaMW at 60613.9 kD
NOV14a,MLPGAWLLWTSLLLLARPAQPCPMGCDCFVQEVFCSDEELATVPLDIPPYTKNIIFVE
CG59934-01 Protein Sequence
TSFTTLETRAFGSNPNLTKVVFLNTQLCQFRPDAFGGLPRLEDLEVTGSSFLNLSTNI
|
FSNLTSLGKLTLNFNMLEALPEGLFQHLAALESLHLQGNQLQALPRRLFQPLTHLKTL
|
NLAQNLLAQLPEELFHPLTSLQTLKLSNNALSGLPQGVFGKLGSLQELFLDSNNISEL
|
PPQVFSQLFCLERLWLQRNAITHLPLSIFASLGNLTFLSLQWNMLRVLPAGLFAHTPC
|
LVGLSLTHNQLETVAEGTFAHLSNLRSLMLSYNAITHLPAGIFRDLEELVKLYLGSNN
|
LTALHPALFQNLSKLELLSLSKNQLTTLPEGIFDTNYNLFNLALHGNPWQCDCHLAYL
|
FNWLQQYTDRLLNIQTYCAGPAYLKGQVVPALNEKQLVCPVTRDHLGFQVTWPDESKA
|
GGSWDLAVQERAARSQCTYSNPEGTVVLACDQAQCRWLNVQLSPWQGSLGLQYNASQE
|
WDLRSSCGSLRLTVSIEARAAGP
|
SEQ ID NO: 3911719 bp
NOV14b,CCATGCTGTGTGATGATCAAGTTTCTCTTCCTGTTCTCTCTCCCTCCTCGAATCTAGA
CG59934-02 DNA Sequence
AGATGCTCCCTGGAGCCTGGCTGCTCTGGACCTCCCTCCTGCTCCTGGCCAGGCCTGC
|
CCAGCCCTGTCCCATGGGTTGTGACTGCTTCGTCCAGGAGGTGTTCTGCTCAGATGAG
|
GAGCTTGCCACCGTCCCACTGGACATCCCGCCATATACGAAAAACATCATCTTTGTGG
|
AGACCTCGTTCACCACATTGGAAACCAGAGCTTTTGGCAGTAACCCCAACTTGACCAA
|
GGTGGTCTTCCTCAACACTCAGCTCTGCCAGTTTAGGCCGGATGCCTTCAGGGGGCTG
|
CCCAGGCTGGAGGACCTGGAGGTCACAGGCAGTAGCTTCTTGAACCTCAGCACCAACA
|
TCTTCTCCAACCTGACCTCGCTGGGCAAGCTCACCCTCAACTTCAACATGCTGGAGGC
|
TCTGCCCGAGGGTCTTTTCCAGCACCTGGCTGCCCTGGAGTCCCTCCACCTGCAGGGG
|
AACCAGCTCCAGGCCCTGCCCAGGAGGCTCTTCCAGCCTCTGACCCATCTGAAGACAC
|
TCAACCTGGCCCAGAACCTCCTGGCCCAGCTCCCGGAGGAGCTGTTCCACCCACTCAC
|
CAGCCTGCAGACCCTGAAGCTGAGCAACAACGCGCTCTCTGGTCTCCCCCAGGGTGTG
|
TTTGGCAAACTGGGCAGCCTGCAGGAGCTCTTCCTGGACAGCAACAACATCTCGGAGC
|
TGCCCCCTCAGGTGTTCTCCCAGCTCTTCTGCCTAGAGAGGCTGTGGCTGCAACGCAA
|
CGCCATCACGCACCTGCCGCTCTCCATCTTTGCCTTCCTGGGTAATCTGACCTTTCTG
|
AGCTTGCAGTGGAACATGCTTCGGGTCCTGCCTGCCGGCCTCTTTGCCCACACCCCGT
|
GCCTGGTTGGCCTGTCTCTGACCCATAACCAGCTGGAGACTGTCGCTGAGGGCACCTT
|
TGCCCACCTGTCCAACCTGCGTTCCCTCATGCTCTCATACAATGCCATTACCCACCTC
|
CCAGCTGGCATCTTCAGAGACCTGGAGGAGTTGGTCAAACTCTACCTGGGCAGCAACA
|
ACCTTACGGCGCTGCACCCAGCCCTCTTCCAGAACCTGTCCAAGCTGGAGCTGCTCAG
|
CCTCTCCAAGAACCAGCTGACCACACTTCCGGAGGGCATCTTCGACACCAACTACAAC
|
CTGTTCAACCTGGCCCTGCACGGTAACCCCTGGCAGTGCGACTGCCACCTGGCCTACC
|
TCTTCAACTGGCTGCAGCAGTACACCGATCGGCTCCTGAACATCCAGACCTACTGCGC
|
TGGCCCTGCCTACCTCAAAGGCCAGGTGGTGCCCGCCTTGAATGAGAAGCAGCTGGTG
|
TGTCCCGTCACCCGGGACCACTTGGGCTTCCAGGTCACGTGGCCGGACGAAAGCAAGG
|
CAGGGGGCAGCTGGGATCTGGCTGTGCAGGAAAGGGCAGCCCGGAGCCAGTGCACCTA
|
CAGCAACCCCGAGGGCACCGTGGTGCTCGCCTGTGACCAGGCCCAGTGTCGCTGGCTG
|
AACGTCCAGCTCTCTCCTCAGCAGGGCTCCCTGGGACTGCAGTACAATGCTAGTCAGG
|
AGTGGGACCTGAGGTCGAGCTGCGGTTCTCTGCGGCTCACCGTGTCTATCGAGGCTCG
|
GGCAGCAGGGCCCTAGTAGCAGCGCATACAGGAGCTG
|
ORF Start: ATG at 61ORF Stop: TAG at 1696
SEQ ID NO: 40545 aaMW at 60715.1 kD
NOV14b,MLPGAWLLWTSLLLLARPAQPCPMGCDCFVQEVFCSDEELATVPLDIPPYTKNIIFVE
CG59934-02 Protein Sequence
TSFTTLETRAFGSNPNLTKVVFLNTQLCQFRPDAFRGLPRLEDLEVTGSSFLNLSTNI
|
FSNLTSLGKLTLNFNMLEALPEGLFQHLAALESLHLQGNQLQALPRRLFQPLTHLKTL
|
NLAQNLLAQLPEELFHPLTSLQTLKLSNNALSGLPQGVFGKLGSLQELFLDSNNISEL
|
PPQVFSQLFCLERLWLQRNAITHLPLSIFAFLGNLTFLSLQWNMLRVLPAGLFAHTPC
|
LVGLSLTHNQLETVAEGTFAHLSNLRSLMLSYNAITHLPAGIFRDLEELVKLYLGSNN
|
LTALHPALFQNLSKLELLSLSKNQLTTLPEGIFDTNYNLFNLALHGNPWQCDCHLAYL
|
FNWLQQYTDRLLNIQTYCAGPAYLKGQVVPALNEKQLVCPVTRDHLGFQVTWPDESKA
|
GGSWDLAVQERAARSQCTYSNPEGTVVLACDQAQCRWLNVQLSPQQGSLGLQYNASQE
|
WDLRSSCGSLRLTVSIEARAAGP
|
[0385] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 14B.
72TABLE 14B
|
|
Comparison of NOV14a against NOV14b.
ProteinNOV14a Residues/Identities/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV14b1..545492/545 (90%)
1..545492/545 (90%)
|
[0386] Further analysis of the NOV14a protein yielded the following properties shown in Table 14C.
73TABLE 14C
|
|
Protein Sequence Properties NOV14a
|
|
PSort0.8493 probability located in lysosome (lumen); 0.3700
analysis:probability located in outside; 0.1325 probability
located in microbody (peroxisome); 0.1000
probability located in endoplasmic reticulum (membrane)
SignalPCleavage site between residues 22 and 23
analysis:
|
[0387] A search of the NOV14a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 14D.
74TABLE 14D
|
|
Geneseq Results for NOV14a
NOV14aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABB53264Human polypeptide #4 - Homo22..439148/444 (33%)4e−56
sapiens, 581 aa. [WO200181363-A1,25..468213/444 (47%)
01 Nov. 2001]
AAY69183Amino acid sequence of murine 1..427150/455 (32%)2e−50
glycoprotein V (GP V) - Mus sp, 566 1..454205/455 (44%)
aa. [WO200008137-A2,
17 Feb. 2000]
AAY69184Amino acid sequence of a human 1..449152/500 (30%)3e−47
glycoprotein V (GP V) - Homo 1..476212/500 (42%)
sapiens, 560 aa. [WO200008137-A2,
17 Feb. 2000]
AAR71294Human glycoprotein V - Homo 1..449152/500 (30%)3e−47
sapiens, 560 aa. [WO9502054-A, 1..476212/500 (42%)
19 Jan. 1995]
AAR85889WD-40 domain-contg. rat insulin-like22..444171/564 (30%)3e−41
growth factor binding protein - Rattus41..601221/564 (38%)
rattus, 603 aa. [WO9521252-A2,
10 Aug. 1995]
|
[0388] In a BLAST search of public sequence datbases, the NOV14a protein was found to have homology to the proteins shown in the BLASTP data in Table 14E.
75TABLE 14E
|
|
Public BLASTP Results for NOV14a
NOV14aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P22792Carboxypeptidase N 83 kDa chain23..531459/521 (88%)0.0
(Carboxypeptidase N regulatory 2..511466/521 (89%)
subunit) - Homo sapiens (Human),
536 aa (fragment).
AAH25836SIMILAR TO 1..545368/547 (67%)0.0
CARBOXYPEPTIDASE N,16..562434/547 (79%)
POLYPEPTIDE 2, 83KD - Mus
musculus (Mouse), 562 aa.
(fragment).
Q9DBB91300018K11RIK PROTEIN - Mus 1..545368/547 (67%)0.0
musculus (Mouse), 570 aa.24..570434/547 (79%)
BAB84586LIB - Rattus norvegicus (Rat),22..439148/444 (33%)4e−59
578 aa.25..468210/444 (46%)
BAB84587LIB - Homo sapiens (Human),22..439148/444 (33%)1e−55
581 aa.25..468213/444 (47%)
|
[0389] PFam analysis predicts that the NOV14a protein contains the domains shown in the Table 14F.
76TABLE 14F
|
|
Domain Analysis of NOV14a
NOV14aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
LRRNT:21..4812/31 (39%)0.00049
domain 1 of 119/31 (61%)
LRR:74..97 9/25 (36%)1.9e+02
domain 1 of 1316/25 (64%)
LRR: 98..121 7/25 (28%)1.5e+02
domain 2 of 1316/25 (64%)
LRR:122..145 9/25 (36%)1
domain 3 of 1318/25 (72%)
LRR:146..16910/25 (40%)0.0093
domain 4 of 1319/25 (76%)
LRR:170..19310/25 (40%)0.022
domain 5 of 1318/25 (72%)
LRR:194..21713/25 (52%)0.00021
domain 6 of 1321/25 (84%)
LRR:218..24112/25 (48%)0.0004
domain 7 of 1322/25 (88%)
LRR:242..265 9/25 (36%)0.29
domain 8 of 1318/25 (72%)
LRR:266..28910/25 (40%)0.067
domain 9 of 1320/25 (80%)
LRR:290..313 7/25 (28%)0.79
domain 10 of 1319/25 (76%)
LRR:314..33710/25 (40%)0.00072
domain 11 of 1320/25 (80%)
LRR:338..36111/25 (44%)0.015
domain 12 of 1318/25 (72%)
LRR:362..38510/25 (40%)0.15
domain 13 of 1317/25 (68%)
LRRCT:395..44620/55 (36%)2e−12
domain 1 of 141/55 (75%)
|
Example 15
[0390] The NOV15 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 15A.
77TABLE 15A
|
|
NOV15 Sequence Analysis
|
|
SEQ ID NO: 411068 bp
NOV15a,GCTTGCTTCCCCAACCCTGCACCGGCCATGCGCCCGGCCTTGGCGGTGGGCCTGGTGT
CG88565-01 DNA Sequence
TCGCAGGCTGCTGCAGTAACGTGATCTTCCTAGAGCTCCTGGCCCGGAAGCATCCAGG
|
ATGTGGGAACATTGTGACATTTGCACAATTTTTATTTATTGCTGTGGAAGGCTTCCTC
|
TTTGAAGCTGATTTGGGAAGGAAGCCACCAGCTATCCCAATAAGGTATTATGCCATAA
|
TGGTGACCATGTTCTTCACCGTGAGCGTGGTGAACAACTATGCCCTGAATCTCAACAT
|
TGCCATGCCCCTGCATATGATATTTAGATCCGGTTCTCTAATTGCCAACATGATTCTA
|
GGAATTATCATTTTGAAGAAAAGGTACAGTATATTCAAATATACCTCCATTGCCCTGG
|
TGTCTGTGGGGATATTTATTTGCACTTTTATGTCAGCAAAGCACGTGACTTCCCAGTC
|
CAGCTTGAGTGAGAATGATGGATTCCAGGCATTTGTGTGGTGGTTACTAGGCATTGGG
|
GCATTGACTTTTGCTCTTCTGATGTCAGCAAGGATGGGCGTATTCCAAGAGACTCTCT
|
ACAAACGATTTGGGAAACACTCCAAGGAGGCTTTGTTTTATAATCACGCCCTTCCACT
|
TCCCGGTTTCGTCTTCTTGGCTTCTGATATTTATGACCATGCAGTTCTATTCAATAAG
|
TCTGAGTTATATGAAATTCCCGTCATCGGAGTGACCCTGCCCATCATGTGGTTCTACC
|
TCCTCATGAACATCATCACTCAGTATGTGTGCATCCGGGGTGTGTTTATCCTCACCAC
|
AGAATGCGCCTCCCTCACCGTCACGCTCGTCGTGACCCTACGCAAATTTGTGAGCCTC
|
ATCTTTTGCATCTTGTACTTCCAGAACCCCTTCACCCTGTGGCACTGGCTGGGCACCT
|
TGTTTGTCTTCATTGGGACCTTAATGTACACAGAGGTGTGGAACAACCTAGGGACCAC
|
AAAAAGTGAGCCTCAGAAGGACAGCAAGAAGAACTGAGGCCTGTCTGGAGTACGTAGA
|
CCAGTGTCGTCGTGAGGGTGGGAC
|
ORF Start: ATG at 28ORF Stop: TGA at 1021
SEQ ID NO: 42331 aaMW at 37423.1 kD
NOV15a,MRPALAVGLVFAGCCSNVIFLELLARKHPGCGNIVTFAQPLFIAVEGFLFEADLGRKP
CG88565-01 Protein Sequence
PAIPIRYYAIMVTMFFTVSVVNNYALNLNIAMPLHMIFRSGSLIANMILGIIILKKRY
|
SIFKYTSIALVSVGIFICTFMSAKQVTSQSSLSENDGFQAPVWWLLGIGALTFALLMS
|
ARMGIFQETLYKRFGKHSKEALFYNHALPLPGFVFLASDIYDHAVLFNKSELYEIPVI
|
GVTLPIMWFYLLMNIITQYVCIRGVFILTTECASLTVTLVVTLRKFVSLIFSILYFQN
|
PFTLWHWLGTLFVFIGTLMYTEVWNNLGTTKSEPQKDSKKN
|
[0391] Further analysis of the NOV15a protein yielded the following properties shown in Table 15B.
78TABLE 15B
|
|
Protein Sequence Properties NOV15a
|
|
PSort0.6400 probability located in plasma membrane; 0.4600
analysis:probability located in Golgi body; 0.3700 probability
located in endoplasmic reticulum (membrane);
0.1000 probability located in endoplasmic
reticulum (lumen)
SignalPCleavage site between residues 22 and 23
analysis:
|
[0392] A search of the NOV15a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 15C.
79TABLE 15C
|
|
Geneseq Results for NOV15a
NOV15aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB88341Human membrane or secretory1..331331/331 (100%)0.0
protein clone PSEC0055 - Homo1..331331/331 (100%)
sapiens, 331 aa.
[EP1067182-A2,
10 Jan. 2001]
AAB74727Human membrane associated protein1..331331/331 (100%)0.0
MEMAP-33 - Homo sapiens, 331 aa.1..331331/331 (100%)
[WO200112662-A2, 22 Feb. 2001]
AAB94391Human protein sequence SEQ ID1..331331/331 (100%)0.0
NO:14953 - Homo sapiens, 331 aa.1..331331/331 (100%)
[EP1074617-A2, 07 Feb. 2001]
AAM96349Human reproductive system related29..255 212/227 (93%)e−120
antigen SEQ ID NO: 5007 - Homo7..233220/227 (96%)
sapiens, 241 aa. [WO200155320-A2,
02 Aug. 2001]
AAB93332Human protein sequence SEQ ID69..250 182/182 (100%)e−100
NO:12434 - Homo sapiens, 185 aa.1..182182/182 (100%)
[EP1074617-A2, 07 Feb. 2001]
|
[0393] In a BLAST search of public sequence datbases, the NOV15 a protein was found to have homology to the proteins shown in the BLASTP data in Table 15D.
80TABLE 15D
|
|
Public BLASTP Results for NOV15a
NOV15aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q969S0MRNA, SIMILAR TO YEAST1..331331/331 (100%)0.0
YEA4P COMPLETE CDS1..331331/331 (100%)
(UNKNOWN) (PROTEIN FOR
MGC:14552) (CDNA FLJ14697 FIS,
CLONE NT2RP2005812) Homo
sapiens (Human), 331 aa.
Q95KB4HYPOTHETICAL 37.4 KDA1..331327/331 (98%)0.0
PROTEIN - Macaca fascicularis1..331330/331 (98%)
(Crab eating macaque)
(Cynomolgus monkey), 331 aa.
Q9JJF7BRAIN CDNA, CLONE MNCB-4414,1..331301/331 (90%)e-178
SIMILAR TO NM_005827 UDP-1..331316/331 (94%)
GALACTOSE TRANSPORTER
RELATED (HOMO SAPIENS) -
Mus musculus (Mouse), 331 aa.
Q9W429CG3774 PROTEIN - Drosophila1..317174/324 (53%)e-101
melanogaster (Fruit fly), 352 aa.5..328237/324 (72%)
Q96K33CDNA FLJ14820 FIS, CLONE69..250 182/182 (100%)e-100
OVARC1000335 - Homo sapiens1..182182/182 (100%)
(Human), 185 aa.
|
[0394] PFam analysis predicts that the NOV15a protein contains the domains shown in the Table 15E.
81TABLE 15E
|
|
Domain Analysis of NOV15a
Identities/
NOV15aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
cytochrome_b_C:257..288 9/38 (24%)7.2
domain 1 of 124/38 (63%)
DUF6: domain 1 of 1136..312423/177 (13%)3.1
110/177 (62%)
CbiM: domain 1 of 1183..323 28/199 (14%)9.1
93/199 (47%)
|
Example 16
[0395] The NOV16 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 16A.
82TABLE 16A
|
|
NOV16 Sequence Analysis
|
|
SEQ ID NO: 431455 bp
NOV16a,CAGCCAGGAGGAAAAAGCTAGGCGTCCACTTTCCGCAGCCATGCTCAAACAGAGTGAG
CG88623-01 DNA Sequence
AGGAGACGGTCCTGGAGCTACAGGCCCTGGAACACGACGGAGCTGAATGTGAACGTGG
|
GTGGCCACAGCTACCAGCTGGACTACTGCGAGCTGGCCGGCTTCCCCAAGACGCGCCT
|
AGGTCGCCTGGCCACCTCCACCAGCCGCAGCCGCCAGCTAAGCCTGTGCGACGACTAC
|
GAGGAGCAGACAGACGAATACTTCTTCGACCGCGACCCGGCCGTCTTCCAGCTGGTCT
|
ACAATTTCTACCTGTCCGGGGTGCTGCTGGTGCTCGACGGGCTGTGTCCGCGCCGCTT
|
CCTGGAGGAGCTGGGCTACTGGGGCGTGCGGCTCAAGTACACGCCACGCTGCTGCCGC
|
ATCTGCTTCGAGGAGCGGCGCGACGAGCTGAGCGAACGGCTCAAGATCCAGCACGAGC
|
TGCGCGCGCAGGCGCAGGTCGAGGAGGCGGAGGAACTCTTCCGCGACATGCGCTTCTA
|
CGGCCCGCAGCGGCGCCGCCTCTGGAACCTCATGGAGAAGCCATTCTCCTCGGTGGCC
|
GCCAAGGCCATCGGGGTGGCCTCCAGCACCTTCGTGCTCGTCTCCGTGGTGGCGCTGG
|
CGCTCAACACCGTGGAGGAGATGCAGCAGCACTCGGGGCAGGGCGAGGGCGGCCCAGA
|
CCTGCGGCCCATCCTGGAGCACGTGGACATGCTGTGCATGGGCTTCTTCACGCTCGAG
|
TACCTGCTGCGCCTAGCCTCCACGCCCGACCTGAGGCGCTTCGCGCGCAGCGCCCTCA
|
ACCTGGTGGACCTGGTGGCCATCCTGCCGCTCTACCTTCAGCTGCTGCTCGAGTGCTT
|
CACGGGCGAGGGCCACCAACGCGGCCAGACGGTGGGCAGCGTGGGTAAGGTGGGTCAG
|
GTGTTGCGCGTCATGCGCCTCATGCGCATCTTCCGCATCCTCAAGCTGGCGCGCCACT
|
CCACCGGACTGCGTGCCTTCGGCTTCACGCTGCGCCAGTGCTACCAGCAGGTGGGCTG
|
CCTGCTGCTCTTCATCGCCATGGGCATCTTCACTTTCTCTGCGGCTGTCTACTCTGTG
|
GAGCACGATGTGCCCAGCACCAACTTCACTACCATCCCCCACTCCTGGTGGTGGGCCG
|
CGGTGAGCATCTCCACCGTGGGCTACGGAGACATGTACCCAGAGACCCACCTGGGCAG
|
GTTTTTTGCCTTCCTCTGCATTGCTTTTGGGATCATTCTCAACGGGATGCCCATTTCC
|
ATCCTCTACAACAAGTTTTCTGATTACTACAGCAAGCTGAAGGCTTATGAGTATACCA
|
CCATACGCAGGGAGAGGGGAGAGGTGAACTTCATGCAGAGAGCCAGAAAGAAGATAGC
|
TGAGTGTTTGCTTGGAAGCAACCCACAGCTCACCCCAAGACAAGAGAATTAGTATTTT
|
ATAGG
|
ORF Start: ATG at 41ORF Stop: TAG at 1442
SEQ ID NO: 44467 aaMW at 53892.6 kD
NOV16a,MLKQSERRRSWSYRPWNTTELNVNVGGHSYQLDYCELAGFPKTRLGRLATSTSRSRQL
CG88623-01 Protein Sequence
SLCDDYEEQTDEYFFDRDPAVFQLVYNFYLSGVLLVLDGLCPRRFLEELGYWGVRLKY
|
TPRCCRICFEERRDELSERLKIQHELRAQAQVEEAEELFRDMRFYGPQRRRLWNLMEK
|
PFSSVAAKAIGVASSTFVLVSVVALALNTVEEMQQHSGQGEGGPDLRPILEHVEMLCM
|
GFFTLEYLLRLASTPDLRRFARSALNLVDLVAILPLYLQLLLECFTGEGHQRGQTVGS
|
VGKVGQVLRVMRLMRIFRILKLARHSTGLRAFGFTLRQCYQQVGCLLLFIAMGIFTFS
|
AAVYSVEHDVPSTNFTTIPHSWWWAAVSISTVGYGDMYPETHLGRFFAFLCIAFGIIL
|
NGMPISILYNKFSDYYSKLKAYEYTTIRRERGEVNFMQRARKKIAECLLGSNPQLTPR
|
QEN
|
[0396] Further analysis of the NOV16a protein yielded the following properties shown in Table 16B.
83TABLE 16B
|
|
Protein Sequence Properties NOV16a
|
|
PSort0.6000 probability located in plasma membrane; 0.4120
analysis:probability located in mitochondrial inner membrane;
0.4000 probability located in Golgi body; 0.3000
probability located in endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0397] A search of the NOV16a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 16C.
84TABLE 16C
|
|
Geneseq Results for NOV16a
NOV16aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAE13614Human Kv10.1 protein variant #421..467447/447 (100%)0.0
(Q77E) - Homo sapiens, 545 aa.99..545447/447 (100%)
[WO200179455-A1, 25 Oct. 2001]
AAE13610Human KvlO.1 protein - Homo21..467447/447 (100%)0.0
sapiens, 545 aa. [WO200179455-A1,99..545447/447 (100%)
25 Oct. 2001]
AAE13613Human Kv10.1 protein variant #321..467446/447 (99%)0.0
(V518M) - Homo sapiens, 545 aa.99..545447/447 (99%)
[WO200179455-A1, 25 Oct. 2001]
AAE13612Human Kv10.1 protein variant #221..467446/447 (99%)0.0
(M285L) - Homo sapiens, 545 aa.99..545447/447 (99%)
[WO200179455-A1, 25 Oct. 2001]
AAE13611Human Kv10.1 protein variant #121..467446/447 (99%)0.0
(L99V) - Homo sapiens, 545 aa.99..545447/447 (99%)
[WO200179455-A1, 25 Oct. 2001]
|
[0398] In a BLAST search of public sequence datbases, the NOV16a protein was found to have homology to the proteins shown in the BLASTP data in Table 16D.
85TABLE 16D
|
|
Public BLASTP Results for NOV16a
NOV16aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
AAL83910VOLTAGE-GATED POTASSIUM21..467447/447 (100%)0.0
CHANNEL KV11.1 - Homo sapiens99..545447/447 (100%)
(Human), 545 aa.
Q95L11VOLTAGE-GATED POTASSIUM21..436169/416 (40%)1e-92
CHANNEL ALPHA SUBUNIT37..438266/416 (63%)
KV2.2 - Oryctolagus cuniculus
(Rabbit), 911 aa.
Q63099Voltage-gated potassium channel21..436167/416 (40%)5e-92
protein Kv2.2 (CDRK) - Rattus37..438266/416 (63%)
norvegicus (Rat), 802 aa.
Q9BXD3POTASSIUM VOLTAGE-GATED21..436167/416 (40%)2e-91
CHANNEL, SHAB-RELATED37..438266/416 (63%)
SUBFAMILY, MEMBER 2 - Homo
sapiens (Human), 911 aa.
Q92953Voltage-gated potassium channel21..436167/416 (40%)
protein Kv2.2 - Homo sapiens37..438
(Human), 806 aa.
|
[0399] PFam analysis predicts that the NOV16a protein contains the domains shown in the Table 16E.
86TABLE 16E
|
|
Domain Analysis of NOV16a
Identities/
NOV16aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
K_tetra: 19..12541/112 (37%)4.8e-18
domain 1 of 172/112 (64%)
ion_trans:223..41449/226 (22%)1.8e-26
domain 1 of 1156/226 (69%)
|
Example 17
[0400] The NOV17 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 17A.
87TABLE 17A
|
|
NOV17 Sequence Analysis
|
|
SEQ ID NO: 451651 bp
NOV17a,ATGGCGGCCAACATGTACCGGGTCGGATCGGGCCGAAAGTCCCCCGGGCGGCCAGCCA
CG88645-01 DNA Sequence
TGACCTTCGGGCGCAGCGGGGCGGCCTCGGTGGTGCTGAACGTGGGCGGCGCCCGGTA
|
TTCGCTGTCCCGGGAGCTGCTGAAGGACTTCCCGCTGCGCCGCGTGAGCCGGCTGCAC
|
GGCTGCCGCTCCGAGCGCGACGTGCTCGAGGTGTGCGACGACTACGACCGCGAGCGCA
|
ACGAGTACTTCTTCGACCGGCACTCGGAGGCCTTCGGCTTCATCCTGCTCTACGTGCG
|
CGGCCACGGCAAGCTGCGCTTCGCGCCGCGGATGTGCGAGCTCTCCTTCTACAACGAG
|
ATGATCTACTGGGGCCTGGAGGGCGCGCACCTCGAGTACTGCTGCCAGCGCCGCCTCG
|
ACGACCGCATGTCCGACACCTACACCTTCTACTCGGCCGACGAGCCGGGCGTGCTGGG
|
CCGCGACGAGGCGCGCCCCGGCGCGCGAGGCGGCTCCCTCCAGGCGCTGGCTGGAGCG
|
CATGCGGCGGACCTTCGAGGAGCCCACGTCGTCGCTGGCCGCAGGCGCTGGCTGGAGC
|
GCATGCGGCGGACCTTCGAGGAGCCCACGTCGTCGCTGGCCGCGCAGATCCTGGCTAG
|
CGTGTCGGTGGTGTTCGTGATCGTGTCCATGGTGGTGCTGTGCGCCAGCACGTTGCCC
|
GACTGGCGCAACGCAGCCGCCGACAACCGCAGCCTGGATGACCGGAGCAGGTACTCCG
|
CCGGCCCTGGGAGGGAGCCCTCCGGGTGTTCTCTTGACAGGATAATTGAAGCTATCTG
|
CATAGGTTGGTTCACTGCCGAGTGCATCGTGAGGTTCATTGTCTCCAAAAACAAGTGT
|
GAGTTTGTCAAGAGACCCCTGAACATCATTGATTTACTGGCAATCACGCCGTATTACA
|
TCTCTGTGTTGATGACAGTGTTTACAGGCGAGAACTCTCAACTCCAGAGGGCTGGAGT
|
CACCTTGAGGGTACTTAGAATGATGAGGATTTTTTGGGTGATTAAGCTTGCCCGTCAC
|
TTCATTGGTCTTCAGACACTCGGTTTGACTCTCAAACGTTGCTACCGAGAGATGGTTA
|
TGTTACTTGTCTTCATTTGTGTTGCCATGGCAATCTTTAGTGCACTTTCTCAGCTTCT
|
TGAACATGGGCTGGACCTGGAAACATCCAACAAGGACTTTACCAGCATTCCTGCTGCC
|
TGCTGGTGGGTGATTATCTCTATGACTACAGTTGGCTATGGAGATATGTATCCTATCA
|
CAGTGCCTGGAAGAATTCTTGGAGGAGTTTGTGTTGTCAGTGGAATTGTTCTATTGGC
|
ATTACCTATCACTTTTATCTACCATAGCTTTGTGCAGTGTTATCATGAGCTCAAGTTT
|
AGATCTGCTAGGGGCCCACCGGTGGAGCAGCTGCCCCCAGACCCCTTGACCCGGTGGT
|
GCTTCCACCCTGCCGGAAGCACCTTGTGTGGCCCCGCCAACAGCATGGCGGTTGCATC
|
CCCAGGAAGCAGGCCCGCAGCGCCCGGAGGGGGTTTCCTGAGGACAGAGGCCCTTGTC
|
CTGATTGTCGCAGCAGGCCCTGTCGATGGACTTAACTGTGAAAATCACCCTTTCAGGG
|
GTGGATGCAAGGACTTCTGAGGGCGGA
|
ORF Start: ATG at 1ORF Stop: TGA at 1642
SEQ ID NO: 46547 aaMW at 60771.7 kD
NOV 17a,MAANMYRVGSGRKSPGRPAMTFGRSGAASVVLNVGGARYSLSRELLKDFPLRRVSRLH
CG88645-01 Protein Sequence
GCRSERDVLEVCDDYDRERNEYFFDRHSEAFGFILLYVRGHGKLRFAPRMCELSFYNE
|
MIYWGLEGAHLEYCCQRRLDDRMSDTYTFYSADEPGVLGRDEARPGARGGSLQALAGA
|
HAADLRGAHVVAGRRRWLERMRRTFEEPTSSLAAQILASVSVVFVIVSMVVLCASTLP
|
DWRNAAADNRSLDDRSRYSAGPGREPSGCSLDRIIEAICIGWFTAECIVRFIVSKNKC
|
EFVKRPLNIIDLLAITPYYISVLMTVFTGENSQLQRAGVTLRVLRMMRIFWVIKLARH
|
FIGLQTLGLTLKRCYREMVMLLVFICVAMAIFSALSQLLEHGLDLETSNKDFTSIPAA
|
CWWVIISMTTVGYGDMYPITVPGRILGGVCVVSGIVLLALPITFIYHSFVQCYHELKF
|
RSARGPPVEQLPPDPLTRWCFHPAGSTLCGPANSMAVASPGSRPAAPGGGFLRTEALV
|
LIVAAGPVDGLNCENHPFRGGCKDF
|
[0401] Further analysis of the NOV 17a protein yielded the following properties shown in Table 17B.
88TABLE 17B
|
|
Protein Sequence Properties NOV17a
|
|
PSort0.6000 probability located in plasma membrane; 0.4000
analysis:probability located in Golgi body; 0.3000 probability
located in endoplasmic reticulum (membrane);
0.3000 probability located in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0402] A search of the NOV17a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 17C.
89TABLE 17C
|
|
Geneseq Results for NOV17a
NOV17aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Indentifier[Patent #, Date]ResiduesRegionValue
|
ABG11444Novel human diagnostic protein23..473188/453 (41%)6e-90
#11435 - Homo sapiens, 466 aa.12..426271/453 (59%)
[WO200175067-A2, 11 Oct. 2001]
ABG11444Novel human diagnostic protein23..473188/453 (41%)6e-90
#11435 - Homo sapiens, 466 aa.12..426271/453 (59%)
[WO200175067-A2, 11 Oct. 2001]
AAY50341Human Kv6.2 protein - Homo sapiens,23..473188/453 (41%)6e-90
466 aa. [DE19841413-C1,12..426271/453 (59%)
23 Sep. 1999]
AAY44568Mouse Voltage-gated Potassium30..463180/446 (40%)9e-89
channel monomer, Kv6.2 variant #3 -60..463263/446 (58%)
Mus sp, 506 aa. [WO200001811-A1,
13 Jan. 2000]
AAY44566Mouse Voltage-gated Potassium30..463180/446 (40%)9e-89
channel monomer, Kv6.2 variant #1 -60..463263/446 (58%)
Mus sp, 506 aa. [WO200001811-A1,
13 Jan. 2000]
|
[0403] In a BLAST search of public sequence datbases, the NOV17a protein was found to have homology to the proteins shown in the BLASTP data in Table 17D.
90TABLE 17D
|
|
Public BLASTP Results for NOV17a
NOV17aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
BAB85520VOLTAGE-GATED K+20..468422/449 (93%)0.0
CHANNEL 6.3 - Homo sapiens 1..425422/449 (93%)
(Human), 436 aa.
BAB85521VOLTAGE-GATED K+108..468 319/361 (88%)e-178
CHANNEL 6.3 - Rattus norvegicus 1..334325/361 (89%)
(Rat), 345 aa.
Q9UJ96CARDIAC POTASSIUM23..473188/453 (41%)2e-89
CHANNEL SUBUNIT (KV6.2) -12..426271/453 (59%)
Homo sapiens (Human), 466 aa.
Q9QYU3CARDIAC POTASSIUM19..482190/470 (40%)5e-89
CHANNEL SUBUNIT (KV6.2) -18..448275/470 (58%)
Rattus rattus (Black rat), 480 aa.
AAL83911VOLTAGE-GATED POTASSIUM30..463182/448 (40%)2e-87
CHANNEL KV6.3 - Homo sapiens61..465262/448 (57%)
(Human), 519 aa.
|
[0404] PFam analysis predicts that the NOV 17a protein contains the domains shown in the Table 17E.
91TABLE 17E
|
|
Domain Analysis of NOV17a
Identities/
NOV17aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
K_tetra:28..13542/112 (38%)4.6e-24
domain 1 of 178/112 (70%)
ion_trans:264..45149/226 (22%)2.3e-24
domain 1 of 1141/226 (62%)
|
Example 18
[0405] The NOV18 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 18A.
92TABLE 18A
|
|
NOV18 Sequence Analysis
|
|
SEQ ID NO: 471178 bp
NOV18a,CCGGTGCGTCCGCCTAGCCCCGCTCCGCCTGAGGCCGTCAGGGCTCCCGAGGATGGAA
CG88738-01 DNA Sequence
GATTCCCAGGAGACATCGCCGTCCTCCAACAACTCCTCGGAGGAGCTCAGCTCTGCTC
|
TGCACCTGTCCAAGGGCATGTCGATCTTCCTCGACGTAAGTACAGATGATGATGGAAA
|
ATTATCCTTTGAAGAATTCAAAGCATATTTTGCAGATGGTGTTCTCAGTGGAGAAGAA
|
TTACACGAGCTTTTCCATACCATTGATACACATAATACTAATAATCTTGACACAGAAG
|
AGCTATGTGAATATTTTTCTCAGCACTTGGGCGAGTATGAGAATGTACTAGCAGCACT
|
TGAAGACCTGAATCTTTCCATCCTGAAGGCAATGGGCAAAACAAAGAAAGACTACCAA
|
GAAGCCTCCAATTTGGAACAATTCGTAACTAGATTTTTATTGAAGGAAACCCTGAATC
|
AGCTGCAGTCTCTCCAGAATTCCCTGGAATGTGCCATGGAAACTACTGAGGAGCAAAC
|
CCGTCAAGAAAGGCAAGGGCCAGCCAAGCCAGAAGTCCTGTCGATTCAATGGCCTCGA
|
AAACGATCAAGCCGCCGAGTCCAGAGACACAACAGCTTCTCCCCAAACAGCCCTCAGT
|
TTAATGTCAGCGGTCCAGGCTTATTAGAAGAAGACAACCAGTGGATGACCCAGATAAA
|
TAGACTCCAGAAATTAATTGATAGACTGGAAAAGAAGGATCTCAAACTCGAACCACCA
|
GAAGAAGAAATTATTGAAGGGAATACTAAATCTCACATCATGCTTGTGCAGCGGCAGA
|
TGTCTGTGATAGAAGAGGACCTGGAAGAATTCCAGCTCGCTCTGAAACACTACGTGGA
|
GAGTGCTTCCTCCCAAAGTGGATGCTTGCGTATTTCTATACAGAAGCTTTCAAATGAA
|
TCTCGCTACATGATCTATGAGTTCTGGGAGAATAGTAGTGTATGGAATAGCCACCTTC
|
AGACAAATTATAGCAAGACATTCCAAAGAAGTAATGTGGATTTCTTGGAAACTCCAGA
|
ACTCACATCTACAATGCTAGTTCCTGCTTCGTGGTGGATCCTGAACAACTAGATGTTC
|
CTAGACATTTTCTTTATGGTTCCAAGTGCAAAACAGGTGTTCTTATCTAAAACGTCAA
|
TTAGAAAATTATCTGCGG
|
ORF Start: ATG at 53ORF Stop: TAG at 1094
SEQ ID NO: 48347 aaMW at 40006.0 kD
NOV18a,MEDSQETSPSSNNSSEELSSALHLSKGMSIFLDVSTDDDGKLSFEEFKAYFADGVLSG
CG88738-01 Protein Sequence
EELHELFHTIDTHNTNNLDTEELCEYFSQHLGEYENVLAALEDLNLSILKAMGKTKKD
|
YQEASNLEQFVTRFLLKETLNQLQSLQNSLECAMETTEEQTRQERQGPAKPEVLSIQW
|
PGKRSSRRVQRHNSFSPNSPQFNVSGPGLLEEDNQWMTQINRLQKLIDRLEKKDLKLE
|
PPEEEIIEGNTKSHIMLVQRQMSVIEEDLEEFQLALKHYVESASSQSGCLRISIQKLS
|
NESRYMIYEFWENSSVWNSHLQTNYSKTFQRSNVDFLETPELTSTMLVPASWWILNN
|
[0406] Further analysis of the NOV18a protein yielded the following properties shown in Table 18B.
93TABLE 18B
|
|
Protein Sequence Properties NOV18a
|
|
PSort0.4500 probability located in cytoplasm; 0.3000 probability
analysis:located in microbody (peroxisome): 0.1000 probability
located in mitochondrial matrix space: 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0407] A search of the NOV18a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 18C.
94TABLE 18C
|
|
Geneseq Results for NOV18a
NOV18aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM43573Human polypeptide SEQ ID NO 251 -27..347157/329 (47%)2e-79
Homo sapiens, 385 aa.60..384215/329 (64%)
[WO200155308-A2, 02 Aug. 2001]
AAM40140Human polypeptide SEQ ID NO89..342108/255 (42%)5e-48
3285 - Homo sapiens, 260 aa. 1..254148/255 (57%)
[WO200153312-A1, 26 Jul. 2001]
AAU19958Novel human calcium-binding27..193 92/175 (52%)9e-41
protein #67 - Homo sapiens, 250 aa.
[WO200155304-A2, 02 Aug. 2001]
AAM43645Human polypeptide SEQ ID NO 323 -27..193 92/175 (52%)9e-41
Homo sapiens, 250 aa.60..234121/175 (68%)
[WO200155308-A2, 02 Aug. 2001]
AAB95504Human protein sequence SEQ ID42..156 74/115 (64%)2e-34
NO:18060 - Homo sapiens, 149 aa. 2..116 92/115 (79%)
[EP1074617-A2, 07 Feb. 2001]
|
[0408] In a BLAST search of public sequence datbases, the NOV18a protein was found to have homology to the proteins shown in the BLASTP data in Table 18D.
95TABLE 18D
|
|
Public BLASTP Results for NOV18a
NOV18aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9ESB5NEURONAL CALCIUM BINDING 1..347330/351 (94%)0.0
PROTEIN NECAB1 - Rattus 1..351336/351 (95%)
norvegicus (Rat), 352 aa.
Q9HBW8NEURONAL CALCIUM BINDING93..347255/255 (100%)e-145
PROTEIN NECAB1 - Homo sapiens 1..255255/255 (100%)
(Human), 255 aa (fragment).
Q96P71NIP1 - Homo sapiens (Human),27..347157/329 (47%)5e-79
362 aa.37..361215/329 (64%)
Q9HBW7NEURONAL CALCIUM BINDING27..347156/329 (47%)3e-78
PROTEIN NECAB3 - Homo sapiens37..361214/329 (64%)
(Human), 362 aa.
Q9D6J42900010M17RIK PROTEIN - Mus27..347148/326 (45%)7e-71
musculus (Mouse), 329 aa.28..328202/326 (61%)
|
[0409] PFam analysis predicts that the NOV18a protein contains the domains shown in the Table 18E.
96TABLE 18E
|
|
Domain Analysis of NOV18a
NOV18aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
efhand: domain 1 of 226..5412/29 (41%)1.2
22/29 (76%)
efhand: domain 2 of 260..88 7/29 (24%)0.99
20/29 (69%)
DUF176:248..33923/105 (22%)0.25
domain 1 of 153/105 (50%)
|
Example 19
[0410] The NOV19 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 19A.
97TABLE 19A
|
|
NOV19 Sequence Analysis
|
|
SEQ ID NO: 493050 bp
NOV19a,AAAATATTAATTTTTAACTTCTGTGCTTATATTGTCATTTCAACTCCTTGCTTAGTAA
CG88902-01 DNA Sequence
CTACAAAACCATTGCAGATCAGTGTGTGAGGGAACTGCCATCATGAGGTCTGACAAGT
|
CAGCTTTGGTATTTCTGCTCCTGCAGCTCTTCTGTGTTGGCTGTGGATTCTGTGGGAA
|
AGTCCTGGTGTGGCCCTGTGACATGAGCCATTGGCTTAATGTCAAGGTCATTCTAGAA
|
GAGCTCATAGTGAGAGGCCATGAGGTAACAGTATTGACTCACTCAAAGCCTTCGTTAA
|
TTGACTACAGGAAGCCTTCTGCATTGAAATTTGAGGTGGTCCATATGCCACAGGACAG
|
AACAGAAGAAAATGAAATATTTGTTGACCTAGCTCTGAATGTCTTGCCAGGCTTATCA
|
ACCTGGCAATCAGTTATAAAATTAAATGATTTTTTTGTTGAAATAAGAGGAACTTTAA
|
AAATGATGTGTGAGAGCTTTATCTACAATCAGACGCTTATGAAGAAGCTACAGGAAAC
|
CAACTACGATGTAATGCTTATAGACCCTGTGATTCCCTGTGGAGACCTGATGGCTGAG
|
TTGCTTGCAGTCCCTTTTGTGCTCACACTTAGAATTTCTGTAGGAGGCAATATGGAGC
|
GAAGCTGTGGGAAACTTCCAGCTCCACTTTCCTATGTACCTGTGCCTATGACAGGACT
|
AACAGACAGAATGACCTTTCTGGAAAGAGTAAAAAATTCAATGCTTTCAGTTTTGTTC
|
CACTTCTGGATTCAGGATTACGACTATCATTTTTGGGAAGAGTTTTATAGTAAGGCAT
|
TAGGTAGACCCACTACATTATGTGAGACTGTGGGAAAAGCTGAGATATGGCAAATACG
|
AACATATTGGGATTTTGAATTTCCTCAACCATACCAACCTAACTTTGAGTTTGTTGGA
|
GGATTGCACTGTAAACCTGCCAAAGCTTTGCCTAAGGAAATGGAAAATTTTGTCCAGA
|
GTTCAGGGGAAGATGGTATTGTGGTGTTTTCTCTGGGGTCACTGTTTCAAAATGTTAC
|
AGAAGAAAAGGCTAATATCATTGCTTCAGCCCTTGCCCAGATCCCACAGAAGGTGTTA
|
TGGAGGTACAAAGGAAAAAAACCATCCACATTAGGAGCCAATACTCGGCTGTATGATT
|
GGATACCCCAGAATGATCTTCTTGGTCATCCCAAAACCAAAGCTTTTATCACTCATGG
|
TGGAATGAATGGGATCTATGAAGCTATTTACCATGGGGTCCCTATGGTGGGAGTTCCC
|
ATATTTGGTGATCAGCTTGATAACATAGCTCACATGAAGGCCAAAGGAGCAGCTGTAG
|
AAATAAACTTCAAAACTATGACAAGCGAAGATTTACTGAGGGCTTTGAGAACAGTCAT
|
TACCGATTCCTCTTATAAAGAGAATGCTATGAGATTATCAAGAATTCACCATGATCAA
|
CCTGTAAAGCCCCTAGATCGAGCAGTCTTCTGGATCGAGTTTGTCATGCGCCACAAAG
|
GAGCCAAGCACCTGCGATCAGCTGCCCATGACCTCACCTGGTTCCAGCACTACTCTAT
|
AGATGTGATTGGGTTCCTGCTGGCCTGTGTGGCAACTGCTATATTCTTGTTCACAAAA
|
TGTTTTTTATTTTCCTGTCAAAAATTTAATAAAACTAGAAAGATAGAAAAGAGGGAAT
|
AGATCTTTCCAAATTCAAGAAAGACCTGATGGGGTAATCCTGTTAATTCCAGCCACAT
|
AGAATTTGGTGAAAACCTTGCTATTTTCATATTATCTATTCTGTTATTTTATCTTAGC
|
TATATAGCCTAGAATTCCACGATCATGAGGTTGTGAGTATATCTCATTCTTTCGTTGT
|
ATTTTCCTAGGTGTCTTTACTCTCTTCTCTCACTTTGTGACACAAGGACATGAATACA
|
TCTAAATTTTCCTATTTCTGATATGACTGTTTTGATGATGTCATTACTTCTATAACCT
|
TAAGTGATAGGGTGACATGCAATATGATTATTCCTGGTGTGCGCCCAAACACATGGAT
|
ATAAAGAGGTAAAAAACTTAAAATTCACAAAATTCAGTAAACCACACAAATCAGGTAA
|
GTGTTCTATGAGATTAGCTGGCTATGAGAAACATAATGATGTTTCTTTTTCAATTTAA
|
ATAAGCCCTTCTACATAGCCAGCATCAGTGATCTCAGAAAATAAATTGCTAATAATGA
|
TGACATGGCATTATGCTTAGAAAAGTTTGCTGTATTTCCATAGACCTCATCTAGATGT
|
CATGGCCTACATTTCTGCCATCACTCAACCAATACTTTTTTCTGTTTTCTTGATGATA
|
AAAAGACCTTTCTCATGATTGCCATCAAATAACAAAAGAAACTATTTTTTTTCTCACA
|
TAGAGAACATGTCAGTAAGATATTCAAGGTGAACAGATTATTTTTGGGATTAGTAACT
|
ATTTGAAATATGTGGTGATAATTACTGAGTTTATAAAATTTATTTGATAGTACACTTA
|
AAGAAGATTTATATGTTTATTCTTTAAAAATGATGAATACTCATAATTCTTATCTCTA
|
TAATCAAAAGTATAATTTACTGTAGAAAAATAAAGAGATGCTTGTTCTGAAAGTAAGA
|
TCAGTGAACTGCTTTTCAGTCTCAATCTTTGAGAATTGTAAATTCATCAAATAATTGC
|
TTACATAGTAAAAATTTAAGGTATTAGAAAACCTGCATAACAAATAGTATTATATATT
|
AAATATTTTGATATGTAAAGCTCTACACAAAGCTAAATATAGTGTAATAATGTTTACA
|
CTAATAAGCAAATATGTTAATCTTCTCATTTTTTTACTGTCATATAATCTTAGTGATA
|
TGCCTATTAATAGTTTTAAATAAATAAATTGGCTCATCTGGCTTTTTGAAAATTTTGA
|
AATTCTTACAGATGTTGATTAGGTATATCTACAAATTAATTTCAATTTTAAAATGATG
|
ATATAAAAATAAATATAAGTATTTTTCTTGTGTATGTATACAATAAATATAAATAAAA
|
TTGTTTACTGTTTTGAAAAAAAAAAAAAAAAAAA
|
ORF Start: ATG at 101ORF Stop: TAG at 1682
SEQ ID NO: 50527 aaMW at 60268.8 kD
NOV19a,MRSDKSALVFLLLQLFCVGCGFCGKVLVWPCDMSHWLNVKVILEELIVRGHEVTVLTH
CG88902-01 Protein Sequence
SKPSLIDYRKPSALKFEVVHNPQDRTEENEIFVDLALNVLPGLSTWQSVIKLNDFFVE
|
IRGTLKMMCESFIYNQTLMKKLQETNYDVMLIDPVIPCGDLMAELLAVPFVLTLRISV
|
GGNMERSCGKLPAPLSYVPVPMTGLTDRMTFLERVKNSMLSVLFHFWIQDYDYHFWEE
|
FYSKALGRPTTLCETVGKAEIWQIRTYWDFEFPQPYQPNFEFVGGLHCKPAKALPKEM
|
ENFVQSSGEDGIVVFSLGSLFQNVTEEKANIIASALAQIPQKVLWRYKGKKPSTLGAN
|
TRLYDWIPQNDLLGHPKTKAFITHGGMNGIYEAIYHGVPMVGVPIFGDQLDNIAHMKA
|
KGAAVEINFKTMTSEDLLRALRTVITDSSYKENAMRLSRIHHDQPVKPLDRAVFWIEF
|
VMRHKGAKHLRSAAHDLTWFQHYSIDVIGFLLACVATAIFLFTKCFLFSCQKFNKTRK
|
IEKRE
|
SEQ ID NO: 511705 bp
NOV19b,ACTACAAAACCATTGCAGATCAGTGTGTGAGGGAACTGCCATCATGAGGTCTGACAAG
CG88902-02 DNA Sequence
TCAGCTTTGGTATTTCTGCTCCTGCAGCTCTTCTGTGTTGGCTGTGGATTCTGTGGGA
|
AAGTCCTGGTGTGGCCCTGTGACATGAGCCATTGGCTTAATGTCAAGGTCATTCTAGA
|
AGAGCTCATAGTGAGAGGCCATGAGGTAACAGTATTGACTCACTCAAAGCCTTCGTTA
|
ATTGACTACAGGAAGCCTTCTGCATTGAAATTTGAGGTGGTCCATATGCCACAGGACA
|
GAACAGAAGAAAATGAAATATTTGTTGACCTAGCTCTGAATGTCTTGCCAGGCTTATC
|
AACCTGGCAATCAGTTATAAAATTAAATGATTTTTTTGTTGAAATAAGAGGAACTTTA
|
AAAATGATGTGTGAGAGCTTTATCTACAATCAGACGCTTATGAAGAAGCTACAGGAAA
|
CCAACTACGATGTAATGCTTATAGACCCTGTGATTCCCTGTGGAGACCTGATGGCTGA
|
GTTGCTTGCAGTCCCTTTTGTGCTCACACTTAGAATTTCTGTAGGAGGCAATATGGAG
|
CGAAGCTGTGGGAAACTTCCAGCTCCACTTTCCTATGTACCTGTGCCTATGACAGGAC
|
TAACAGACAGAATGACCTTTCTGGAAAGAGTAAAAAATTCAATGCTTTCAGTTTTGTT
|
CCACTTCTGGATTCAGGATTACGACTACCATTTTTGGGAAGAGTTTTATAGTAAGGCA
|
TTAGGAAGGCCCACTACATTATGTGAGACTGTGGGAAAAGCTGAGATATGGCTAATAC
|
GAACATATTGGGATTTTGAATTTCCTCAACCATACCAACCTAACTTTGAGTTTGTTGG
|
AGGATTGCACTGTAAACCTGCCAAAGCTTTGCCTAAGGAAATGGAAAATTTTGTCCAG
|
AGTTCAGGGGAAGATGGTATTGTGGTGTTTTCTCTGGGGTCACTGTTTCAAAATGTTA
|
CAGAAGAAAAGGCTAATATCATTGCTTCAGCCCTTGCCCAGATCCCACAGAAGGTGTT
|
ATGGAGGTACAAAGGAAAAAAACCATCCACATTAGGAGCCAATACTCGGCTGTATGAT
|
TGGATACCCCAGAATGATCTTCTTGGTCATCCCAAAACCAAAGCTTTTATCACTCATG
|
GTGGAATGAATGGGATCTATGAAGCTATTTACCATGGGGTCCCTATGGTGGGAGTTCC
|
CATATTTGGTGATCAGCTTGATAACATAGCTCACATGAAGGCCAAAGGAGCAGCTGTA
|
GAAATAAACTTCAAAACTATGACAAGCGAAGATTTACTGAGGGCTTTGAGAACAGTCA
|
TTACCGATTCCTCTTATAAAGAGAATGCTATGAGATTATCAAGAATTCACCATGATCA
|
ACCTGTAAAGCCCCTAGATCGAGCAGTCTTCTGGATCGAGTTTGTCATGCGCCACAAA
|
GGAGCCAAGCACCTGCGATCAGCTGCCCATGACCTCACCTGGTTCCAGCACTACTCTA
|
TAGATGTGATTGGGTTCCTGCTGACCTGTGTGGCAACTGCTATATTCTTGTTCACAAA
|
ATGTTTTTTATTTTCCTGTCAAAAATTTAATAAAACTAGAAAGATAGAAAAGAGGGAA
|
TAGATCTTTCCAAATTCAAGAAAGACCTGATGGGGTAATCCTGTTAATTCCAGCCACA
|
TAGAATTTGGTGAAAACCTTGCT
|
ORF Start: ATG at 44ORF Stop: TAG at 1625
SEQ ID NO: 52527 aaMW at 60283.8 kD
NOV19b,MRSDKSALVFLLLQLFCVGCGFCGKVLVWPCDMSHWLNVKVILEELIVRGHEVTVLTH
CG88902-02 Protein Sequence
SKPSLIDYRKPSALKFEVVHMPQDRTEENEIFVDLALNVLPGLSTWQSVIKLNDFFVE
|
IRGTLKMMCESFIYNQTLMKKLQETNYDVMLIDPVIPCGDLMAELLAVPFVLTLRISV
|
GGNMERSCGKLPAPLSYVPVPMTGLTDRMTFLERVKNSMLSVLFHFWIQDYDYHFWEE
|
FYSKALGRPTTLCETVGKAEIWLIRTYWDFEFPQPYQPNFEFVGGLHCKPAKALPKEM
|
ENFVQSSGEDGIVVFSLGSLFQNVTEEKANIIASALAQIPQKVLWRYKGKKPSTLGAN
|
TRLYDWIPQNDLLGHPKTKAFITHGGMNGIYEAIYHGVPMVGVPIFGDQLDNIAHMKA
|
KGAAVEINFKTMTSEDLLRALRTVITDSSYKENAMRLSRIHHDQPVKPLDRAVFWIEF
|
VMRHKGAKHLRSAAHDLTWFQHYSIDVIGFLLTCVATAIFLFTKCFLFSCQKFNKTRK
|
IEKRE
|
[0411] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 19B.
98TABLE 19B
|
|
Comparison of NOV19a against NOV19b.
NOV19a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV19b1..527525/527 (99%)
1..527525/527 (99%)
|
[0412] Further analysis of the NOV 19a protein yielded the following properties shown in Table 19C.
99TABLE 19C
|
|
Protein Sequence Properties NOV19a
|
|
PSort0.4600 probability located in plasma membrane; 0.3700
analysis:probability located in endoplasmic reticulum
(membrane); 0.3000 probability located in lysosome
(membrane); 0.1840 probability located in microbody
(peroxisome)
SignalPCleavage site between residues 24 and 25
analysis:
|
[0413] A search of the NOV 19a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 19D.
100TABLE 19D
|
|
Geneseq Results for NOV19a
NOV19aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier]Patent #, Date]ResiduesRegionValue
|
AAE15434Human drug metabolising enzyme1..527525/527 (99%)0.0
(DME)-1 - Homo sapiens, 527 aa.1..527525/527 (99%)
[WO200179468-A2, 25 Oct. 2001]
AAU29284Human PRO polypeptide sequence1..527525/527 (99%)0.0
#261 - Homo sapiens, 527 aa.1..527525/527 (99%)
[WO200168848-A2, 20 Sep. 2001]
AAB28677Human carbohydrate-modifying3..527367/528 (69%)0.0
enzyme Incyte IDNo:2912330CD1 -2..529414/528 (77%)
Homo sapiens, 529 aa.
[WO200063351-A2, 26 Oct. 2000]
AAW47126Uridine diphospho-9..527326/523 (62%)0.0
glucuronosyltransferase 2B178..530400/523 (76%)
(UGT2B17) enzyme - Homo sapiens,
530 aa. [WO9744466-A1,
27 Nov. 1997]
AAY78933Human UDP-glucuronosyltransferase3..527332/528 (62%)0.0
2B4 amino acid sequence - Homo2..528393/528 (73%)
sapiens, 528 aa. [WO200006776-A1,
10 Feb. 2000]
|
[0414] In a BLAST search of public sequence datbases, the NOV19a protein was found to have homology to the proteins shown in the BLASTP data in Table 19E.
101TABLE 19E
|
|
Public BLASTP Results for NOV19a
NOV19aIdentities/
ProteinResidues/Similaritie
AccessionMatchfor the MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9H6S4CDNA: FLJ21934 FIS, CLONE79..527 448/449 (99%)0.0
HEP04364 - Homo sapiens1..449448/449 (99%)
(Human), 449 aa.
Q9R110UDP1..527377/530 (71%)0.0
GLUCURONOSYLTRANSFERASE1..530435/530 (81%)
UGT2A3 - Cavia porcellus
(Guinea pig), 530 aa.
AAH25795RIKEN CDNA 2010321J07 GENE -1..527358/533 (67%)0.0
Mus musculus (Mouse), 534 aa.1..531427/533 (79%)
Q9D8112010321J07RIK PROTEIN -1..527357/533 (66%)0.0
Mus musculus (Mouse), 534 aa.1..531427/533 (79%)
O75795UDP-glucuronosyltransferase 2B179..527326/523 (62%)0.0
precursor, microsomal (EC 2.4.1.17)8..530400/523 (76%)
(UDPGT) (C19-steroid specific UDP-
glucuronosyltransferase) -
Homo sapiens
(Human), 530 aa.
|
[0415] PFam analysis predicts that the NOV19a protein contains the domains shown in the Table 19F.
102TABLE 19F
|
|
Domain Analysis of NOV19a
Identities/
NOV19aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
UDPGT:24..525305/507 (60%)1.1e-284
domain 1 of 1426/507 (84%)
|
Example 20
[0416] The NOV20 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 20A.
103TABLE 20A
|
|
NOV20 Sequence Analysis
|
|
SEQ ID NO: 531855 bp
NOV20a,GGTGCCATGGAGATGGAGAGCGCGGCGGCCTCCACACGTTTCCACCAGCCTCACATGG
CG89048-01AGAGGAAGATGAGTGCGATGGCCTGTGAGATCTTCAACGAGCTTAGGCTAGAGGGCAA
DNA Sequence
GCTCTGCGACGTGGTCATCAAGGTCAATGGCTTTGAGTTCAGTGCCCATAAGAGCGTC
|
CTCTGTAGCTGCAGTTCCTACTGTAGGGCTTTGTTTACAAGTGGCTGGAACAACACTG
|
AAAAGAAGGTATACAACATCCCTGGCATTTCTCCCGACATGATGAAGCTAATCATTGA
|
GTATGCATACACCCGGACCGTGCCTATCACACCGGACAATGTGGAGAAACTGCTTGCT
|
GCTGCAGACCAGTTTAACATCATGGGTATCGTCAGGGGTTGCTGCGAGTTCCTCAAGT
|
CAGAGCTGTGCTTGGATAATTGTATCGGCATCTGTAAGTTCACGGACTACTACTACTG
|
TCCTGAGCTGAGGCAGAAGGCCTACATGTTCATACTGCACAACTTTGAGGAGATGGTG
|
AAAGTCTCGGCAGAATTTTTAGAGCTCTCGGTCACTGAACTTAAGGATATCATTGAGA
|
AAGATGAGCTCAATGTCAAACAGGAAGATGCTGTATTTGAGGCCATTTTAAAGTGGAT
|
TTCTCATGACCCCCAAAATAGAAAGCAGCACATTTCAATTTTGCTTCCTAAGGTCCGC
|
CTGGCCCTAATGCATGCTGAGTACTTCATGAACAATGTTAAGATGAATGACTATGTCA
|
AAGACAGTGAGGAATGCAAACCAGTCATCATTAATGCCCTAAAGGCCATGTATGACCT
|
CAACATGAATGGACCCTCTAATTCTGATTTCACCAACCCACTCACCAGACCACGCTTG
|
CCCTATGCCATCCTCTTTGCAATTGGTGGCTGGAGTGGTGGGAGCCCCACCAATGCCA
|
TTGAGGCATATGACGCTCGGGCAGACAGATGGGTGAATGTTACTTGTGAGGAAGAGAG
|
TCCCCGTGCCTACCATGGGGCAGCCTATTTGAAAGGCTATGTGTATATCATTGGGGGG
|
TTTGATAGTGTAGACTATTTCAATAGTGTTAAGCGTTTTGACCCAGTCAAGAAAACTT
|
GGCATCAGGTGGCCCCGATGCACTCCAGACGTTGCTATGTCAGTGTGACAGTCCTCGG
|
CAATTTTATTTATGCCATGGGAGGATTTGATGGCTACGTGCGTCTAAACACTGCTGAA
|
CGTTATGAGCCAGAGACCAATCAATGGACACTCATCGCCCCCATGCACGAACAGAGGA
|
GTGATGCAAGCGCCACAACACTTTATGGGAAGGTATACATATGTGGTGGGTTTAATGG
|
AAACGAGTGCCTGTTCACAGCAGAAGTGTATAACACTGAAAGTAATCAGTGGACAGTC
|
ATAGCACCCATGAGAAGCAGGAGGAGTGGAATAGGCGTGATTGCTTATGGAGAACATG
|
TATATGCGGTAGGTGGCTTTGATGGAGCTAATCGACTTAGGAGTGCCGAAGCCTACAG
|
CCCTGTGGCTAACACTTGGCGCACAATCCCCACTATGTTTAATCCTCGTAGCAATTTT
|
GGCATCGAGGTGGTGGATGACCTCTTGTTTGTGGTGGGTGGCTTTAATGGTTTTACCA
|
CCACCTTTAATGTTGAGTGCTATGATGAAAAGACCGATGAGTGGTATGATGCTCATGA
|
CATGAGTATATACCGCAGTGCTCTGAGCTGCTGTGTAGTACCAGGGCTGGCCAATGTT
|
GAGGAATATGCAGCTAGACGGGACAACTTCCCAGGATTAGCACTGCGAGATGAAGTAA
|
AATATTCTGCTTCGACAAGTACCCTACCTGTATGAGCCTCTTCATTTAGCTAATAAA
|
ORF Start: ATG at 7ORF Stop: TGA at 1831
SEQ ID NO: 54608 aaMW at 68855.6 kD
NOV20a,MEMESAAASTRFHQPHMERKMSAMACEIFNELRLEGKLCDVVIKVNGFEFSAHKSVLC
CG89048-01SCSSYCRALFTSGWNNTEKKVYNIPGISPDMMKLIIEYAYTRTVPITPDNVEKLLAAA
Protein Sequence
DQFNIMGIVRGCCIFLKSELCLDNCIGICKFTDYYYCPELRQKAYMFILHNFEEMVKV
|
SAEFLELSVTELKDIIEKDELNVKQEDAVFEAILKWISHDPQNRKQHISILLPKVRLA
|
LMHAEYFMNNVKMNDYVKDSEECKPVIINALKAMYDLNMNGPSNSDFTNPLTRPRLPY
|
AILFAIGGWSGGSPTNAIEAYDARADRWVNVTCEEESPRAYHGAAYLKGYVYIIGGFD
|
SVDYFNSVKRFDPVKKTWHQVAPMHSRRCYVSVTVLGNFIYAMGGFDGYVRLNTAERY
|
EPETNQWTLIAPMHEQRSDASATTLYGKVYICGGFNGNECLFTAEVYNTESNQWTVIA
|
PMRSRRSGIGVIAYGEHVYAVGGFDGANRLRSAEAYSPVANTWRTIPTMFNPRSNFGI
|
EVVDDLLFVVGGFNGFTTTFNVECYDEKTDEWYDAHDMSIYRSALSCCVVPGLANVEE
|
YAARRDNFPGLALRDEVKYSASTSTLPV
|
SEQ ID NO: 551875 bp
NOV20b,GGTGCCATGGAGATGGAGAGCGCGGCGGCCTCCACACGTTTCCACCAGCCTCACATGG
CG89048-02AGAGGAAGATGAGTGCGATGGCCTGTGAGATCTTCAACGAGCTTAGACTAGAGGGCAA
DNA Sequence
GCTCTGCGACGTGGTCATCAAGGTCAATGGCTTTGAGTTCAGTGCCCATAAGAACATC
|
CTCTGTAGCTGCAGTTCCTACTTTAGAGCTTTGTTTACAAGTGGCTGGAACAACACTG
|
AAAAGAAGGTATACAACATCCCTGGCATTTCTCCTGACATGATGAAGCTAATCATTGA
|
GTATGCATACACCCGGACCGTGCCTATCACACCGGACAATGTGGAGAAACTGCTTGCT
|
GCTGCAGACCAGTTTAACATCATGGGTATCGTCAGGGGTTGCTGCGAGTTCCTCAAGT
|
CAGAGCTGTGCTTGGATAATTGTATCGGCATCTGTAAGTTCACGGACTACTACTACTG
|
TCCTGAGCTGAGGCAGAAGGCCTACATGTTCATACTGCACAACTTTGAGGAGATGGTG
|
AAAGTCTCGGCAGAATTTTTAGAGCTCTCGGTCACTGAACTTAAGGATATCATTGAGA
|
AAGATGAGCTCAATGTCAAACAGGAAGATGCTGTATTTGAGGCCATTTTAAAGTGGAT
|
TTCTCATGACCCCCAAAATAGAAAGCAGCACATTTCAATTTTGCTTCCTAAGGTTCGC
|
CTGGCCCTAATGCATGCTGAGTACTTCATGAACAATGTTAAGATGAATGACTATGTCA
|
AAGACAGTGAGGAATGCAAACCAGTCATCATTAATGCCCTAAAGGCCATGTATGACCT
|
CAACATGAATGGACCCTCTAATTCTGATTTCACCAACCCACTCACCAGACCACGCTTG
|
CCCTATGCCATCCTCTTTGCAATTGGTGGCTGGAGTGGTGGGAGCCCCACCAATGCCA
|
TTGAGGCATATGACGCTCGGGCAGACAGATGGGTGAATGTTACTTGTGAGGAAGAGAG
|
TCCCCGTGCCTACCATGGGGCAGCCTATTTGAAAGGCTATGTGTATATCATTGGGGGG
|
TTTGATAGTGTAGACTATTTCAATAGTGTTAAGCGTTTTGACCCAGTCAAGAAAACTT
|
GGCATCAGGTGGCCCCGATGCACTCCAGACGTTGCTATGTCAGTGTGACAGTCCTCGG
|
CAATTTTATTTATGCCATGGGAGGATTTGATGGCTACGTGCGTCTAAACACTGCTGAA
|
CGTTATGAGCCAGAGACCAATCAATGGACACTCATCGCCCCCATGCACGAACAGAGGA
|
GTGATGCAAGCGCCACAACACTTTATGGGAAGGTCTACATATGTGGTGGGTTTAATGG
|
AAACGAGTGCCTGTTCACAGCAGAAGTGTATAAAACTGAAAGTAATCAGTGGACAGTC
|
ATAGCACCCATGAGAAGCAGGAGGAGTGGAATAGGCGTGATTGCTTATGGAGAACATG
|
TATATGCGGTAGGTGGCTTTGATGGAGCTAATCGACTTAGGAGTGCCGAAGCCTACAG
|
CCCTGTGGCTAACACTTGGCGCACAATCCCCACTATGTTTAATCCTCGTAGCAATTTT
|
GGCATCGAGGTGGTGGATGACCTCTTGTTTGTGGTGGGTGGCTTTAATGGTTTTACCA
|
CCACCTTTAATGTTGAGTGCTATGATGAAAAGACCGATGAGTGGTATGATGCTCATGA
|
CATGAGTATATACCGCAGTGCTCTGAGCTGCTGTGTAGTACCAGGGCTGGCCAATGTT
|
GAGGAATATGCAGCTAGACGGGACAACTTCCCAGGATTAGCACTGCGAGATGAAGTAA
|
AATATTCTGCTTCGACAAGTACCCTACCTGTATGAGCCTCTTCATTTAGCTAAGGGCG
|
AATTCCAGCACATGTGGGC
|
ORF Start: ATG at 7ORF Stop: TGA at 1831
SEQ ID NO: 56608 aaMW at 68954.8 kD
NOV20b,MEMESAAASTRFHQPHMERKMSAMACEIFNELRLEGKLCDVVIKVNGFEFSAHKNILC
CG89048-02
Protein SequenceSCSSYFRALFTSGWNNTEKKVYNIPGISPDMMKLIIEYAYTRTVPITPDNVEKLLAAA
|
DQFNIMGIVRGCCEFLKSELCLDNCIGICKFTDYYYCPELRQKAYMFILHNFEEMVKV
|
SAEFLELSVTELKDIIEKDELNVKQEDAVFEAILKWISHDPQNRKQHISILLPKVRLA
|
LMHAEYFMNNVKMNDYVKDSEECKPVIINALKAMYDLNMNGPSNSDFTNPLTRPRLPY
|
AILFAIGGWSGGSPTNAIEAYDARADRWVNVTCEEESPRAYHGAAYLKGYVYIIGGFD
|
SVDYFNSVKRFDPVKKTWHQVAPMHSRRCYVSVTVLGNFIYAMGGFDGYVRLNTAERY
|
EPETNQWTLIAPMHEQRSDASATTLYGKVYICGGFNGNECLFTAEVYKTESNQWTVIA
|
PMRSRRSGIGVIAYGEHVYAVGGFDGANRLRSAEAYSPVANTWRTIPTMFNPRSNFGI
|
EVVDDLLFVVGGFNGFTTTFNVECTDEKTDEWYDAHDMSIYRSALSCCVVPGLANVEE
|
YAARRDNFPGLALRDEVKYSASTSTLPV
|
[0417] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 20B.
104TABLE 20B
|
|
Comparison of NOV20a against NOV20b.
ProteinNOV20a Residues/Identities/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV20b1..608604/608 (99%)
1..608606/608 (99%)
|
[0418] Further analysis of the NOV20a protein yielded the following properties shown in Table 20C.
105TABLE 20C
|
|
Protein Sequence Properties NOV20a
|
|
PSort0.4500 probability located in cytoplasm; 0.1000 probability
analysis:located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen);
0.0923 probability located in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0419] A search of the NOV20a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 20D.
106TABLE 20D
|
|
Geneseq Results for NOV20a
NOV20aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABG03507Novel human diagnostic protein15..554203/544 (37%)e-101
#3498 - Homo sapiens, 816 aa.221 . . . 758 291/544 (53%)
[WO200175067-A2, 11 Oct. 2001]
ABG03507Novel human diagnostic protein15 . . . 554203/544 (37%)e-101
#3498 - Homo sapiens, 816 aa.221 . . . 758 291/544 (53%)
[WO200175067-A2, 11 Oct. 2001]
AAB92953Human protein sequence SEQ ID15 . . . 554201/544 (36%)2e-99
NO:11635 - Homo sapiens, 609 aa.44 . . . 581288/544 (51%)
[EP1074617-A2, 07 Feb. 2001]
AAM38956Human polypeptide SEQ ID NO25 . . . 554193/535 (36%)3e-99
2101 - Homo sapiens, 587 aa.36 . . . 564297/535 (55%)
[WO200153312-A1, 26 Jul. 2001]
AAM78511Human protein SEQ ID NO 1173 - 3 . . . 554201/562 (35%)6e-97
Homo sapiens, 593 aa.19 . . . 570301/562 (52%)
[WO200157190-A2, 09 Aug. 2001]
|
[0420] In a BLAST search of public sequence datbases, the NOV20a protein was found to have homology to the proteins shown in the BLASTP data in Table 20E.
107TABLE 20E
|
|
Public BLASTP Results for NOV20a
NOV20aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96MC0CDNA FLJ32662 FIS, CLONE1 . . . 608604/608 (99%)0.0
TESTI1000064, WEAKLY SIMILAR1 . . . 608606/608 (99%)
TO FUGU RUBRIPES SEX
COMB ON MIDLEG-LIKE 2
PROTEIN (SCML2)
GENE - Homo sapiens
(Human), 614 aa.
Q9D5V24921517C11RIK PROTEIN - Mus1 . . . 608598/608 (98%)0.0
musculus (Mouse), 608 aa.1 . . . 608602/608 (98%)
Q9DA074921517C11RIK PROTEIN - Mus1 . . . 608597/608 (98%)0.0
musculus (Mouse), 608 aa.1 . . . 608602/608 (98%)
Q9W6R2KETCH PROTEIN - Fugu rubripes66 . . . 578 255/513 (49%)e-152
(Japanese pufferfish) (Takifugu1 . . . 511353/513 (68%)
rubripes), 518 aa (fragment).
Q9W6R3KELCH PROTEIN - Fugu rubripes65 . . . 589 251/525 (47%)e-141
(Japanese pufferfish) (Takifugu1 . . . 523333/525 (62%)
rubripes), 531 aa (fragment).
|
[0421] PFam analysis predicts that the NOV20a protein contains the domains shown in the Table 20F.
108TABLE 20F
|
|
Domain Analysis of NOV20a
Identities/
NOV20aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
K_tetra: 41 . . . 13416/112 (14%)7.6
domain 1 of 149/112 (44%)
BTB: domain 1 of 1 23 . . . 13635/143 (24%)1.2e-25
91/143 (64%)
Kelch: domain 1 of 5328 . . . 37314/47 (30%)68e-10
40/47 (85%)
Kelch: domain 2 of 5375 . . . 42015/47 (32%)4e-10
38/47 (81%)
Colipase_C:422 . . . 438 9/17 (53%)5.5
domain 1 of 110/17 (59%)
Kelch: domain 3 of 5422 . . . 46715/47 (32%)1.2e-09
37/47 (79%)
Kelch: domain 4 of 5469 . . . 51416/47 (34%)5.5e-08
36/47 (77%)
Kelch: domain 5 of 5516 . . . 56113/47 (28%)8.5e-09
37/47 (79%)
|
Example 21
[0422] The NOV21 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 21A.
109TABLE 21A
|
|
NOV21 Sequence Analysis
|
|
SEQ ID NO:57903 bp
NOV21a,CCGGGCCTGGCGGGGGGACCATGGGCGCCTCGGTCTCCAGGGGCCGGGCCGCCCGGGT
CG89098-01 DNA Sequence
CCCCGCGCCGGAGCCGGAACCCGAAGAGGCGCTGGACCTGAGCCAACTACCCCCAGAG
|
CTGCTTCTGGTGGTGCTGAGCCACGTCCCCCCGCGCACGCTGCTCGGGCGCTGCCGCC
|
AAGTGTGCCGGGGCTGGCGAGCCCTGGTGGACGGCCAGGCCCTGTGGCTGCTGATCCT
|
GGCCCGCGACCACGGCGCCACCGGCCGCGCGCTGCTGCACCTCGCCCGCAGCTGCCAG
|
TCTCCCGCCCGTAACGCCAGGCCTTGCCCCCTGGGCCGCTTCTGCGCGCGCAGACCCA
|
TCGGACGCAACCTTATTCGCAACCCCTGCGGCGAAGGCCTCCGAAAGTGGATGGTGCA
|
ACACGGTGGGGACGGCTGGGTGGTGGAGGAAAACAGGACAACCGTGCCTGGGGCCCCT
|
TCTCAGACGTGCTTCGTGACTTCATTCAGCTGGTGTTGCAAGAAGCAGGTCTTGGACC
|
TAGAGGAGGAGGGTCTGTGGCCAGAACTGCTGGATAGTGGCAGGATTGAGATTTGTGT
|
CTCTGACTGGTGGGGAGCCCGACACGACAGCGGCTGTATGTACAGACTCCTCGTCCAA
|
CTTCTAGACGCCAACCAGACTGTTCTAGATAAATTCTCTGCTGTGCCTGATCCCATCC
|
CGCAGTGGAACAACAATGCCTGCCTTCACGTGACCCACGTGTTCTCCAACATCAAGAT
|
GGGCGTCCGCTTTGTGTCTTTCGAACACCGGGGCCAGGACACACAGTTCTGGGCTGGC
|
CACTATGGAGCCCGTGTGACCAACTCCAGTGTGATCGTGCGAGTCCGTCTGTCCTAGT
|
CCAGCACTACCCTTCTTGCAAGACAGCCTGACT
|
ORF Start: ATG at 21ORF Stop: TAG at 867
SEQ ID NO:58282 aaMW at 31494.9 kD
NOV21a,MGASVSRGRAARVPAPEPEPEEALDLSQLPPELLLVVLSHVPPRTLLGRCRQVCRGWR
CG89098-01 Protein Sequence
ALVDGQALWLLILARDHGATGRALLHLARSCQSPARNARPCPLGRFCARRPIGRNLIR
|
NPCGEGLRKWMVQHGGDGWVVEENRTTVPGAPSQTCFVTSFSWCCKKQVLDLEEEGLW
|
PELLDSGRIEICVSDWWGARHDSGCMYRLLVQLLDANQTVLDKFSAVPDPIPQWNNNA
|
CLHVTHVFSNIKMGVRFVSFEHRGQDTQFWAGHYGARVTNSSVIVRVRLS
|
[0423] Further analysis of the NOV21 a protein yielded the following properties shown in Table 21B.
110TABLE 21B
|
|
Protein Sequence Properties NOV21a
|
|
PSort0.4500 probability located in cytoplasm; 0.2436 probability
analysis:located in lysosome (lumen); 0.1644 probability located in
microbody (peroxisome); 0.1000 probability located in
mitochondrial matrix space
SignalPCleavage site between residues 13 and 14
analysis:
|
[0424] A search of the NOV21 a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 21C.
111TABLE 21C
|
|
Geneseq Results for NOV21a
NOV21aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABG23890Novel human diagnostic protein121..259 123/151 (81%)1e-64
#23881 - Homo sapiens, 194 aa.27 . . . 176126/151 (82%)
[WO200175067-A2, 11 Oct. 2001]
ABG23890Novel human diagnostic protein121 . . . 259 123/151 (81%)1e-64
#23881 - Homo sapiens, 194 aa.27 . . . 176126/151 (82%)
[WO200175067-A2, 11 Oct. 2001]
AAY91629Human secreted protein sequence18 . . . 282111/272 (40%)2e-49
encoded by gene 23 SEQ ID NO:302 -42 . . . 300151/272 (54%)
Homo sapiens, 331 aa.
[WO200006698-A1, 10 Feb. 2000]
AAM39365Human polypeptide SEQ ID NO 2510 -10 . . . 277117/284 (41%)4e-47
Homo sapiens, 295 aa.26 . . . 293155/284 (54%)
[WO200153312-A1, 26 Jul. 2001]
AAY83046F-box protein FBP-6 - Homo sapiens 2 . . . 282113/295 (38%)9e-47
338 aa [WO200012679-A1,26 . . . 307155/295 (52%)
06 Mar. 2000]
|
[0425] In a BLAST search of public sequence datbases, the NOV2la protein was found to have homology to the proteins shown in the BLASTP data in Table 21D.
112TABLE 21D
|
|
Public BLASTP Results for NOV21a
NOV21aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96C87SIMILAR TO F-BOX ONLY1 . . . 282282/283 (99%)e-170
PROTEIN 17 - Homo sapiens1 . . . 283282/283 (99%)
(Human), 283 aa.
Q9N0C8UNNAMED PROTEIN PRODUCT -1 . . . 282271/284 (95%)e-160
Macaca fascicularis1 . . . 280272/284 (95%)
(Crab eating macaque)
(Cynomolgus monkey), 280 aa.
Q96EF6SIMILAR TO F-BOX ONLY1 . . . 282165/286 (57%)5e-91
PROTEIN 17 - Homo sapiens1 . . . 278201/286 (69%)
(Human), 278 aa.
Q96LQ4CDNA FLJ25205 FIS, CLONE1 . . . 282165/286 (57%)5e-91
REC05844, HIGHLY SIMILAR TO10 . . . 287 201/286 (69%)
MUS MUSCULUS F-BOX PROTEIN
FBX17 MRNA - Homo sapiens
(Human), 287 aa.
Q9QZM8F-BOX PROTEIN FBX17 - Mus24 . . . 233 132/222 (59%)1e-73
musculus (Mouse), 242 aa.16 . . . 237 160/222 (71%)
|
[0426] PFam analysis predicts that the NOV21a protein contains the domains shown in the Table 21E.
113TABLE 21E
|
|
Domain Analysis of NOV21a
NOV21aIdentities/
MatchSimilaritiesExpect
Pfam DomainRegionfor the Matched RegionValue
|
F-box:24 . . . 7216/49 (33%)1.5e-07
domain 1 of 137/49 (76%)
|
Example 22
[0427] The NOV22 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 22A.
114TABLE 22A
|
|
NOV22 Sequence Analysis
|
|
SEQ ID NO: 591634 bp
NOV22a,ACTGCTCCTGACAGAAGGATGCCACAGCTGAGCCTGTCCTCGCTGGGCCTTTGGCCAA
CG89126-01
DNA SequenceTGGCAGCATCCCCGTGGCTGCTCCTGCTGCTGGTTGGGGCCTCCTGGCTCCTGGCCCG
|
CATCCTGGCCTGGACCTACACCTTCTATGACAACTGCTGCCGCCTCCAGTGTTTCCCA
|
CAGCCCCCAAAACAGAACTGGTTTTGGGGTCACCTGGGCCTGGTCACCCCCACGGAAG
|
AGGGCATGAAGACATTGACCCAGCTGGTGGCCACATATCCCCAGGGCTTTAAGGTCTG
|
GCTGGGTCCCATCATCCCCTTCATCGTTTTATGCCACCCTGACACCATCCGGTCTATC
|
ACCAATGCCTCAGCTGCCATTGTACCCAAGGATAATCTCTTCTACAGCTTCCTGAAGC
|
CCTGGCTGGGGGATGGGCTCCTGCTGAGTGGCGGTGACAAGTGGAGCCGCCACCGTCG
|
GATGCTGACGCCCGCCTTCCATTTCAACATCCTGAAGTCCTATATAACGATCTTCAAC
|
AAGAGTGCAAACATCATGCTTGACAAGTGGCAGCACCTGGCCTCAGAGGGCAGCAGTC
|
GTCTGGACATGTTTGAGCACATCAGCCTCATGACCTTGGACAGTCTGCAGAAATGTGT
|
CTTCAGCTTTGAAAGCAATTGTCAGGAGAAACCCAGTGAATATATTGCCGCCATCTTG
|
GAGCTCAGTGCCTTTGTAGAAAAGAGAAACCAGCAGATTCTCTTGCATATTGACTTCC
|
TGTATTATCTCACCCCTGATGGGCAGCGCTTCCACAGGGCCTGCAGACTGGTGCACGA
|
CTTCACAGATGCCGTCATCCAGGAGCGGCGCCGCACCCTCCCTAGCCAGGGTGTTGAT
|
GATTTCCTCCAAGCCAAGGCCAAGTCCAAGACTTTAGACTTCATTGATGTGCTTCTGC
|
TGAGCAAGGATGAAGACGGGAAGAAGTTATCTGATGAGGACATAAGAGCAGAAGCTGA
|
CACCTTTATGTTTGAGGGCCATGACACCACAGCCAGTGGTCTCTCCTGGGTCCTGTAC
|
CACCTTGCAAAGCACCCAGAATACCAGGAGCGCTGCCGGCAAGAAGTGCAAGAGCTTC
|
TGAAGGACCGTGAACCTATAGAGATTGAATGGGACGACCTGGCCCAGCTGCCCTTCCT
|
GACCATGTGCATTAAGGAGAGCCTGCGGTTGCATCCCCCAGTCCCGGTCATCTCCCGC
|
GGCTGCACCCAGGACTTTGTGCTCCCAGACGGCCGCGTCATCCCCAAAGGCATTATCT
|
GCCTCATCAGTGTTTTTGGAACCCATCACAACCCAGCTGTGTGGCCGGACCCTGAGGT
|
CTACGACCCCTTCCGCTTTGACCCAGAGAACATCAAGGAGAGGTCACCTCTGGCTTTT
|
ATTCCCTTCTCGGCAGGGCCCAGGAACTGCATCGGGCAGACGTTCGCGATGGCGGAGA
|
TGAAGGTGGTCCTGGCGCTCACGCTGCTGCGCTTCCGCGTCCTGCCTGACCACACCGA
|
GCCCCGCAGGAAGCCGGAGCTGGTCCTGCGCGCAGAGGGCGGACTTTGGCTGCGGGTG
|
GAGCCCCTGAGCTGAGTTCTGCAGAGACCCACTCTGACCCCACTAAAATGACCCCTGA
|
TTCATCAAAA
|
ORF Start: ATG at 19ORF Stop: TGA at 1579
SEQ ID NO: 60520 aaMW at 59618.3 kD
NOV22a,MPQLSLSSLGLWPMAASPWLLLLLVGASWLLARILAWTYTFYDNCCRLQCFPQPPKQN
CG89126-01
Protein SequenceWFWGHLGLVTPTEEGMKTLTQLVATYPQGFKVWLGPIIPFIVLCHPDTIRSITNASAA
|
IVPKDNLFYSFLKPWLGDGLLLSGGDKWSRHRRMLTPAFHFNILKSYITIFNKSANIM
|
LDKWQHLASEGSSRLDMFEHISLMTLDSLQKCVFSFESNCQEKPSEYIAAILELSAFV
|
EKRNQQILLHIDFLYYLTPDGQRFHRACRLVHDFTDAVIQERRRTLPSQGVDDFLQAK
|
AKSKTLDFIDVLLLSKDEDGKKLSDEDIRAEADTPMFEGHDTTASGLSWVLYHLAKHP
|
EYQERCRQEVQELLKDREPIEIEWDDLAQLPFLTMCIKESLRLHPPVPVISRGCTQDF
|
VLPDGRVIPKGIICLISVFGTHHNPAVWPDPEVYDPFRFDPENIKERSPLAFIPFSAG
|
PRNCIGQTFAMAEMKVVLALTLLRFRVLPDHTEPRRKPELVLRAEGGLWLRVEPLS
|
[0428] Further analysis of the NOV22a protein yielded the following properties shown in Table 22B.
115TABLE 22B
|
|
Protein Sequence Properties NOV22a
|
|
PSort0.8200 probability located in outside; 0.4362 probability
analysis:located in lysosome (lumen); 0.2522 probability located
in microbody (peroxisome); 0.1000 probability located in
endoplasmic reticulum (membrane)
SignalPCleavage site between residues 37 and 38
analysis:
|
[0429] A search of the NOV22a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 22C.
116TABLE 22C
|
|
Geneseq Results for NOV22a
NOV22aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAB85779Human drug metabolizing enzyme (ID1 . . . 519467/519 (89%)0.0
No. 6825202CD1) - Homo sapiens,1 . . . 519488/519 (93%)
524 aa. [WO200159127-A2, 16-AUG-
2001]
ABG28261Novel human diagnostic protein1 . . . 520468/520 (90%)0.0
#28252 - Homo sapiens, 728 aa.209 . . . 728 487/520 (93%)
[WO200175067-A2, 11-OCT-2001]
ABG28261Novel human diagnostic protein1 . . . 520468/520 (90%)0.0
#28252 - Homo sapiens, 728 aa.209 . . . 728 487/520 (93%)
[WO200175067-A2, 11-OCT-2001]
AAB65229Human PRO1129 (UNQ568) protein1 . . . 520448/520 (86%)0.0
sequence SEQ ID NO: 264 - Homo1 . . . 520476/520 (91%)
sapiens, 524 aa. [WO200073454-A1,
07-DEC-2000]
AAB87552Human PRO1129 - Homo sapiens,1 . . . 520448/520 (86%)0.0
524 aa. [WO200116318-A2, 08-MAR-1 . . . 520476/520 (91%)
2001]
|
[0430] In a BLAST search of public sequence datbases, the NOV22a protein was found to have homology to the proteins shown in the BLASTP data in Table 22D.
117TABLE 22D
|
|
Public BLASTP Results for NOV22a
NOV22 aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
O60634CYTOCHROME P-450 - Homo sapiens1..520474/520 (91%)0.0
(Human), 520 aa.1..520494/520 (94%)
Q96AQ5CYTOCHROMEP 450, SUBFAMILY1..519466/519 (89%)0.0
IVF, POLYPEPTIDE 11 - Homo1..519488/519 (93%)
sapiens (Human), 524 aa.
Q9HBI6CYP4F11 - Homo sapiens 1..519465/519 (89%)0.0
(Human), 524 aa.
P78329Cytochrome P450 4F2 (EC 1.14.13.30)1..520468/520 (90%)0.0
(CYPIVF2) (Leukotriene-B4 omega-1..520487/520 (93%)
hydroxylase) (Leukotriene-B4 20-
monooxygenase) (Cytochrome P450-
LTB-omega) - Homo sapiens (Human),
520 aa.
Q9HCS2CYTOCHROME P4501..520448/520 (86%)0.0
(CYTOCHROME P450 ISOFORM1..520476/520 (91%)
4F12) - Homo sapiens (Human), 524 aa.
|
[0431] PFam analysis predicts that the NOV22a protein contains the domains shown in the Table 22E.
118TABLE 22E
|
|
Domain Analysis of NOV22a
Identities/
NOV22aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
ATP-synt_8: 1..6217/66 (26%)5.6
domain 1 of 133/66 (50%)
p450: domain 1 of 1 52..519162/515 (31%)3.2e − 145
381/515 (74%)
|
Example 23
[0432] The NOV23 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 23A.
119TABLE 23A
|
|
NOV23 Sequence Analysis
|
|
SEQ ID NO:61 3764 bp
|
NOV23a,ACGGTGCTGGTCTGAGCTGGACCTTGTCTGATGGCTTCCTCCAACCCTCCTCCACAGC
CG89367-01 DNA SequenceCTGCCATAGGAGATCAGCTGGTTCCAGGAGTCCCAGGCCCCTCCTCTGAGGCAGAGGA
CGACCCAGGAGAGGCGTTTGAGTTTGATGACAGTGATGATGAAGAGGACACCAGCGCA
GCCCTGGGCGTCCCCAGCCTTGCTCCTGAGAGGGACACAGACCCCCCACTGATCCACT
TGGACTCCATCCCTGTCACTGACCCAGACCCAGCAGCTGCTCCACCCGGCACAGGGGT
GCCAGCCTGGGTGAGCAATGGGGATGCTGCGGATTCAGCCTTCTCCGGGGCCCGGCAC
TCCAGCTGGAAGCGGAAGAGTTCCCGTCGCATTGACCGGTTCACTTTCCCCGCCCTGG
AAGAGGATGTGATTTATGACGACGTCCCCTGCGAGAGCCCAGATGCGCATCAGCCCGG
GGCAGAGAGGAACCTGCTCTACGAGGATGCGCACCGGGCTGGGGCCCCTCGGCAGGCG
GAGGACCTAGGCTGGAGCTCCAGTGAGTTCGAGAGCTACAGCGAGGACTCGGGGGAGG
AGGCCAAGCCGGAGGTCGAGGTCGAGCCCGCCAAGCACCGAGTGTCCTTCCAGCCCAA
GATGACCCAGCTCATGAAGGCCGCCAAGAGCGGGACCAAGGATGGGCTGGAGAAGACA
CGGATGGCCGTGATGCGCAAAGTCTCCTTCCTGCACAGGAAGGACGTCCTCGGTGACT
CGGAGGAGGAGGACATGGGGCTCCTGGAGGTCAGCGTTTCGGACATCAAGCCCCCAGC
CCCAGAGCTGGGCCCCATGCCAGAGGGCCTGAGCCCTCAGCAGGTGGTCCGGAGGCAT
ATCCTGGGCTCCATCGTGCAGAGCGAAGGCAGCTACGTGGAGTCTCTGAAGCGGATAC
TCCAGGACTACCGCAACCCCCTGATGGAGATGGAGCCCAAGGCGCTGAGCGCCCGCAA
GTGCCAGGTGGTGTTCTTCCGCGTGAAGGAGATCCTGCACTGCCACTCCATGTTCCAG
ATCGCCCTGTCCTCCCGCGTGGCTGAGTGGGATTCCACCGAGAAGATCGGGGACCTCT
TCGTGGCCTCGTTTTCCAAGTCCATGGTGCTAGATGTGTACAGTGACTACGTGAACAA
CTTCACCAGTGCCATGTCCATCATCAAGAAGGCCTGCCTCACCAAGCCTGCCTTCCTC
GAGTTCCTCAAGCGACGGCAGGTGTGCAGCCCAGACCGTGTCACCCTCTACGGGCTGA
TGGTCAAGCCCATCCAGAGGTTCCCACAGTTCATACTCCTGCTTCAGGACATGCTGAA
GAACACCCCCAGGGGCCATCCGGACAGGCTGTCGCTGCAGCTGGCCCTCACAGAGCTG
GAGACGCTGGCTGAGAAGCTGAACGAGCAGAAGCGGCTGGCTGACCAGGTGGCTGAGA
TCCAGCAGCTGACCAAGAGCGTCAGTGACCGCAGCAGCCTCAACAAGCTGTTGACCTC
AGGCCAGCGGCAGCTGCTCCTGTGTGAGACGTTGACGGAGACCGTGTACGGTGACCGA
GGGCAGCTAATTAAGTCCAAGGAGCGTCGGGTCTTCCTGCTCAACGACATGCTTGTCT
GTGCCAACATCAACTTCAAGCCTGCCAACCACAGGGGCCAGCTGGAGATTAGCAGCCT
GGTGCCCCTGGGGCCCAAGTATGTGGTGAAGTGGAACACGGCGCTGCCCCAGGTGCAG
GTGGTGGAGGTGGGCCAGGACGGTGGCACCTATGACAAGGACAATGTGCTCATCCAGC
ACTCAGGCGCCAAGAAGGCCTCTGCCTCAGGGCAGGCTCAGAATAAGGTGTACCTCGG
CCCCCCACGCCTCTTCCAGGAGCTGCAGGACCTGCAGAAGGACCTGGCCGTGGTGGAG
CAGATCACGCTTCTCATCAGCACGCTGCACGGCACCTACCAGAACCTGAACATGACTG
TGGCTCAAGACTGGTGCCTGGCCCTGCAGAGGCTGATGCGGGTGAAGGAGGAAGAGAT
CCACTCGGCCAACAAGTGCCGTCTCAGGCTCCTGCTTCCTGGGAAACCCGACAAGTCC
GGCCGCCCCATTAGCTTCATGGTGGTTTTCATCACCCCCAACCCCCTGAGCAAGATTT
CCTGGGTCAACAGGTTACATTTGGCCAAAATCGGACTCCGGGAGGAGAACCAGCCAGG
CTGGCTATGCCCGGATGAGGACAAGAAGAGCAAAGCCCCATTCTGGTGCCCGATCCTG
GCCTGCTGCATCCCTGCCTTCTCCTCCCGGGCACTCAGCCTGCAGCTTGGGGCCCTGG
TCCACAGTCCTGTCAACTGTCCCCTGCTGGGTTTCTCAGCAGTCAGCACCTCCCTTCC
ACAGGGCTACCTCTGGGTCGGGGGCGGACAGGAAGGCGCAGGGGGCCAGGTGGAAATC
TTTTCCTTGAACCGGCCCTCGCCCCGCACCGTCAAGTCCTTCCCACTGGCAGCCCCTG
TGCTCTGCATGGAGTATATCCCGGAGCTGGAGGAGGAGGCGGAGAGCAGAGACGAGAG
CCCGACAGTTGCTGACCCCTCGGCCACGGTGCATCCAACCATCTGCCTCGGGCTCCAG
GATGGCAGCATCCTCCTCTACAGCAGTGTGGACACTGGCACCCAGTGCCTGGTGAGCT
GCAGGAGCCCAGGTCTGCAGCCTGTGCTCTGCCTGCGACACAGCCCCTTCCACCTGCT
CGCTGGCCTGCAGGATGGGACCCTTGCTGCTTACCCTCGGACCAGCGGAGGTGTCCTG
TGGGACCTGGAGAGCCCTCCCGTGTGCCTGACTGTGGGGCCCGGGCCTGTCCGCACCC
TGTTGAGCCTGGAGGATGCCGTGTGGGCCAGCTGTGGGCCCCGGGTCACTGTCCTGGA
AGCCACCACCCTGCAGCCTCAGCAAAGCTTCGAGGCGCACCAGGACGAGGCAGTGAGC
GTGACACACATGGTGAAGGCGGGCAGCGGCGTCTGGATGGCCTTCTCCTCCGGCACCT
CCATCCGCCTCTTCCACACTGAGACCCTGGAGCATCTGCAAGAGATCAACATCGCCAC
CAGGACCACCTTCCTCCTGCCAGGCCAGAAGCACTTGTGTGTCACCAGCCTCCTGATC
TGCCAGGGTCTGCTCTGGGTGGGCACTGACCAGGGTGTCATCGTCCTGCTGCCCGTGC
CTCGGCTGGAAGGCATCCCCAAGATCACAGGGAAAGGCATGGTCTCACTCAATGGGCA
CTGTGGGCCTGTGGCCTTCCTGGCTGTGGCTACCAGCATCCTGGCCCCTGACATCCTG
CGGAGTGACCAGGAGGAGGCTGAGGGGCCCCGGGCTGAGGAGGACAAGCCAGACGGGC
AGGCACACGAGCCCATGCCCGACAGCCACGTGGGCCGAGAGCTGACCCGCAAGAAGGG
CATCCTCTTGCAGTACCGCCTGCGCTCCACCGCACACCTCCCGGGCCCGCTGCTCTCC
ATGCGGGAGCCGGCGCCTGCTGATGGCGCAGCTTTGGAGCACAGCGAGGAGGACGGCT
CCATTTACGAGATGGCCGACGACCCCGACGTCTGGGTGCGCAGCCGGCCCTGCGCCCG
CGACGCCCACCGCAAGGAGATTTGCTCTGTGGCCATCATCTGGGGCGGGCAGGGCTAC
CGCAACTTTGGCAGCGCTCTGGGCAGCAGTGGGAGGCAGGCCCCGTGTGGGGAGACGG
ACAGCACCCTCCTCATCTGGCAGGTGCCCTTGATGCTATAGCGCCTCCCCTC
|
|
ORF Start: ATG at 31 ORF Stop: TAG at 3751
SEQ ID NO: 62 1240 aa MW at 135723.2 kD
|
NOV23a,MASSNPPPQPAIGDQLVPGVPGPSSEAEDDPGEAFEFDDSDDEEDTSAALGVPSLAPE
G89367-01 Protein SequenceRDTDPPLIHLDSIPVTDPDPAAAPPGTGVPAWVSNGDAADSAFSGARHSSWKRKSSRR
IDRFTFPALEEDVIYDDVPCESPDAHQPGAERNLLYEDAHRAGAPRQAEDLGWSSSEF
ESYSEDSGEEAKPEVEVEPAKHRVSFQPKMTQLMKAAKSGTKDGLEKTRMAVMRKVSF
LHRKDVLGDSEEEDMGLLEVSVSDIKPPAPSLGPMPEGLSPQQVVRRHILGSIVQSEG
SYVESLKRILQDYRNPLMEMEPKALSARKCQVVFFRVKEILHCHSMFQIALSSRVAEW
DSTEKIGDLFVASFSKSMVLDVYSDYVNNFTSAMSIIKKACLTKPAFLEFLKRRQVCS
PDRVTLYGLMVKPIQRFPQFILLLQDMLKNTPRGHPDRLSLQLALTELETLAEKLNEQ
KRLADQVAEIQQLTKSVSDRSSLNKLLTSGQRQLLLCETLTETVYGDRGQLIKSKERR
VFLLNDMLVCANINFKPANHRGQLEISSLVPLGPKYVVKWNTALPQVQVVEVGQDGGT
YDKDNVLIQHSGAKKASASGQAQNKVYLGPPRLFQELQDLQKDLAVVEQITLLISTLH
GTYQNLNMTVAQDWCLALQRLMRVKEEEIHSANKCRLRLLLPGKPDKSGRPISFMVVF
ITPNPLSKISWVNRLHLAKIGLREENQPGWLCPDEDKKSKAPFWCPILACCIPAFSSR
ALSLQLGALVHSPVNCPLLGFSAVSTSLPQGYLWVGGGQEGAGGQVEIFSLNRPSPRT
VKSFPLAAPVLCMEYIPELEEEAESRDESPTVADPSATVHPTICLGLQDGSILLYSSV
DTGTQCLVSCRSPGLQPVLCLRHSPFHLLAGLQDGTLAAYPRTSGGVLWDLESPPVCL
TVGPGPVRTLLSLEDAVWASCGPRVTVLEATTLQPQQSFEAHQDEAVSVTHMVKAGSG
VWMAFSSGTSIRLFHTETLEHLQEINIATRTTFLLPGQKHLCVTSLLICQGLLWVGTD
QGVIVLLPVPRLEGIPKITGKGMVSLNGHCGPVAFLAVATSILAPDILRSDQEEAEGP
RAEEDKPDGQAHEPMPDSHVGRELTRKKGILLQYRLRSTAHLPGPLLSMREPAPADGA
ALEHSEEDGSIYEMADDPDVWVRSRPCARDAHRKEICSVAIISGGQGYRNFGSALGSS
GRQAPCGETDSTLLIWQVPLML
|
[0433] Further analysis of the NOV23a protein yielded the following properties shown in Table 23B.
120TABLE 23B
|
|
Protein Sequence Properties NOV23a
|
|
PSort0.7600 probability located in nucleus; 0.3000 probability
analysis:located in microbody (peroxisome); 0.1000 probability
located in mitochondrial matrix space; 0.1000
probabilty located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0434] A search of the NOV23a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 23C.
121TABLE 23C
|
|
Geneseq Results for NOV23a
NOV23aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB92909Human protein sequence SEQ ID433..1022583/590 (98%)0.0
NO:11539 - Homo sapiens, 596 aa. 1..585583/590 (98%)
[EP1074617-A2, 07 Feb. 2001]
ABG08838Novel human diagnostic protein150..1239510/1143 (44%)0.0
#8829 - Homo sapiens, 1129 aa. 14..1127691/1143 (59%)
[WO200175067-A2, 11 Oct. 2001]
ABG08838Novel human diagnostic protein150..1239510/1143 (44%)0.0
#8829 - Homo sapiens, 1129 aa. 14..1127691/1143 (59%)
[WO200175067-A2, 11 Oct. 2001]
ABG08836Novel human diagnostic protein211..1239479/1042 (45%)0.0
#8827 - Homo sapiens, 988 aa. 1..986645/1042 (60%)
[WO200175067-A2, 11 Oct. 2001]
ABG08836Novel human diagnostic protein211..1239479/1042 (45%)0.0
#8827 - Homo sapiens, 988 aa. 1..986645/1042 (60%)
[WO200175067-A2, 11 Oct. 2001]
|
[0435] In a BLAST search of public sequence datbases, the NOV23a protein was found to have homology to the proteins shown in the BLASTP data in Table 23D.
122TABLE 23D
|
|
Public BLASTP Results for NOV23a
NOV23aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9HCE6KIAA1626 PROTEIN - Homo1..12401239/1279 (96%)0.0
sapiens (Human), 1284 aa6..12841240/1279 (96%)
(fragment).
AAH21843HYPOTHETICAL 77.6 KDA524..1240 664/720 (92%)0.0
PROTEIN - Mus musculus1..720 688/720 (95%)
(Mouse), 720 aa (fragment).
Q9NVT3CDNA FLJ10521 FIS, CLONE433..1022 583/590 (98%)0.0
NT2RP2000841 - Homo sapiens1..585 583/590 (98%)
(Human), 596 aa.
Q9GMY0HYPOTHETICAL 63.8 KDA208..773 563/566 (99%)0.0
PROTEIN - Macaca fascicularis1..566 564/566 (99%)
(Crab eating macaque)
(Cynomolgus monkey), 567 aa.
O15013KIAA0294 PROTEIN - Homo150..1239 510/1143 (44%)0.0
sapiens (Human), 1121 aa.6..1119691/1143 (59%)
|
[0436] PFam analysis predicts that the NOV23a protein contains the domains shown in the Table 23E.
123TABLE 23E
|
|
Domain Analysis of NOV23a
Identities/
NOV23aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
RhoGEF:281..463 60/209 (29%)2.8e −
domain 1 of 1135/209 (65%)33
|
Example 24
[0437] The NOV24 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 24A.
124TABLE 24A
|
|
NOV24 Sequence Analysis
|
|
SEQ ID NO: 63750 bp
NOV24aGTTATAAGCGCCATGGCTATGACTAGTGTCAAATTGCTTGCCATTGTTTTAAGAAAGC
CG89645-01
DNA SequenceCAGATACCTGGATTGGACTCTGGGGTGTTCTCCGAGGGACACCTTCGTCACACAAACT
|
CTGTACTTCCTGGAATCGATATAGCACTAAGTTATGTGCACCAAATTATAAAAGACTT
|
ATCCATAATATTTTCTCACTGAAATTCTCAGGGCTTTTAATATCTCCAGAATATATTT
|
TTCCATTTTCCATAAGACTCAAAAGTAATAAAAGCTCTAATAAATCTACTACAAAGTC
|
ACTGCAAAAAGTAGAAGATGAAGAGGACTCTGAGGAAGACAGCAATCATGATGAGATG
|
AGTGAGCAGGAAGAGGAGCTTGAGGATGACCCTACGATAGTCAAAGACTATCAAGACC
|
TGGAAAAAGCAGTGCAGTCTTTTTGGTATGACGATGTCCTGAAGACAGGCCTAGATAT
|
TGGGAGAAACAAAGTGGAAGATGCGTTCTACAAAGGTGAACTCAGGCTGAATGGGGAA
|
AAGTTATGGAAGAAAAACAGAACGGTCAAAGTGGGAGATACACTGGATCTTCTCATTG
|
GAGAGGATAAAGAAGGAGGAACCGAGACAGTGATGAGGATTCTCCTGAAAAAAGTGTT
|
TGAAGACAAAGTGAAAAACACAGTGGTGTTACGGCGGTGGAAAAACTTAAAGTTGCCT
|
AGGAAGAGAATGTCTAAATAAACGGATTGCTTTTTGCAATACAGCTGCTTTCTA
|
ORF Start: ATG at 13ORF Stop: TAA at 715
SEQ ID NO: 64234 aaMW at 27161.9 kD
NOV24a,MAMTSVKLLAIVLRKPDTWIGLWGVLRGTPSSHKLCTSWNRYSTKLCAPNYKRLIHNI
CG89645-01
Protein SequenceFSLKFSGLLISPEYIFPFSIRLKSNKSSNKSTTKSLQKVEDEEDSEEDSNHDEMSEQE
|
EELEDDPTIVKDYQDLEKAVQSFWYDDVLKTGLDIGRNKVEDAFYKGELRLNGEKLWK
|
KNRTVKVGDTLDLLIGEDKEGGTETVMRILLKKVFEDKVKNTVVLRRWKNLKLPRKRM
|
SK
|
[0438] Further analysis of the NOV24a protein yielded the following properties shown in Table 24B.
125TABLE 24B
|
|
Protein Sequence Properties NOV24a
|
|
PSort0.6213 probability located in mitochondrial matrix space;
analysis:0.3222 probability located in mitochondrial inner
membrane; 0.3222 probability located in mitochondrial
intermembrane space; 0.3222 probability located in
mitochondrial outer membrane
SignalPCleavage site between residues 15 and 16
analysis:
|
[0439] A search of the NOV24a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 24C.
126TABLE 24C
|
|
Geneseq Results for NOV24a
NOV24aIdentities
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM01599Peptide #281 encoded by probe for1..211211/228 (92%)e − 117
measuring human breast gene18..245 211/228 (92%)
expression - Homo sapiens, 245 aa.
[WO200157270-A2, 09 Aug. 2001]
AAM26263Peptide #300 encoded by probe for1..211211/228 (92%)e − 117
measuring placental gene expression -18..245 211/228 (92%)
Homo sapiens, 245 aa.
[WO200157272-A2, 09 Aug. 2001]
AAM13856Peptide #290 encoded by probe for1..211211/228 (92%)e − 117
measuring cervical gene expression -18..245 211/228 (92%)
Homo sapiens, 245 aa.
[WO200157278-A2, 09 Aug. 2001]
AAM65987Human bone marrow expressed probe1..211211/228 (92%)e − 117
encoded protein SEQ ID NO: 26293 -18..245 211/228 (92%)
Homo sapiens, 245 aa.
[WO200157276-A2, 09 Aug. 2001]
AAM53608Human brain expressed single exon1..211211/228 (92%)e − 117
probe encoded protein SEQ ID NO:18..245 211/228 (92%)
25713 - Homo sapiens, 245 aa.
[WO200157275-A2, 09 Aug. 2001]
|
[0440] In a BLAST search of public sequence datbases, the NOV24a protein was found to have homology to the proteins shown in the BLASTP data in Table 24D.
127TABLE 24D
|
|
Public BLASTP Results for NOV24a
NOV24aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9P0P8HSPC230 (SIMILAR TO 1 . . . 234198/241 (82%) e−106
HYPOTHETICAL PROTEIN) - 1 . . . 240213/241 (88%)
Homo sapiens (Human), 240 aa.
Q9CQF41700021F05RIK PROTEIN (RIKEN 1 . . . 234167/241 (69%)3e−89
CDNA 1700021F05 GENE) - Mus 1 . . . 240194/241 (80%)
musculus (Mouse), 240 aa.
Q9VAX9CG4884 PROTEIN - Drosophila99 . . . 231 44/134 (32%)1e−11
melanogaster (Fruit fly), 189 aa.47 . . . 176 78/134 (57%)
Q9U7C9PUTATIVE CALMODULIN-80 . . . 174 29/99 (29%)0.003
BINDING PROTEIN CAM-BP38 -99 . . . 190 51/99 (51%)
Dictyostelium discoideum (Slime
mold), 340 aa.
O67632Tyrosyl-tRNA synthetase (EC94 . . . 193 23/100 (23%)0.14
6.1.1.1) (Tyrosine-tRNA ligase)291 . . . 385 48/100 (48%)
(TYRRS) - Aquifex aeolicus, 392 aa.
|
[0441] PFam analysis predicts that the NOV24a protein contains the domains shown in the Table 24E.
128TABLE 24E
|
|
Domain Analysis of NOV24a
Identities/
Similarities
NOV24afor the
Pfam DomainMatch RegionMatched RegionExpect Value
|
No Significant Matches Found
|
Example 25
[0442] The NOV25 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 25A.
129TABLE 25A
|
|
NOV25 Sequence Analysis
|
|
SEQ ID NO: 651835 bp
NOV25a,TCATCTCCCCGATTCCCTTCCCCTCTCCTCCCTCCTCTCCTTCCTCTCCTCCTTCCTC
CG89677-01
DNA SequenceTTCTCCTTTCCCCATCCCGGTCAGGGGCGGGCGCTGGAGCCAGCAGGCCTCCCCACAT
|
CATCTTCATCCTCACGGACGACCAAGGCTACCACGACGTGGGCTACCATGGTTCAGAT
|
ATCGAGACCCCTACGCTGGACAGGCTGGCGGCCAAGGGGGTCAAGTTGGAGAATTATT
|
ACATCCAGCCCATCTGCACGCCTTCGCGGAGCCAGCTCCTCACTGGCAGGTACCAGAT
|
CCACACAGGACTCCAGCATTCCATCATCCGCCCACAGCAGCCCAACTGCCTGCCCCTG
|
GACCAGGTGACACTGCCACAGAAGCTGCAGGAGGCAGGTTATTCCACCCATATGGTGG
|
GCAAGTGGCACCTGGGCTTCTACCGGAAGGAGTGTCTGCCCACCCGTCGGGGCTTCGA
|
CACCTTCCTGGGCTCGCTCACGGGCAATGTGGACTATTACACCTATGACAACTGTGAT
|
GGCCCAGGCGTGTGCGGCTTCGACCTGCACGAGGGTGAGAATGTGGCCTGGGGGCTCA
|
GCGGCCAGTACTCCACTATGCTTTATGCCCAGCGCGCCAGCCATATCCTGGCCAGCCA
|
CAGCCCTCAGCGTCCCCTCTTCCTCTATGTGGCCTTCCAGGCAGTACACACACCCCTG
|
CAGTCCCCTCGTGAGTACCTGTACCGCTACCGCACCATGGGCAATGTGGCCCGGCGGA
|
AGTACGCGGCCATGGTGACCTGCATGGATGAGGCTGTGCGCAACATCACCTGGGCCCT
|
CAAGCGCTACGGTTTCTACAACAACAGTGTCATCATCTTCTCCAGTGACAATGGTGGC
|
CAGACTTTCTCGGGGGGCAGCAACTGGCCGCTCCGAGGACGCAAGGGCACTTATTGGG
|
AAGGTGGCGTGCGGGGCCTAGGCTTTGTCCACAGTCCCCTGCTCAAGCGAAAGCAACG
|
GACAAGCCGGGCACTGATGCACATCACTGACTGGTACCCGACCCTGGTGGGTCTGGCA
|
GGTGGTACCACCTCAGCAGCCGATGGGCTAGATGGCTACGACGTGTGGCCGGCCATCA
|
GCGAGGGCCGGGCCTCACCACGCACGGAGATCCTGCACAACATTGACCCACTCTACAA
|
CCATGCCCAGCATGGCTCCCTGGAGGGCGGCTTTGGCATCTGGAACACCGCCGTGCAG
|
GCTGCCATCCGCGTGGGTGAGTGGAAGCTGCTGACAGGAGACCCCGGCTATGGCGATT
|
GGATCCCACCGCAGACACTGGCCACCTTCCCGGGTAGCTGGTGGAACCTGGAACGAAT
|
GGCCAGTGTCCGCCAGGCCGTGTGGCTCTTCAACATCAGTGCTGACCCTTATGAACGG
|
GAGGACCTGGCTGGCCAGCGGCCTGATGTGGTCCGCACCCTGCTGGCTCGCCTGGCCG
|
AATATAACCGCACAGCCATCCCGGTACGCTACCCAGCTGAGAACCCCCGGGCTCATCC
|
TGACTTTAATGGGGGTGCTTGGGGGCCCTGGGCCAGTGATGAGGAAGAGGAGGAAGAG
|
GAAGGGAGGGCTCGAAGCTTCTCCCGGGGTCGTCGCAAGAAAAAATGCAAGATTTGCA
|
AGCTTCGATCCTTTTTCCGTAAACTCAACACCAGGCTAATGTCCCAACGGATCTGATG
|
GTGGGGAGGGAGAAAACTGTCCTTTAGAGGATCTTCCCCACTCCGGCTTGGCCCTGCT
|
GTTTCTCAGGGAGAAGCCTGTCACATCTCCATCTACAGGGAGTTGGAGGGTGTAGAGT
|
CCCTTGGTTGAACAGGGTAGGGAGCCTGGATAGGAGT
|
ORF Start: at 1ORF Stop: TGA at 1678
SEQ ID NO: 66559 aaMW at 62814.5 kD
NOV25a,SSPRFPSPLLPPLLPLLLPLLLSPSRSGAGAGASRPPHIIFILTDDQGYHDVGYHGSD
CG89677-01
Protein SequenceIETPTLDRLAAKGVKLENYYIQPICTPSRSQLLTGRYQIHTGLQHSIIRPQQPNCLPL
|
DQVTLPQKLQEAGYSTHMVGKWHLGFYRKECLPTRRGFDTFLGSLTGNVDYYTYDNCD
|
GPGVCGFDLHEGENVAWGLSGQYSTMLYAQRASHILASHSPQRPLFLYVAFQAVHTPL
|
QSPREYLYRYRTMGNVARRKYAAMVTCMDEAVRNITWALKRYGFYNNSVIIFSSDNGG
|
QTFSGGSNWPLRGRKGTYWEGGVRGLGFVHSPLLKRKQRTSRALMHITDWYPTLVGLA
|
GGTTSAADGLDGYDVWPAISEGRASPRTEILHNIDPLYNHAQHGSLEGGFGIWNTAVQ
|
AAIRVGEWKLLTGDPGYGDWIPPQTLATFPGSWWNLERMASVRQAVWLFNISADPYER
|
EDLAGQRPDVVRTLLARLAEYNRTAIPVRYPAENPRAHPDFNGGAWGPWASDEEEEEE
|
EGRARSFSRGRRKKKCKICKLRSFFRKLNTRLMSQRI
|
[0443] Further analysis of the NOV25a protein yielded the following properties shown in Table 25B.
130TABLE 25B
|
|
Protein Sequence Properties NOV25a
|
|
PSort0.8650 probability located in lysosome (lumen); 0.4419
analysis:probability located in outside; 0.2799 probability
located in microbody (peroxisome); 0.1000 probability
located in endoplasmic reticulum (membrane)
SignalPCleavage site between residues 30 and 31
analysis:
|
[0444] A search of the NOV25a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 25C.
131TABLE 25C
|
|
Geneseq Results for NOV25a
NOV25aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB85482Human 25278 sulfatase polypeptide -31..559525/529 (99%)0.0
Homo sapiens, 569 aa.41..569526/529 (99%)
[WO200155411-A2, 02 Aug. 2001]
AAB85483Human 23553 sulfatase polypeptide -37..539321/505 (63%)0.0
Homo sapiens, 599 aa.76..580406/505 (79%)
[WO200155411-A2, 02 Aug. 2001]
AAB51184Human sulfatase protein B SEQ ID 1..513296/535 (55%)e-164
NO:13 - Homo sapiens, 533 aa. 8..532367/535 (68%)
[US6153188-A, 28 Nov. 2000]
AAU29061Human PRO polypeptide sequence37..384244/349 (69%)e-155
#38 - Homo sapiens, 515 aa.76..424288/349 (81%)
[WO200168848-A2, 20 Sep. 2001]
AAB44257Human PRO708 (UNQ372) protein37..384244/349 (69%)e-155
sequence SEQ ID NO:114 - Homo76..424288/349 (81%)
sapiens, 515 aa. [WO200053756-A2,
14 Sep. 2000]
|
[0445] In a BLAST search of public sequence datbases, the NOV25a protein was found to have homology to the proteins shown in the BLASTP data in Table 25D.
132TABLE 25D
|
|
Public BLASTP Results for NOV25a
NOV25aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
CAC60190SEQUENCE 6 FROM PATENT31..559 525/529 (99%)0.0
WO0155411 - Homo sapiens41..569 526/529 (99%)
(Human), 569 aa.
CAC60191SEQUENCE 8 FROM PATENT37..539 321/505 (63%)0.0
WO0155411 - Homo sapiens76..580 406/505 (79%)
(Human), 599 aa.
P33727Arylsulfatase B precursor (EC1..513297/532 (55%)e-169
3.1.6.12) (ASB) (N-8..534370/532 (68%)
acetylgalactosamine-4-sulfatase)
(G4S) - Felis silvestris catus (Cat),
535 aa.
P15848Arylsulfatase B precursor (EC1..513297/535 (55%)e-165
3.1.6.12) (ASB) (N-8..532369/535 (68%)
acetylgalactosamine-4-sulfatase)
(G4S) - Homo sapiens (Human),
533 aa.
KJHUABN-acetylgalactosamine-4-sulfatase1..513296/535 (55%)e-165
(EC 3.1.6.12) precursor [validated] -8..532369/535 (68%)
human, 533 aa.
|
[0446] PFam analysis predicts that the NOV25a protein contains the domains shown in the Table 25E.
133TABLE 25E
|
|
Domain Analysis of NOV25a
NOV25aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
Sulfatase: domain 1 of 137..461147/543 (27%)1.4e-96
322/543 (59%)
|
Example 26
[0447] The NOV26 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 26A.
134TABLE 26A
|
|
NOV26 Sequence Analysis
|
|
SEQ ID NO: 67862 bp
NOV26a,GCTGGGTCAGGAAAGCCTGCATCCCGCTCCCCTGGGCCCCGACGGGCGGGCGCACTGC
CG89697-01
DNA SequenceGCGGGGCCCACCGGCCGCAGACCTGCGTGGCCGTCCAGGGCGTCGCCATGTCCTCGGT
|
GTTTGGAAAACCCCGCGCGGGCAGCGGGCCTCAGAGCGCGCCCCTCGAGGTCAACCTG
|
GCCATCCTGGGGCGCCGCGGGGCTGGCAAGTCTGAGGACACCTACAGCTCCGAGGAGA
|
CTGTGGACCACCAGCCTGTCCACCTGAGGGTCATGGACACTGCAGACCTGGACACCCC
|
CAGGAACTGCGAGCGCTACCTGAACTGGGCCCATGCCTTCCTGGTGGTGTACAGCGTC
|
GACAGCCGCCAGAGCTTTGATAGCAGCAGCAGCTACCTGGAGCTGCTTGCCTTGCACG
|
CGAAGGAGACACAGCGCAGCATCCCTGCCCTGCTGCTGGGCAACAAGCTGGACATGGC
|
TCAGTACAGGCAAGTCACCAAGGCAGAGGGTGTGGCTTTGGCAGGCAGGTTTGGGTGC
|
CTGTTTTTCGAGGTCTCTGCCTGTCTGGACTTTGAGCACGTGCAGCATGTCTTCCACG
|
AGGCAGTGCGAGAGGCACGGCGGGAGCTGGAGAAGAGCCCCCTGACCCGGCCCCTCTT
|
CATCTCCGAGGAGAGGGCCCTGCCCCACCAGGCCCCGCTCACCGCGCGGCATGGGCTG
|
GCCAGCTGCACCTTCAACACGCTCTCCACCATCAACCTGAAGGAGATGCCCACTGTGG
|
CCCAGGCCAAGCTGGTCACCGTGAAGTCATCCCGGGCCCAGAGCAAGCGCAAGGCGCC
|
TACCCTGACTCTCCTGAAGGGCTTCAAGATCTTCTGAGGCCCCCTCCCCA
|
ORF Start: ATG at 106ORF Stop: TGA at 847
SEQ ID NO: 68247 aaMW at 27424.0kD
NOV26a,MSSVFGKPRAGSGPQSAPLEVNLAILGRRGAGKSEDTYSSEETVDHQPVHLRVMDTAD
CG89697-01LDTPRNCERYLNWAHAFLVVYSVDSRQSFDSSSSYLELLALHAKETQRSIPALLLGNK
Protein Sequence
LDMAQYRQVTKAEGVALAGRFGCLFFEVSACLDFEHVQHVFHEAVREARRELEKSPLT
|
RPLFISEERALPHQAPLTARHGLASCTFNTLSTINLKEMPTVAQAKLVTVKSSRAQSK
|
RKAPTLTLLKGFKIF
|
SEQ ID NO: 691101 bp
NOV26b,GCTGGGTCAGGAAAGCCTGCATCCCGCTCCCCTGGGCCCCGACGGGCGGGCGCACTGC
CG89697-02
DNA SequenceGCGGGGCCCACCGGCCGCAGACCTGCGTGGCCGTCCAGGGCGTCGCCATGTCCTCGGT
|
GTTTGGAAAACCCCGCGCGGGCAGCGGGCCTCAGAGCGCGCCCCTCGAGGTCAACCTG
|
GCCATCCTGGGGCGCCGCGGGGCTGGCAAGTCTGAGGACACCTACAGCTCCGAGGAGA
|
CTGTGGACCACCAGCCTGTCCACCTGAGGGTCATGGACACTGCAGACCTGGACACCCC
|
CAGGAACTGCGAGCGCTACCTGAACTGGGCCCATGCCTTCCTGGTGGTGTACAGCGTC
|
GACAGCCGCCAGAGCTTTGATAGCAGCAGCAGCTACCTGGAGCTGCTTGCCTTGCACG
|
CGAAGGAGACACAGCGCAGCATCCCTGCCCTGCTGCTGGGCAACAAGCTGGACATGGC
|
TCAGTACAGGCAAGTCACCAAGGCAGAGGGTGTGGCTTTGGCAGGCAGGTTTGGGTGC
|
CTGTTTTTCGAGGTCTCTGCCTGTCTGGACTTTGAGCACGTGCAGCATGTCTTCCACG
|
AGGCAGTGCGAGAGGCACGGCGGGAGCTGGAGAAGAGCCCCCTGACCCGGCCCCTCTT
|
CATCTCCGAGGAGAGGGCCCTGCCCCACCAGGCCCCGCTCACCGCGCGGCATGGGCTG
|
GCCAGCTGCACCTTCAACACGCTCTCCACCATCAACCTGAAGGAGATGCCCACTGTGG
|
CCCAGGCCAAGCTGGTCACCGTGAAGTCATCCCGGGCCCAGAGCAAGCGCAAGGCGCC
|
TACCCTGACTCTCCTGAAGGGCTTCAAGATCTTCTGAGGCCCCCTCCCCAGGAAGCCT
|
AGGCTCGGTGGCTGGACAGGACTGCAGCAGGACAGGGACTGGCTTCTCACCACCAGCC
|
TTTCCGTCTGATGGACAGCAGACCCCGCCTCCAGCACCAAGCAGTGTCCCTACACTCA
|
ATTCACTGTGTGAGCTGGAGTGGCAGGCAGGGATGCTGCTTCTGTTCCCTCCAGGCCT
|
TGGTCTTCATGGTAACCACCGCGTCTGTGGCCGTAGAGGGAAACAGCCACTGTGAAC
|
ORF Start: ATG at 106ORF Stop: TGA at 847
SEQ ID NO: 70247 aaMW at 27424.0kD
NOV26b,MSSVFGKPRAGSGPQSAPLEVNLAILGRRGAGKSEDTYSSEETVDHQPVHLRVMDTAD
CG89697-02
Protein SequenceLDTPRNCERYLNWAHAFLVVYSVDSRQSFDSSSSYLELLALHAKETQRSIPALLLGNK
|
LDMAQYRQVTKAEGVALAGRFGCLFFEVSACLDFEHVQHVFHEAVREARRELEKSPLT
|
RPLFISEERALPHQAPLTARHGLASCTFNTLSTINLKEMPTVAQAKLVTVKSSRAQSK
|
RKAPTLTLLKGFKIF
|
[0448] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 26B.
135TABLE 26B
|
|
Comparison of NOV26a against NOV26b.
ProteinNOV26a Residues/Identities/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV26b1..247234/247 (94%)
1..247234/247 (94%)
|
[0449] Further analysis of the NOV26a protein yielded the following properties shown in Table 26C.
136TABLE 26C
|
|
Protein Sequence Properties NOV26a
|
|
PSort0.4500 probability located in cytoplasm; 0.3200 probability
analysis:located in microbody (peroxisome); 0.1000 probability
located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0450] A search of the NOV26a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 26D.
137TABLE 26D
|
|
Geneseq Results for NOV26a
NOV26aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABB17201Human nervous system related35..172125/138 (90%)4e-67
polypeptide SEQ ID NO 5858 - Homo 5..142127/138 (91%)
sapiens, 142 aa. [WO200159063-A2,
16 Aug. 2001]
ABG20939Novel human diagnostic protein123..247 124/125 (99%)4e-65
#20930 - Homo sapiens, 1048 aa.924..1048124/125 (99%)
[WO200175067-A2, 11 Oct. 2001]
ABG20939Novel human diagnostic protein123..247 124/125 (99%)4e-65
#20930 - Homo sapiens, 1048 aa.924..1048124/125 (99%)
[WO200175067-A2, 11 Oct. 2001]
ABB57198Mouse ischaemic condition related20..16758/168 (34%)2e-19
protein sequence SEQ ID NO:484 -20..18488/168 (51%)
Mus musculus, 217 aa.
[WO200188188-A2, 22 Nov. 2001]
AAM40661Human polypeptide SEQ ID NO 5592 -20..16654/166 (32%)2e-18
Homo sapiens, 227 aa.34..19780/166 (47%)
[WO200153312-A1, 26 Jul. 2001]
|
[0451] In a BLAST search of public sequence datbases, the NOV26a protein was found to have homology to the proteins shown in the BLASTP data in Table 26E.
138TABLE 26E
|
|
Public BLASTP Results for NOV26a
NOV26aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9NYN1RIS - Homo sapiens (Human), 1..247247/266 (92%) e-135
266 aa. 1..266247/266 (92%)
AAH18060RIC (DROSOPHILA)-LIKE,20..16758/168 (34%)1e-19
EXPRESSED IN NEURONS -20..18490/168 (53%)
Homo sapiens (Human), 217 aa.
Q99578RIN PROTEiN (RIBA) - Homo20..16758/168 (34%)1e-19
sapiens (Human), 217 aa.20..18490/168 (53%)
Q92964RIN - Homo sapiens (Human),20..16759/170 (34%)3e-19
217 aa.20..18491/170 (52%)
Q9QWX5GTP-BINDING PROTEIN ROC2 -20..16758/168 (34%)4e-19
Mus musculus (Mouse), 217 aa.20..18488/168 (51%)
|
[0452] PFam analysis predicts that the NOV26a protein contains the domains shown in the Table26F.
139TABLE 26F
|
|
Domain Analysis of NOV26a
Identities/
Similarities
Pfam DomainNOV26a Match Regionfor the Matched RegionExpect Value
|
ras: domain 1 of 122 . . . 20356/216 (26%)1.5e−14
126/216 (58%)
|
Example 27
[0453] The NOV27 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 27A.
140TABLE 27A
|
|
NOV27 Sequence Analysis
|
|
SEQ ID NO: 71500 bp
NOV27a,CCATGGTCAACCCCACCGTGTCCTTCAACATCACTGTCAATGGTGAGCCCTTGGGCTG
CG90001-01
DNA SequenceTGTCTCCTTCAAGCTGTTTGCAGACAAGTTTCCAAAGACAGCAGAAAACTTTCGTGCT
|
CTGAGCACTGGAGAGAAAGGATTTGATTATAAATGTTCCTCCTTTCACAGAATTATTC
|
CAGGGTTTATGTGTCAGGGTGGTGACTTCACACGCCATAATGGCACTGGTGGCAAGTC
|
CATCTACGGGGAGAAATTTGATGATGAGAACTTCATCCTAAAGCATACAGGTCCTGGC
|
ATCTTGTCCATGGCAAATGCTGGACCCAACACAAATGATTCCCAGTTTTTCATCTGCA
|
CTGCCAAGACTGAGTGGTTGGATGGCAAGCCCATGGTCTTTGGCAAGGTGAAAGATGG
|
CATGAATATTGTGGAGGCCATGGAGCACTTCGGGTCTGGGAATGGCAAGACCATCAAG
|
AAGATCACCATCGCTGACTGTACACAACTTGACTAA
|
ORF Staff: ATG at 3ORF Stop: TAA at 498
SEQ ID NO: 72165 aaMW at 18022.4 kD
NOV27a,MVNPTVSFNITVNGEPLGCVSFKLFADKFPKTAENFRALSTGEKGFDYKCSSFHRIIP
CG90001-01
Protein SequenceGFMCQGGDFTRHNGTGGKSIYGEKFDDENFILKHTGPGILSMANAGPNTNDSQFFICT
|
AKTEWLDGKPMVFGKVKDGMNIVEAMEHFGSGNGKTIKKITIADCTQLD
|
[0454] Further analysis of the NOV27a protein yielded the following properties shown in Table 27B.
141TABLE 27B
|
|
Protein Sequence Properties NOV27a
|
|
PSort0.6400 probability located in microbody (peroxisome); 0.4500 probability
analysis:located in cytoplasm; 0.1000 probability located in mitochondrial matrix space;
0.1000 probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0455] A search of the NOV27a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 27C.
142TABLE 27C
|
|
Geneseq Results for NOV27a
Protein/NOV27aIdentities/
Organism/Residues/Similarities for
GeneseqLengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABG29337Novel human diagnostic protein1 . . . 165151/165 (91%)6e−87
#29328 - Homo sapiens, 162 aa.1 . . . 162156/165 (94%)
[WO200175067-A2, 11 OCT. 2001]
ABG12722Novel human diagnostic protein1 . . . 165151/165 (91%)6e−87
#12713 - Homo sapiens, 162 aa.1 . . . 162156/165 (94%)
[WO200175067-A2, 11 OCT. 2001]
ABG29337Novel human diagnostic protein1 . . . 165151/165 (91%)6e−87
#29328 - Homo sapiens, 162 aa.1 . . . 162156/165 (94%)
[WO200175067-A2, 11 OCT. 2001]
ABG12722Novel human diagnostic protein1 . . . 165151/165 (91%)6e−87
#12713 - Homo sapiens, 162 aa.1 . . . 162156/165 (94%)
[WO200175067-A2, 11 OCT. 2001]
AAU01195Human cyclophilin A protein - Homo1 . . . 165145/165 (87%)7e−82
sapiens, 165 aa. [WO200132876-A2,1 . . . 165152/165 (91%)
10 MAY 2001]
|
[0456] In a BLAST search of public sequence datbases, the NOV27a protein was found to have homology to the proteins shown in the BLASTP data in Table 27D.
143TABLE 27D
|
|
Public BLASTP Results for NOV27a
NOV27aIdentities/
ProteinProtein/Residues/Similarities for
AccessionOrganism/Matchthe MatchedExpect
NumberLengthResiduesPortionValue
|
CAC39529SEQUENCE 26 FROM PATENT1 . . . 165145/165 (87%)2e−81
WO0132876 - Homo sapiens1 . . . 165152/165 (91%)
(Human), 165 aa.
Q9CWJ5PEPTIDYLPROLYL ISOMERASE A -1 . . . 164144/164 (87%)7e−81
Mus musculus (Mouse), 164 aa.1 . . . 164151/164 (91%)
Q9BRU4PEPTIDYLPROLYL ISOMERASE A1 . . . 165144/165 (87%)7e−81
(CYCLOPHILIN A) - Homo sapiens1 . . . 165151/165 (91%)
(Human), 165 aa.
P05092Peptidyl-prolyl cis-trans isomerase A2 . . . 165144/164 (87%)7e−81
(EC 5.2.1.8) (PPIase) (Rotamase)1 . . . 164151/164 (91%)
(Cyclophilin A) (Cyclosporin A-
binding protein) - Homo sapiens
(Human),, 164 aa.
P04374Peptidyl-prolyl cis-trans isomerase A2 . . . 164144/163 (88%)7e−81
(EC 5.2.1.8) (PPIase) (Rotamase)1 . . . 163150/163 (91%)
(Cyclophilin A) (Cyclosporin A-
binding protein) - Bos taurus
(Bovine), and, 163 aa.
|
[0457] PFam analysis predicts that the NOV27a protein contains the domains shown in the Table 27E.
144TABLE 27E
|
|
Domain Analysis of NOV27a
Identities/
NOV27aSimilarities
PfamMatchfor the MatchedExpect
DomainRegionRegionValue
|
pro_isomerase:5 . . . 165109/180 (61%)3.8e−97
domain 1 of 1147/180 (82%)
|
Example 28
[0458] The NOV28 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 28A.
145TABLE 28A
|
|
NOV28 Sequence Analysis
|
|
SEQ ID NO: 731388 bp
NOV28a,AAATGGTGGCCTCCATGTTCTTCCGCCCGCTGTTGGTGGCCGCCACCCTTCGGACCAC
CG90011-01
DNA SequenceACTGCGGGCTGCTGCTCAGGTTCTGGGAAGTTCTGGATTGTTTAATAACCATGGACTC
|
CAAGTACAGCAGCAACAGCAAAGGAATCTCTCACTACATGAATACATGAGTATGGAAT
|
TATTGCAAGAAACTGGTGTCTCTGTTCCCAAAGGATATGTGGCAAAGAGACCAGATGA
|
AGCTTATGCAATTGCCAAAAAATTAGGTTCAAAAGATGTTGTGATGAAGGCACAGGTT
|
TTAGCTGGTGGTAGAGGAAAAGGAACATTTGAAAGTGGCCTCAAAGGAGGAGTGAAGA
|
TGGTTTTCTCTCCAGAAGAAGCAAAAGCTGTTCCTTCACAAATGATTAGGAAACAGTT
|
GTTTACCAAGCAAATGGGAGAAAAGGGCAGAATATGCAATCAGGTATTGGTCTGTGAG
|
AGAAAATATCCCAAGAGAGAGTGCTACTTTGCAATAACAATGGAAAGGTCATTTCAAG
|
GTCTTGTATTAATAGGAAGTTTACATAGTGGGGCCAACATTGAAGATGTTGCTGCTGA
|
GACTCCTGAAGCAATAATTAAAGTACCTATTGATATTGTAGAAGGTATCAAAGAGGAA
|
CAAGCTCTCCAGCTTGCACAGAAGATGGGATTTCCATCTAATATTGTGGCTTCAGCAG
|
CAGAAAACATGATCAAGCTTTACAGCCTTTTTCTGAAATACGATGCAACCATGATAGA
|
AATAAATTCAATGGTGGAAGATTCAGATGGAGCTGCATTGTGTAAGGATGCAAAGATC
|
AATTTTGACTCTAATTCAGCCTATCGCCAAAAGAAAATGTTTGATCTACAGGACTGGA
|
CCCAGGAAGATGAAAGGAACAAAGATGCTGCTAAGGCAGATCTCAACTACACTGGCCT
|
CGATGGAAGTATAGGCTGCCTAGTAAATGGTGCTGGTTTGGCTATGGCCACAATGGAT
|
ATAATAAAACTTCATGGAGAGACTCCAGCTAATTTCCTTGTTGGTGGTGGTGCTACAG
|
TCCATCAAGTAACAGAAGCATTTAAGCCTATCACTTCAGATAAAAAGGTACTGGCTAT
|
TCTGGTCAACATTTGTGGAGGAATCATGCACTGTGATATTACAGCAAAGGGTATAGTC
|
ATGGCAGTAAAAAGTTTGGAAATTAAAATACCTGTTGTGGTACAGTTACAAGGTACAC
|
AAGTTGATGATGTTAAGGCACTAAAAGCAGACAGTGGACTTAAAATACTTGCTTGTGA
|
TGATTTGGTGGAAGCTGCTAGAGTGCTTGTAAAGCTCTCTGAAATAGTGAAGCAAGCA
|
AAGCAAGCGCATGTGGATGTGAAATTTCAATTGCCAATATGATCTGAAAACCCA
|
ORF Start: ATG at 3ORF Stop: TGA at 1374
SEQ ID NO: 74457 aaMW at 49653.4 kD
NOV28a.MVASMFFRPLLVAATLRTTLRAAAQVLGSSGLFNNHGLQVQQQQQRNLSLHEYMSMEL
CG90011-01
Protein SequenceLQETGVSVPKGYVAKRPDEAYAIAKKLGSKDVVMKAQVLAGGRGKGTFESGLKGGVKM
|
VFSPEEAKAVPSQMIRKQLFTKQMGEKGRICNQVLVCERKYPKRECYFAITMERSFQG
|
LVLIGSLHSGANIEDVAAETPEAIIKVPIDIVEGIKEEQALQLAQKMGFPSNIVASAA
|
ENMIKLYSLFLKYDATMIEINSMVEDSDGAALCKDAKINFDSNSAYRQKKMFDLQDWT
|
QEDERNKDAAKADLNYTGLDGSIGCLVNGAGLAMATMDIIKLHGETPAMFLVGGGATV
|
HQVTEAFKPITSDKKVLAILVNICGGIMHCDITAKGIVMAVKSLEIKIPVVVQLQGTQ
|
VDDVKALDADSGLKILACDDLVEAARVLVKLSEIVKQAKQAHVDVKFQLPI
|
[0459] Further analysis of the NOV28a protein yielded the following properties shown in Table 28B.
146TABLE 28B
|
|
Protein Sequence Properties NOV28a
|
|
PSort0.7464 probability located in mitochondrial inner membrane;
analysis:0.6000 probability located in mitochondrial matrix space;
0.6000 probability located in mitochondrial intermembrane
space; 0.6000 probability located in mitochondrial outer
membrane
SignalPCleavage site between residues 25 and 26
analysis:
|
[0460] A search of the NOV28a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 28C.
147TABLE 28C
|
|
Geneseq Results for NOV28a
Protein/NOV28aIdentities/
Organism/Residues/Similarities for
GeneseqLengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB70163DNA encoding human synthetase 1 . . . 457403/463 (87%)0.0
#13 - Homo sapiens, 463 aa. 1 . . . 463421/463 (90%)
[WO200107628-A2, 01 FEB. 2001]
AAG66501ATP-specific succinyl-CoA 1 . . . 457403/463 (87%)0.0
synthetase 51 - Unidentified, 463 aa. 1 . . . 463421/463 (90%)
[WO200155402-Al, 02 AUG. 2001]
AAM41474Human polypeptide SEQ ID NO 1 . . . 457403/463 (87%)0.0
6405 - Homo sapiens, 485 aa.23 . . . 485421/463 (90%)
WO200153312-Al, 26 JUL. 2001]
AAM39688Human polypeptide SEQ ID NO 1 . . . 457403/463 (87%)0.0
2833 - Homo sapiens, 463 aa. 1 . . . 463421/463 (90%)
[WO200153312-Al, 26 JUL. 2001]
AAB93393Human protein sequence SEQ ID 1 . . . 457401/463 (86%)0.0
NO: 12573 - Homo sapiens, 463 aa. 1 . . . 463421/463 (90%)
[EP1074617-A2, 07 FEB. 2001]
|
[0461] In a BLAST search of public sequence datbases, the NOV28a protein was found to have homology to the proteins shown in the BLASTP data in Table 28D.
148TABLE 28D
|
|
Public BLASTP Results for NOV28a
NOV28aIdentities/
ProteinProtein/Residues/Similarities for
AccessionOrganism/Matchthe MatchedExpect
NumberLengthResiduesPortionValue
|
Q9P2R7ATP SPECIFIC SUCCINYL COA 1 . . . 457402/463 (86%)0.0
SYNTHETASE BETA SUBUNIT - 1 . . . 463421/463 (90%)
Homo sapiens (Human), 463 aa.
Q9NV21CDNA FLJ10985 FIS, CLONE 1 . . . 457401/463 (86%)0.0
PLACE1001817, HIGHLY SIMILAR 1 . . . 463421/463 (90%)
TO HOMO SAPIENS ATP-SPECIFIC
SUCCINYL-COA SYNTHETASE BETA
SUBUNIT MRNA - Homo sapiens
(Human), 463 aa.
O95194ATP-SPECIFIC SUCCINYL-COA33 . . . 457380/426 (89%)0.0
SYNTHETASE BETA SUBUNIT - 1 . . . 426396/426 (92%)
Homo sapiens (Human), 426 aa
(fragment).
Q9NVP7CDNA FLJ10596 FIS, CLONE 1 . . . 457382/458 (83%)0.0
NT2RP2004799, MODERATELY 1 . . . 441402/458 (87%)
SIMILAR TO PROBABLE
SUCCINYL-COA LIGASE -
Homo sapiens (Human), 441 aa.
O97580ATP-SPECIFIC SUCCINYL-COA34 . . . 457366/425 (86%)0.0
SYNTHETASE BETA SUBUNIT - 1 . . . 425393/425 (92%)
Sus scrofa (Pig), 425 aa (fragment).
|
[0462] PFam analysis predicts that the NOV28a protein contains the domains shown in the Table 28E.
149TABLE 28E
|
|
Domain Analysis of NOV28a
Identities/
NOV28aSimilarities
PfamMatchfor the MatchedExpect
DomainRegionRegionValue
|
ATP-grasp: 76 . . . 252 70/191 (37%)7.9e−44
domain 1 of 1144/191 (75%)
ligase-CoA:300 . . . 436 84/150 (56% 8e−57
domain 1 of 1120/150 (80%)
|
Example 29
[0463] The NOV29 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 29A.
150TABLE 29A
|
|
NOV29 Sequence Analysis
|
|
SEQ ID NO: 753170 bp
NOV29a,GAAGGCAGGGGGGCTCGGAGAAGACGGACTCTGCTTTCGCTCCCCCTTTCTTCCCCAT
CG90204-01
DNA SequenceCCCTAACATGGGCTTTGCCCTGGAGCGCTTCGCAGAAGCCGTGGACCCGGCTCTGGAG
|
TGCAAACTGTGCGGCCAGGTGCTTGAAGAGCCCCTGTGCACGCCGTGCGGGCACGTCT
|
TCTGCGCCAGCTGCCTGTTGCCCTGGGCGGTGCGGAGGCGCCGGTGCCCGCTGCAGTG
|
CCAGCCCTTGGCGCCCGGCGAGCTGTACCGGGTGCTGCCGCTGCGCAGCCTCATCCAG
|
AAGCTGCGAGTCCAGTGCGACTACCGCGCCCGCGGCTGCGGCCACTCGGTCAGGCTGC
|
ACGAGCTGGAGGCGCACGTCGAGCACTGCGACTTCGGCCCTGCCCGCCGGCTCCGCAG
|
CCGCGGGGGCTGCGCTTCGGGGCTGGGCGGTGGTGAGGTGCCCGCGCGGGGGGGCTGC
|
GGTCCGACACCCAGGGCTGGCCGGGGCGGGGGCGCGCGCGGGGGGCCGCCGGGCGGCC
|
GCTGGGGCCGCGGGCGGGGACCCGGGCCTCGGGTCCTCGCCTGGAGGCGGCGCGAGAA
|
GGCGCTGCTGGCGCAGCTCTGGGCGCTGCAGGGCGAGGTGCAGCTCACGGCGCGCAGG
|
TACCAGGAGAAGTTCACCCAATACATGGCTCACGTCCGCAACTTCGTCGGCGACCTCG
|
GTGGCGGCCACCGCAGGGATGGAGAGCATAAGCCATTCACTATTGTGTTAGAAAGAGA
|
AAATGACACTTTGGGATTCAATATTATAGGAGGTCGACCAAATCAGAATAATCAGGAA
|
GGAACATCGACTGAAGGAATTTACGTTTCAAAAATTTTAGAAAATGGACCTGCTGACA
|
GAGCAGATGGCCTGGAGATTCATGACAAAATCATGGAGGTCAATGGGAAGGATCTTTC
|
AAAGGCCACTCATGAAGAGGCAGTGGAAGCTTTTCGCAATGCCAAGGAGCCCATTGTG
|
GTGCAGGTGTTAAGGCGAACACCTCTTAGTAGACCAGCCTATGGGATGGCTTCAGAAG
|
TGCAGCTTATGAATGCCAGCACTCAGACGGACATCACCTTCGAACACATCATGGCTCT
|
GGCCAAGCTTCGTCCACCTACCCCTCCACATGAATTTTATGAGGACAATGAGTATATT
|
TCCAGCTTGCCTGCTGATGCAGACAGAACAGAAGACTTTGAATATGAGGAGGTCGAGT
|
TGTGTCGTGTTAGCAGTCAAGAGAAGCTGGGCCTGACAGTCTGTTACCGAACAGATGA
|
TGAAGAAGACACCAGCATTTATGTCAGCGAGGTTGACCCAAATAGCATTGCTGCCAAA
|
GACGGCCGGATTCGAGAAGGGGATCGGATTTTGCAAATAAATGGGGAAGATGTCCAGA
|
ATCGAGAAGAAGCAGTGGCCTTGCTGTCTAACGATGAGTGTAAGAGAATCGTGCTGCT
|
TGTTGCAAGGCCAGAGATTCAGCTGGATGAAGGCTGGCTGGAAGATGAAAGGAATGAA
|
TTCTTAGAGGAGTTAAACTTGGAGATGTTGGAAGAAGAGCATAATGAAGCAATGCAGC
|
CCACTGCCAATGAGGTGGAGCAGCCAAAAAAGCAAGAAGAAGAAGAAGGCACAACAGA
|
CACTGCAACATCCTCATCCAACAACCATGAGAAGGACAGTGGAGTAGGACGTACAGAT
|
GAAAGCTTGCGAAATGATGAGAGCTCAGAGCAGGAGAATGCAGCCGAGGACCCCAATA
|
GCACATCTTTGAAGAGCAAGAGAGACCTGGGGCAGAGCCAAGACACTCTGGGAAGTGT
|
TGAACTTCAGTACAATGAGAGCCTCGTATCTGGTGAATACATTGACTCAGACTGCATT
|
GGCAACCCAGATGAGGACTGTGAAAGATTCAGGCAGCTCTTGGAGCTCAAATGCAAGA
|
TTCGAAATCATGGAGAGTATGACCTGTATTACTCAAGCAGCACAATTGAATGCAATCA
|
AGGGGAGCAAGAGGGAGTGGAGCATGAGCTACAGTTGCTTAATGAAGAACTGAGAAAC
|
ATTGAGCTTGAGTGTCAGAATATCATGCAGGCTCACAGGCTCCAGAAAGTGACAGACC
|
AGTATGGAGACATCTGGACATTGCATGATGGAGGATTCCGGAATTATAACACCAGCAT
|
AGATATGCAAAGGGGAAAGCTAGATGACATCATGGAGCATCCAGAAAAGTCTGACAAG
|
GACAGTTCTAGTGCTTACAACACAGCTGAGAGCTGCAGAAGTACTCCGCTCACTGTAG
|
ACCGTTCCCCTGACAGTTCCCTTCCAAGGGTGATCAACCTCACCAATAAGAAAAACCT
|
GAGAAGCACAATGGCAGCCACCCAGTCCTCTTCCGGACAGAGCAGTAAAGAGTCGACC
|
TCCACCAAAGCCAAAACCACTGAGCAAGGTTGTAGCGCTGAAAGCAAGGAGAAGGTTT
|
TAGAAGGCAGCAAGCTTCCTGATCAAGAGAAGGCAGTCAGCGAACACATCCCTTACCT
|
CTCTCCTTACCACAGCTCCTCATATAGATATGCAAACATCCCAGCACACGCCCGGCAT
|
TATCAAAGCTACATGCAGTTAATTCAACAGAAATCTGCAGTCGAGTATGCTCAGAGTC
|
AGCTCAGCTTGGTGAGCATGTGCAAGGAGTCTCAGAAGTGTTCAGAGCCCAAGATGGA
|
ATGGAAGGTGAAAATTAGGAGCGACGGGACACGGTACATCACAAAGAGACCCGTGCGA
|
GACCGAATCCTGAAGGAACGTGCCTTAAAGATCAAGGAAGAGCGGAGTGGCATGACCA
|
CAGACGATGACACCATGAGCGAGATGAAAATGGGGCGCTACTGGAGCAAAGAGGAGAG
|
AAAGCAGCACCTGGTTAGGGCCAAAGAGCAGCGCCGTCGCCGTGAGTTCATGATGCGA
|
AGCAGGTTAGAGTGTCTCAAGGAGAGCCCTCAGAGCGGCAGTGAGGGCAAGAAGGAGA
|
TCAATATCATTGAACTGAGTCACAAAAAGATGATGAAAAAGAGAAACAAGAAAATTTT
|
GGACAACTGGATGACAATCCAAGAACTGATGACCCATGGGGCCAAGTCTCCAGATGGC
|
ACGAGAGTCCATAATGCCTTCTTGTCGGTGACCACTGTATGACCGAATGAATGGAATG
|
CATGCGACTGATTTTAGGAGGATGCTACCAGTTTCGGT
|
ORF Start: ATG at 66ORF Stop: TGA at 3114
SEQ ID NO: 761016 aaMW at 114909.3 kD
NOV29a,MGFALERFAEAVDPALECKLCGQVLEEPLCTPCGHVFCASCLLPWAVRRRRCPLQCQP
CG90204-01
Protein SequenceLAPGELYRVLPLRSLIQKLRVQCDYRARGCGHSVRLHELEAHVEHCDFGPARRLRSRG
|
GCASGLGGGEVPARGGCGPTPRAGRGGGARGGPPGGRWGRGRGPGPRVLAWRRREKAL
|
LAQLWALQGEVQLTARRYQEKFTQYMAHVRNFVGDLGGGHRRDGEHKPFTIVLEREND
|
TLGFNIIGGRPNQNNQEGTSTEGIYVSKILENGPADRADGLEIHDKIMEVNGKDLSKA
|
THEEAVEAFRNAKEPIVVQVLRRTPLSRPAYGMASEVQLMNASTQTDITFEHIMALAK
|
LRPPTPPHEFYEDNEYISSLPADADRTEDFEYEEVELCRVSSQEKLGLTVCYRTDDEE
|
DTSIYVSEVDPNSIAAKDGRIREGDRILQINGEDVQNREEAVALLSNDECKRIVLLVA
|
RPEIQLDEGWLEDERNEFLEELNLEMLEEEHNEAMQPTANEVEQPKKQEEEEGTTDTA
|
TSSSNNHEKDSGVGRTDESLRNDESSEQENAAEDPNSTSLKSKRDLGQSQDTLGSVEL
|
QYNESLVSGEYIDSDCEGNPDEDCERFRQLLELKCKIRNHGEYDLYYSSSTIECNQGE
|
QEGVEHELQLLNEELRNIELECQNIMQAHRLQKVTDQYGDIWTLHDGGFRNYNTSIDM
|
QRGKLDDIMEHPEKSDKDSSSAYNTAESCRSTPLTVDRSPDSSLPRVINLTNKKNLRS
|
TMAATQSSSGQSSKESTSTKAKTTEQGCSAESKEKVLEGSKLPDQEKAVSEHIPYLSP
|
YHSSSYRYANIPAHARHYQSYMQLIQQKSAVEYAQSQLSLVSMCKESQKCSEPKMEWK
|
VKIRSDGTRYITKRPVRDRILKERALKIKEERSGMTTDDDTMSEMKMGRYWSKEERKQ
|
HLVRAKEQRRRREFMMRSRLECLKESPQSGSEGKKEINIIELSHKKMMKKRNKKILDN
|
WMTIQELMTHGAKSPDGTRVHNAFLSVTTV
|
[0464] Further analysis of the NOV29a protein yielded the following properties shown in Table 29B.
151TABLE 29B
|
|
Protein Sequence Properties NOV29a
|
|
PSort0.9800 probability located in nucleus; 0.3000 probability
analysis:located in microbody (peroxisome); 0.1000 probability located
in mitochondrial matrix space; 0.1000 probability located in
lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0465] A search of the NOV29a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 29C.
152TABLE 29C
|
|
Geneseq Results for NOV29a
NOV29aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABB11704Human semaphorin domain-1..1016610/1076 (56%)0.0
associated protein homologue, SEQ33..1098780/1076 (71%)
ID NO:2074—Homo sapiens, 1098
aa. [WO200157188-A2, 09-AUG-2001]
AAM25720Human protein sequence SEQ ID1..1016608/1076 (56%)0.0
NO:1235—Homo sapiens, 1098 aa.33..1098778/1076 (71%)
[WO200153455-A2, 26-JUL-2001]
AAU17405Novel signal transduction pathway376..742 231/370 (62%)e-132
protein, Seq ID 970—Homo sapiens,27..395 303/370 (81%)
439 aa. [WO200154733-A1, 02-AUG-2001]
AAM25485Human protein sequence SEQ ID641..838 196/198 (98%)e-110
NO:1000—Homo sapiens. 206 aa9..206 196/198 (98%)
[WO200153455-A2, 26-JUL-2001]
AAM93242Human polypeptide, SEQ ID NO:376..602 148/230 (64%)1e-78
2673—Homo sapiens, 320 aa.4..232 190/230 (82%)
[EP1130094-A2, 05-SEP-2001]
|
[0466] In a BLAST search of public sequence datbases, the NOV29a protein was found to have homology to the proteins shown in the BLASTP data in Table 29D.
153TABLE 29D
|
|
Public BLASTP Results for NOV29a
NOV29aIdentities/
ProteinResidues/Similarites for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9UPQ7KIAA1095 PROTEIN—Homo1..1016611/1076 (56%)0.0
sapiens (Human), 1098 aa (fragment).33..1098781/1076 (71%)
Q9QY55SEMAPHORIN CYTOPLASMIC1..1016610/1077 (56%)0.0
DOMAIN-ASSOCIATED PROTEIN1..1063776/1077 (71%)
3A—Mus musculus (Mouse), 1063
aa.
Q9QY54SEMAPHORIN CYTOPLASMIC1..1016597/1050 (56%)0.0
DOMAIN-ASSOCIATED PROTEIN1..1011759/1050 (71%)
3B—Mus musculus (Mouse), 1011
aa.
Q91Z03SIMILAR TO SEMAF376..1016 412/657 (62%)0.0
CYTOPLASMIC DOMAIN4..654 521/657 (78%)
ASSOCIATED PROTEIN
3—Mus musculus (Mouse), 654 aa.
Q9QY39HYPOTHETICAL 77.1 KDA344..1016 334/710 (47%)e-166
PROTEIN—Mus musculus (Mouse),1..686 446/710 (62%)
686 aa.
|
[0467] PFam analysis predicts that the NOV29a protein contains the domains shown in the Table 29E.
154TABLE 29D
|
|
Domain Analysis of NOV29a
NOV29aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
FYVE: domain 1 of 133 . . . 46 6/14 (43%)3.5
11/14 (79%)
zf-C3HC4: domain 1 of 118 . . . 5615/55 (27%)2.7e-06
32/55 (58%)
PDZ: domain 1 of 2224 . . . 31331/93 (33%)2.8e-20
80/93 (86%)
prion: domain 1 of 1111 . . . 33357/849 (23%)4
79/249 (32%)
PDZ: domain 2 of 2382 . . . 46629/87 (33%)3.9e-15
66/87 (76)
|
Example 30
[0468] The NOV30 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 30A.
155TABLE 30A
|
|
NOV30 Sequence Analysis
|
|
SEQ ID NO: 774071 bp
NOV30a,ATGTCCCACACTCAGGCCGACCTGGCCCTGCGGCCCCCGCCTCCTCTTGCCACCGCGG
CG90385-01 Protein Sequence
GGCAGCCCCGCCTCCGGCCCCCTCCTCGCCGAGCGCGCCGCTTCTCCGGGAAGGCTGA
|
GCCCCGGCCGCGCTCTTCTCGTCTCAGCCGCCGTAGCTCAGTCGACTTGGGGCTGCTG
|
AGCTCTTGGTCCCTGCCAGCCTCACCCGCTCCGGACCCCCCCGATCCTCCGGACTCCG
|
CTGGACCCAGCGAGCCCCTCTGAAGGCTGCGGAAGACTCCGCGCGTCCCGAGCTCCCG
|
GTGGACCGAGGGAGCCCCTGTGAAGGCTGCGGAAGACTCCGCGCGTCCCGAGCTCCCG
|
GACTCTGCAGTGGCCCCCGGGTCCAGGGAGCCGCTAAGCGTCCCTGAACCTGTGGCCC
|
TAGAGCGGCGCCGGGAGCAGGAAGAAAAGCAGGACATGGAGACCCAGGCTGTGGCAAC
|
GTCCCCCGATGGCCGATACCTCAAGTTTGACATCGAGATTCGACGTGCCTCCTTCAAG
|
ACGGTGTATCOAGGGCTAGACACCGACACCACAGTGGAGGTAGCCTGGTGTGACCTCC
|
AGACTCGGAAACTGTCTAGAGCTGAGCGGCAGCGCTTCTCAGACGACGTGCAGATGCT
|
CAAGGGGCTGCAGCACCCCAACATCGTCCGCTTCTATGATTCGTGGAAGTCGGTGCTG
|
AGGGGCCAGGTTTGCATCGTGCTGGTCACCGAACTCATGACCTCCGGCACGCTCAAGA
|
CACATAGGCTAGGAGAATGCTGGCAGAAGATGCGCAGGCGGCAGCAGGGTGCGGCAGG
|
GGGGAACTTCCCAGTGGGGGGCTCCTTCCCGGAGGACGTGTCCCCCCACCAGGACTCT
|
GGCTATGCGCCCTCCCCCAGGTACCTGAGGCGGTTCCCGGAGATGAAGCCGCGGGTCC
|
TTCAGCGCTGGAGCCGCCAAATCCTGCGGGGACTTCATTTCCTACACTCCCGGGTTCC
|
TCCCATCCTCCTCCGGGATCTCAAGTGCGACAATGTCTTTATCACGGGACCTACTGCC
|
TCTGTCAAAATCGGGGACCTGGACCTGGCCACGCTCAAGCGCGCCTCCTTTGCCAAGA
|
GTGTCATCGGCACCCCGGAATTCATGGCCCCCGAGATGTACGACGAAAAGTACGATGA
|
GGCCGTGCACGTCTACGCGTTCGGCATGTGCATGCTGGAGATGGCCACCTCTGAGTAC
|
CCGTACTCCGAGTGCCAGAATGCCGCGCAAATCTACCGCAAGGTCACTTCGGGCAGAA
|
AGCCGAACAGCTTCCACAAGGTGAAGATACCCGAGGTCAAGGAGATCATTGAAGGCTG
|
CATCCGCACCGATAAGAACGAGACCTTCACCATCCAGCACCTCCTGGCCCACGCCTTC
|
TTCCGCGAGGAGCGCGGTGTGCACCTGCAACTAGCCGACGACGACGACGGCGACAAGC
|
CGGGCCTCAAGCTCTGCCTGCGCATGGACGACGCCCGGCGCGCGGGGCGCCCACGGGA
|
CAACCAGGCCATCGAGTTCCTGTTCCAGCTGGGCCGGGACGCGGCCGAGGAGGTGGCA
|
CAGGAGATGGTGGCTCTCGGCTTGGTCTCTGAAGCCGATTACCAGCCACTGGCCCGTG
|
CAGTACGTGAACGCGTTGCTGCCATCCAGCGAAAGCGTGAGAAGCTGCGTAAAGCAAG
|
GGAATTGGAGGCACTCCCACCAGAGCCAGGACCTCCACCAGCAACTGTGCCCATGGCC
|
CCCGGTCCCCCCAGTCTCTTCCCCCCTCAGCCTGAGGAOCCAGAGGCAGACCAGCACC
|
AGCCCTTCCTTTTCCGCCACCCCACCTACTCATCTACCACTTCGCATTCCGAGACTGA
|
TGGCTACCTCAGCTCCTCCGCCTTCCTGGATCCCTCAGACCCTGCCCTTCAGCCCCCT
|
GGGGGGGTGCCATCCACCCTGGCTGAGTCCCATCTCTGCCTCCCCTCGGCTTTTGCCC
|
TATCCATTCCACGTTCTGGCCCTGGAAGTGACTTTTCCCCCGGGGACAGCTATGCCTC
|
AGATGCAGCTTCAGGCCTTAGCGATGTGGGAGAAGGGATGGGACAAATGAGGAGACCC
|
CCACCGAGGAATCTCCGGCGCAGACCCCGATCCCGGCTGCGGGTCACTAGTGTCTCAG
|
ACCAGAATGACAGAGTGGTTGAGTGCGAGCTACAGACCCATAACAGCAAGATCGTGAC
|
CTTCCGATTTCATCTGGATGGGGACAGCCCGCAAGACATTGCAGCTGCCATGGTATAT
|
AACGAGTTCATTCTGCCTTCGCAGCGAGATGGATTTCTCAGACCGATTCGGGAGATTA
|
TCCAGCGACTGGAGACCCTCTTGAACAGAGACACTCGCCCCATGGAGCCTCCTGAAGA
|
CACCCTAAGCCCCCAGCACACCCGAGTTTCCCGTCCCACTCTCTCAGTGTCCCTGCAG
|
TTCTCTCCCCACGACTTCTCCACCTACGTTCTCTCCCACTTGTTCTCAGGGGAAGCCA
|
GGCTGGCGCCCATCTCTGAAGAGGGAAAGCCGCAGCTTCTTCGGCGTTTCCAAGTGAC
|
TTCATCCAAGCAACCCGCTGAGCCTCTTCCCTTGCAGCCAACATCCCCCACTCTCTCT
|
GGTTCTCCAAAACCTTCAACCCCTCAGCTCACTTCAGAGAGCTCAGATACACAGGACA
|
GTCCTGCAGCCGCCCCAGAGACCAGGGAAGCTCTGGCTGAGAGCCACCGTGCAGCTGA
|
GGGTCTGGGGGCTGGAGTTGAGGAGGAAGGAGATGATGGGAAGGAACCCCAAGTTGGG
|
GGCAGCCCCCAACCCCTGAGCCATCCCAGCCCAGTGTGGATGAACTACTCCTACAGCA
|
GCCTGTGTTTGAGCAGCGAGGAGTCAGAAAGCAGTGGGGAAGATGAGGAGTTCTGGGC
|
TGAGCTGCAGAGTCTTCGGCAGAAGCACTTGTCAGAGGTCGAAACACTACAGACACTA
|
CAGAAAAAAGAAATTGAAGATTTGTACAGCCCGCTGGGGAAGCAGCCCCCACCGGGTA
|
TTGTCCCCCCAGCTGCTATGCTGTCCAGCCGCCAGCGCCGCCTCTCCAACGCCAGCTT
|
CCCCACCTCCCGCCGCAACAGCCTACAGCGCTCTGAGCCCCCAGGCCCTGGCATCATG
|
CGAAGGAACTCTCTGAGTGGCAGCAGCACCGGCTCCCAGGAGCAGCGCGCAAGCAAGG
|
GGGTGACATTCGCCGGGGATGTTGGCAGGATGTTTGGAGTTGTCGCCACAGAGACGAT
|
TGAAGACGCCCTGCTTCACTTGGCCCAGCAGAATGACCAAGCACTCAGGGAGGCTTCG
|
GGGCGGCTGGCCCCCTTCAGCGAGCCCCAGATCGTGGAATTTGTTTTTCTCCTGTCTG
|
AACAATGGTGTCTGGAGAAATCTGTGAGCTACCAGGCTGTAGAAATCCTAGAAAGGTT
|
TATCGTAAAACACGCAGAGAAGATCTGCAGGCAAGCCACAATCAGGATAACCCCTAAT
|
AAGAGAGAGTCTCAGAATTGGAGGGCTCTGAAACAGCAGCTTCTCAACAAGTTTACTC
|
TCCGTCTTGTGTCAGGTCTTCAGCTGGCCAGCAAACTTTCCTTCCGAAACAAAATAAT
|
CAGCAACATTACAGTCTTGAATTTCCTCCAGGCTCTAGGCTATCTACACACTAAAGAA
|
GAACTGCTGGAATCAGAGCTTGATGTTTTGAAGTCCTTGAACTTCCGAATTAATCTGC
|
CCACTCCCCTGGCATATGTGGAGACGCTCCTAGAGGTTTTAGGATACAATGGCTCTTT
|
GGTTCCAGCCATGAGGCTGCATGCAACCTGCCTGACACTGCTCGACCTGGTCTATCTT
|
CTGCATGAACCCATATATCAGAGCCTGTTGACGGCTTCAATTGAGAACTCCACTCCCA
|
GTCAGCTGCAAGGGGAAAAGTTTACTTCAGTGAAGGAAGACTTCATGCTGTTGCCAGT
|
AGGAATCATTGCAGCAAGTGCTTTCATCCAAAACCATGAGTGTTGGAGCCAGGTATGC
|
ACCACTGAGCAGGACCAGCATGAGAGAGTTAAGGTGCACCAGCATGCCCTTTTTGTCA
|
TATCAGCCTGA
|
ORF Start: ATG at 1ORF Stop: TGA at 4069
SEQ ID NO: 781356 aaMW at 150401.9kD
NOV30a, MSQTEADLALRPPPPLGTAGQPRLGPPPRRARRFSGKAEPRPRSSRLSRRSSVDLGLL
CG90385-01 Protein Sequence
SSWSLPASPAPDPPDPPDSAGPGPARSPPPSSKEPPEGTWTEGAPVKAAEDSARPELP
|
DSAVGTGSRETLRVTEAVALERRREGEEKEDMETQAVATSPDGRYLKFDIEIGRGSFK
|
TVYRGLDTDTTVEVAWCELQTRKLSRAERQRFSEEVEMLKGLQHPNIVRFYDSWKSVL
|
RGQVCIVLVTELMTSGTLKTHRLGECWQKMRRRQQGAAGGNFPVGGSFPEDVSPHQDS
|
GYAPSPRYLRRFREMKPRVLQRWSRQILRGLHFLHSRVPPILHRDLKCDNVFITGPTG
|
SVKIGDLGLATLKRASFAKSVIGTTEFMATEMYEEKYDEAVDVYAFGMCMLEMATSEY
|
PYSECQNAAQIYRKVTSGRKPNSFHKVKIPEVKEIIEGCIRTDKNERFTIQDLLAHAF
|
FREERGVHVELAEEDDGEKPGLKLWLRMEDARRGGRPRDNQAIEFLFQLGRDAAEEVA
|
QEMVALGLVCEADYQPVARAVRERVAAIQRKREKLRKARELEALPPEPGPPPATVPMA
|
PGPPSVFPPEPEEPEADQHQPFLFRHASYSSTTSDCETDGYLSSSGFLDASDPALQPP
|
GGVPSSLAESHLCLPSAFALSIPRSGPGSDFSPGDSYASDAASGLSDVGEGMGQMRRP
|
PGRNLRRRPRSRLRVTSVSDQNDRVVECQLQTHNSKMVTFRFDLDGDSPEEIAAAMVY
|
TGTTSVFTTETEETEADGHGTFLFRHASYSSTTSDCETDGYLSSSGFLDASDTALGTT
|
NEFILPSERDGFLRRIREIIQRVETLLKRDTGPMEAAEDTLSPQHTRVSGPTLSVSLE
|
FSPHDFSTYVLSHLFSGEARLAPISEEGKPQLVGRFQVTSSKEPAEPLPLQPTSPTLS
|
GSPKPSTPQLTSESSDTEDSAGGGPETREALAESDRAAEGLGAGVEEEGDDGKEPQVG
|
GSPQPLSHPSPVWMNYSYSSLCLSSEESESSGEDEEFWAELQSLRQKHLSEVETLQTL
|
QKKEIEDLYSRLGKQPPPGIVAPAAMLSSRQRRLSKGSFPTSRRNSLQRSEPPGPGIM
|
RRNSLSGSSTGSQEQRASKGVTFAGDVGRMFGVVATETIEDALLHLAQQNEQAVREAS
|
GRLGRFREPQIVEFVFLLSEQWCLEKSVSYQAVEILERFMVKQAENICRQATIRITPN
|
KRESQNWRALKQQLVNKFTLRLVSCVQLASKLSFRNKIISNITVLNFLQALGYLHTKE
|
ELLESELDVLKSLNFRINLPTPLAYVETLLEVLGYNGCLVPAMRLHATCLTLLDLVYL
|
LHEPIYESLLRASIENSTPSQLQGEKFTSVKEDFMLLAVGIIAASAFIQNHECWSQVC
|
TTEQDQHERVKDEHALFVISA
|
SEQ ID NO: 794019 bp
NOV30b,AATTTTCTACCCTTCGGCGCCCTGCTCTTTCCTCATGTTGGCAATCCCCGGCCACGGA
CG90385-02 DNA Sequence
GACCACCGTCCTCATGTCCCAGACTGAGGCCGACCTGGCCCTGCGGCCCCCGCCTCCT
|
CTTGGCACCCCGGGGCAGCCCCGCCTCGCGCCCCCTCCTCGCCGAGCGCGCCGCTTCT
|
CCGGGAAGGCTGAGCCCCCGCCGCGCTCTTCTCCTCTCAGCCGCCGTAGCTCACTCGA
|
CTTGGGGCTGCTGAGCTCTTGGTCCCTCCCACCCTCACCCGCTCCGGACCCCCCCGAT
|
CCTCCGGACTCCGCTGCTCCTGGCCCCCCGAGGACCCCACCGCCTAGCTCCAAAGAAC
|
CCCCCGAGGGCACGTGGACCGAGGGAGCCCCTGTGAAGGCTGCGGAAGACTCCGCGCG
|
TCCCGACCTCCCGCACTCTGCACTCCGCCCGGCGTCCAGGGACCCGCTAAGGGTCCCT
|
GAAGCTCTGGCCCTAGAGCGGCGGCGCGAGCAGGAAGAAAAGSAGGACATGGAGACCC
|
AGGCTGTGGCAACGTCCCCCGATGGCCGATACCTCAAGTTTGACATCGAGATTGGACG
|
TGGCTCCTTCAAGACGGTGTATCGAGGGCTAGACACCGACACCACAGTGGAGGTGGCC
|
TGGTGTGAGCTGCACGTGCGGACTCGGAAACTCTCTAGACCTGAGCCGCAGCGCTTCT
|
CAGAGGAGGTGGAGATGCTCAAGGGGCTGCAGCACCCCAACATCGTCCGCTTCTATGA
|
TTCGTGGAAGTCGGTGCTGAGGGCCCAGGTTTGCATCCTGCTGGTCACCGAACTCATG
|
ACCTCGGGCACGCTCAAGACGTACCTGAGGCGGTTCCGGGAGATGAAGCCGCGGGTCC
|
TTCAGCGCTCGACCCGCCAAATCCTGCGGGGACTTCATTTCCTACACTCCCCGGTTCC
|
TCCCATCCTGCACCGGGATCTCAAGTGCGACAATGTCTTTATCACGGGACCTACTGGC
|
TCTGTCAAAATCCGCGACCTGGCCCTGGCCACGCTCAAGCGCGCCTCCTTTGCCAAGA
|
GTGTCATCGGTACCCCGGAATTCATGGCCCCCGACATGTACCAGGAAAAGTACGATGA
|
GGCCGTCGACGTGTACGCGTTCGGCATGTGCATGCTGGAGATGGCCACCTCTGAGTAC
|
CCGTACTCCCAGTGCCAGAATGCCGCGCAAATCTACCGCAAGGTCACTTCGGTGAGAG
|
GGATGGGGCTCGCCGGAAAGGCAATTCCAGACGTGAAGGACATCATTGAACCCTGCAT
|
CCGCACGGATAAGAACGAGAGGTTCACCATCCAGGACCTCCTGGCCCACGCCTTCTTC
|
CGCGAGGAGCCCGGTGTGCACGTCGAACTAGCGGAGGACGACGACGGCGAGAAGCCGG
|
GCCTCAAGCTCTGGCTGCGCATGGAGGACGCGCGGCGCGGGGGGCGCCCACGGGACAA
|
CCAGGCCATCGAGTTCCTGTTCCAGCTGGGCCGGGACGCCGCCGAGGAGCTGGCACAG
|
GAGATGGTGAGCCGAGGATTGGTCTGTGAAGCCGATTACCAGCCAGTGGCCCGTGCAG
|
TACGTGAACGGGTTGCTGCCATCCAGCGAAAGCGTCAGAAGCTGCGTAAACCAACGGA
|
ACGGGTTGCTGCCATCCAGCGAAAGCGTGAGAAGCTGCGTAAAGCAAGGGAATTGGAG
|
GCACTCCCACCAGAGCCAGGACCTCCACCAGCAACTGTGCCCATGGCCCCCGGTCCCC
|
CCAGTCTCTTCCCCCCTGAGCCTGAGGAGCCAGAGGCAGACCAGCACCAGCCCTTCCT
|
TTTCCCCCACGCCAGCTACTCATCCTCTCCCTCCAACAGCTATGCCTCAGATGCAGCT
|
TCAGGCCTTAGCGATGTGGCAGAAGGGATGGGACAAATGAGGAGACCCCCTGGGAGGA
|
ATCTCCGGCGCAGACCCCGATCCCGGCTGCGGGTCACTAGTCAGCATGATGAAGAGGG
|
CTTGACCTTCTCCCCTCTGCTGACTTTGAATCTCAAGGTCTCAGACCAGAATGACAGA
|
GTGGTTGAGTGCCAGCTACAGACCCATAACAGCAAGATGCTGACCTTCCGATTTGATC
|
TGGATCGGGACAGCCCGGAAGAGATTGCAGCTGCCATGGTGTATAACCAGTTCATTCT
|
GCCTTCGGAGCGAGATGGATTTCTCACACGGATTCGGGAGATTATCCAGCGAGTGGAG
|
ACCCTGTTGAAGAGAGACCCCATCACTTCTCCCCCATGTCATCCCAGCCCCTCCCCAT
|
TCTCCCCCATTTCTTCCCAGGTCTCCTCAAATCCCTCTCCACACCCCACCAGCTCTCC
|
ACTTCCATTCTCCTCCAGCACACCCGAGTTTCCCGTCCCACTCTCTCAGTGTCCCTGG
|
AGTTCTCTCCCCACGACTTCTCCACCTACGTTCTCTCCCACTTGTTCTCAGCTCACTC
|
TTAGTTCCCCTTTCTTTCCTCCGTGCCCCTCCACTTCTTCCTTCCCCTCCACCACAGC
|
AGCCCCTCTCCTTTCTCTCGCTAGTGCCTTCTCACTGCCTGTGATCACTGTGGCCCAG
|
TCCCTGCTGGCCTTCTCCACCTCCTCATCTTCTCCTGGAACTCCTTTGTCTCGTCGAA
|
ACCCATTTTCCCCTGGAACCCCCATTTCCCCAGGTCCCATCTTCCCCATCACTTCTCC
|
CCCATGTCATCCCAGCCCCTCCCCATTCTCCCCCATTTCTTCCCAGGTCTCCTCAAAT
|
CCCTCTCCACACCCCACCAGCTCTCCACTTCCATTCTCCTCCAGCACACCCGAGTTTC
|
CGGTCCCACTCTCTCAGTGTCCCTGGAGTTCTCTCCCCACGACTTCTCCACCTACGTT
|
CTCTCCCACTTGTTCTCAGGTCGGGACTGTCGTCTCAACATACCCTCCTTCCCAGAAG
|
AGCTCCAGAGCAGCACCTCCCTGGAGCACACGAGCTGGACAGCCTTCTCCACCTCCTC
|
ATCTTCTCCTGGAACTCCTTTGTCTCCTGGAAACCCATTTTCCCCTGGAACCCCCATT
|
TCCCCACGTCCCATCTTCCCCATCACTTCTCCCCCATGTCATCCCAGCCCCTCCCCAT
|
TCTCCCCCATTTCTTCCCAGGTCTCCTCAAATCCCTCTCCACACCCCACCAGCTCTCC
|
ACTTCCATTCTCCTCCAGCACACCCGAGTTTCCGGTCCCACTCTCTCAGTGTCCCTGG
|
AGTTCTCTCCCCACGACTTCTCCACCTACGTTCTCTCGTTGGGCCTTTCCAAGTGACT
|
TCATCCAAGGAACCGCCTGAGCCTCTTCCCTTGCAGCCAACATCCCCCACTCTCTCTG
|
GTTCTCCAAAACCTTCAACCCCTCAGCTCACTTCAGAGAGCTCAGATACAGAGGACAG
|
TGCTGGAGGCGGGCCAGAGAGGCACTTGTCAGAGGTGGAAACACTACAGACACTACAG
|
AAAAAAGAAATTGAAGATTTGTACAGCCGGCTGGCGAAGCAGCCCCCACCGGGTATTG
|
TGGCCCCACCTGCTATGCTGTCCAGCCGCCAGCCCCGCCTCTCCAAGGCCAGCTTCCC
|
CACCTCCCGCCGCAACAGCCTACAGCGCTCTGAGCCCTGCTGCGTGCCTGCAATCCCA
|
GCTACTCAGCAGGCTGAGGCAGGAGAATTTCTTGAGCCTGGGAGGCCCACGTTGCAGA
|
ATCATTCTCCCTCTGCCCTTCGACAGAATTCCTCTTCTCCTCCTTCCCATAACCTGTT
|
TTTAACAGCATCCAACCCACCCATTCTTTCTGCAACCTCAAGATGCTGCATGAGCTTC
|
CATACCTCACTGGGAAGTTGGATCTTCATTCTTAAGCCTCCCATGTATACATGTTAAA
|
TACATTTGTAATCTTTTTCTCATATTAATTAATCTGCCTTATCTCAGTCACTTTTCAG
|
CGAACTTGTAGGTAGTCACTCACATAAGGATCCTCCAGTTTACATAGTTTTGTAAATG
|
TCAGGTCACCCTGTCCCACTAGACCACCAAGCTCCTGCAGGACACCTGGGATTTAACT
|
TTTTTTTTTTTTTTTTT
|
ORF Start: ATG at 72ORF Stop: TAA At 3825
SEQ ID NO: 801251 aaMW at 137141.7kD
NOV30b, MSQTEADLALRPPPPLGTAGQPRLGPPPRRARRFSGKAEPRPRSSRLSRRSSVDLGLL
CG90385-02 Protein Sequence
SSWSLPASPAPDPPDPPDSAGPGPARSPPPSSKEPPEGTWTEGAPVKAAEDSARPELP
|
DSAVGTGSRETLRVTEAVALERRREGEEKEDMETGAVATSTDGRYLKFDIEIGRGSFK
|
TVYRGLDTDTTVEVAWCELGVRTRKLSRAERGRFSEEVSMLKGLGHTNIVRFYDSWKS
|
VLRGQVCIVLVTELMTSGTLKTYLRRFREMKPRVLQRWSRQILRGLHFLHSRVPPILH
|
RDLKCDNVFITGPTGSVKIGDLGLATLKRASFAKSVIGTPEFMAPEMYEEKYDEAVDV
|
YAFGMCMLEMATSEYPYSECQNAAQIYRKVTSVRGMGLAGKAIPEVKEIIEGCIRTDK
|
NERFTIQDLLAHAFFREERGVHVELAEEDDGEKPGLKLWLRMEDARRGGRPRDNQAIE
|
FLFQLGRDAAEEVAQEMVSGGLVCEADYQPVARAVRERVAAIQRKREKLRKARERVAA
|
IGRKREKLRKARELEALTTETGTTTATVTMATCTTSVFTTETEETEADGHGTFLFRHA
|
SYSSSTSNSYASDAASGLSDVGEGMGGMRRTTGRNLRRRTRSRLRVTSGHDESGLTFS
|
PLLTLNLKVSDQNDRVVECQLQTHNSKMVTFRFDLDGDSPEEIAAAMVYNEFILPSER
|
DGFLRRIREIIGRVETLLKRDTITSTTCHTSTSTTSTISSGVSSNTSTHTTSSTLTFS
|
SSTPEFPVPLSQCPWSSLPTTSPPTFSPTCSQVTLSSPFFFPCPSTSSFPSTTAAPLL
|
SLASAFSLAVMTVAGQSLLAFSTSSSSPGTPLSPGNPFSPGTPISPGIFPITSPPCHP
|
SPSPFSPISSQVSSNPSPHPTSSPLPFSSSTPEFPVPLSQCPWSSLPTTSPPTFSPTC
|
SQVGTVVSTYPPSQKSSRAAPPWSTGACQPSPPPHLLLELLCLLETHFPLEPPFPQVP
|
SSPSLLPHVIPAPPHSPPFLPRSPQIPLHTPPALHFHSPPAHPSFRSHSLSVPGVLSP
|
RLLHLRSLVGRFQVTSSKEPAEPLPLQPTSPTLSGSPKPSTPQLTSESSDTEDSAGGG
|
PERHLSEVETLQTLQKKEIEDLYSRLGKQPPPGIVAPAAMLSSRQRRLSKGSFPTSRR
|
NSLQRSEPWCVPAIPATQEAEAGEFLEPGRRRLQNHSPSALRQNSSSPPSHNLFLTGS
|
KPPILSATSRWWMSFHTSLGSWIFILKPPMYTC
|
[0469] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 30B.
156TABLE 30B
|
|
Comparison of NOV30a against NOV30b.
NOV30a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV30b1 . . . 551419/553 (75%)
1 . . . 507420/553 (75%)
|
[0470] Further analysis of the NOV30a protein yielded the following properties shown in Table 30C.
157TABLE 30C
|
|
Protein Sequence Properties NOV30a
|
|
PSort0.8500 probability located in endoplasmic reticulum
analysis:(membrane); 0.8200 probability located in nucleus; 0.4400
probability located in plasma membrane; 0.3000 probability
located in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0471] A search of the NOV30a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 30D.
158TABLE 30D
|
|
Geneseq Results for NOV30a
NOV30aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAU03531Human protein kinase #31 - Homo 1 . . . 895781/898 (86%)0.0
sapiens, 1513 aa. [WO200138503- 1 . . . 850788/898 (86%)
A2, 31 MAY 2001]
AAB65656Novel protein kinase, SEQ ID NO: 20 . . . 630336/638 (52%)e-169
183 - Homo sapiens, 1920 aa. 97 . . . 667402/638 (62%)
[WO200073469-A2, 07 DEC. 2000]
AAM40523Human polypeptide SEQ ID NO 67 . . . 556299/491 (60%)e-164
5454 - Homo sapiens, 470 aa. 34 . . . 467357/491 (71%)
[WO200153312-A1, 26 JUL. 2001]
AAM38736Human polypeptide SEQ ID NO161 . . . 630298/497 (59%)e-158
1881 - Homo sapiens, 502 aa. 4 . . . 451350/497 (69%)
[WO200153312-A1, 26 JUL. 2001]
AAM38737Human polypeptide SEQ ID NO298 . . . 630223/360 (61%)e-117
1882 - Homo sapiens, 591 aa.185 . . . 540265/360 (72%)
[WO200153312-A1, 26 JUL. 2001]
|
[0472] In a BLAST search of public sequence datbases, the NOV30a protein was found to have homology to the proteins shown in the BLASTP data in Table 30E.
159TABLE 30E
|
|
Public BLASTP Results for NOV30a
NOV30aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96DT8SERINE/THREONINE PROTEIN 1 . . . 895781/898 (86%)0.0
KINASE (EC 2.7.1.37) - Homo 1 . . . 850788/898 (86%)
sapiens (Human), 1231 aa.
Q96J92HYPOTHETICAL 134.7 KDA 1 . . . 895781/898 (86%)0.0
PROTEIN - Homo sapiens (Human), 13 . . . 862788/898 (86%)
1243 aa.
Q9H4A3HYPOTHETICAL 250.8 KDA 20 . . . 686349/701 (49%)e-170
PROTEIN - Homo sapiens (Human), 97 . . . 730423/701 (59%)
2382 aa.
Q9JTH7PROTEIN KINASE WNK1 - Rattus 38 . . . 630344/654 (52%)e-170
norvegicus (Rat), 2126 aa. 68 . . . 668416/654 (63%)
Q9D9955430417M23RIK PROTEIN1075 . . . 1333226/259 (87%)e-125
(RIKEN CDNA 5430417M23 19 . . . 277243/259 (93%)
GENE) - Mus musculus (Mouse),
334 aa.
|
[0473] PFam analysis predicts that the NOV3 Oa protein contains the domains shown in the Table 30F.
160TABLE 30F
|
|
Domain Analysis of NOV30a
Identities/
Similarities
NOV30a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
pkinase: domain 1 of 2162 . . . 253 28/92 (30%)8.7e-09
64/92 (70%)
pkinase: domain 2 of 2297 . . . 465 56/208 (27%)1.5e-40
136/208 (65%)
Ca_channel_B:678 . . . 732 10/55 (18%)4
domain 1 of 1 33/55 (60%)
cyclin: domain 1 of 11112 . . . 1236 31/152 (20%)0.4
76/152 (50%)
|
Example 31
[0474] The NOV31 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 31A.
161TABLE 31A
|
|
NOV31 Sequence Analysis
|
|
SEQ ID NO:817961 bp
NOV31a,CTGTCTAATTCTTCCTTTCTCTCAATATAGGTATGGCATCACAGCTGCAAGTGTTTTC
CG90635-01 DNA SEQUENCE
GCCCCCATCAGTGTCGTCGAGTGCCTTCTGCAGTGCGAACAAACTGAAAATACAGCCC
|
TCTGGCTQGGATGTTTCAGQACAGAGTAGCAACGACAAATATTATACCCACAGCAAAA
|
CCCTCCCAGCCACACAAGGGCAAGCCAACTCCTCTCACCAGGTACCAAATTTCAACAT
|
CCCTGCTTACGACCAGGGCCTCCTCCTCCCAGCTCCTGCAGTGGAGCATATTGTTGTA
|
ACAGCCGCTGATAGCTCGGGCAGTGCTGCTACATCAACCTTCCAAAGCAGCCAGACCC
|
TGACTCACAGAAGCAACGTTTCTTTGCTTGAGCCATATCAAAAATGTGGATTGAAACG
|
AAAAACTGAGGAAGTTGACAGCAACGGTAGTGTGCAGATCATAGAAQAACATCCCCCT
|
CTCATGCTGCAAAACAGGACTGTCGTGGGTGCTGCTGCCACAACCACCACTGTGACCA
|
CAAAGAGTAGCAGTTCCAGCGGAGAAGGCGATTACCAGCTGGTCCAGCATGAGATCCT
|
TTGCTCTATGACCAATAGCTATGAAGTCTTGGAGTTCCTAGGCCGGGGGACATTTGGA
|
CAGGTCGCTAAGTGCTGGAAGAGGAQCACCAACGAAATTGTGGCTATTAAAATCTTGA
|
AGAACCACCCCTCCTATGCCAGACAAGGACAGATTGAAGTGACCATCCTTTCCCGCCT
|
AAGCAGTGAAAATGCTGATGAGTATAATTTTGTCCGTTCATACGAGTGCTTTCAGCAT
|
AAGCAGTGAAAATGCTGATGAGTATAATTTTGTCCGTTCATACGAGTGCTTTCAGCAT
|
AAGAATCACACCTGCCTTGTTTTTGAAATGTTGGAGCAGAACTTATATGATTTTCTAA
|
AGCAAAACAAATTTAGCCCACTGCCACTCAAGTACATCAGACCAATCTTGCAGCAGGT
|
GGCCACAGCCTTGATGAAGCTCAAGAGTCTTGGTCTGATCCACGCTGACCTTAAGCCT
|
GAAAACATCATGCTGGTTGATCCAGTTCGCCAGCCCTACCGAGTGAAGGTCATTGACT
|
TTGGTTCTGTCAGTCACGTTTCCAAAGCTGTGTGCTCAACCTACTTACAGTCACGTTA
|
CTACAGAGCTCCTGAAATTATCCTTCGATTACCATTCTGTGAAGCTATTGACATGTGG
|
TCACTGGCCTQTGTAATAGCTGAGCTGTTCCTGCGATGGCCTCTTTATCCTGGTGCTT
|
CAGAATACGAGCACATTCGTTATATTTCACAAACACAAGGCTTGCCAGCTCAATATCT
|
TCTCAGTGCCGGAACAAAAACAACCAGCTTTTTCAACAGAGATCCTAATTTGGGGTAC
|
CCACTGTGGAGGCTTAACACACCTCAAGAACATGAACTGCAGACTGGAATAAAATCAA
|
AAGAAGCTCGGAAGTACATTTTTAATTGCTTAGATGACATGGCTCAGGTGAATATGTC
|
TACAGACCTGGAGGGAACAGACATGTTGGCAGACAAGGCACACCCAAGAGAATACATT
|
GATCTGTTAAAGAAAATGCTCACAATTGATGCAGATAACAGAATTACCCCTCTAAAAA
|
CTCTTAACCATCAGTTTGTGACAATGACTCACCTTTTGGATTTTCCACATAGCAATCA
|
GAGTGTTAAGTCTTGTTTTCACAACATQGAGATCTGCAAGCGGAGGCTTCACATGTAT
|
GATACAGTGAGTCACATCAAGAGTCCCTTCAGTACACATGTTGCCCCAAATACAAGCA
|
CAAATCTAACCATGAGCTTCAGCAATCAGCTCAATACAGTGCACAATCAGGCCAGTGT
|
TCTAGCTTCCAGTTCTACTGCAGCAGCTGCTACTCTTTCTCTGGCTAATTCAGATGTC
|
TCACTACTAAACTACCAGTCAGCTTTGTACCCATCATCTGCTGCACCAGTTCCTGGAG
|
TTCCCCAGCAGCGTGTTTCCTTGCACCCTGGAACCACCCAGATTTGCACTCACACAGA
|
TCCATTCCAACAGACATTTATAGTATGTCCACOTGCCTTTCAAAGTGGACTACAACCA
|
ACAACAAAGCATTCTGGATTCCCTGTGACOATGGATAATGCTGTACCGATTGTACCCC
|
AGGCACCAGCTCCTCAGCCACTACAGATTCAGTCAGGAGTTCTCACGCAGGGAAGCTG
|
TACACCACTAATGGTACCAACTCTCCACCCTCAAGTAGCCACCATCACACCGCAGTAT
|
GCGGTGCCCTTTACTCTGAGCTGCGCAGCCGGCCGGCCGGCGCTGGTTGAACAGACTG
|
CCGCTGTACTGCAGGCCTGGCCTGGAGGGACTCAGCAAATTCTCCTGCCTTCAACTTG
|
GCAACAGTTGCCTGGGGTACCTCTACACAACTCTGTCCAGCCCACAGCAATGATTCCA
|
GAGGCCATGGCGAGTGGACAGCAGCTACCTGACTCGAGGAGGAATGCCCACTCTCATG
|
GCAACCAGTACAGCACTATCATGCAGCAGCCATCCTTGCTGACTAACCATGTQACATT
|
GGCCACTGCTCAGCCTCTGAATGTTGGTGTTGCCCATGTTGTCAGACAACAACAATCC
|
AGTTCCCTCCCTTCGAAGAAGAATAAGCAGTCAGCTCCAGTCTCTTCCAAGTCCTCTC
|
TAGATGTTCTGCCTTCCCAAGTCTATTCTCTGGTTGGGAGCAGTCCCCTCCGCACCAC
|
ATCTTCTTATAATTCCTTCGTCCCTGTCCAAGATCAGCATCAGCCCATCATCATTCCA
|
GATACTCCCAGCCCTCCTGTGAGTGTCATCACTATCCGAAGTGACACTGATGAGCAAG
|
AGGACAACAAATACAAGCCCAGTAGCTCTGGACTGAAGCCAAGGTCTAATGTCATCAT
|
TTATGTCACTGTCAATGATTCTCCACACTCTGACTCTTCTTTGAGCAGCCCTTATTCC
|
ACTGATACCCTGAGTGCTCTCCCAGGCAATAGTGGATCCGTTTTGGAGGGGCCTGGCA
|
GAGTTGTGGCAGATGGCACTGGCACCCCCACTATCATTCTGCCTCCACTGAAAACTCA
|
GCTTGGTGACTGCACTGTAGCAACCCAGCCCTCAGGTGGTCTCCTGAGCAATAAGACT
|
AAGCCACTCGCTTCAGTCAGTGCGCAGTCATCTCGATOCTGTATCACCCCCACAGGGT
|
ATCGAGCTCAACGCGGGGGCACCAGTGCAGCACAACCACTCAATCTTAGCCAGAACCA
|
GCAGTCATCGCCGGCTCCAACCTCACAGGAGAGAAGCAGCAACCCACCCCCCCGCAGG
|
CAGCAGGCGTTTGTGGCCCCTCTCTCCCAAGCCCCCTACACCTTCCAGCATGGCAGCC
|
CGCTACACTCGACAGGGCACCCACACCTTGCCCCGGCCCCTGCTCACCTGCCAAGCCA
|
GGCTCATCTGTATACGTATGCTGCCCCGACTTCTCCTGCTGCACTGGGCTCAACCAGC
|
TCCATTGCTCATCTTTTCTCCCCACACGGTTCCTCAAGOCATGCTGCAGCCTATACCA
|
CTCACCCTAGCACTTTGGTGCACCAGGTCCCTGTCAGTGTTGGGCCCAGCCTCCTCAC
|
TTCTGCCAGCGTGGCCCCTGCTCAGTACCAACACCAGTTTGCCACCCAATCCTACATT
|
GGGTCTTCCCGAGGCTCAACAATTTACACTGGATACCCGCTGAGTCCTACCAAGATCA
|
GCCAGTATTCCTACTTATAGTTGGTGAGCATGAGGGAGGAGGAATCATGGCTACCTTC
|
TCCTGGCCCTGCGTTCTTAATATTGGGCTATGGAGAGATCCTCCTTTACCCTCTTGAA
|
ATTTCTTAGCCAGCAACTTGTTCTGCAGGGGCCCACTGAAGCAGAAGGTTTTTCTCTG
|
GGGGAACCTGTCTCAGTGTTGACTGCATTGTTGTAGTCTTCCCAAAGTTTGCCCTATT
|
TTTAAATTCATTATTTTTGTCACAGTAATTTTGGTACTTGGAAGAGTTCAGATGCCCA
|
TCTTCTGCAGTTACCAAGGAAGAGAGATTGTTCTGAAGTTACCCTCTGAAAAATATTT
|
TGTCTCTCTGACTTGATTTCTATAAATGCTTTTAAAAACAAGTGAAGCCCCTCTTTAT
|
TTCATTTTGTGTTATTGTGATTGCTGGTCAGGAAAAATGCTGATAGAAGGAGTTGAAA
|
TCTGATGACAAAAAAAGAAAAATTACTTTTTGTTTGTTTATAAACTCAGACTTGCCTA
|
TTTTATTTTAAAAGCGGCTTACACAATCTCCCTTTTGTTTATTGGACATTTAAACTTA
|
CAGAGTTTCAGTTTTGTTTTAATGTCATATTATACTTAATGGGCAATTGTTATTTTTG
|
CAAAACTCGTTACGTATTACTCTGTCTTACTATTGAGATTCTCTCAATTCCTCCTGTG
|
TTTGTTATAAAGTAGTGTTTAAAAGGCAGCTCACCATTTGCTCGTAACTTAATGTGAG
|
AGAATCCATATCTGCGTGAAAACACCAAGTATTCTTTTTAAATGAAOCACCATGAATT
|
CTTTTTTAAATTATTTTTTAAAAGTCTTTCTCTCTCTGATTCACCTTAAATTTTTTTA
|
TCGAAAAAGCCATTAAGGTGGTTATTATTACATGGTGGTGGTCGTTTTATTATATGCA
|
AAATCTCTGTCTATTATCAGATACTGGCATTGATGAGCTTTGCCPAAAGATTAGTATG
|
AATTTTCACTAATACACCTCTGTTTTGCTCATCTCTCCCTTCTGTTTTATGTGATTTG
|
TTTGGGGAQAAAGCTAAAAAAACCTCAAACCAGATAAGAACATTTCTTGTGTATAGCT
|
TTTATACTTCAAAGTAGCTTCCTTTGTATGCCAGCAGCAAATTGAATGCTCTCTTATT
|
AAGACTTATATAATAAGTGCATGTAGGAATTCCAAAAAATATTTTAAAAATTTATTAC
|
TGAATTTAAAAATATTTPAGAAGTTTTGTAATGGTCGTCTTTTAATATTTTACATAAT
|
TAAATATGTACATATTGATTAGAAAAATATAACAAGCAATTTTTCCTGCTAACCCAAA
|
ATGTTATTTGTAATCAAATGTGTAGTGATTACACTTGAATTGTCTACTTAGTCTCTAT
|
GTGATCCTCCAGTGTTATCCCGGACATGGATTGATGTCTCCATTGTATTTAAACCAAA
|
ATGAACTGATACTTGTTGGAATGTATGTGAACTAATTGCAATTATATTAGAGCATATT
|
ACTGTAGTGCTGAATGAGCAGGGGCATTGCCTGCAAGGAGAGGAGACCCTTGGAATTG
|
TTTTGCACAGGTGTGTCTGGTGAGGAGTTTTTCAGTCTGTGTCTCTTCCTTCCCTTTC
|
TTCCTCCTTCCCTTATTGTAGTGCCTTATATGATAATGTAGTGGTTAATAGACTTTAC
|
AGTGAGCTTGCCTTAGGATGGACCAGCAAGCCCCCGTGGACCCTAAGTTGTTCACCGG
|
GATTTATCACAACAGCATTAGTAGCTGTATTGTGTAATGCATTGTTCTCAGTTTCCCT
|
GCCAACATTGAAAAATAAAAACAGCAGCTTTTCTCCTTTACCACCACCTCTACCCCTT
|
TCCATTTTGGATTCTCGGCTGAGTTCTCACAGAAGCATTTTCCCCATGTGGCTCTCTC
|
ACTGTGCGTTGCTACCTTGCTTCTCTGAGAATTCAGGAAGCAGGTGAGAGGAGTCAAG
|
CCAATATTAAATATGCATTCTTTTAAAGTATGTGCAATCACTTTTAGAATGAATTTTT
|
TTTTCCTTTTCCCATQTGGCACTCCTTCCTGCACATAGTTGACATTCCTAGTAAAATA
|
TTTGCTTGTTGAAAAAAACATGTTAACAGATGTGTTTATACCAAAGAGCCTCTTGTAT
|
TGCTTACCATGTCCCCATACTATGACGAGAAGTTTTCTGGTGCCGCTGGTGACAAGGA
|
ACTCACAGAAAQGTTTCTTAGCTGCTGAAGAATATAGACAAGOAACCAAAGCCTGTTC
|
AGTCATTGAGGCTTTTGAGGTTTCTTTTTTAACAGCTTGTATAGTCTTGGGGCCCTTC
|
AAGCTGTGAAATTGTCCTTGTACTCTCAGCTCCTGCATOGATCTGGGTCAACTACAAG
|
GTACTGGGGATGGGGACATTCCTGCCCATAAAGGATTTGGGGAAAGAAGATTAATCCT
|
AAAATACAGGTGTGTTCCATCCGAATTGAAAATGATATATTTGACATATAATTTTAGG
|
ACTGGTTCTGTGTAGATAGACATGGTGTCAAGGAGGTCCACCATGGAGATCGGAGATT
|
TCATGGAGCCTGGTCAGCCAGCTCTGTACCAGGTTGAACACCGAGGAGCTGTCAAAGT
|
ATTTGGAGTTTCTTCATTGTAAGCAGTAAGGGCTTCCAAGATGGGGCAGGTAGTCCGT
|
ACAGCCTACCAGGAACATGTTGTGTTTTCTTTATTTTTTAAAATCATTATATTGAGTT
|
GTGTTTTCAGCACTATATTGGTCAACATACCCAAGCAGTTTGTATAATTTCTCTCACT
|
AGTGTCATACAGTTTTCTGCTCAACATGTGTGATCTTTGTGTCTCCTTTTTGCCAAGC
|
ACATTCTGATTTTCTTGTTGGAACACAGCTCTAGTTTCTAAAGGACAAATTTTFTGTT
|
CCTTGTCTTTTTTCTGTAAGCGACAAGATTTGTTGTTTTTGTAAGAAATGAGATGCAG
|
GAAAGAAAACCAAATCCCATTCCTGCACCCCAGTCCAATAAGCAGATACCACTTAAGA
|
TAGGAGTCTAAACTCCACAGAAAAGGATAATACCAAGAGCTTGTATTGTTACCTTAGT
|
CACTTCCCTAGCAGTGTGTCCCTTTAAAAACTAGAGATTTTTCAGTCTTAGTCTCCAA
|
ACTGGCATTTCCGATTTTCCAGCATAAAAATCCACCTGTGTCTGCTGAATCTGTATGT
|
ATGTGCTCACTGTGGCTTTAGATTCTGTCCCTGGGGTTACCCCTGTTCGCCCTGACAG
|
GAAGGGAGGAAGCCTGGTGAATTTAGTGAGCAGCTGGCCTGGGTCACAGTGACCTGAC
|
CTCAPACCAGCTTAAGGCTTTAAGTCCTCTCTCAGAACTTGGCATTTCCAACTTCTTC
|
CTTTCCGGGTGAGAGAAGAAGCGGAGAAGGGTTCAGTGTAGCCACTCTGGGCTCATAG
|
GGACACTTGOTCACTCCACAGTTTTTAATAGCTCCCAGGAGGTGATATTATTTTCAGT
|
GCTCAGCTGAAATACCAACCCCAGGAATAAGAACTCCATTTCAAACAGTTCTGGCCAT
|
TCTGAGCCTGCTTTTGTCATTGCTCATCCATTGTCCTCCACTAGAGGGGCTAAGCTTG
|
ACTGCCCTTAGCCAGGCAACCACAGTAATGTGTGTTTTCTTCAGCATTATTATGCAAA
|
AATTCACTAGTTGAGATGGTTTGTTTTAGGATAGGAAATGAAATTGCCTCTCAGTCAC
|
AGGAGTGGCCCGAGCCTGCTTCCTATTTTGATTTTTTTTTTTTTTAACTGATAGATGG
|
TGCAGCATGTCTACATGGTTGTTTGTTGCTAAACTTTATATAATGTGTGGTTTCAATT
|
CAGCTTGAAAAATAATCTCACTACATGTAGCAGTACATTATATGTACATTATATGTAA
|
TCTTAGTATTTCTGCTTTGAATCCTTGATATTGCAATGGAATTCCTACTTTATTAAAT
|
GTATTTCATATGCTAGTTATTCTGTCCGATTTAAACTTTTTTTGCTTTCTCCCTTTTT
|
TTGGTTGTGCGCTTTCTTTTACAACAAGCCTCTAGAAACAGATAGTTTCTGAGAATTA
|
CTCAGCTATGTTTGTAATGCAGATGTACTTAGGGAGTATGTAAAATAATCATTTTAAC
|
AAAAGAAATAGATATTTAAAATTTAATACTAACTATGGCAAAAGGGTCCATTGTCTAA
|
AACATAGTTTATCTTTGGATTCAATGTTTGTCTTTCGTTTTACAAAGTAGCTTGTATT
|
TTCAGTATTTTCTACATAATATGGTAAAATGTAGAGCAATTGCAATGCATCAATAAAA
|
TGGGTAAATTTTCTG
|
ORF Start: ATG at 33ORF Stop: TAG at 3672
SEQ ID NO: 821213 aaMW at 131132.5kD
NOV31a, MASQLQVFSPPSVSSSAFCSAKKLKIEPSGWDVSGQSSNDKYYTHSKTLPATQGQANS
CG90635-01 Protein Sequence
SHQVANFNIPAYDQGLLLPAPAVEHIVVTAADSSGSAATSTFQSSQTLTHRSNVSLLE
|
PYQKCGLKRKSEEVDSNGSVQIIEEHPPLMLQNRTVVGAAATTTTVTTKSSSSSGEGD
|
YQLVQHEILCSMTNSYEVLEFLGRGTFGQVAKCWKRSTKEIVAIKILKNHPSYARQGQ
|
IEVSILSRLSSENADEYNFVRSYECFQHKNHTCLVFEMLEQNLYDFLKQNKFSPLPLK
|
YIRPILQQVATALMKLKSLGLIHADLKPENIMLVDPVRQPYRVKVIDFGSASHVSKAV
|
CSTYLQSRYYRAPEIILGLPFCEAIDMWSLGCVIAELFLGWPLYPGASEYEQIRYISQ
|
TQGLPAEYLLSAGTKTTRFFNRDPNLGYPLWRLKTPEEHELETGIKSKEARKYIFNCL
|
DDMAQVNMSTDLEGTDMLAEKADRREYIDLLKKMLTIDADKRITPLKTLNHQFVTMTH
|
LLDFPHSNQSVKSCFQNMEICKRRVHMYDTVSQIKSPFTTHVAPNTSTNLTMSFSNQL
|
NTVHNQASVLASSSTAAAATLSLANSDVSLLNYQSALYPSSAAPVPGVAQQGVSLQPG
|
TTQICTQTDPFQQTFIVCPPAFQSGLQATTKHSGFPVRMDNAVPIVPQAPAAQPLQIQ
|
SGVLTQGSCTPLMVATLHPQVATITPQYAVPFTLSCAAGRPALVEQTAAVLQAWPGGT
|
QQILLPSTWQQLPGVALHNSVQPTAMIPEAMGSGQQLADWRRNAHSHGNQYSTIMQQP
|
SLLTNHVTLATAQPLNVGVAHVVRQQQSSSLPSKKNKQSAPVSSKSSLDVLPSQVYSL
|
VGSSPLRTTSSYNSLVPVQDQHQPIIIPDTPSPPVSVITIRSDTDEEEDNKYKPSSSG
|
LKPRSNVISYVTVNDSPDSDSSLSSPYSTDTLSALRGNSGSVLEGPGRVVADGTGTRT
|
IIVPPLKTQLGDCTVATQASGGLLSNKTKPVASVSGQSSGCCITPTGYRAQRGGTSAA
|
QPLNLSQNQQSSAAPTSQERSSNPAPRRQQAFVAPLSQAPYTFQHGSPLHSTGHPHLA
|
PAPAHLPSQAHLYTYAAPTSAAALGSTSSIAHLFSPQGSSRHAAAYTTHPSTLVHQVP
|
VSVGPSLLTSASVAPAQYQHQFATQSYIGSSRGSTIYTGYPLSPTKISQYSYL
|
[0475] Further analysis of the NOV31 a protein yielded the following properties shown in Table 31B.
162TABLE 31C
|
|
Protein Sequence Properties NOV31a
|
|
PSort0.4974 probability located in mitochondrial matrix space;
analysis:0.3000 probability located in microbody (peroxisome); 0.2147
probability located in mitochondrial inner membrane; 0.2147
probability located in mitochondrial intermembrane space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0476] A search of the NOV31a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 31C.
163TABLE 31C
|
|
Geneseq Results for NOV31a
NOV31aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAE11767Human kinase (PKIN)-1 protein-1..12131206/1213 (99%)0.0
Homo sapiens, 1210 aa.1..12101208/1213 (99%)
[WO200181555-A2, 01-NOV-2001]
AAB65661Novel protein kinase, SEQ ID NO:1..1213 771/1259 (61%)0.0
188—Homo sapiens, 1171 aa.8..1171 893/1259 (70%)
[WO200073496-A2, 07-DEC-2000]
AAY53013Human secreted protein clone574..1213 617/663 (93%)0.0
co155_12 protein sequence SEQ ID1..654 624/663 (94%)
NO:32—Homo sapiens, 654 aa.
[WO9957132-A1, 11-NOV-1999]
AAM25563Human protein sequence SEQ ID196..850 468/656 (71%)0.0
NO:1078—Homo sapiens, 590 aa.1..575 513/656 (77%)
[WO200153455-A2, 26-JUL-2001]
AAW00215Drug resistance-associated protein10..1186 563/1268 (44%)0.0
kinase—Homo sapiens, 1160 aa.6..1160 718/1268 (56%)
[WO9627015-A2, 06-SEP-1996]
|
[0477] In a BLAST search of public sequence datbases, the NOV31a protein was found to have homology to the proteins shown in the BLASTP data in Table 31 D.
164TABLE 31D
|
|
Public BLASTP Results for NOV31a
NOV31aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9QUQ8NUCLEAR BODY ASSOCIATED1..12131179/1213 (97%)0.0
KINASE 2B - Mus musculus1..12101195/1213 (98%)
(Mouse), 1210 aa.
O88904HOMEODOMAIN-INTERACTING1..12131177/1213 (97%)0.0
PROTEIN KINASE 1 - Mus musculus1..12091194/1213 (98%)
(Mouse), 1209 aa.
Q9QZR3NUCLEAR BODY ASSOCIATED1..12131134/1213 (93%)0.0
KINASE 2A - Mus musculus1..11651150/1213 (94%)
(Mouse), 1165 aa.
Q9QZR5Homeodomain-interacting protein1..1213 792/1258 (62%)0.0
kinase 2 (EC 2.7.1.-) (Nuclear body8..1196 918/1258 (72%)
associated kinase 1) (Sialophorin tail
associated nuclear serine/threonine
kinase) - Mus musculus (Mouse),
1196 aa.
Q99P45HOMEODOMAIN-INTERACTING1..1213 791/1258 (62%)0.0
PROTEIN KINASE 2B - Mus8..1196 917/1258 (72%)
musculus (Mouse),
1196 aa.
|
[0478] PFam analysis predicts that the NOV31 a protein contains the domains shown in the Table 31E.
165TABLE 31E
|
|
Domain Analysis of NOV31a
NOV31aIdentities/
MatchSimilaritiesExpect
Pfam DomainRegionfor the Matched RegionValue
|
pkinase: domain 1 of 2190..411 85/235 (36%)1.9e-48
170/235 (72%)
pkinase: domain 2 of 2492..51813/30 (43%)0.013
20/30 (67%)
|
Example 32
[0479] The NOV32 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 32A.
166TABLE 32A
|
|
NOV32 Sequence Analysis
|
|
SEQ ID NO: 833157 BP
NOV32a, TGGTTCCCGGGAGCGGTAGAGCTGGACCCGGACCCAAGGCAGTCCAGGCTCCCCGGGG
CG90729-01 DNA Sequence
CTGCACACATGGAGGGCCAGAGCAGCAGGCGCAGCAGGAGGCCACGGACCCGCGCTGG
|
CCTGGGTTCCCTGCCCATGCCCCACGGTGTTGCCCAAACTGCGGCACCCTCCAAGGTG
|
GACTCAAGTTTTCAGCTCCCAGCAAAGAAGAACGCAGCCCTAGGACCCTCGGAACCAA
|
GGTTGCCTCTGGCACCTGTAGGGCCACGGGCAGCTATGTCAGCTTCCTCGGAAGGACC
|
GAGGCTGCCTCTGGCATCTCCCCGACCAATCCTGGCTCCACTGTGTACCCCTGAAGGG
|
CAGAAAACAGCTACTGCCCACCGCAGCTCCAGCCTGGCCCCAACATCTGTGGGCCAGC
|
TGCTGATGTCTGCCTCACCTGGACCAAAGCCTCCCCCAGCGACCACAQGCTCAGTTCT
|
GGCTCCGACGTCCCTGGCGCTGGTGATGCCTGCCTCAGCAGGGCCAACATCTCCCCCA
|
GTCACCCTGGGGCCCAATCTGGCCCCAACCTCCAGAGACCAGAAGCAGGAGCCACCTG
|
CCTCCGTGGGACCCAAGCCAACACTGGCAGCCTCTGGCCTGAGCCTCGCCCTGGCTTC
|
TGAGGAGCAGCCCCCAGAACTCCCCTCCACCCCTTCCCCGGTGCCCAGTCCAGTTCTG
|
TCTCCAACTCAGGAACAGGCCCTGGCTCCAGCATCCACCGCATCAGGCGCAGCCTCTG
|
TGGGACAGACATCAGCTAGAAAGAGCGATGCCCCAGCCCCTAGACCTCTCCCTGCTTC
|
TGAGCGGCATCTCCAGCCTCCAGCTCAGACATCTGCTCCTACAGGCTCCCCACCCTGC
|
ATCCAAACCTCCCCAGACCCTCGGCTCTCCCCCTCCTTCCGAGCCCGGCCTGAGGCCC
|
TCCACAGCAGCCCTCAGGATCCTCTTTTGCCACGGCCACCCCAGACCTTGCCCTTGGA
|
TGTGGGCCACGGTCCTTCAGACCCTCGCACTCACTCCCCTQGACTTCTGTCCCCCACC
|
TTCCGGCCTCGGGCCCCCTCAGGCCAGACTCTGCCCCCACCTCTGCCCAAGCCACCCC
|
GATCACCCAGCCGTTCCCCAAGCCACTCCCCGAATCGCTCTCCCTGTGTTCCCCCAGC
|
CCCTCACATGGCCCTCCCAACGCTTGGCACACAGAGTACAGGGCCTGGCAGGTGCCTG
|
AGCCCCAACCTTCAGGCCCAAGAAGCCCCAGCCCCAGTCACCACCTCCTCTTCTACAT
|
CCACCCTGTCATCCTCCCCTTGGTCAGCTCAGCCTACCTGGAACAGCGACCCCGGCTT
|
CCGGATCACTGTGGTCACATGGAACGTGGGCACTGCCATGCCCCCAGACGATGTCACA
|
TCCCTCCTCCACCTGCGCCCTGCTGACGACACCGACGCCGCAGACATGATCGCCATAC
|
GGTTGCACGAAGTGAACTCCATGCTCAACAAGCGACTCAAGGACGCCCTCTTCACGGA
|
CCAGTGGAGTGAGCTGTTCATGGATGCGCTAGGGCCCTTCAACTTCCTGCTGGTGAGT
|
TCGGTGAGGATGCAGGGTCTCATCCTGCTGCTGTTCCCCAAGTACTACCACCTGCCCT
|
TCCTGCGAGACGTGCAGACCGACTGCACGCGCACTGGCCTGGGCGGCTACTGGGGTAA
|
CAAGGGTGGCGTGAGCGTGCGCCTGGCGGCCTTCGGGCACATGCTCTGCTTCCTGAAC
|
TGCCACTTCCCTGCGCATATGGACAAGGCGGAGCAGCGCAAAGACAACTTCCAGACCA
|
TCCTCAGCCTCCAGCAGTTCCAAGGGCCGGGCGCACAGGGCATCCTGGATCATGACCT
|
CGTGTTCTGGTTCGGGGACCTGAACTTCCGCATTGACAGCTATCACCTGGACTTTGTC
|
AAGTTTGCCATCGACAGTGACCAGCTCCAPCAGCTCTGGGAGAAGGACCAGCTCAACA
|
TGGCCAAGAACACCTGGCCCATTCTGAAGGGCTTTCACGACGGGCCCCTCAACTTCCC
|
TCCCACCTTCAAGTTTGATGTGGGTACCAACAAATACGATACCAGTGCCAAGAAACGG
|
AAGCCACCTTCGACAGACCGTATCCTATCCAACGTCAACCCTCCAGGTCGGGGTCCCA
|
GCCCCTCAGGACGGAAGAGCCACCGACTCCAGGTGACGCACCACAGCTACCGCAGCCA
|
CATGGAATACACAGTCAGCCACCACAAGCCTGTGGCTGCCCAGTTCCTCCTGCACTTT
|
GCCTTCAGGGACGACATGCCACTGGTGCGGCTCGACQTCGCAGATGAGTGGGTGCCGC
|
CCGAGCAGGCGGTGCTGAGGTACCCCATGGAAACAGTGTTCGCCCGCAGCTCCTCCGA
|
CTGGATCGGCTTATACCGGGTGGGTTTCCCCCATTCCAAGGACTATGTGCCTTATGTC
|
TGGGCCAAACATGAAGATCTGGATGGGAATACCTACCAGGTAACATTCAGTGAGGAAT
|
CACTCCCCAAGGGCCATGGAGACTTCATCCTGGGCTACTATAGTCACAACCACAGCAT
|
CCTGATCGGCATCACTGAACCCTTCCAGATCTCQCTGCCTTCCTCGGAGTTGGCCAGC
|
AGCAGCACAGACAGCTCAGGCACCAGCTCAGAGGGAGAGGATGACAGCACACTGGAGC
|
TCCTTGCACCCAAGTCCCGCAGCCCCAGTCCTGGCAAGTCCAAGCGACACCGCAGCCG
|
CAGCCCGGGACTGGCCAGGTTCCCTGGGCTTGCCCTACCGCCCTCATCCCGTGAACGC
|
CGTGGTGCCAGCCGTACCCCCTCACCCCAGAGCCGCCGCCTGTCCCGAGTGGCTCCTG
|
ACAGGAGCAGTAATGGCAGCAGCCGGGGCAGTAGTGAAGAGGQGCCCTCTGGGTTGCC
|
TGCCCCCTGGGCCTTCCCACCAGCTGTGCCTCGAAGCCTGCGCCTGTTGCCCCCCTTG
|
CCCCTACAGACTGTAGACCCTGCTGGTGGTGGCTCCTGGGGACCTGATCGGGAGGCCC
|
TGGCGCCCAACAGCCTGTCTCCTAGTCCCCAGGGCCATCGGGGGCTGGAGCAAGGGGG
|
CCTGGGGCCCTGAGGGTGGGGTAGCCAGATGGGCCAAGGTCACCACCATTCTGCCTCA
|
ATCTTTTGCAAGCCCACCTCCCTCT
|
ORF Start: ATG at 67 ORF Stop: TGA at 3085
SEQ ID NO: 841006 aaMW at 107195.5kd
NOV32a, MEGQSSRGSRRPGTRAGLGSLPMPQGVAQTGAPSKVDSSFQLPAKKNAALGPSEPRLA
CG90729-01 Protein Sequence
LAPVGPRAAMSASSEGPRLALASPRPILAPLCTPEGQKTATAHRSSSLAPTSVGQLVM
|
SASAGPKPPPATTGSVLAPTSLGLVMPASAGPRSPPVTLGPNLAPTSRDQKQEPPASV
|
GPKPTLAASGLSLALASEEQPPELPSTPSPVPSFVLSPTQEQALAPASTASGAASVGQ
|
TSARKRDAPAPRPLPASEGHLQPPAQTSGPTGSPPCIQTSPDPRLSPSFRARPEALHS
|
SPEDPVLPRPPQTLFLDVCQGPSEPGTHSPCLLSFTFRPGAPSGQTVPPPLPKPPRSP
|
SRSPSHSPNRSPCVPPAPDMALPRLGTQSTGPGRCLSPNLQAQEAPAPVTTSSSTSTL
|
SSSPWSAQPTWKSDPGFRITVVTWNVGTAMPPDDVTSLLHLGGGDDSDGADMIAIGLQ
|
EVNSMLNKRLKDALFTDQWSELFMDALGPFNFVLVSSVRMQGVILLLFAKYYHLPFLR
|
DVQTDCTRTGLGGYWGNKGGVSVRLAAFGHMLCFLNCHLPAHMDKAEQRKDNFQTILS
|
LQQFQGPGAQGILDHDLVFWFGDLNFRIESYDLHFVKFAIDSDQLHQLWEKDQLNMAK
|
NTWPILKGFQEGPLNFAPTFKFDVGTNKYDTSAKKRKPAWTDRILWKVKAPGGGPSPS
|
GRKSHRLQVTQHSYRSHMEYTVSDHKPVAAQFLLQFAFRDDMPLVRLEVADEWVRPEQ
|
AVVRYRMETVFARSSWDWIGLYRVGFRHCKDYVAYVWAKHEDVDGNTYQVTFSEESLP
|
KGHGDFILGYYSHNHSILIGITEPFQISLPSSELASSSTDSSGTSSEGEDDSTLELLA
|
PKSRSPSPGKSKRHRSRSPGLARFPGLALRPSSRERRGASRSPSPQSRRLSRVAPDRS
|
SNGSSRGSSEEGPSGLPGPWAFPPAVPRSLGLLPALRLETVDPGGGGSWGPDREALAP
|
NSLSPSPQGHRGLEEGGLGP
|
[0480] Further analysis of the NOV32a protein yielded the following properties shown in Table 32B.
167TABLE 32B
|
|
Protein Sequence Properties NOV32a
|
|
PSort0.8500 probability located in endoplasmic reticulum
analysis:(membrane); 0.8200 probability located in nucleus;
0.4400 probability located in plasma membrane;
0.1297 probability located in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0481] A search of the NOV32a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 32C.
168TABLE 32C
|
|
Geneseq Results for NOV32a
NOV32aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResideusRegionValue
|
AAB27845Sequence homologous to protein613..993381/381 (100%)0.0
fragment encoded by gene 4 - Homo
sapiens, 381 aa. [WO200055199-A1,
21 Sep. 2000]
AAG73981Human colon cancer antigen protein481..735253/255 (99%)e-153
SEQ ID NO:4745 - Homo sapiens, 3..257254/255 (99%)
279 aa. [WO200122920-A2,
05 Apr. 2001]
AAB27846Protein fragment encoded by gene 4 -613..993260/382 (68%)e-145
Homo sapiens, 382 aa. 1..382262/382 (68%)
[WO200055199-A1,
21 Sep. 2000]
AAB27797Human secreted protein #4 - Homo504..735230/232 (99%)e-139
sapiens, 255 aa. [WO200055199-A1, 1..232231/232 (99%)
21 Sep. 2000]
AAW97094Phosphatidylinositol 4,5-488..839167/355 (47%)2e-92
bisphosphate 5-phosphatase - Homo 1..355221/355 (62%)
sapiens, 372 aa. [WO9900507-A1,
07 Jan. 1999]
|
[0482] In a BLAST search of public sequence datbases, the NOV32a protein was found to have homology to the proteins shown in the BLASTP data in Table 32D.
169TABLE 32D
|
|
Public BLASTP Results for NOV32a
NOV32aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9UDT9WUGSC:H_DJ412A9.2 PROTETN - 1..10061005/1044 (96%)0.0
Homo sapiens (Human), 1056 aa 13..10561006/1044 (96%)
(fragment).
Q9JMC1PROLINE-RICH INOSITOL 1..1006 842/1008 (83%)0.0
POLYPHOSPHATE 5- 1..1001 884/1008 (87%)
PHOSPHATASE - Rattus norvegicus
(Rat), 1001 aa.
Q15735PHOSPHATIDYLINOSITOL613..1006 394/394 (100%)0.0
(4,5)BISPHOSPHATE 5- 4..397 394/394 (100%)
PHOSPHATASE HOMOLOG -
Homo sapiens (Human), 397 aa
(fragment).
O09040PUTATIVE PHOSPHATASE425..843 201/423 (47%)e-108
(PUTATIVE PHOSPHOINOSITIDE 34..454 262/423 (61%)
5-PHOSPHATASE TYPE II) - Mus
musculus (Mouse), 468 aa.
Q9BT40SKIP FOR SKELETAL MUSCLE425..839 196/418 (46%)e-107
AND KIDNEY ENRICHED 16..431 258/418 (60%)
INOSITOL PHOSPHATASE - Homo
sapiens (Human), 448 aa.
|
[0483] PFam analysis predicts that the NOV32a protein contains the domains shown in the Table 32E.
170TABLE 32E
|
|
Domain Analysis of NOV32a
NOV32aIdentities/
MatchSimilaritiesExpect
Pfam DomainRegionfor the Matched RegionValue
|
IPPc: domain 1 of 2 56..4126/357 (2%)5.9e+04
249/357 (70%)
IPPc: domain 2 of 2421..736128/424 (30%)8.9e−108
249/424 (59%)
|
Example 33
[0484] The NOV33 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 33A.
171TABLE 33A
|
|
NOV33 Sequence Analysis
|
|
SEQ ID NO: 856261 bp
NOV33a,AAATTCAAACCAGTCAGCTTCATCTGGGCCTCTGATTCATCTTTATTCCCTCCATCAT
CG90760-01 DNA Sequence
CTACACTTGATTTTATTTGTACCAAGGAGATGCGTGTCTAATGTTTTTCTTTCTTCTA
|
TTTCTAGGAGGGCTGTTGGCCTGCTGCTGTGCTGCTGAACAGTATGCAGTCCTTTCGG
|
GAGCAAAGCAGTTACCACGGAAACCAGCAAAGCTACCCACAGGAGGTACACGGCTCAT
|
CCCGGCTAGAAGAGTTCAGCCCTCGTCAGGCCCAGATGTTCCAGAATTTTGGAGGTAC
|
AGGTGGCAGTAGTGCCAGCAGTCGCAGTGGCAGTCGTCGTGOACGACGAGGAGCAGCA
|
GCTGCTGCGGCAGCGATGGCTAGCGACACCTCTGGCCATCAAGGTTACCAGGGTTTCA
|
GGAAAGAGGCTGGAGATTTTTACTACATGGCAGGCAACAAAGACCCCGTGACTACAGG
|
AACCCCACAGCCTCCTCAGCGAAGGCCTTCTGGGCCTCTCCACAGCTATGGACCCCCC
|
CAGGGGAGCAGCTTTGGCAATCAGTATGGGAGTGAGGGTCATGTGGGCCAGTTTCAAG
|
CACAGCACTCTGGCCTTGGCGGTGTGTCACATTATCAGCAGGATTACACTGGGCCTTT
|
CTCTCCAGGGAGTCCTCAGTACCAACACCACGCTTCCAGCCACCAGCACCACCAGCAA
|
GTCCAGCAGTTGAGACAACAGCTTTACCAGTCCCATCAGCCCCTQCCACAGGCCACTG
|
GCCAACCAGCATCCAGCTCATCCCATCTACAGCCAATCCAGCGGCCCTCAACTCTGCC
|
ATCCTCTGCTCCTGGTTACCAGTTAAGAGTCGGTCAGTTTGGCCAACACTATCAGTCT
|
TCTGCTTCCTCCTCCTCCTCCTCCTCCTTCCCTTCACCACAGCGTTTTAGCCAGTCTG
|
GACAGAGCTATGATGGCAGTTACAATGTGAATGCTGGATCTCAGTATGAAGGACACAA
|
TGTGGGTTCTAATGCACAGGCTTATGGAACACAATCCAATTACAOCTATCAGCCTCAA
|
TCTATGAAGAATTTTGAACAGGCAAAGATTCCACAAGGGACCCAACAGGGGCAGCAGC
|
AGCAGCAACCGCAGCAACAACAACACCCTTCTCAGCATGTGATGCAGTATACTAACGC
|
TGCCACCAAGCTGCCCCTGCAAAGCCAACTGGOGCAGTACAACCAGCCTGAGGTTCCT
|
GTGAGGTCCCCCATGCAGTTTCACCAGAACTTCAGCCCCATTTCTAACCCTTCTCCAG
|
CTGCCTCTGTCGTTCAGTCTCCAAGCTGTAGTTCTACCCCATCTCCTCTCATGCAGAC
|
TGGGGAGAATCTCCAGTGTGCGCAAGGCAGTGTGCCTATCCGTTCCAGAAACAGAATT
|
TTACAGTTAATGCCTCAACTCAGTCCAACCCCATCAATGATGCCCAGTCCTAATTCTC
|
ATGCTGCAGGCTTCAAAGGGTTTGGACTAGAACGCGTACCAGAAAAGCCACTGACAGA
|
TCCTGGGTTCAGTACTTTGAGTGCTCTCAGTACTCAAGTGGCCAATCTTCCTAACACT
|
GTCCAGCACATGTTACTTTCTGATGCCCTGACTCCTCAGAAGAAGACCTCCAAGAGGC
|
CCTCATCTTCCAAGAAACCAGAPAGCTGCACAAATTCTGAACGCTCCTCACAACCTGA
|
AGAACAGCTGAACTCCCCTATGGCAGAGTCATTAGATGGAGGCTGCTCCAGCAGTTCA
|
GAGGATCAAGGCCAGAGAGTGCGGCAACTAAGTGGCCAGACCACCAGCTCTCACACCA
|
CCTACAAGGGTGGAGCCTCTGAGAAAGCTGGCTCCTCACCGGCACAAGGTGCTCAGAA
|
TGAACCCCCCAGACTCAATGCTAGTCCTGCCGCAAGAGAAGAGGCCACCTCACCAGGC
|
GCTAAGCACATGCCATTGTCATCCGACGCGAACCCAAAGCTTAATGACAACACTGTTG
|
GGGTGATTGTCTCCCGGGAAGCCATGACAGGTCGGGTAGAAAAGCCTGGTGGACAAGA
|
TAAAGGCTCCCAAGAGGAPGATCCTGCAGCCACTCAAAGGCCACCTAGCAATGGTGCG
|
GCAAAGGAAACCAGTCATGCATCACTTCCCCAGCCAGAGCCTCCAGGAGGAGGAGGGA
|
GCAAAGCAAACAAOAATGGCGATAACAACTCCAACCATAATGGACAAGGAAATGGCCA
|
GAGTGGCCACTCTGCAGCGGGCCCTGGTTTTACGAGCAGAACTGAGCCTAGCAAATCT
|
CCTGGAAGTCTGCGCTATAGTTACAAAGATAGTTTCCGGTCAGCCGTGCCACGAAATC
|
TCAGTGGCTTTCCTCAGTATCCTACAGGGCAAGAAAAGGGAGATTTCACTGGCCATGG
|
GGAACGAAAGGGTAGAAATGAAAAATTCCCAAGCCTCCTGCAGCAAGTGCTTCAGGGT
|
TACCACCACCACCCTGACAGQAGATATTCTAGGAGTACTCAAGAGCATCAGGGGATGG
|
CTGGTAGCCTAQAACGAACCACAAGGCCCAATGTCTTGGTTAGTCAAACCAATGAATT
|
AGCTAGCAGGGGCCTTCTGAACAAAAGCATTGCGTCTCTATTAGAAAATCCCCACTCG
|
GGCCCCTGGGAAAGGAAATCAAGCAGCACAGCTCCTGAAATCAAACAGATCAATTTGA
|
CTGACTATCCAATTCCCAGAAAGTTTGAAATAGAGCCTCAGTCATCAGCACATOAGCC
|
TGGGGGTTCCCTCTCTGAAACAAGATCAGTGATCTCTGATATTTCTCCACTAAGACAG
|
ATTGTCAGGGACCCAGGGGCTCACTCACTGGGACACATGAGTGCCGACACCAGAATTG
|
GGAGGAATGACCGTCTCAATCCAACTTTAAGTCAGTCGGTCATTCTTCCTGGTGGTTT
|
GGTGTCCATGGAAACCAAGCTGAAATCCCAGAGCGCGCAGATAAAAGAGGAAGACTTT
|
GAACAGTCTAAATCTCAAQCTAGTTTCAACAACAAGAAATCPGCAGACCACTGCCATC
|
CTCCTAGCATCAACCATGAGTCTTACCGCGGCAATGCCAGCCCTGGAGCACCAACCCA
|
TGATTCCCTTTCAGACTATGGCCCGCAAGACAGCAGACCCACGCCAATGCGGCGCGTC
|
CCTGGCAGAGTrGGTGCTCCGGAGGGCATGAGGGGTCGGTCCCCTTCTCAATATCATC
|
ACTTTGCAGAAAAATTGAAAATGTCTCCTCGGCGGAGCAGAGGCCCAGGGGGAGACCC
|
TCATCACATGAATCCACACATGACCTTTTCAGAGAGGGCTAACCGGAGTTCTTTACAC
|
ACTCCCTTTTCTCCCAACTCAGAAACCCTCGCCTCTGCTTATCATGCAAATACTCGCG
|
CTCATGCTTATCGGGACCCTAACGCACCTTTQAATTCTCAGCTGCATTATAAGAGACA
|
GATGTACCAACAGCAACCAGAGGAGTATAAAGACTCQAGCAGCCOTTCTGCTCACGGA
|
GTAATTGCTGCAGCACAGCACACGCAGGAGGGGCCACCGAAGAGTCCAAGGCAGCAGC
|
AGTTTCTTGACAGAGTACGGAGCCCTCTGAAAAATQACAAAGATGGTATGATGTATGG
|
CCCACCAGTGGGGACTTACCATGACCCCAGTGCCCAGGAGGCTGGGCGCTGCCTAATG
|
TCTACTGATGGTCTCCCTAACAAGGCCATGGAATTAAAGCATGGCPCCCAGAAGTTAC
|
AAGAATCCTCTTGGCATCTTTCTCGGCAAACTTCTCCAGCCAAAAGCAGCGGTCCTCC
|
AGGAATGTCCAGTCAAAAAAGCTATGGGCCGCCCCATGAGACTGATGGACATGGACTA
|
GCTGAGGCTACACAGTCATCCAAACCTCGTAGTGTTATCCTGACACTTCCAGGCCAGG
|
AGGATCATTCTTCTCAAAACCCCTTAATCATGAGGAGGCGTGTTCQTTCTTTTATCTC
|
TCCCATTCCCAGTAACAGACAGTCACAAGATGTAAAGAACAGTAGCACTGAAGATAAA
|
GGTCGCCTCCTTCACTCATCAAAAGAAGCCGCTGATAAAGCATTCAATTCCTATCCCC
|
ATCTTTCTCACAGTCAGGATATCAAGTCTATCCCTAAGAGAGATTCCTCCAAGGACCT
|
TCCAAGTCCAGATAGTAGAAACTGCCCTCCTCTTACCCTCACAAGCCCTGCTAAGACC
|
AAAATACTGCCCCCACGGAAAGGACGGGGATTGAAATTGGAAGCTATACTTCACAACA
|
TTACATCCCCAAATATTAGGAGGAGCGCATCTTCGAACAGTGCCGAGGCTGCGGCAGA
|
CACGGTTACGCTTGATGATATACTGTCTTTGAAGAGTGGTCCTCCTGAAGGTGGGAGT
|
GTTGCTGTTCAGGATGCTGACATAGAGAAGAGAAAAGGTGAOGTCGCTTCGCACCTAC
|
TCAGTCCAGCAAACCAGGAGTTGCACGTAGAGAAACCTCTTCCAAGGTCTTCAGAAGA
|
GTGGCGTGGCACCGTGGATGACAAAGTGAAGACAGAGACACATGCAGAAACAGTTACT
|
GCCGGAAAGGAACCCCCTGGTGCCATGACATCCACAACCTCACAGAAGCCTGGTAGTA
|
ACCAAGGGAGACCAGATGGTTCCCTGGGTGGAACAGCACCTTTAATCTTTCCAGACTC
|
AAAGAATGTACCTCCAGTGGGCATATTGGCCCCTGAGGCAAACCCCAAGGCTGAAGAG
|
AAGGAGAACGATACAGTGACGATTTCACCGAAGCAAGAGGGTTTCCCTCCAAAGGGAT
|
ATTTCCCATCAGGAAAGAACAAGGGGAGACCCATTGGTAGTGTGAATAAGCAAAAGAA
|
ACAGCAGCACCCACCGCCTCCACCCCCTCAGCCCCCACAGATACCAGAAGGTTCTGCA
|
GATGGAGAGCCAAAGCCAAAAAAACAGAGGCAAAGGAGGGACAGAACGAAGCCTGGGG
|
CCCAGCCGAGGAAGCGAAAAACCAAACAAGCAGTTCCCATTGTGGAACCCCAAGAACC
|
TGAGATCAAACTAAAATATGCCACCCAGCCACTGGATAAAACTGATGCCAAGAACAAG
|
TCTTTTTACCCTTACATCCATGTAGTAAATAAGTGTGAACTTGGAGCCGTTTGTACAA
|
TCATCAATGCTGAGGAAGAAGAACAGACCAAATTAGTGAGGGGCAGGAAGGGTCAGAG
|
GTCACTGACCCCTCCACCTAGCAGCACTGAAAGCAAGGCGCTCCCGGCCTCGTCCTTT
|
ATGCTGCAGGGACCTCTTGTGACAGAGTCTTCGCTTATGCGGCACCTGCTTTGCTGTC
|
TGTGTGGCAAGTGGGCCAGTTACCGGAACATCGGTGACCTCTTTGGACCTTTPTATCC
|
CCAAGATPATGCAGCCACTCTCCCGAAGAATCCACCTCCTAAGAGGGCCACAGAAATG
|
CAGAGCAAACTTAACGTACGGCACAAAAGTGCTTCTAATCGCTCCAAGACCGACACTG
|
AGGAGGAGGAAGAGCAGCACCACCAGCAGAAQGAGCAGAGAAGCCTCCCCCCACACCC
|
CACGTTTAAGCGGCGCCACCGCTCGGAAGACTGTGGTGGAGGCCCTCGGTCCCTGTCC
|
AGGGGCCTCCCTTCTAAAAAAGCAGCCACTGAGGGCAGCAGTGAAAACACTGTTTTGG
|
ACTCGAAGCCCTCCGTGCCCACCACTTCAGAAGGTGGCCCTGACCTGGAGTTACAAAT
|
CCCTGAACTACCTCTTGACAGCAATGAATTTTGGGTCCATGAGGGTTGTATTCTCTGG
|
GCCAATGGAATCTACCTGGTTTGTGGCAGGCTCTATGGCCTGCAGGAAGCGCTGGAAA
|
TGACAAGGTCGTCGACACTCGTGATGGAATCCAAATCCTCCTACCCTGCACCCACACC
|
CAACAAAGGCTGCTCCTTCCGATACCATTACCCGTGTGCCATTGATGCAGATTGTTTG
|
CTACATGAGGAGAACTTCTCGGTGAGGTGCCCTAAGCAACAAGGTGAGACTGTGGAGA
|
TGAGAAGGTGGTGGACACTCGTGATGGAATGGAAATCGTCCTACCGTGCAGCCACACC
|
CTGCCCTGCCCCCCCCCGCCCCGCCCCCGTGCCTGCCCATGCCAGCACTTCCTTAACT
|
TCTCACATCACACTCAAACCAGTGACACCACAGGAAAGAAAGACCCAAGACGTTGCAA
|
TCGCTGTTTCCATCCACACAATCTCCATAGTGACAATCTCCGCGCAGGCCCCACGCCT
|
GGGATGATGGCGAAAGCGTCGGCGGATTAAAAGGGACGGATAAATATATATATAT
|
ORF Start: ATG at 160ORF Stop: TGA at 6211
SEQ ID NO: 862017 aaMW at 218079.6kD
NOV33a, MQSFREQSSYHGNQQSYPQEVHGSSRLEEFSPRQAQMFQNFCGTCGSSCSSCSCSCGG
CG90760-01 Protein Sequence
RRGAAAAAAAMASETSGHQGYQGFRKEAGDFYYMAGNKDPVTTGTPQPPQRRPSGPVQ
|
SYGPPQGSSPCNQYGSEGEVCQFQAQHSGLCGVSHYQQDYTGPESPCSAQYQQQASSQ
|
QQQQQVQQLRQQLYQSHQPLPQATGQPASSSSHLQPMQRPSTLPSSAAGYQLRVCQFC
|
QHYQSSASSSSSSSFPSPQRFSQSGQSYDGSYNVNAGSQYEGHNVGSNAQAYGTQSNY
|
SYQPQSMKNFEQAKIPQGTQQGQQQQQPQQQQHPSQHVMQYTNAATKLPLQSQVGQYN
|
QPEVFVRSPMQFHQNFSPISNPSPAASVVQSPSCSSTPSPLMQTGENLQCCQGSVPMG
|
SRNRILQLMPQLSPTPSMMPSPNSEAACFKGFCLEGVPEKRLTDPCLSSLSALSTQVA
|
NLPNTVQHMLLSDALTPQKKTSKRPSSSKKADSCTNSEGSSQPEEQLKSPMAESLDGG
|
CSSSSEDQGERVRQLSGQSTSSDTTYKGGASEKAGSSPAQCAQNEPPRLNASPAAREE
|
ATSPGAKDMPLSSDCNPKVNEKTVGVIVSREAMTGRVEKFCCQDKCSQEDDPAATQRP
|
PSNGGAKETSHASLPQPEPPGGGGSKGNKNGDNNSNHNGEGNGQSGHSAAGPGFTSRT
|
EPSKSPGSLRYSYKDSFGSAVPRNVSGFPQYPTGQEKGDFTCHGERKGRNEKFPSLLQ
|
EVLQGYHHHPDRRYSRSTQEHQGMAGSLEGTTRPNVLVSQTNELASRGLLNKSIGSLL
|
ENPHWGPWERKSSSTAPEMKQINLTDYPIPRKFEIEPQSSAHEPGGSLSERRSVICDI
|
SPLRQIVRDPGAHSLGHMSADTRIGRNDRLNPTLSQSVILPCGLVSNETKLKSQSGQI
|
KEEDFEQSKSQASFNNKKSGDHCHPPSIKHESYRGNASPGAATHDSLSDYGPQDSRPT
|
PMRRVPGRVGGREGMRGRSPSQYHDFAEKLKMSPGRSRGPGGDPHHMNPHMTFSERAN
|
RSSLHTPFSPNSETLASAYHANTRAHAYGDPNAGLNSQLHYKRQMYQQQPEEYKDWSS
|
GSAGGVIAAAQHRQEGPRKSPRQQQFLDRVRSPLKNDKDGMMYGPPVGTYHDPSAQEA
|
GRCLMSSDGLPNKGMELKHGSQKLQESCWDLSRQTSPAKSSGPPGMSSQKRYGPPHET
|
DGHGLAEATQSSKPGSVMLRLPGQEDHSSQNPLIMRRRVRSFISPIPSKRQSQDVKNS
|
STEDKCRLLESSKEGADKAFNSYAHLSHSQDIKSIPKRDSSKDLPSPDSRNCPAVTLT
|
SPAKTKILPPRKGRGLKLEAIVQKITSPNIRRSASSNSAEAGGDTVTLDDILSLKSGP
|
PEGGSVAVQDADIEKRKGEVASDLVSPANQELHVEKPLPRSSEEWRGSVDDKVKTETH
|
AETVTASKEPPCAMTSTTSQKPGSNQGRPDGSLGGTAPLIFPDSKNVPPVGILAPEAN
|
PKAEEKENDTVTISPKQECPPPKGYFPSGKKKCRPIGSVNKQKKQQQPPPPPPQPPQI
|
PEGSADGEPKPKKQRQRRERRKPGAQPRKRKTKQAVPIVEPQEPEIKLKYATQPLDKT
|
DAKNKSFYPYIHVVNKCELGAVCTIINAEEEEQTKLVRGRKGQRSLTPPPSSTESKAL
|
PASSPNLQGPVVTESSVMCHLVCCLCCKWASYRNMGDLPCPPYPQDYAATLPKNPPPK
|
RATEMQSKVKVRHKSASNGSKTDTEEEEEQQQQQKEQRSLAAHPRFKRRHRSEDCGGG
|
PRSLSRGLPCKKAATEGSSEKTVLDSKPSVPTTSEGGPELELQIPELPLDSNEFWVHE
|
GCILWANGIYLVCGRLYGLQEALEIAREMKCSHCQEAGATLGCYNKGCSFRYHYPCAI
|
DADCLLHEENFSVRCPKQQGETVEMRRWWTLVMEWKSSYRAATPCPAPPRPARVPAHA
|
STSLSSHITLKPVTPQERKTQDVGMAVSMDTISIVTMWGEGGGVG
|
[0485] Further analysis of the NOV33a protein yielded the following properties shown in Table 33B.
172TABLE 33B
|
|
Protein Sequence Properties NOV33a
|
|
PSort0.9700 probability located in nucleus; 0.3000
analysis:probability located in microbody (peroxisome);
0.1000 probability located in mitochondrial
matrix space; 0.1000 probability located in
lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0486] A search of the NOV33a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 33C.
173TABLE 33C
|
|
Geneseq Results for NOV33a
NOV33aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB40646Human ORFX ORF410 polypeptide1559..1933133/416 (31%)2e-48
sequence SEQ ID NO:820 - Homo 98..491196/416 (46%)
sapiens, 494 aa. [WO200058473-A2,
05 Oct. 2000]
AAY51611Human HSGT1 protein - Homo 1..594179/625 (28%)8e-36
sapiens, 1798 aa. [WO200008143- 1..522248/625 (39%)
A2, 17 Feb. 2000]
ABG19553Novel human diagnostic protein161..592106/451 (23%)4e-11
#19544 - Homo sapiens, 901 aa.407..801158/451 (34%)
[WO200175067-A2, 11 Oct. 2001]
ABG19553Novel human diagnostic protein161..592106/451 (23%)4e-11
#19544 - Homo sapiens, 901 aa.407..801158/451 (34%)
[WO200175067-A2, 11 Oct. 2001]
AAE13839Human lung tumour-specific protein1851..193030/81 (37%)9e-10
SCC2-29 - Homo sapiens, 4019 aa.3537..361645/81 (55%)
[WO200172295-A2, 04 Oct. 2001]
|
[0487] In a BLAST search of public sequence datbases, the NOV33a protein was found to have homology to the proteins shown in the BLASTP data in Table 33D.
174TABLE 33D
|
|
Public BLASTP Results for NOV33a
Nov33a
ProteinResiduesIdentities/
AccessionMatchSimilarities for theExpect
NumberProtein/Organism/LengthResiduesMatched PortionValue
|
Q9EPQ8STROMELYSIN-1 PDGF-1..19331781/1965 (90%)0.0
RESPONSIVE ELEMENT1..19601828/1965 (92%)
BINDING PROTEIN - Mus
musculus (Mouse), 1965 aa.
O14528KIAA0292 (DJ257I20.4) - Homo245..1963 1692/1719 (98%)0.0
sapiens (Human), 1716 aa1..16981694/1719 (98%)
(fragment).
Q9UGU0DJ597B2.1 (TRANSCRIPTION245..1885 1641/1641 (100%)0.0
FACTOR 20 (AR1, KIAA0292)) -1..1641 1641/1641 (100%)
Homo sapiens (Human), 1641 aa
(fragment).
Q13078AR1 - Homo sapiens (Human), 9351083..201 7 935/935 (100%)0.0
aa (fragment).1..935 935/935 (100%)
Q60792STROMELYSIN PDGF988..1902 839/915 (91%)0.0
RESPONSIVE ELEMENT1..913 863/915 (93%)
BINDING PROTEIN
TRANSCRIPTION FACTOR -
Mus musculus (Mouse),937 aa.
|
[0488] PFam analysis predicts that the NOV33a protein contains the domains shown in the Table 33E.
175TABLE 33E
|
|
Domain Analysis of NOV33a
Identities/
NOV33aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Flu_PB1:920..935 7/16 (44%)3.4
domain 1 of 113/16 (81%)
Integrase_Zn:1886..18944/9 (44%)6.9
domain 1 of 1 9/9 (100%)
PHD: domain 1 of 11886..193212/53 (23%)0.81
24/53 (45%)
|
Example 34
[0489] The NOV34 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 34A.
176TABLE 34A
|
|
NOV34 Sequence Analysis
|
|
SEQ ID NO: 87549 bp
NOV34a,GGGCCGGCGGCAGTCTGGGAGCGGCGCGCCATGTACACCATCATCAACCGGCCCAGCA
CG90770-01 DNA SEQUENCE
AGTTGGTCGCGCAGCGCCGCACAGGTCTCACGCAGCAGCAGGTGAAGGGCCAGCTCCA
|
GGAGCTCCTGAAAAGCCGGCAGCCCGCCCCGCCGACCTTCCAGCCCCAGCGGGCGCAG
|
CCCTTCGCGCAGCCGCTCGGACCCTGGCCCCTGTCGAGTGCAGGGCCAAGGCTTGTGT
|
TCAATCGTGTGAATCGCCCGCGGGACCCCTCCAAGTCCCCATCCCTCCAGGGGACCCA
|
GGAGACCTACACACTGCCCCACAAGGAGAATGTCCCCTTTGTGTCCGAAGCCTGGCAC
|
CAGGTGCCGCAGCAGCTGGATGGTGGCCCAGCCCGTGAGGGCGGGACAAGGCCTCTGC
|
AGTGGGTGGAGAGQATCCCCAATCCCCGGCTGCAGAACTTCGTGCCCATTGACTTGGA
|
CGAGTCGTGGGCGCACCACTTCCTGGCTAGAATCACCAGCTGTTCCTAGCGGCTGCTG
|
GGAGGGAGCGCTGCTATGGTCTACCTA
|
ORF Start: ATG at 31ORF Stop: TAG at 511
SEQ ID NO: 88160 aaMW at 18139.5kD
NOV34a,MYTIINGPSKLVAQRRTGLTQQQVKGQLQELLKSRQPAPPTLQPQRAQPFAQPLGWP
CG90770-01 Protein Sequence
LSSAGPRLVFNRVNRRDPSKSPSLQGTQETYTLAHKENVRFVSEAWQQVRQQLDGGP
|
AGEGGTRPVQWVERIPNPRLQNFVPIDLDEWWAQHFLARITSCS
|
[0490] Further analysis of the NOV34a protein yielded the following properties shown in Table 34B.
177TABLE 34B
|
|
Protein Sequence Properties NOV34a
|
|
PSort0.8127 probability located in lysosome (lumen);
analysis:0.4687 probability located in microbody (peroxisome);
0.3600 probability located in mitochondrial matrix
space; 0.0000 probability located in endoplasmic
reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0491] A search of the NOV34a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 34C.
178TABLE 34C
|
|
Geneseq Results for NOV34a
NOV34aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAB43129Human ORFX ORF2893 polypeptide 1 . . . 160123/162 (75%)4e−63
sequence SEQ ID NO: 5786-Homo 1 . . . 159127/162 (77%)
sapiens, 159 aa. [WO200058473-A2,
05-OCT-2000]
AAB59006Breast and ovarian cancer associated40 . . . 160 97/129 (75%)3e−51
antigen protein sequence SEQ ID 714-72 . . . 200103/129 (79%)
Homo sapiens, 200 aa.
[WO200055173-A1, 21-SEP-2000]
ABG06662Novel human diagnostic protein #6653-85 . . . 14656/62 (90%)9e−28
Homo sapiens, 63 aa. 1 . . . 6260/62 (96%)
[WO200175067-A2, 11-OCT-2001]
ABG06662 Novel human diagnostic protein #6653-85 . . . 14656/62 (90%)9e−28
-Homo sapiens, 63 aa. 1 . . . 6260/62 (96%)
[WO200175067-A2, 11-OCT-2001]
AAM82721Human immune/haematopoietic22 . . . 9953/80 (66%)3e−21
antigen SEQ ID NO: 10314-Homo13 . . . 8759/80 (73%)
sapiens, 101 aa. [WO200157182-A2,
09-AUG-2001]
|
[0492] In a BLAST search of public sequence datbases, the NOV34a protein was found to have homology to the proteins shown in the BLASTP data in Table 34D.
179TABLE 34D
|
|
Public BLASTP Results for NOV34a
NOV34aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96KV8C349E10.1.1 (NOVEL PROTEIN, 1 . . . 160137/160 (85%)1e−76
ISOFORM 1)-Homo sapiens 1 . . . 160144/160 (89%)
(Human), 160 aa.
Q9BUT9HYPOTHETICAL 22.4 KDA 1 . . . 160137/160 (85%)1e−76
PROTEIN-Homo sapiens (Human),48 . . . 207144/160 (89%)
207 aa (fragment).
Q9CQB29530058B02RIK PROTEIN-Mus 1 . . . 160121/160 (75%)1e−66
musculus (Mouse), 160 aa. 1 . . . 160134/160 (83%)
Q96J99HTB-Homo sapiens (Human),83 . . . 148 58/66 (87%)2e−30
113 aa. 1 . . . 66 64/66 (96%)
Q969E9C349E10.1.2 (NOVEL PROTEIN, 1 . . . 61 51/61 (83%)4e−22
ISOFORM 2) (SIMILAR TO RIKEN 1 . . . 61 53/61 (86%)
CDNA 9530058B02 GENE)-Homo
sapiens (Human), 75 aa.
|
[0493] PFam analysis predicts that the NOV34a protein contains the domains shown in the Table 34E.
180TABLE 34E
|
|
Domain Analysis of NOV34a
Identities/
NOV34aSimilaritiesExpect
Pfam DomainMatch Regionfor the Matched RegionValue
|
No Significant Matches Found
|
Example 35
[0494] The NOV35 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 35A.
181TABLE 35A
|
|
NOV35 Sequence Analysis
|
|
SEQ ID NO: 891255 bp
NOV35a,GCGAGTGTTTCCTGCAATATTGGATTGGCAATGACAGGCTGGAGCTGCCTTGTGACAG
CG91002-01 DNA Sequence
GAGCAGGAGGGTTTCTGGGTCAGAGGATCATCCGCCTCTTGGTGGAGGAGAAGGAGCT
|
GAAGGAGATCAGGGCCTTGGACAAGGCCTTCAGACCAAAGCTGAGCGAGGAATTTTCT
|
GAGCTCCAGAACAAGACCAAGCTGACAGTGCTGGAAGGAGACATTCTGGATGAGCCAT
|
TCCTCAAGAGAGCCTGCCACGACATGTCGGTCATCATCCACACTGCCTCTATCATATA
|
TGTCATCGGTGTCACTCACAGAGACTCCATCATGAATGTCAATGTGAAAGGTACACAG
|
CTTCTGTTGGACCCCTGTGTCCAAGCTACAGTGCCAGTCTTCATCTACACCAGTACCC
|
CAGAGGTACCCGGQCCCAATTCCTACAAGGAAATCATCCAGAACAGTCACGAAGAACA
|
GCCTCTGCAAAACACATGGTACCCTCCATACCCACACAGCAAAAAGCTTGCTGAGAAG
|
GCTGTGCTCGCGGCTAATGGGTGGACTCTGAAAAACGGTGGCACCTTGTACACTTGTG
|
CCTTAAGACCAATGTTTATCTATGGGGAAGGAACCCCAATCCTTTCTGCCGGTATAAA
|
TGAGGCCCTAAACAACCATCGGATCCTGTCAAGTTTCAGCAAGTTCTCCAGAGTCAAC
|
CCACTCTATATTGGAAACATGGCCTCGGCCCACATTCTGGCCTTGAGGCCCTGCGGGG
|
AGCCCAAGAAGGCCCCAAGTGTCCGAGGACAGTTCTACTACATCTCAGATGACATGCC
|
TCACCAAAGCTATGATAACGTTAATTACATCCTGAGCAAAGAGTTCGGCCTCTGCCTT
|
GATTCTAGATCGACCCTTCCTTTATCTCTGATGTACTGGATTGGCTTCCTGCTGGAAA
|
TAGTGAGCTTCCTGCTGAGCCCAATTTACACCTATCGACACCCCTTCAACCACCACAG
|
AGTGACATTGTCAAATAGCGTGTTCACCTTCTCTTACAAGAAGGCTCAGCAAGATCTG
|
GCATATAAGCCACTTTACAGCTGGGAGGAAGCCAAGAAGAATGTACCTGAGTCTGTTA
|
GAACCGTGGAGTGGGTTTCCCTTGTGGACTGGCACAAGGAGACCGTGAAGTCCAAGAC
|
TCAGCGATTTAAGGATGACAGAGATGTGCATGTGGGTATTGTTAGGTGATGTCATCAA
|
GCTCCATCCTCCTGGCTTCATACAGAAGGTGACAATG
|
ORF Start: ATG at 31ORF Stop: TGA at 1207
SEQ ID NO: 90392 aaMW at 44837.2kD
NOV35A, MTGWSCLVTGAGGFLGQRIIRLLVEEKELKEIRALDKAFRPKLREEFSELQNKTKLTV
CG91002-02 DNA Sequence
LEGDTLDEPPLKRACQDMSVIINTASIIYVIGVTHRESIMNVNVKGTQLLLEACVQAT
|
VPVFIYTSTPEVAGPNSYKEIIQNSHEEEPLENTWYAPYPHSKKLAEKAVLAANGWTL
|
HILALRALREPKKAPSVRGQFYYISDDMPHQSYDNLNYILSKEFGLCLDSRWSLPLSL
|
MYWIGFLLEIVSFLLRPIYTYRHPFNHHRVTLSNSVFTFSYKKAQQDLAYKPLYSWEE
|
AKKNVPESVRTVEWVSLVDWHKETVKSKTQRFKDDRDVHVGIVR
|
SEQ ID NO: 911125 bp
NOV35b, GAGTGTGTTGTCTGCTACTTTGGATTGGCCATGACAGGCTGGAGCTGCCTTGTGACAG
CG91002-02 DNA Sequence
GAGCAGGAGGGTTTCTGGGTCAGAGTATCATCCGCCTCTTGGTGGAGGAGAAGGAGCT
|
GAAGGAGATCAGGGCCTTGGACAAGGCCTTCAGACCGGAATTGAGGAAGGAATTTTCT
|
GAGCTCCAGAACAAGACCAAGCTGACGGTGCTGGAAGGAGACATTCTGGATGAGTCAT
|
GCCTGAAGAGAGCCTGCCAGGACATGTCGCTCATCATCCACACCACCTCCATCATAGA
|
CATCATCGGTGTCACTCACAGAGAGTCCATCATGAACATCAATGTGAAACGTACCCAG
|
CTTCTGTTGGAQGCCTGTGTCCAACCTACAGTGCCAATCTTCATCTACACCAGTACCC
|
TAGAGGTAGCCAGGCCCATTCCTACAAGGAAATCATTCAGAACGGTCATGAAGGAAGA
|
GCCTCTGGAAAACACATGGTACGCTCCATACCCACACAGCAAAAAGCTTGCTGAGAAG
|
GCTGTGCTGGCGGCTAATGCGCTGACTCTGAAAAACGGTCGCACCTTGTACACTTCTG
|
CCGTAAGACCAATGTTTATCTATGGGGAAGGAAGCCCAATCCTTTCTGCOGGTATAAA
|
TGAGCCCCTAAACAACCATCGAATCCTGTCAACTTTCAGCAAGTTCTCCAGAGTCAAC
|
CCACTCTATGTTGGCAACATAGCCTGGGCCCACATTCTGGCCTTGAGGGCCCTGCGGG
|
AGCCCAAGAAGGCCCCAAGTGTCCGAGGACAGTTCTACTATATCTCAGATGACACGCC
|
TCACCAAAGCTATGATAACCTTAGTTACACCTTGAGCAAAGAGTTCGGCCTCTGCCTT
|
GATTCCAGTTGGAGCCTGCCTTTATCCCTGACGTACTGGATTGGCTTCCTGCTGGAAA
|
TAGTGAGCTTCCTGCCGAGGCCAGTTTACACCTGTCGACCGCCCTTCAACCACCACAG
|
AGTGACATTGTCAAATAGCGTGTTCACCTTCTCTTACAAGAAGGCTCAGCAAGATCTG
|
GCATATAAGTCACTTTACAGCTGCGAGGAACCCAAGCACAAAACCATCGAGTCCGTTG
|
GTTCCCTTGTGGACTGGCACAAGGAGACCCTGAAGTCCAAGACTCAGTGATCGAAGGA
|
TGACAGACATGTGCATCTGGGTATTCTTACGAGATGTCGTCAACCTCCATCCTCCTGC
|
CTTCATACAGGTGACAAGGGCAAAAGTCCAGGT
|
ORF Start: ATG at 31ORF Stop: TGA at 1150
SEQ ID NO: 92373 aaMW at 42150.9kD
NOV35b, MTGWSCLVTGAGGFLGQSIIRLLVEEKELKEIRALDKAFRPELRKEFSELQNKTKLTV
CG91002-02 Protein Sequence
LEGDILDESCLKRACQDMSVIIHTTSIIDIIGVTHRESIMNINVKGTQLLLEACVQAT
|
VPIFIYTSTLEVARPNSYKEIIQNGHEEEPLENTWYAPYPHSKKLAEKAVLAANGLTL
|
KNGGTLYTCAVRPMFIYGEGSPILSAGINEALNNHGILSSFSKFSRVNPVYVGNIAWA
|
HILALRALREPKKAPSVRGQFYYISDDTPHQSYDNLSYTLSKEFGLCLDSSWSLPLSL
|
TYWIGFLLEIVSFLPRPVYTCRPPFNHHRVTLSNSVFTFSYKKAQQDLAYKSLYSWEE
|
AKQKTMEWVGSLVDWHKETLKSITQ
|
[0495] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 35B.
182TABLE 35B
|
|
Comparison of NOV35a against NOV35b.
Identities/
NOV35a Residues/Similarities for
Protein SequenceMatch Residuesthe Matched Region
|
NOV35b1 . . . 378333/379 (87%)
1 . . . 373346/379 (90%)
|
[0496] Further analysis of the NOV35a protein yielded the following properties shown in Table 35C.
183TABLE 35C
|
|
Protein Sequence Properties NOV35a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body;
0.3000 probability located in endoplasmic reticulum
(membrane); 0.1000 probability located in mitochondrial
inner membrane
SignalPCleavage site between residues 17 and 18
analysis:
|
[0497] A search of the NOV35a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 35D.
184TABLE 35D
|
|
Geneseq Results for NOV35a
NOV35aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAE15156Human HSD3B1 protein-Homo 1 . . . 378332/379 (87%)0.0
sapiens, 373 aa. [WO200179552-A1, 1 . . . 373349/379 (91%)
25-OCT-2001]
AAU10556Human HSD3B2 polypeptide- 3 . . . 378324/377 (85%)0.0
Homo sapiens, 372 aa. 2 . . . 372346/377 (90%)
[WO200177126-A2, 18-OCT-2001]
ABG02527Novel human diagnostic protein116 . . . 378226/264 (85%)e−129
#2518-Homo sapiens, 259 aa. 2 . . . 259239/264 (89%)
[WO200175067-A2, 11-OCT-2001]
ABG02527Novel human diagnostic protein116 . . . 378226/264 (85%)e−129
#2518-Homo sapiens, 259 aa. 2 . . . 259239/264 (89%)
[WO200175067-A2, 11-OCT-2001]
AAU30804Novel human secreted protein #1295- 3 . . . 311234/313 (74%)e−126
Homo sapiens, 373 aa. 36 . . . 326251/313 (79%)
[WO200179449-A2, 25-OCT-2001]
|
[0498] In a BLAST search of public sequence datbases, the NOV35a protein was found to have homology to the proteins shown in the BLASTP data in Table 35E.
185TABLE 35E
|
|
Public BLASTP Results for NOV35a
NOV35aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
A543253beta-hydroxy-Delta5-Steroid1 . . . 378331/379 (87%)0.0
dehydrogenase (EC 1.1.1.145)/delta5-1 . . . 373353/379 (92%)
delta4 isomerase (EC 5.3.3.-)-rhesus
macaque, 373 aa.
P273653 beta-hydroxysteroid2 . . . 378330/378 (87%)0.0
dehydrogenase/delta 5-->4-isomerase1 . . . 372352/378 (92%)
(3Beta-HSD) [Includes: 3-beta-hydroxy-
delta(5)-steroid dehydrogenase (EC
1.1.1.145) (3-beta-hydroxy-5-ene steroid
dehydrogenase) (Progesterone reductase)
Steroid delta-isomerase (EC 5.3.3.1)
(Delta-5-3-ketosteroid isomerase)]-
Macaca mulatta (Rhesus macaque), 372
aa.
CAC27312SEQUENCE 39 FROM PATENT1 . . . 378332/379 (87%)0.0
WO0102600-Homo sapiens (Human),1 . . . 373349/379 (91%)
373 aa.
P140603 beta-hydroxysteroid2 . . . 378331/378 (87%)0.0
dehydrogenase/delta 5-->4-isomerase1 . . . 372348/378 (91%)
type I (3Beta-HSD I) (Trophoblast
antigen FDO161G) [Includes: 3-beta-
hydroxy-delta(5)-steroid dehydrogenase
(EC 1.1.1.145) (3-beta-hydroxy-5-ene
steroid dehydrogenase) (Progesterone
reductase); Steroid delta-isomerase (EC
5.3.3.1) (Delta-5-3-ketosteroid
isomerase)]-Homo sapiens (Human),
372 aa.
DEHUH23beta-hydroxy-Delta5-steroid3 . . . 378324/377 (85%)0.0
dehydrogenase multifunctional protein II-2 . . . 372346/377 (90%)
human, 372 aa.
|
[0499] PFam analysis predicts that the NOV35a protein contains the domains shown in the Table 35F.
186TABLE 35F
|
|
Domain Analysis of NOV35a
Identities/
Similarities
NOV35a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Epimerase: domain6 . . . 351 74/432 (17%) 2e−05
1 of 1214/432 (50%)
3Beta_HSD: domain1 . . . 361184/425 (43%)3.6e−193
1 of 1330/425 (78%)
|
Example 36
[0500] The NOV36 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 36A.
187TABLE 36A
|
|
NOV36 Sequence Analysis
|
|
SEQ ID NO: 931540 bp
NOV36a,AGGTCACCATGGCTGTTATTGGCTCCCTTGCTGTCCCCAGCTGTTCCCCACCTCACCT
CG91298-01 DNA Sequence
CCCCCCCTTGCTGCCTGTGTCCAGAAGGCGTGCACCGGTTCCAGTGCATCAGAAACCT
|
GGTTCCAGAATTTGGAGTCTCCAGTTCTCACGTTAGGGTGCTTTCTTCCCCGGCAGAC
|
TTTTTCGAGCTCATGAAGCGGCAGATAAGAGTACCCAAGAGGCGCGTCCTGATGGCAT
|
CCCTCTACCTGGGGACAGGTCCTTTGGAACACGAGCTGGTAGACTGCCTGGAAAGTAC
|
TCTAGAAAAGTCACTCCAAGCAAACTTTCCTTCAAATCTCAACOTCTCCATTCTCTTA
|
GACTTCACGCGGGGCTCACGAGGTAGGAAGAACTCCCGCACAATGCTGCTCCCACTCC
|
TGCGGAGGTTCCCAGAGCAGGTCCGAGTCTCCCTCTTTCACACGCCGCACCTCCGTGG
|
GCTGCTTCGGCTCCTCATCCCTGACCCCTTCAACGAGACCATCCGCCTCCAGCACATT
|
AAGGTGTACCTCTTCGACAACACCGTCATCTTGAGCGGGGCAAACTTGAGTGACTCCT
|
ACTTCACCAACCCCCACGACCGCTACGTGTTCCTGCACCACTGTGCCGAGATTCCCGA
|
CTTCTTCACGGAGCPGGTGGACGCGGTGGGGGATGTCTCCCTGCAGCTGCAGGGGGAC
|
GACACGGTGCAGGTGGTGGATGGGATGGTGCATCCTTACAAAGGTGACCGGGCCGAGT
|
ACTGCAAGGCAGCCAATAAGAGGGTCATGGATGTGATCAACTCAGCCAGGACCCGCCA
|
CCAGATGCTGCATGCCCAGACCTTCCACAGCAACTCTCTTTTGACCCAGGAAGATGCA
|
GCAGATGCTGCATGCCCAGACCTTCCACAGCAACTCTCTTTTGACCCAGGAAGATGCA
|
GCAGCTGCTGGGGATCGCAGACCAGCCCCTGACACCTGGATTTATCCGCTGATTCAGA
|
TGAAGCCCTTCGAGATTCAAATCGATGAGATTGTCACTGACACCCTGTTGACTGAGGC
|
GGAGCGCGGGGCAAAGGTCTACCTCACCACTGGCTATTTCAACCTGACCCAGGCCTAC
|
ATGGACCTGGTCTTGGGCACTCGGGCTGAGTACCAGATCCTGCTGGCCTCACCAGACG
|
TGAATGGCTTCTTTGGGGCCAAGGGGGTGGCCGGCGCCATCCCAGCGGCCTATGTGCA
|
CATCGAGCGACAGTTCTTCAGTGAGGTGTGCAGCCTGGGACAGCAGGAGCGGGTCCAG
|
CTTCAGGAGTACTGGCGGAGGGCCTGGACGTTCCACOCCAAAGGTCTCTGGCTGTACC
|
TGGCAGGGAGCAGCCTGCCCTGTCTCACGCTGATTGGCTCTCCTAATTTTGGGTACAG
|
GTCAGTTCACCGGGACCTGGAGGCCCAGATTGCGATCGTGACGGAGAACCAGGCCCTG
|
CAGCAGCAGCTTCACCAGGAGCAAGAGCAGCTCTACCTGAGGTCACGTGTGGTGTCCT
|
CTGCCACCTTCCAGCAGCCGAGTCGCCAGGTGAAGCTGTGGGTGAAGATGGTCACTCC
|
ACTGATCAAGAACTTCTTCTGAGGACAGACAG
|
ORF Start: at 1ORF Stop: TGA at 1528
SEQ ID NO: 94509 aaMW at 57653.7kD
NOV36a, RSPNLLLAPLLSPAVPQVTSPPCCLCPEGVHRFQWIRNLVPEFGVSSSHVRVLSSPAE
CG91298-01 Protein Sequence
FFELMKGQIRVAKRRVVMASLYLGTGPLEQELVDCLESTLEKSLQAKFPSNLKVSILL
|
DFTRGSRGRKNSRTMLLPLLRRFPEQVRVSLFHTPHLRGLLRLLIPERFNETIGLQHI
|
KVYLFDNSVILSGANLSDSYFTNRQDRYVFLQDCAEIADFFTELVDAVCDVSLQLQGD
|
DTVQVVDGMVHPYKGDRAEYCKAANKRVMDVINSARTRQQMLHAQTFHSNSLLTQEDA
|
AAAGDRRPAPDTWIYPLIQMKPPEIQIDEIVTETLLTEAERGAKVYLTTGYFNLTQAY
|
MDLVLGTRAEYQILLASPEVNGFFGAKGVAQAIPAAYVWTERQFFSEVCSLCQQERVQ
|
LQEYWRRGWTFHAKGLWLYLAGSSLPCLTLIGSPNFGYRSVHRDLEAQIAIVTENQAL
|
QQQLHQEQEQLYLRSGVVSSATFEQPSRQVKLWVKMVTPLIKNFF
|
[0501] Further analysis of the NOV36a protein yielded the following properties shown in Table 36B.
188TABLE 36B
|
|
Protein Sequence Properties NOV36a
|
|
PSort0.5500 probability located in lysosome (lumen);
analysis:0.3700 probability located in outside; 0.1000
probability located in endoplasmic reticulum (membrane);
0.1000 probability located in endoplasmic reticulum (lumen)
SignalPCleavage site between residues 21 and 22
analysis:
|
[0502] A search of the NOV36a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 36C.
189TABLE 36C
|
|
Geneseq Results for NOV36a
NOV36aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAU08803Human phosphatidyl glycerol 1 . . . 509509/509 (100%)0.0
phosphate (PGP) synthase-Homo 48 . . . 556509/509 (100%)
sapiens, 556 aa. [WO200164895-A2,
07-SEP-2001]
AAG83315P patens lipid metabolism related325 . . . 439 49/128 (38%)1e−16
protein #33-Physcomitrella patens, 9 . . . 136 73/128 (56%)
137 aa. [WO200138541-A1, 31-
MAY-2001]
AAG80868Lipid biosynthesis protein sequence325 . . . 439 49/128 (38%)1e−16
#26-Physcomitrella patens, 137 aa. 9 . . . 136 73/128 (56%)
[WO200138484-A2, 31-MAY-2001]
AAW55550H. pylori ORE 65 . . . 200 35/139 (25%)0.69
01ce21104_33203250_c3_87 secreted 71 . . . 195 65/139 (46%)
protein-Helicobacter pylori, 502 aa.
[WO9737044-A1, 09-OCT-1997]
AAW55452H. pylori ORF 65 . . . 200 35/139 (25%)0.69
02ae11612_33203250_cl_51 secreted 71 . . . 195 65/139 (46%)
protein-Helicobacter pylori, 502 aa.
[WO9737044-A1, 09-OCT-1997]
|
[0503] In a BLAST search of public sequence datbases, the NOV36a protein was found to have homology to the proteins shown in the BLASTP data in Table 36D.
190TABLE 36D
|
|
Public BLASTP Results for NOV36a
NOV36aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
CAD10244SEQUENCE 1 FROM PATENT 1 . . . 509509/509 (100%)0.0
WO0164895-Homo sapiens (Human), 556 48 . . . 556509/509 (100%)
aa.
AAL87040SILENCER-ASSOCIATED FACTOR- 3 . . . 509482/507 (95%)0.0
Mus musculus (Mouse), 513 aa. 7 . . . 513503/507 (99%)
Q9Z2Z7PHOSPHATIDYLGLYCEROPHOSPHATE 1 . . . 509478/509 (93%)0.0
SYNTHASE-Cricetulus griseus (Chinese 45 . . . 553500/509 (97%)
hamster), 553 aa.
Q96A75UNKNOWN (PROTEIN FOR 1 . . . 280280/280 (100%)e−160
IMAGE: 3623672) (HYPOTHETICAL 37.3 49 . . . 328280/280 (100%)
KDA PROTEIN)-Homo sapiens (Human),
331 aa (fragment).
Q9NPW7HYPOTHETICAL 30.8 KDA PROTEIN-239 . . . 509271/271 (100%)e−156
Homo sapiens (Human), 271 aa (fragment). 1 . . . 271271/271 (100%)
|
[0504] PFam analysis predicts that the NOV36a protein contains the domains shown in the Table 36E.
191TABLE 36E
|
|
Domain Analysis of NOV36a
Identities/
NOV36aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
ART: domain 1 of 1 1 . . . 16 9/16 (56%)7.7
14/16 (88%)
PLDc: domain 1 of 2168 . . . 194 8/34 (24%)0.036
21/34 (62%)
PLDc: domain 2 of 2413 . . . 446 7/35 (20%)50
24/35 (69%)
|
Example 37
[0505] The NOV37 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 37A.
192TABLE 37A
|
|
NOV37 Sequence Analysis
|
|
SEQ ID NO: 951458 bp
NOV37a,GCCCGGCCATGGAGCGCCAGGTCCTACGGCTTCGCCAGGCGTTCCGGTCCGGCCGATC
CG91383-01 DNA Sequence
TCGGCCGCTGCGCTTCCGATTGCAGCAGCTTGAGGCCCTGCGGAGGATGGTGCAAGAG
|
CGCGAGAAGGAAATCTTAGCAGCCATCGCGGCAGACCTGAGCAAAAGTGAACTCAATG
|
CATACAGTCATGAAGTCATTACCATCCTTGGAGAGATTGATTTTATGTTGGGGAATCT
|
TCCTGAATTGGCTTCTGCGAGACCGGCTAAGAAGAACCTGCTAACCATGATGGATGAG
|
GCCTACGTTCAGCCCGAGCCTCTGGGAGTCGTACTGATTATTGGAGCTTGGAATTACC
|
CTTTCGTTCTTACCATGCAACCGCTGGTGGGAGCCATTGCTGCAGGTAATGCTGCCAT
|
TGTTAAGCCCTCAGAACTCAGTGAAAACACGGCCAAGATCTTGGCTGAACTCCTCCCT
|
CAGTACTTAGACAAGGACCTGTATGCGATTGTTAATGGCGGTATCCCGGAAACCACGG
|
AGCTTCTGAAGCAGCGGTTTGACCACATTCTCTATACAGGGAACACTGCAGTTGGAAA
|
AATTGTCATGGAAGCTGCTGCCAAGCATCTGACCCCTGTGACCCTGGAACTCGGCGGG
|
AAAAGCCCTTGTTACATTGACAGAGACTGTGATCTGGACGTGGCTTGCAGGCGCGTAG
|
CCTGGGGAAAGTACATGAATTGTGGTCAAACCTGCATTGCTCCTGACTATATCCTGTG
|
CGAAGCCTCCCTCCAGAATCAAATCGTACAGAAGATTAAGGAAACGGTGAAGGACTTT
|
TATGGGGAAAACATAAAGGCTTCTCCTGACTATGAAAGGATCATCAATCTTCGTCACT
|
TTAAGAGGTTACAAAGTCTGCTTAAAGGCCAGAAAATAGCTTTCGGTGGAGAGATGGA
|
TGAGGCCACACGCTACTTAGCCCCAACCATACTTACAGATGTTGATCCTAACTCCAAG
|
ACGAAGCCATAAATTTCATAAATGACCGTGAAAAGCCCCTGGCTCTCTACGTATTTTC
|
ACGAAGCCATAAATTTCATAAATGACCGTGAAAACCCCCTGGCTCTCTACCTATTTTC
|
TCGTAACAATAAGGTAATCAAACGGGTGATAGATGAGACCTCCAGTGGTGGAGTCACC
|
GGCAATGATGTCATCATGCACTTCACTGTTAATTCTCTGCCCTTTGGAGGTGTGGGTG
|
CCAGTGGAATGGGGGCGTATCATGGAAAATACAGTTTTGATACCTTTTCTCATCAGCG
|
CCCCTGCTTGTTAAAAGGGTTAAAGGGGGAAAGCGTCAACAAGCTCAGGTACCCACCC
|
TTTATTCTTAAATAGATATTTTTAGAGAAGACTTAGCTTCACATGCAAACACACACAC
|
ACACACAC
|
ORF STart: ATG AT 9ORF Stop: TAA at 1401
SEQ ID NO: 96464 aaMW at 51606.2kD
NOV37,aMERQVLRLRQAFRSGRSRPLRFRLQQLEALRRMVQEREKEILAAIAADLSKSELNAYS
CG91383-01 Protein Sequence
HEVITILGEIDFMLGNLPELASARPAKKNLLTMMDEAYVQPEPLGVVLIIGAWNYPFV
|
LTMQPLVGAIAAGNAAIVKPSELSENTAKILAELLPQYLDKDLYAIVNGGIPETTELL
|
KQRFDHILYTGNTAVGKIVMEAAAKHLTPVTLELGGKSPCYIDRDCDLDVACRRVAWG
|
KYMNCGQTCIAPDYILCEASLQNQIVQKIKETVKDFYGENIKASPDYERIINLRHFKR
|
LQSLLKGQKIAFGGEMDEATRYLAPTILTDVDPNSKVMQEEIFGPILPIVSVKNVDEA
|
INFINDREKPLALYVFSRNNKVIKRVIDETSSGGVTGNDVIMHFTVNSLPFGGVGASG
|
MGAYHGKYSFDTFSHQRPCLLKGLKGESVNKLRYPPTPTPLPTHSPFLALAFPCCLYS
|
[0506] Further analysis of the NOV37a protein yielded the following properties shown in Table 37B.
193TABLE 37B
|
|
Protein Sequence Properties NOV37a
|
|
PSort0.7694 probability located in mitochondrial matrix space;
analysis:0.4477 probability located in mitochondrial inner membrane;
0.4477 probability located in mitochondrial intermembrane
space; 0.4477 probability located in mitochondrial
outer membrane
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0507] A search of the NOV37a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 37C.
194TABLE 37C
|
|
Geneseq Results for NOV37a
NOV37aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAB58156Lung cancer associated polypeptide45 . . . 428211/384 (54%)e−124
sequence SEQ ID 494-Homo sapiens,28 . . . 411276/384 (70%)
430 aa. [WO200055180-A2, 21-SEP-
2000]
AAG82730S. epidermidis open reading frame12 . . . 442195/433 (45%)e−111
protein sequence SEQ ID NO: 2554-15 . . . 445286/433 (66%)
Staphylococcus epidermidis, 459 aa.
[WO200134809-A2, 17-MAY-2001]
AAG82076S. epidermidis open reading frame12 . . . 442195/433 (45%)e−111
protein sequence SEQ ID NO: 1246-15 . . . 445286/433 (66%)
Staphylococcus epidermidis, 459 aa.
[WO200134809-A2, 17-MAY-2001]
AAG21988Arabidopsis thaliana protein fragment 5 . . . 442203/443 (45%)e−110
SEQ ID NO: 24747-Arabidopsis16 . . . 456286/443 (63%)
thaliana, 484 aa. [EP1033405-A2, 06-
SEP-2000]
AAG11789Arabidopsis thaliana protein fragment 5 . . . 442203/443 (45%)e−110
SEQ ID NO: 10644-Arabidopsis16 . . . 456286/443 (63%)
thaliana, 484 aa. [EP1033405-A2, 06-
SEP-2000]
|
[0508] In a BLAST search of public sequence datbases, the NOV37a protein was found to have homology to the proteins shown in the BLASTP data in Table 37D.
195TABLE 37D
|
|
Public BLASTP Results for NOV37a
NOV37aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q99L64ALCOHOL DEHYDROGENASE1 . . . 456441/456 (96%)0.0
FAMILY 3, SUBFAMILY A2-Mus1 . . . 456446/456 (97%)
musculus (Mouse), 484 aa.
Q99PH3FATTY ALDEHYDE1 . . . 456441/456 (96%)0.0
DEHYDROGENASE-Mus musculus1 . . . 456445/456 (96%)
(Mouse), 484 aa.
Q99PH4FATTY ALDEHYDE1 . . . 456441/456 (96%)0.0
DEHYDROGENASE VARIANT1 . . . 456445/456 (96%)
FORM-Mus musculus (Mouse), 507
aa.
P47740Fatty aldehyde dehydrogenase (EC1 . . . 456440/456 (96%)0.0
1.2.1.3) (Aldehyde dehydrogenase,1 . . . 456444/456 (96%)
microsomal) (ALDH class 3)-Mus
musculus (Mouse), 484 aa.
P30839Fatty aldehyde dehydrogenase (EC1 . . . 456421/456 (92%)0.0
1.2.1.3) (Aldehyde dehydrogenase,1 . . . 456438/456 (95%)
microsomal) (ALDH class 3)-Rattus
norvegicus (Rat), 484 aa.
|
[0509] PFam analysis predicts that the NOV37a protein contains the domains shown in the Table 37E.
196TABLE 37E
|
|
Domain Analysis of NOV37a
Identities/
NOV37aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
aldedh: domain 1 of 11 . . . 429174/492 (35%)1.3e−192
389/492 (79%)
|
Example 38
[0510] The NOV38 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 38A.
197TABLE 38A
|
|
NOV38 Sequence Analysis
|
|
SEQ ID NO: 975571 bp
NOV38a,CTCAGGCCCCCTCCCCCCGCCGCCCCCGCCCCCGGGGAAGGCAGGCGCCGAGCTGAGC
CG91403-01 DNA Sequence
CGGGGCCGATGCAGCTGAGCCGCGCCGCCGCCGCCGCCGCCGCCGCCCCTGCGGAGCC
|
CCCGGAGCCGCTGTCCCCCGCGCCGGCCCCGGCCCCGGCCCCCCCCGGCCCCCTCCCG
|
CGCAGCGCGGCCGACGGGGCTCCGGCGGGGGGGAAGGGGGGGCCGGGGCGCCGCCGCG
|
CGGAGTCCCCGGGCGCTCCGTTCCCCGGCGCGAGCGGCCCCGGCCCGGGCCCCGGCGC
|
GGGGATGGACGGCCCCGGGGCCAGCGCCGTGGTCGTGCGCGTCGGCATCCCGGACCTG
|
CAGCAGACGAAGTGCCTGCGCCTGGACCCGGCCGCGCCCGTGTGGGCCGCCAAGCAGC
|
GCGTGCTCTGCGCCCTCAACCACAGCCTCCAGGACGCGCTCAACTATGGGCTTTTCCA
|
GCCGCCCTCCCGGGGCCGCGCCGGCAAGTTCCTGGATGAGGAGCGGCTCCTGCACCAG
|
TACCCGCCCAACCTGGACACGCCCCTGCCCTACCTGGAGTTTCGATACAAGCGGCGAG
|
TTTATGCCCAGAACCTCATCGATGATAAGCAGTTTGCAAAGCTTCACACAAAGGCGAA
|
CCTGAAGAAGTTCATGGACTACGTCCAGCTGCATAGCACGGACAAGGTGGCACGCCTG
|
TTGGACAAGGGGCTGGACCCCAACTTCCATGACCCTGACTCAGGAGAGTGCCCCCTGA
|
GCCTCGCAGCCCAGCTGGACAACGCCACGGACCTGCTAAAGGTGCTGAAGAATGGTGG
|
TGCCCACCTGGACTTCCGCACTCGCGATGGGCTCACTGCCGTGCACTGTGCCACACGC
|
CAGCGGAATGCGGCAGCACTGACGACCCTGCTGGACCTGGGGGCTTCACCTGACTACA
|
AGGACAGCCGCGGCTTGACACCCCTCTACCACAGCGCCCTCGCGGGTGGGGATCCCCT
|
CTGCTGTGAGCTGCTTCTCCACGACCACGCTCAGCTGGGGATCACCGACCAGAATGGC
|
TGGCAGGAGATCCACCAGGCCTGCCGCTTTGGGCACGTGCAGCATCTGGAGCACCTGC
|
TGTTCTATGGGGCAGACATGGGGGCCCAGAACGCCTCGGGGAACACAGCCCTGCACAT
|
CTGTGCCCTCTACAACCAGGAGAGCTGTGCTCGTGTCCTGCTCTTCCGTGGAGCTAAC
|
AGGGATGTCCGCAACTACAACAGCCAGACAGCCTTCCAGGTGGCCATCATCGCAGGGA
|
ACTTTGAGCTTGCAGAGGTTATCAAGACCCACAAAGACTCGGATGTTGTACCATTCAG
|
GGAAACCCCCAGCTATGCGAAGCCGCGGCGACTGGCTGGCCCCAGTGGCTTGGCATCC
|
CCTCCGCCTCTGCAGCGCTCAGCCACCGATATCAACCTGAAGGGGGAGGCACAGCCAG
|
CAGCTTCTCCTGGACCCTCGCPGACAAGCCTCCCCCACCAGCTGCTGCTCCAGCGGCT
|
GCAAGAGGAGAAAGATCGTGACCGGGATGCCGACCAGGAGAGCAACATCAGTGGCCCT
|
TTAGCACGCACCGCCCGCCAAAGCAAGATCACCGAGCCTGGCGCGCCCAGGAGCTGTA
|
TTCGAATTCCAGCTCGGTTCCCCGCCCCCCCTGCCCCCCCCGCACCGCCGCCCCGGGG
|
CCCGAAGCGGAAACTTTACAGCGCCGTCCCCGGCCGCAAGTTCATCGCCGTGAAGGCG
|
CACAGCCCCCAGGGTGAAGGCGAGATCCCGCTGCACCGCGGCGAGGCCGTGAAGGTGC
|
TCAGCATTGCCGAGGGCCGTTTCTGCGAGGGAACCGTGAAAGGCCGCACGGGCTGGTT
|
CCCGGCCGACTGCGTGGAGGAAGTGCAGATGAGGCAGCATGACACACGGCCTGAAACG
|
CGGGAGGACCGGACGAAGCGGCTCTTTCGGCACTACACAGTGGGCTCCTACGACAGCC
|
TCACCTCACACAGCGATTATGTCATTGATGACAAAGTGGCTGTCCTGCAGAAACGGGA
|
CCACGAGGGCTTTGGTTTTGTGCTCCGGGGAGCCAAAGCAGAGACCCCCATCGAGGAG
|
TTCACGCCCACGCCAGCCTTCCCGGCGCTGCAGTATCTCGAGTCGGTGGACGTGGAGG
|
GTGTGGCCTGGAGGGCCGGGCTGCGCACGGGAGACTTCCTCATCGAGGTGAACGGGGT
|
GAACGTGGTGAAGGTCGGACACAAGCAGGTGGTGGCTCTGATTCGCCAGGGTGGCAAC
|
CGCCTCGTCATGAAGGTTGTGTCTGTGACAAGGAAGCCAGAAGAGGACGGGGCTCGGC
|
GCAGAGCCCCACCGCCCCCCAAGAGGGCCCCCAGCACCACACTGACCCTGCGCTCCAA
|
GTCCATGACAGCTGAGCTCGAGGAACTTGCTGAGATGGAGCCTCCTTGCTGTGCAGAG
|
AAGCTGGACGAGATGCTGGCAGCCGCCGCAGAGCCAACGCTGCGGCCAGACATCGCAG
|
ACGCAGACTCCAGAGCCGCCACCGTCAAACAGAGGCCCACCAGTCGGAGGATCACACC
|
CGCCGAGATTAGCTCATTGTTTGAACGCCAGGGCCTCCCAGGCCCAGAGAAGCTGCCG
|
GGCTCCTTGCGGAAGGGGATTCCACGGACCAAGTCTGTACGGGACCACGAGAAGCTGG
|
CGTCCCTGCTGGAACGGCGCTTCCCGCGGAGCACCTCGATGCAAGACCCGGTGCGCGA
|
GGGTCGCGGCATCCCGCCCCCGCCGCAGACCGCGCCGCCTCCCCCGCCCGCGCCCTAC
|
TACTTCCACTCGGGGCCGCCCCCCGCCTTCTCGCCGCCCCCCCCGCCCGGCCGCGCCT
|
ACGACACGGTGCGCTCCAGCTTCAAGCCCGGCCTGGAGGCGCGCCTGGGCGCGGGCGC
|
TGCCGGCCTGTACGAGCCGGGCGCGGCCCTCGGCCCGCTGCCGTATCCCGAGCGGCAG
|
AAGCGCGCGCGCTCCATGATCATCCTGCAGGACTCGGCGCCCGAGTCGGGCGACGCCC
|
CTCGACCCCCGCCCGCGGCCACCCCGCCCGAGCGACCCAAGCGCCGGCCGCGGCCGCC
|
CGGCCCCGACAGCCCCTACGCCAACCTCCGCGCCTTCAGCGCCAGCCTCTTCGCTCCG
|
TCCAAGCCGCAGCGCCGCAAGAGCCCCCTGGTGAAGCAGCTGCAGGTGGAGGACGCGC
|
AGGAGCGCGCGGCCCTGGCCGTGGGCAGCCCCGGTCCCGGCGGCGGCAGCTTCGCCCG
|
CGAGCCCTCCCCGACCCACCGCGGTCCGCGCCCGGGTGGCCTCGACTACGGCGCGGGC
|
GATGGCCCGGGGCTCGCGTTCGGCGGCCCGGGCCCGGCCAAGGACCGGCGGCTGGAGG
|
AGCGGCGCCCCTCCACTGTCTTCCTGTCCGTGGCGGCCATCGAGGGCAGCGCCCCCGG
|
CGCGGATCTGCCATCCCTACAGCCCTCCCGCTCCATCCACGAGCGCCTCCTGGGGACC
|
GCCCCCACCGCCGGCCGCGACCTGCTGCTGCCCTCCCCGGTGTCTGCCCTGAAGCCGT
|
TGGTCAGCGGCCCGAGCCTGGGGCCCTCGGGTTCCACCTTCATCCACCCACTCACCGG
|
CAAACCCCTGGACCCCAGCTCACCCCTGGCCCTTGCCCTGGCTGCCCGAGAGCGAGCT
|
CTGGCCTCCCAGGCGCCCTCCCGGTCCCCCACACCCGTGCACAGTCCCGACGCCGACC
|
GCCCCGGACCCCTGTTTGTCGATGTACAGOCCCGGGACCCAGAGCGAGGCTCCCTGGC
|
TTCCCCGGCTTTCTCCCCACGGAGCCCAGCCTGGATTCCTGTGCCTGCTCGCAGGGAG
|
GCAGAGAAGGTCCCCCGGGAGGAGCGGAAGTCACCCGAGGACAAGAAGTCCATGATCC
|
TCACCGTCCTGGACACATCCCTGCAGCGCCCAGCTGGCCTCATCGTTGTGCACGCCAC
|
CAGCAACGGGCAGGAGCCCAGCAGGCTGGGGGGGGCCGAAGAGGAGCGCCCGGGCACC
|
CCGGAGTTGGCCCCGGCCCCCATGCAGTCAGCGGCTGTGGCAGAGCCCCTGCCCAGCC
|
CCCGGGCCCAGCCCCCTGGTGGCACCCCGGCAGACGCCGGGCCAGGCCAGGGCAGCTC
|
AGAGCAAGAGCCACAGCTGCTGTTTGCTGTGAACCTGCCACCTGCCCAGCTGTCGTCC
|
AGCGATGAGGAGACCAGGGAGGAGCTGGCCCGAATTGGGTTGGTGCCACCCCCTGAAG
|
AGTTTGCCAACGGGGTCCTGCTGGCCACCCCACTCGCTGGCCCGGGCCCCTCGCCCAC
|
CACGGTGCCCAGCCCGGCCTCAGGGAAGCCCAGCAGTGAGCCACCCCCTGCCCCTGAG
|
TCTGCAGCCGACTCTGGGGTGGAGGAGGCTGACACACGCAGCTCCAGCGACCCCCACC
|
TGGAGACCACAAGCACCATCTCCACGGTGTCCAGCATGTCCACCTTGAGCTCGGAGAG
|
CGGGGAACTCACTGACACCCACACCTCCTTCGCTGACGGACACACTTTTCTACTCGAG
|
AAGCCACCAGTGCCTCCCAAGCCCAAGCTCAAGTCCCCGCTGGGGAAGGGGCCGGTGA
|
CCTTCAGGGACCCGCTGCTGAAGCAGTCCTCGGACAGCGAGCTCATGGCCCAGCAGCA
|
CCACGCCGCCTCTGCCGGGCTGGCCTCTGCCGCCGGCCCTGCCCGCCCTCCCTACCTC
|
TTCCAGAGAAGGTCCAAGCTATGGGGGGACCCCGTGGAGAGCCGGGGGCTCCCTGGGC
|
CTGAAGACGACAAACCAACTGTGATCAGTGAGCTCAGCTCCCGCCTGCAGCAGCTGAA
|
CAAGGACACGCGTTCCCTGGGGGAGGAACCAGTTGGTGGCCTGGGCAGCCTGCTGGAC
|
CCTCCCAAGAAGTCGCCCATCGCAGCAGCTCGGCTCTTCAGCAGCCTCGGTGAGCTGA
|
GCTCCATTTCAGCGCAGCGCAGCCCCGGGGGCCCGGGCGGCGGGGCCTCGTACTCGGT
|
GACGCCCAGTGGCCGCTACCCCCTGGCGAGACGCGCCCCGAGCCCGGTGAAGCCCGCG
|
TCGCTGGAGCGGGTGGACGGGCTCGGGGCGGGCGCGGGGGGCGCAGGGCGGCCCTTCG
|
GCCTCACGCCCCCCACCATCCTCAAGTCGTCCAGCCTCTCCATCCCGCACGAGCCCAA
|
GGAGGTGCGCTTCGTGGTGCGCAGCGTGAGCGCGCGCAGTCGCTCCCCCTCGCCGTCG
|
CCGCTGCCCTCGCCCGCGTCCGGCCCCGGCCCCGGCGCCCCCGGCCCACGCCGACCCT
|
TCCAGCAGAAGCCGCTGCAGCTCTGGAGCAAGTTCGACGTGGGCGACTGGCTGGAGAG
|
CATCCACCTAGGCGAGCACCGCGACCGCTTCGAGGACCATGAGATAGAAGGCGCGCAC
|
CTACCCGCGCTTACCAAGGACGACTTCGTGGAGCTGGGCGTCACGCGCGTGGGCCACC
|
GCATGAACATCGAGCGCGCGCTCAGGCAGCTGGACGGCAGCTGACGCCCCACCCCCAC
|
TCCCGCCCCGGCCGTGCCCTGCCGGCAGCGCCCCCCACCCCCACCCCGCGCCCCGGGC
|
TCG
|
ORF Start: ATG at 67 ORF Stop: TGA at 5494
SEQ ID NO: 98 1809 aa MW at 192126.1kD
NOV38a, MQLSRAAAAAAAAPAEPPEPLSPAPAPAPAPPGPLPRSAADGAPAGGKGGPGRRRAES
CG91403-01 Protein Sequence
PGAPFPGASGPGPGPGAGMDGPGASAVVVRVGIPDLQQTKCLRLDPAAPVWAAKQRVL
|
CALNHSLQDALNYGLFQPPSRGRAGKFLDEERLLQEYPPNLDTPLPYLEFRYKRRVYA
|
QNLIDDKQFAKLHTKANLKKFMDYVQLHSTDKVARLLDKGLDPNFHDPDSGECPLSLA
|
AQLDNATDLLKVLKNGGAHLDFRTRDGLTAVHCATRQRNAAALTTLLDLGASPDYKDS
|
RGLTPLYHSALGGCDALCCELLLHDEAQLGITDEMGWQEIEQACRFGEVQHLEHLLFY
|
GADMGAQNASGNTALHICALYNQESCARVLLFRGANRDVRNYNSQTAFQVAIIAGNFE
|
LAEVIKTHKDSDVVPFRETPSYAKRRRLAGPSGLASPRPLQRSASDINLKGEAQPAAS
|
PGPSLRSLPHQLLLQRLQEEKDRDRDADQESNISGPLAGRAGQSKISEPGAPRSCIRI
|
RARPPAPPAPPAPPPRGPKRKLYSAVPGRKFIAVKAHSPQGEGETPLHRGEAVKVLSI
|
GEGGFWEGTVKGRTGWFPADCVEEVQMRQHDTRPETREDRTKRLFRHYTVGSYDSLTS
|
HSDYVIDDKVAVLQKRDHEGFGPVLRGAKAETPIEEFTPTPAFPALQYLESVDVEGVA
|
WRAGLRTGDFLIEVNGVNVVKVGHKQVVALIRQGGNRLVMKVVSVTRRPEEDGARRRA
|
PPPPKRAPSTTLTLRSKSMTAELEELAEMEPPCCAEKLDEMLAAAAEPTLRPDIADAD
|
SRAATVKQRPTSRRITPAEISSLFERQGLPGPEKLPGSLRKGIPRTKSVGEDEKLASL
|
LEGRFPRSTSMQDFVREGRGIPPPPQTAPPPPPAPYYFDSGPPPAFSPPPPRGRAYDT
|
VRSSFKPGLEARLGAGAAGLYEPGAALGPLPYPERQKRARSMIILQDSAPESGDAPRP
|
PPAATPPERPKRRPRPPGPDSPYANLGAFSASLFAPSKPQRRKSPLVKQLQVEDAQER
|
AALAVGSPGPGGGSFAREPSPTHRGPRPGGLDYGAGDGPGLAFGGPGPAKDRRLEERR
|
RSTVFLSVGAIEGSAPGADLPSLQPSRSIDERLLGTGPTAGRDLLLPSPVSALKPLVS
|
GPSLGPSGSTFIHPLTGKPLDPSSPLALALAARERALASQAPSRSPTPVHSPDADRPG
|
PLFVDVQARDPERGSLASPAFSPRSPAWIPVPARREAEKVPREERKSPEDKKSMTLSV
|
LDTSLQRPAGLIVVHATSNGQEPSRLGGAEEERPGTPELAPAPMQSAAVAEPLPSPRA
|
QPPGGTPADAGPGQGSSEEEEPELVFAVNLPPAQLSSSDEETREELARIGLVPPEEFA
|
NGVLLATPLAGPGPSPTTVPSPASGKPSSEPPPAPESAADSGVEEADTRSSSDPHLET
|
TSTISTVSSMSTLSSESGELTKTHTSFADGHTFLLEKPPVPPKPKLKSPLGKGPVTFR
|
DPLLKQSSDSELMAQQHHAASAGLASAAGPARPRYLFQRRSKLWGDPVESRGLPGPED
|
DKPTVISELSSRLQQLNKDTRSLGEEPVGGLGSLLDPAKKSPIAAARLFSSLGELSSI
|
SAQRSPGGPGGGASYSVRPSGRYPVARRAPSPVKPASLERVBGLGAGAGGAGRPFGLT
|
PPTILKSSSLSIPHEPKEVRFVVRSVSARSRSPSPSPLPSPASGPGPGAPGPRRPFQQ
|
KPLQLWSKFDVGDWLESIHLGEHRDRFEDHEIEGAHLPALTKDDFVELGVTRVGHRMN
|
IERALRQLDGS
|
[0511] Further analysis of the NOV38a protein yielded the following properties shown in Table 38B.
198TABLE 38B
|
|
Protein Sequence Properties NOV38a
|
|
PSort0.7000 probability located in nucleus;
analysis:0.3000 probability located in microbody
(peroxisome); 0.1681 probability located
in lysosome (lumen); 0.1000 probability
located in mitochondrial matrix space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0512] A search of the NOV38a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 38C.
199TABLE 38C
|
|
Geneseq Results for NOV38a
NOV38aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAY83017Rat shank 3a-Rattus rattus, 1740 77 . . . 18091624/1745 (93%)0.0
aa. [WO200011204-A2, 02-MAR- 1 . . . 17401654/1745 (94%)
2000]
AAB31517Amino acid sequence of the rat 77 . . . 18091623/1745 (93%)0.0
Shank3a polypeptide-Rattus sp, 1 . . . 17401653/1745 (94%)
1740 aa. [WO200078921-A2, 28-
DEC-2000]
AAB31516Amino acid sequence of the rat 85 . . . 1804 822/2129 (38%)0.0
Shank 1a polypeptide-Rattus sp, 1 . . . 20821038/2129 (48%)
2087 aa. [WO200078921-A2, 28-
DEC-2000]
AAB12000Rat p3103 protein-Rattus sp, 2091 38 . . . 1727 781/2055 (38%)0.0
aa. [JP2000184884-A, 04-JUL- 24 . . . 2024 989/2055 (48%)
2000]
AAB31518Amino acid sequence of the rat420 . . . 1806 605/1531 (39%)0.0
Shank2 polypeptide-Rattus sp, 13 . . . 1467 791/1531 (51%)
1470 aa. [WO200078921-A2, 28-
DEC-2000]
|
[0513] In a BLAST search of public sequence datbases, the NOV38a protein was found to have homology to the proteins shown in the BLASTP data in Table 38D.
200TABLE 38D
|
|
Public BLASTP Results for NOV38a
NOV38aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9WUY7PROLINE RICH SYNAPSE 1 . . . 18091689/1812 (93%)0.0
ASSOCIATED PROTEIN 2- 1 . . . 18061717/1812 (94%)
Ruttus norvegicus (Rat), 1806 aa.
Q9JLU4SHANK POSTSYNAPTIC 77 . . . 18091624/1745 (93%)0.0
DENSITY PROTEIN 3A-Rattus 1 . . . 17401654/1745 (94%)
norvagicus (Rat), 1740 aa.
Q9BYB0KIAA1650 PROTEIN-Homo1013 . . . 1809 797/797 (100%)0.0
sapiens (Human), 797 aa 1 . . . 797 797/797 (100%)
(fragment).
Q9WV47SPANK-2-Rattus norvegicus 307 . . . 1204 796/904 (88%)0.0
(Rat), 905 aa (fragment). 1 . . . 897 817/904 (90%)
BAB84864FLJ00090 PROTEIN-Homo1040 . . . 1809 770/770 (100%)0.0
sapiens (Human), 770 aa 1 . . . 770 770/770 (100%)
(fragment).
|
[0514] PFam analysis predicts that the NOV38a protein contains the domains shown in the Table 38E.
201TABLE 38E
|
|
Domain Analysis of NOV38a
Identities/
NOV38aSimilaritiesExpect
Pfam DomainMatch Regionfor the Matched RegionValue
|
ank: domain 1 of 5 224 . . . 25711/34 (32%)8.9
25/34 (74%)
ank: domain 2 of 5 258 . . . 29011/33 (33%)0.00017
20/33 (61%)
ank: domain 3 of 5 291 . . . 32411/34 (32%)1.7
21/34 (62%)
ank: domain 4 of 5 325 . . . 35712/33 (36%)0.014
23/33 (70%)
ank: domain 5 of 5 358 . . . 39010/33 (30%)4.4e−07
28/33 (85%)
SH3: domain 1 of 1 551 . . . 60515/58 (26%)6.2e−07
44/58 (76%)
PDZ: domain 1 of 1 648 . . . 74122/97 (23%)0.0023
65/97 (67%)
SAM: domain 1 of 11744 . . . 180727/68 (40%)5.7e−20
52/68 (76%)
|
Example 39
[0515] The NOV39 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 39A.
202TABLE 39A
|
|
NOV39 Sequence Analysis
|
|
SEQ ID NO: 991435 bp
NOV3 9a,CCCACCCTCGGGATGGACCCCTTCGAGGACACGCTGCGGCGACTGCGGGAGGCCTTCC
CG91434-01 DNA Sequence
ACGCGGGGCGCACGCGGCCGGCCGAGTTCCGGGCTGCGCAGCTCCAGGGCCTGGGCCA
|
CTTCCTTCAAGAAAACAAGCAGCTTCTGCCCGACGTGCTGGCCCAGGACCTGCATAAG
|
CCAGCTTTCCAGCCACACATATCTGAGCTCATCCTTTGCCAGAACGAGGTTGACTACG
|
CTCTCAAGAACCTCCGGGCCTGGATGAAGGATGAACCACGGTCCACGAACCTGTTCAT
|
GAACCTGCACTCGGTCTTCATCTGCAAGCAGCCCTTTGGCCTGGTCCTCATCATTCCG
|
CCCTGGAACTATCCGCTGAACCTGACGCTGGTGCCCCTCGTGGGAGCCCTCGCTGCAG
|
GGAACTGTGTGGTGCTGAAGCCATCGGACATTAGCAAGAACGTCGAGAACATCCTGGC
|
CGAGGTGCTGCCCCAGTACGTGGACCAGAGCTGCTTTGCTGTGGTGCTGGGCGGACCC
|
CAGGAGACAGGGCAGCTGCTAGAGCACAAGTTCGACTACATCTTCTTCACACGGAGCC
|
CTCGTGTGGGCAAGATTGTCATGACTGCTGCCACCAAGCACCTGACCCCTGTCACCCT
|
GGAGCTGGGGCGCAAGAACCCCTCCTACGTGGACGACAACTGCGACCCCCAGACCGTG
|
GCCAACCGCGTGGCCTGGTTCTGCTACTTCAATGCCGGCCAGACCTGCGTGGCCCCTG
|
ACTATGTCCTCTGCAGCCCCCAGATGCAGGACAGGCTGCTGCCCGCCCTCCAGACCAC
|
CATCACCCGTTTCTATGGCGACCACCCCCAGACCTCCCCAAACCTGGGCCGCATCATC
|
AACCAGAAACAGTTCCAGCGGCTGCGGGCATTGCTGGGCTGCCGCCCTGTGGCCATTG
|
GGGGCCAGACCGATGACAGCGATCGCTACATCGCCCCCACCCTGCTGGTGGATGTGCA
|
GGAGATGCAGCCTGTGATGCAGGAGGAGATCTTCGCGCCCATCCTGCCCATCGTCAAC
|
GTGCAGAGCTTGGACGAGGCCATCGAGTTCATCAACCCGCGCGAGAAGCCCCTGGCCC
|
TGTACGCCTTCTCCAACAGCAGCCAGGTGGTCAAGCGGGTGCTGACCCAGACCAGCAG
|
CGGGGGCTTCTGTGGGAACGACGGCTTCATGCACATGACCCTGGCCAGCCTGCCTTTT
|
GGAGGAGTCGGCCACAGTGGGATGGGCCGGTACCACGGCAAGTTCACCTTCCACACCT
|
TCTCCCACCACCGCACCTGCCTGCTCGCCCCCTCCGGCCTGGAGAAATTAAAGGAGAT
|
CCGCTACCCACCCTATACCGACTGGAACCAGCAGCTGTTACGCTGGGGCATGGCCTCC
|
CAGAGCTGCACCCTCCTGTGAGCGTCCCACCCGCCTCCAACGG
|
ORF Start: ATG at 13ORF Stop: TGA at 1411
SEQ ID NO: 100466 aaMW at 52175.4kd
NOV39a, MDPPEDTLRRLREAFEAGRTRPAEFRAAQLQGLGEFLQENKQLLRDVLAQDLHKPAFE
CG91434-01 Protein Sequence
ADISELILCQNEVDYALKNLRAWMKDEPRSTNLFMKLDSVFIWKEPFGLVLIIAPWNY
|
PLNLTLVPLVGALAAGNCVVLKPSEISKNVEKILAEVLPQYVDQSCFAVVLGGPQETG
|
QLLEHKLDYIFFTGSPRVGKIVMTAATKHLTPVTLELGGKNPCYVDDNCDPQTVANRV
|
AWFCYFNAGQTCVAPDYVLCSPEMQERLLPALQSTITRFYGDDPQSSPNLGRIINQKQ
|
FQRLRALLGCGRVAIGGQSDESDRYIAPTVLVDVQEMEPVMQEEIFGPILPIVNVQSL
|
DEAIEFINRREKPLALYAFSNSSQVVKRVLTQTSSGGFCGNDGFMHMTLASLPFGGVG
|
HSGMGRYHGKFTFDTFSHHRTCLLAPSGLEKLKEIRYPPYTDWNQQLLRWGMGSQSCT
|
LL
|
[0516] Further analysis of the NOV39a protein yielded the following properties shown in Table 39B.
203TABLE 39B
|
|
Protein Sequence Properties NOV39a
|
|
PSort0.7900 probability located in plasma membrane;
analysis:0.3000 probability located in Golgi body;
0.2000 probability located in endoplasmic reticulum
(membrane); 0.1594 probability located in microbody
(peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0517] A search of the NOV39a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 39C.
204TABLE 39C
|
|
Geneseq Results for NOV39a
NOV39aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAB58156Lung cancer associated polypeptide46 . . . 434374/389 (96%)0.0
sequence SEQ ID 494-Homo sapiens,26 . . . 414379/389 (97%)
430 aa. [WO200055180-A2, 21 -SEP-
2000]
AAG82730S. epidermidis open reading frame 1 . . . 446188/448 (41%)e−101
protein sequence SEQ ID NO: 2554- 1 . . . 446276/448 (60%)
Staphylococcus epidermidis, 459 aa.
[WO200134809-A2, 17-MAY-2001]
AAG82076S. epidermidis open reading frame 1 . . . 446188/448 (41%)e−101
protein sequence SEQ ID NO: 1246- 1 . . . 446276/448 (60%)
Staphylococcus epidermidis, 459 aa.
[WO200134809-A2, 17-MAY-2001]
AAG39994Arabidopsis thaliana protein fragment11 . . . 455184/452 (40%)e−100
SEQ ID NO: 49563-Arabidopsis82 . . . 529281/452 (61%)
thaliana, 550 aa. [EP1033405-A2, 06-
SEP-2000]
AAG14814Arabidopsis thaliana protein fragment11 . . . 455184/452 (40%)e−100
SEQ ID NO: 14819-Arabidopsis82 . . . 529281/452 (61%)
thaliana, 550 aa. [EP1033405-A2, 06-
SEP-2000
|
[0518] In a BLAST search of public sequence datbases, the NOV39a protein was found to have homology to the proteins shown in the BLASTP data in Table 39D.
205TABLE 39D
|
|
Public BLASTP Results for NOV39a
NOV39aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P43353Aldehyde dehydrogenase 7 (EC 1 . . . 466413/468 (88%)0.0
1.2.1.5)-Homo sapiens (Human), 1 . . . 468428/468 (91%)
468 aa.
P48448Aldehyde dehydrogenase 8 (EC82 . . . 466357/385 (92%)0.0
1.2.1.5)-Homo sapiens 1 . . . 385372/385 (95%)
385 aa.
Q96IB2SIMILAR TO ALDEHYDE82 . . . 466357/385 (92%)0.0
DEHYDROGENASE 3 FAMILY, 1 . . . 385373/385 (96%)
MEMBER B2-Homo sapiens
(Human), 385 aa.
AAL56246ALDEHYDE DEHYDROGENASE20 . . . 466353/449 (78%)0.0
ALDH3B1 (EC 1.2.1.3)-Mus 1 . . . 449396/449 (87%)
musculus (Mouse), 449 aa
(fragment).
Q90ZZ7ALDEHYDE DEHYDROGENASE- 1 . . . 456244/458 (53%)e−144
Brachydanio rerio (Zebrafish) (Zebra 1 . . . 458324/458 (70%)
danio), 473 aa.
|
[0519] PFam analysis predicts that the NOV39a protein contains the domains shown in the Table 39E.
206TABLE 39E
|
|
Domain Analysis of NOV39a
Identities/
NOV39aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
COLFI: domain 1235 . . . 248 7/18 (39%)9.2
of 1 11/18 (61%)
aldedh: domain 1 1 . . . 432157/493 (32%)3.1e−180
of 1378/493 (77%)
|
Example 40
[0520] The NOV40 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 40A.
207TABLE 40A
|
|
NOV40 Sequence Analysis
|
|
SEQ ID NO:1011026 bp
NOV40a,GTCATTATGGATCTGATCAACTCTACTGATTACCTGATCAATGCCTCTACTTTAGTAA
|
CG91484-01 DNA SequenceGAAACAGCACTCAGTTTCTAGCTCCTGCATCAAAAATGATTATTGCCCTTTCTTTGTA
|
CATTTCATCTATAATTGGTACCATCACCAATGGCCTCTATCTATGGCTCCTAAGATTC
|
AAGATGAAACAGACTGTCAATACTCTCTTATTTTTTCATCTCATTCTCTCTTATTTTA
|
TTTCAACAATGATTCTGCCATTTATGGCCACCTCCCAACTTCAAGACAATCACTGGAA
|
CTTTGGAACTGCCTTGTGCAAGGTCTTCAATGGCACTTTGTCTCTGGGGATGTTCACC
|
TCTGTTTTCTTCCTTTCGGCCATCGGTCTTGATCGTTACCTTCTCACTCTTCACCCAG
|
TGTGGTCCCAGCAGCACCGAACCCCGCGCTGGGCTTCCAGCATTGTCCTGGGAGTCTG
|
GATTTCAGCCGCTGCCCTCAGCATCCCCTATTTGATTTTCAGAGAGACACATCATGAC
|
CGTAAAGGAAAGGTGACTTGCCAAAATAACTATGCTGTGTCTACTAACTGGGAAAGCA
|
AGGAGATGCAAGCATCAAGGCAGTGGATTCATGTGGCCTGTTTCATCAGCCGCTTCTT
|
GCTGGGCTTTCTTCTGCCTTTCTTCATCATCATCTTTTGTTATGAAAGAGTACCCAGC
|
AAGGTGAAAGAGAGGAGCCTGTTTAAATCCAGCAAGCCCTTCAAAGTTATGATGACTG
|
CCATTATCTCTTTCTTTGTGTGTTGGATGCCCTACCATATACACCAGGGCTTACTTCT
|
CACTACGAACCAGTCACTACTTTTAGAGTTGACTTTGATACTTACAGTGCTAACCACT
|
TCTTTCAATACTATCTTTTCTCCCACACTCTACTTATTTGTTGGGGAGAATTTCAAAA
|
AGGTCTTCAAGAAGTCCATTCTTGCTCTGTTTGAGTCAACATTTAGTGAAGATTCTTC
|
TGTAGAAAGGACACAAACCTAAACTCACAAGCCTAAATTT
|
ORF Start: ATG at 7ORF Stop: TAA at 1006
SEQ ID NO:102333 aa MW at 38231.5 kD
NOV40a,MDLINSTDYLINASTLVRNSTQFLAPASKMIIALSLYISSIIGTITNGLYLWVLRFKM
|
CG91484-01 Protein SequenceKQTVNTLLFFHLILSYFISTMILPFMATSQLQDNHWNFGTALCKVFNGTLSLGMFTSV
|
FFLSATGLDRYLLThHPVWSQQERTPRWASSIVLGVWISAAALSIPYLIFRETHHDRK
|
GKVTCQNNYAVSTNWESKEMQASRQWIHVACFISRFLLGFLLPFFIIIFCYERVASKV
|
KERSLFKSSKPFKVMMTAITSFFVCWMPYHIHQGLLLTTNQSLLLELTLILTVLTTSF
|
NTIFSPTLYLFVCENFKVFKKSILALFESTFSEDSSVERTQT
|
[0521] Further analysis of the NOV40a protein yielded the following properties shown in Table 40B.
208TABLE 40B
|
|
Protein Sequence Properties NOV40a
|
|
PSort0.6000 probability located in plasma membrane:
analysis:0.4000 probability located in Golgi body;
0.3620 probability located in mitochondrial
inner membrane; 0.3000 probability located in
endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0522] A search of the NOV40a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 40C.
209TABLE 40C
|
|
Geneseq Results for NOV40a
NOV40aIdentities/
Residues/Similarities for
GeneseqMatchthe MatchedExpect
IdentifierProtein/Organism/Length [Patent #, Date]ResiduesRegionValue
|
AAW09642Rabbit G protein-linked receptor from128 . . . 252 97/125 (77%)1e−55
smooth muscle of the gut- 1 . . . 125110/125 (87%)
Oryctolagus cuniculus, 125 aa.
[JP08283295-A, 29-OCT-1996]
AAR91230Rabbit G-protein coupled receptor128 . . . 252 97/125 (77%)1e−55
protein portion-Oryctolagus 1 . . . 125110/125 (87%)
cuniculus, 125 aa. [WO9605302-A1,
22-FEB-1996]
AAW86323Kidney injury associated molecule 5 . . . 326122/332 (36%)2e−55
HW055 protein-Rattus sp, 372 aa. 18 . . . 341188/332 (55%)
[WO9853071-A1, 26-NOV-1998]
ABB56354Non-endogenous human GPCR 36 . . . 327 98/299 (32%)7e−48
protein, SEQ ID NO: 501-Homo 33 . . . 330163/299 (53%)
sapiens, 351 aa. [WO200177172-A2,
18-OCT-2001]
ABB56354Non-endogenous human GPCR 36 . . . 327 98/299 (32%)7e−48
protein, SEQ ID NO: 501-Homo 33 . . . 330163/299 (53%)
sapiens, 351 aa. [WO200177172-A2,
18-OCT-2001]
|
[0523] In a BLAST search of public sequence datbases, the NOV40a protein was found to have homology to the proteins shown in the BLASTP data in Table 40D.
210TABLE 40D
|
|
Public BLASTP Results for NOV40a
NOV40aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
O88416Probable G protein-coupled receptor 1 . . . 333240/333 (72%)e−140
GPR33-Mus musculus (Mouse). 339 1 . . . 332275/333 (82%)
aa.
P97468Chemokine receptor-like 1 (G-protein25 . . . 326121/310 (39%)2e−54
coupled receptor DEZ)-Mus musculus34 . . . 340177/310 (57%)
(Mouse), 371 aa.
O75748Probable G protein-coupled receptor 8 . . . 326120/326 (36%)9e−54
CHEMR23-Homo sapiens (Human),18 . . . 340186/326 (56%)
371 aa.
Q99788Chemokine receptor-like 1 (G-protein 8 . . . 326120/326 (36%)9e−54
coupled receptor DEZ) (G protein-20 . . . 342186/326 (56%)
coupled receptor ChemR23)-Homo
sapiens (Human), 373 aa.
O35786Chemokine receptor-like 1 (G-protein 5 . . . 326119/334 (35%)1e−50
coupled receptor DEZ) (G-protein18 . . . 340186/334 (55%)
coupled chemoattractant-like receptor)-
Rattus norvegicus (Rat), 371 aa.
|
[0524] PFam analysis predicts that the NOV40a protein contains the domains shown in the Table 40E.
211TABLE 40E
|
|
Domain Analysis of NOV40a
Identities/
NOV40aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
LGT: domain 1 of 122 . . . 290 48/350 (14%)2.5
179/350 (51%)
7tm_1: domain 1 of 146 . . . 299 70/279 (25%)2.6e−41
184/279 (66%)
7tm_2: domain 1 of 125 . . . 306 54/297 (18%)1.4
163/297 (55%)
|
Example 41
[0525] The NOV41 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 41 A.
212TABLE 41A
|
|
NOV41 Sequence Analysis
|
|
SEQ ID NO:103950 bp
NOV41a,GGCGGTCGGGGAGGCCGGTGCCATGGGGTCGCGGAGGGCCCCCAGCCGGGGCTGGGGC
|
CG91514-01 DNA SequenceGCGGGTGGGCGGTCGGGGGCGGGGGGCGACGGTGAGGACGACCGCCCCGTGTGGATCC
|
CCAGCCCAGCCACCCGGAGCTACCTGCTCAGCGTGCGGCCCGAGACCAGCTTATCAAG
|
CAACCGCTTGTCTCACCCCAGCTCTGGAACGAGCACCTTCTGCTCCATCATTGCTCAC
|
CTCACAGAGGAGACCCAGCCGCTATTTGAGACCACGCTCAAGTCCCGGTCTGTGTCCG
|
AGGACAGCGACGTCACGTTCACCTCCATCGTCACAGGATACCCAGACCCAGAGGTGAC
|
CTGGTACAAGGATGATACCGAGCTGGACCGCTACTGTGGCTTGCCAAAATATGAGATC
|
ACTCATCAGGGCAACCGCCACACACTGCAGCTGTACAGGTGTCGAGAAGAAGATGCCG
|
CCATCTACCAGGCCTCTCCCCAGAACAGCAAGGGCATTGTGTCCTGCTCAGGGGTCCT
|
GGAGGTGGGCACCATGACTGAGTACAAGATCCACCAGCGCTGGTTCGCCAAGTTGAAG
|
CGCAAGGTCTGCAGCTAGCCTACTCCCTTTGGAATGCAATAAAGGCAGCATTGTGTGC
|
CCTGCTTGCCCTCATCTGGTGTGGTTGGAGGTCTGTGGAGTCAAGGTCCCCCTCTCCC
|
AGGCAGGCTCTCTGAGGGCATTCTCTAGTCCCACGCCCACTGGAAAAATGAATCTATA
|
TTTTGGTTCCTGGACCGAAGTTCAGTCGCAGCCTTCTGTGGCCACAGAAAGACAGCTT
|
GTGCTGCTTGCACAACTGAGCTGCTGGTCTGTACCCCTTAAGCAGGGTGTCTGGGACT
|
TACGCCTTTGGAATTGCTCTTCATTCAGAAGAGGAACACAAAGGAAGCCACCCAGGAA
|
GGAACCACAGAGCTGGGOGCTC
|
ORF Start: ATG at 23ORF Stop: TAG at 596
SEQ ID NO:104191 aa MW at 21218.4 kD
NOV41a,MGSRRAPSRCWGAGGRSGAGGDGEDDGPVWIPSPASRSYLLSVRPETSLSSNRLSEPS
|
CG91514-01 Protein SequenceSGRSTFCSIIAQLTEETQPLFETTLKSRSVSEDSDVRFTCIVTGYPEPEVTWYKDDTE
|
LDRYCGLPKYEITHQGNRHTLQLYRCREEDAAIYQASAQNSKGIVSCSGVLEVGTMTE
|
YKIHQRWFAKLKRKVCS
|
[0526] Further analysis of the NOV41 a protein yielded the following properties shown in Table 41B.
213TABLE 41B
|
|
Protein Sequence Properties NOV41a
|
|
PSort0.4500 probability located in cytoplasm;
analysis:0.3821 probability located in microbody
(peroxisome); 0.1000 probability located in
mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0527] A search of the NOV41 a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 41C.
214TABLE 41C
|
|
Geneseq Results for NOV41a
NOV41aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAM79811Human protein SEQ ID NO 3457 - 33..17850/169 (29%)2e-11
Homo sapiens, 1621 aa.246..41476/169 (44%)
[WO2000157190-A2, 09-AUG-2001]
AAM78827Human protein SEQ ID NO 1489 - 33.17850/169 (29%)2e-11
Homo sapiens, 1452 aa. 78..24676/169 (44%)
[WO200157190-A2, 09-AUG-2001]
AAB63803Human prostate cancer associated 45..17441/130 (31%)1e-10
antigen protein sequence SEQ ID106..23263/130 (47%)
NO:1165 - Homo sapiens, 242 aa.
[WO200073801-A2, 07-DEC-2000]
AAY70078Human striated muscle preferentially 24..16950/149 (33%)8e-10
expressed partial protein - Homo374..51073/149 (48%)
sapiens, 661 aa. [WO200009689-A2,
24-FEB-2000]
AAW77048Human striated muscle preferentially 24.16950/149 (33%)8e-10
expressed protein - Homo sapiens,374..51073/149 (48%)
661 aa. [WO9835040-A2,
[13-AUG-1998]
|
[0528] In a BLAST search of public sequence datbases, the NOV4 la protein was found to have homology to the proteins shown in the BLASTP data in Table 41D.
215TABLE 41D
|
|
Public BLASTP Results for NOV41a
Identities/
NOV41aSimilarities
ProteinResidues/for the
AccessionMatchMatchedExpect
NumberProtein/Organism/LengthResiduesProtionValue
|
Q96L96MUSCLE ALPHA-KINASE - Homo 1..191187/191 (97%)e-109
sapiens (HUman), 1907 aa.203..393189/191 (98%)
Q924C5MYOCYTIC 1..191174/191 (91%)e-102
INDUCTION/DIFFERENTIATION 1..191181/191 (94%)
ORIGINATOR - Mus musculus (Mouse),
1678 aa.
P29294Myosin light chain kinase, smooth muscle 45.1174 42/130 (32%)2e-11
(EC 2.7.1.117) (MLCK) [Contains:1009..1136 61/130 (46%)
Telokin] - Oryctolagus cuniculus
(Rabbit), 1147 aa.
Q15746Myosin light chain kinase, smooth muscle 77.186 37/110 (33%)2e-11
and non-muscle isozymes (EC 2.7.1.117)414..521 59/110 (53%)
(MLCK) [Contains: Telokin (Kinase
related protein) (KRP)] - Homo sapiens
(Human), 1914 aa.
A41675telokin - rabbit, 155 aa. 45..174 42/130 (32%)3e-11
17.144 61/130 (46%)
|
[0529] PFam analysis predicts that the NOV41a protein contains the domains shown in the Table 41 E.
216TABLE 41E
|
|
Domain Analysis of NOV41a
NOV41aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
ig: domain 1 of 191..15414/67 (21%)7.8e−07
49/67 (73%)
|
Example 42
[0530] The NOV42 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 42A.
217TABLE 42A
|
|
NOV42 Sequence Analysis
|
|
SEQ ID NO:1051680 bp
NOV42aTGCAGGTGCGTTGTACCCTTTTACGTGAGGCGGTGACGGCGGTTCGGAAGTCCTCTGG
|
CG91587-01 DNA SequenceCCTCCCCGCGGCCGCTCGCAGCTTGCTGGCCTCTCCCGCGCCTCACGTCGGACTCCGT
|
CTCCGCGGCACGGAAGCAGCATCCAAGCTGAGGACATCCACCAGGAGTTGACCTGCCC
|
CATCTGCCTCGACTATTTCCACCACCCGGTGTCCATCGACTCCGGCCACAACTTCTGC
|
CGCGGCTGCCTGCACCGCAACTGCGCGCCGGGCGGCGGCCCGTTCCCCTGCCCCGAAT
|
GTCGGCACCCATCGGCGCCCGCCGCCCTGCGACCCAACTGCGCCCTCGCCAGGCTGAC
|
TGACAAGACGCAGCGCCGGCGCCTGGGCCCCGTOCCCCCGGGCCTGTGCGGCCGCCAC
|
TCGCAGCCGCTGCGGCTCTTCTGCCAGGACGACCAGCGaCCAGTGTGCCTGGTGTGCA
|
GGGAGTCCCAGGAGCACCAGACTCACGCCATGGCACCCATCCACGAGGCCTTCGAGAG
|
CTACCGGACAGGTAACTTTGACATCCACGTGGATGAATCCAAGAGAAGACTAATTACC
|
CTGCTCTTGTACCATTCTAAGCAGGACGAGAAACTTCTTAAGTCTCAGCGTAATCTCG
|
TGGCCAAGATGAAGAAAGTCATGCATTTACAGGATGTAGAAGTGAAGAACGCCACACA
|
GTGCAAGGATAAGATAAAGAGTCAGCGAATGAGAATCAGCACGCAGTTTTCAAAGCTG
|
CACAACTTCCTGGTTGAAGAAGAGGACCTCTTTCTTCAGAGATTCAACAAAGAAGAAG
|
AAGAGACGAAGAAGAAGCTGAATGAGAACACGTTAAAACTCAATCAAACTATCGCTTC
|
ATTGAAGAAGCTCATCTTAGAGGTGGGGCAGAAGAGCCAGGCTCCCACCCTGGAGCTG
|
CTTCAGAATCCAAAAGAAGTGTTGACCAGGAGTGAGATCCAGGATGTGAACTATTCCC
|
TTGAACCTGTGAAGGTGAAGACAGTGTGCCAGATACCATTGATGAAGGAAATGCTAAA
|
CCGATTCCAAGTGGCTGTAAACCTAGCTGAAGACACAGCTCATCCCAAACTCGTCTTC
|
TCCCAGGAAGGCAGATACGTGAAAAATACAGCATCAGCCAGTTCTTGGCCAACTGCTT
|
TTGTAGAGAGATTTCAGCACTTACCCTGTGTTCTGGGAAAAAACGTTTTCACCTCAGG
|
GAAACATTACTGGGAAGTTGAGAGTAGACATAGTCTGGACCTTGCTGTTGGGGTGTGT
|
CGGGAGGACGTCATGGGAATTACTGATCGTTCAAAAATGTCCCCAGATGTGGGCATCT
|
GGGCGATTTATTGCAGTGCTQCTCCCTATTGCCCCTTGATAGGCTTCCCTCCAACTCC
|
CACCCAGCAAGAGCCAGCTCTCCACCGAGTGGGGGTTNACCTGGATCGTGGGACTGGG
|
AATGTCTCCTTCTACAGCGCTGTGGACGGAGTGCACCTGCACACCTTTTCTTGTTCTT
|
CTGTCTCACGCCTCCGGCCATTTTTTTGGTTGAGTCCATTAGCATCTTTACTCATTCC
|
ACCACTGACTGATACGAAATGAGGCTTTTCTTCCCCTGACCAAAACTCCTTCCCTGTA
|
GTCCAGCTGAGGGACACACATCCCTGGGCCCTCTTCTGCCCTTCATGTCTCTATCC
|
ORF Staff: at 29ORF Stop: at 1478
SEQ ID NO:106483 aa MW at 55196.9 kD
NOV42aMEAEDIQEELTCPICLDYFQDPVSIECGHNFCRCCLHRNWAPGGGPFPCPECRHPSAP
|
CG91587-01 Protein SequenceAALRPNWALARLTEKTQRRRLGPVPPGLCGRHWEPLRLFCEDDQRPVCLVCRESQEHQ
|
THANAPIDEAFESYRTGHFDIHVDHWKRRLIRLLLYHSKQEEKLLKSQRNLVAKNKKV
|
MHLQDVEVKNATQWKDKIKSQRMRISTEFSKLENFLVEEEDLFLQRLNKEEEETKKKL
|
HEHTLKLNQTIASLKKLILEVGEKSQAPTLELLQNPKEVLTRSEIQDVNYSLEAVKVK
|
TVCQIPLMKEMLKRFQVAVNLAEDTAHPKLVFSQEGRYVKNTASASSWPTAFVERFQH
|
LPCVLGKNVFTSGKHYWEVESRDSLEVAVGVCREDVMGITDRSKMSPDVGIWAIYWSA
|
AGYWPLIGFPGTPTQQEPALHRVGVXLDRGTGNVSFYSAVDGVHLHTFSCSSVSRLRP
|
FFWLSPLASLVIPPVTDRK
|
[0531] Further analysis of the NOV42a protein yielded the following properties shown in Table 42B.
218TABLE 42B
|
|
Protein Sequence Properties NOV42a
|
|
PSort0.5050 probability located in cytoplasm; 0.3117 probability
analysis:located in microbody (peroxisome); 0.1000 probability
located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0532] A search of the NOV42a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 42C.
219TABLE 42C
|
|
Geneseq Results for NOV42a
NOV42aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABB11609Human secreted protein homologue,261..483 221/240 (92%)e−124
SEQ ID NO:1979 - Homo sapiens,6..245221/240 (92%)
326 aa. [WO200157188-A2,
09 Aug. 2001]
AAB42919Human ORFX ORF2683 polypeptide3..480183/501 (36%)2e−76
sequence SEQ ID NO:5366 - Homo7..472264/501 (52%)
sapiens, 477 aa.
[WO200058473-A2, 05 Oct. 2000]
AAR15148Ro/SSA autoantigen - Homo sapiens,8..466173/468 (36%)4e−67
475 aa. [WO9117171-A,12..446 244/468 (51%)
14 Nov. 1991]
AAB43498Human cancer associated protein3..480169/519 (32%)1e−64
sequence SEQ ID NO:943 - Homo74..560 248/519 (47%)
sapiens, 580 aa. [WO200055350-A1,
21 Sep. 2000]
AAW78921Human haemochromatosis protein6..481160/490 (32%)5e−58
RoRet - Homo sapiens, 465 aa.10..465 233/490 (46%)
[WO9814466-A1, 09 Apr. 1998]
|
[0533] In a BLAST search of public sequence datbases, the NOV42a protein was found to have homology to the proteins shown in the BLASTP data in Table 42D.
220TABLE 42D
|
|
Public BLASTP Results for NOV42a
NOV42aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9C037TRIPARTITE MOTIF PROTEIN1..483482/500 (96%)0.0
TRIM4 ISOFORM ALPHA - Homo1..500482/500 (96%)
sapiens (Human), 500 aa.
Q9C036TRIPARTITE MOTIF PROTEIN1..483456/500 (91%)0.0
TRIM4 ISOFORM BETA - Homo1..474456/500 (91%)
sapiens (Human), 474 aa.
Q96F06SIMILAR TO TRIPARTITE1..306280/306 (91%)e−162
MOTIF PROTEIN 4 - Homo sapiens1..280280/306 (91%)
(Human), 294 aa.
JE0343terf protein - rat, 477 aa.3..480198/498 (39%)6e−82
7..472274/498 (54%)
Q9WV59RING FINGER PROTEIN TERF -3..480198/498 (39%)7e−82
Rattus norvegicus (Rat), 477 aa7..472274/498 (54%)
|
[0534] PFam analysis predicts that the NOV42a protein contains the domains shown in the Table 42E.
221TABLE 42E
|
|
Domain Analysis of NOV42a
NOV42aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
zf-C3HC4:12..5218/54 (33%)2.1e−15
domain 1 of 137/54 (69%)
zf-B_box: 82..12319/48 (40%)5.5e−12
domain 1 of 133/48 (69%)
SNAP 25:111..27536/217 (17%)2.3
domain 1 of 185/217 (39%)
SPRY: domain 1 of 1360..48033/157 (21%)3.5e−12
82/157 (52%)
|
Example 43
[0535] The NOV43 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 43A.
222TABLE 43A
|
|
NOV43 Sequence Analysis
|
|
SEQ ID NO:1072295 bp
NOV43a,CACCAGAAATGATCCTTTCCCAANGCTGCTCCTCCCCATCACAAGCCCTTCCTTTCATCC
|
CG91631-01 DNA SequenceCATTCCTGCCGGTTCAGCGANCCGGAGTCGCCCAGGCCTGAACTCCTACCCAGGTCCC
|
CGGCCCCCGCCCCGGCCCCCCAGACACCGGAGCCACCCCCCGGGCTCCNGNAGCCCAG
|
CGCGGCCTAGACTCAAGTCTGGGTTTCAGCTGCCGCCAGCCCTATTGCTGCTGTTGCT
|
GTTCTCTGTCCTTCGCCCAGGGNATGGAGGCCTTTTCCTGACTGATTACTCCACCTGC
|
TCACCCCGCAAGCTGAGTCCTTTCCGCTCCTTTGCCAGCACCGAGCTCTTCCACTTCC
|
ATGTTCCTGAGGACACATTCCTGGCTGTTTGGAACCTCATCATCTTCAAGGAGCAAGG
|
GGGAACTTTTGGGGACCACTGCCCAGACCAAAGTGTAACTGTCTATTTCCGGTCCGGG
|
GCACCCCCTGTCATCAATCCCCTGCATACACACTTCCCAGGGGACACAGCTGTGCCTG
|
GGGTTTTCTCACTGACCCTCAGCTGGACACTGCCCAACCGCACCTCAGGCATOTTTAA
|
CGTCAGCAGCCCCTTACCTGGGGACTGGTTCTTGGCTGCCCACCTTCCCCAGCCCCAC
|
GGCCACATCTCTGTCAAGGGTCTCCAGGATGAGTGTCAGTACCTCCTTCAGCCGCAGC
|
TGATTGTCCGGCGGTTGCTGGACGTCGCTGTGCTCGTTCCTCGCCGGCCCTCACAGCA
|
AACCCTCTCCCCACACAATCGCTCAGCCCTCTACAAGGTCTTTGTGCCCAGCTTCACT
|
TACAGGGTTTCAGCACAGCTGGTGTGTGTGGCCGGCCGTCGOGTATCTGCCTGCCCCC
|
TGTCACTGCGTCTGCGTCCCAAAGCCCCACCCCTGCACAACTCAAGCTCTGTCGCCTG
|
TGGACGTGCCTCAGGATGCCAGCTGGAGCTGGCACTGCCCCCCTGGGGCCACTGGGTC
|
TACGTGCGTGTGGAAACATCATCCCGGGCCCCTGGTAGGACCATCCGCTTCCAGCTGT
|
GTGTGCAGTTGCAAGAGTGCCCACAGCCCGGCCTGCTCCCACCCCTCGTCCCTGGAGC
|
TGCCATGAACATGCCCCAGTCCCTGGGCAACCAGCCACTGCCCCCAGAACCGCCATCC
|
CTTGGAACCCCTGCGGAGGGGCCTGGGACCACGTCCCCACCCGAGCACTGCTGGCCAG
|
TGCGCCCGACTCTGCGCAACGAGCTGGACACCTTCTCTGTCCACTTCTACATCTTCTT
|
TGGCCCAAGTGTCGCCCTTCCCCCTGAGCGCCCACCCGTGTTCGCCATGAGGCTGTTG
|
CCAGTGCTGGACAGTGGACGCGTCCTCACCCTGGAGCTCCAGCTCAATGCGACCTCCG
|
TGCGCCAGGAAAACGTGACGGTGTTTGGATGCTTGACTCACGACGTACCCTTGAGCCT
|
GGGGGATGCAGCAGTGACCTGTTCCAAAGAGTCCCTGGCCGGCTCCTCCTCCTCTGTC
|
AGTGCCACCACCAGGCTTCCCAGCCTGCCAATCCCATTCCCGCAGACGGGGACCTGGT
|
TCCTGGCCCTCCGCTCCCTGTGCGGCGTGGGGCCTCCGTTCGTCCGGTGCCGCAACGC
|
GACGCCCCAGGTGCGCATGCGCACCTTCCTGTCCCCATGCGTGGACGACTGCGGCCCC
|
TACGGCCAGTGCAAGCTGCTGCGCACACACAATTATCTGTACGCAGCCTGCGAGTGCA
|
AGGCCCGGTGGAGACGCTGGGGCTGCACCGACAGTGCAGATGCCCTCACCTATGGATT
|
CCAGCTGCTGTCCACACTCCTCCTCTGCCTGACCAACCTCATCTTTCTGCCACCTGTG
|
GTCCTGGCCATTCGGAGTCGATATGTGCTGGAAGCTGCAGTCTACACCTTCACCATGT
|
TCTTCTCCACCTTCTATCATGCCTGTGACCACCCAGCCATCGTGGTTTTCTGCATCAT
|
GGACTACGATGTGCTCCAGTTCTGTGATTTCCTGGGCTCCTTAATCTCCGTCTCCGTC
|
ACTCTCATTGCCATGGCTCGTTTACAGCCCGTGGTCAAGCAGGTGCTGTATTTCCTGG
|
GAGCTATCCTGCTGTCCATGGCTCTGCAGCTTGACCGACATGGACTCTGGAACCTGCT
|
TGGACCCAGTCTCTTCGCCCTGGGGATCTTGGCCACAGCCTGGAACCCCATAGAGATG
|
AACAAATCATCAGCGCTATTCTCCAGTTGCTCAGCAGGGCAAGGATTTGTGCCTGGCC
|
TTCCACGTACTTTTGCAGTAAGACGATGCTGAG
|
ORF Start: at 258ORF Stop: end of sequence
SEQ ID NO:108761 aa MW at 82335.7 kD
NOV43a,MDPFPXLLPITSPSFHPIPAGSAXRSRPGLNSYPGPRPPPRPARHRSHPPGWXSGARP
|
CG91631-01 Protein SequenceRLKSGFQLPPALLLLLLFSVLGPGXCCLFLTDYSTCSPRKLSPFRSFASTELFHFEVP
|
EDTFLAVNNLITFKEQGGTFGDHCPDQSVTVYFRSGAPPVINPLETEFPGDTAVPGVF
|
SLTLSWTLPNRTSGTFVSSPLPGDWFLAA11LPQAHGHISVKGLQDECQYLLQPQLIV
|
RRLLDVALVPGRPSEQTLSPIHHRSALYKVFVPSFTYRVSAQLVCVGCRGVSACPLSL
|
RLRPKAPPLHMSSSVACGGASGCQLELALFPWCHWVYVRVETSSRGPGRTIRFQLCVQ
|
LQECPQPGLLRALVPGAANNMNPQSLGNQPLPAPPSLGTPAEGPGTTSPPEHCWPVRP
|
TLRNELDTFSVHIFYIFFGPSVALPPERPAVFAMRLPVLDSGGVLSLELQLNASSVRQ
|
ENVTVFGCLTHEVPLSLGDAAVTCSKESLAGFLLSVSATTRVARLRTPFPQTGTWFLA
|
LRSLCGVGPRFVRCRNATAEVRNRTPLSPCVDCGPYGQCKLLRTITNYLYAACECKAG
|
WRGWGCTDSADALTYGFQLLSTLLLCLSNLMFLPPVVLAIRSRYVLEAAVYTPTMFFS
|
TFYHACDQPCIVVFCIMDYDVLQPCDFLGSLMSVWVTVIAMARLQPWKQVVLYLLGAM
|
LLSMALQLDRHGLWNLLGPSLFALGTLATAWNPIEMNKSSALFSSCSAGQGFVPGLPG
|
TFGVRGC
|
[0536] Further analysis of the NOV43a protein yielded the following properties shown in Table 43B.
223TABLE 43B
|
|
Protein Sequence Properties NOV43a
PSort0.6000 probability located in plasma membrane; 0.4000
analysis:probability located in Golgi body; 0.3000 probability
located in endoplasmic reticulum (membrane);
0.3000 probability located in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0537] A search of the NOV43a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 43C.
224TABLE 43C
|
|
Geneseq Results for NOV43a
NOV43aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM93940Human polypeptide, SEQ ID NO:367..639273/273 (100%)e−162
4122 - Homo sapiens, 338 aa. 1..273273/273 (100%)
[EP1130094-A2, 05 Sep. 2001]
ABB11995Human M83 protein homologue, 89..727270/641 (42%)e−144
SEQ ID NO:2365 - Homo sapiens, 3..630381/641 (59%)
728 aa. [WO200157188-A2,
09 Aug. 2001]
AAY21590Human secreted protein (clone C544-536..727114/192 (59%)4e−70
1) - Homo sapiens, 310 aa. 9..200149/192 (77%)
[WO9926973-A1, 03 Jun. 1999]
AAW52985Homo sapiens clone C544_1 protein -536..727113/192 (58%)1e−69
Homo sapiens, 229 aa. 9..200148/192 (76%)
[WO9807859-A2, 26 Feb. 1998]
AAY52590Human prostate growth-associated611..727117/117 (100%)2e−62
membrane protein PGAMP-2 - 1..117117/117 (100%)
Homo sapiens, 410 aa.
[WO9961469-A2, 02 Dec. 1999]
|
[0538] In a BLAST search of public sequence datbases, the NOV43a protein was found to have homology to the proteins shown in the BLASTP data in Table 43D.
225TABLE 43D
|
|
Public BLASTP Results for NOV43a
NOV43aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9P0U7NAG-5 PROTEIN - Homo sapiens1..639639/639 (100%)0.0
(Human), 704 aa.1..639639/639 (100%)
O75539HYPOTHETICAL 63.6 KDA120..639 520/520 (100%)0.0
PROTEIN - Homo sapiens (Human),1..520520/520 (100%)
585 aa (fragment).
Q9HBY2NASOPHARYNGEAL367..639 273/273 (100%)e−162
CARCINOMA RELATED1..273273/273 (100%)
PROTEIN - Homo sapiens
(Human), 338 aa.
Q96S25M83 - Homo sapiens (Human),71..727 277/666 (41%)e−145
771 aa.21..673 390/666 (57%)
AAH21557TRANSMEMBRANE PROTEIN 871..727 275/665 (41%)e−145
(FIVE MEMBRANE-SPANNING21..673 386/665 (57%)
DOMAINS) - Homo sapiens
(Human), 771 aa.
|
[0539] PFam analysis predicts that the NOV43a protein contains the domains shown in the Table 43E.
226TABLE 43E
|
|
Domain Analysis of NOV43a
NOV43aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
No Significant Matches Found
|
Example 44
[0540] The NOV44 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 44A.
227TABLE 44A
|
|
NOV44 Sequence Analysis
|
|
SEQ ID NO: 1091147 bp
|
NOV44a,ATGGCGGCGGCGGCGGCTCAGGGGGGCGGGGGCGGGGAGCCCCGTAGAACCGAGGGGG
CG91643-01 DNA SequenceTCGGCCCGGGGGTCCCGGGGGAGGTGGAGATGGTGAAGGGGCAGCCGTTCGACGTGGG
CCCGCGCTACACGCAGTTGCAGTACATCGGCGAGGGCGCGTACGGCATGGTCAGCTCG
GCCTATGACCACGTGCGCAAGACTCGCGTGGCCATCAAGAAGATCAGCCCCTTCGAAC
ATCAGACCTACTGCCAGCGCACGCTCCGGGAGATCCAGATCCTGCTGCGCTTCCGCCA
TGAGAATGTCATCGGCATCCGAGACATTCTGCGGGCGTCCACCCTGGAAGCCATGAGA
GATGTCTACATTGTGCAGGACCTGATGGAGACTGACCTGTACAAGTTGCTGAAAAGCC
AGCAGCTGAGCAATGACCATATCTGCTACTTCCTCTACCAGATCCTGCGGGGCCTCAA
GTACATCCACTCCGCCAACGTGCTCCACCGAGATCTAAAGCCCTCCAACCTGCTCAGC
AACACCACCTGCGACCTTAAGATTTGTGATTTCGGCCTGGCCCGGATTGCCGATCCTG
AGCATGACCACACCGGCTTCCTGACGGAGTATGTGGCTACGCGCTGGTACCGGGCCCC
AGAGATCATGCTGAACTCCAAGGGCTATACCAAGTCCATCGACATCTGGTCTGTGGGC
TGCATTCTGGCTGAGATGCTCTCTAACCGGCCCATCTTCCCTGGCAAGCACTACCTGG
ATCAGCTCAACCACATTCTGGGCATCCTGGGCTCCCCATCCCAGGAGGACCTGAATTG
TATCATCAACATGAAGGCCCGAAACTACCTACAGTCTCTGCCCTCCAAGACCAAGGTG
GCTTGGGCCAAGCTTTTCCCCAAGTCAGACTCCAAAGCCCTTGACCTGCTGGACCGGA
TGTTAACCTTTAACCCCAATAAACGGATCACAGTGGCCGAGGAGCCCTTCACCTTCGC
CATGGAGCTGGATGACCTACCTAAGGAGCGGCTGAAGGAGCTCATCTTCCAGGAGACA
GCACGCTTCCAGCCCGGAGTGCTGGAGGCCCCCTAGCCCAGACAGACATCTCTGCACC
CTGGGGCCTGGACCTGCCTCCTGCCTGCCCCTCTCCCGCCAGACT
|
|
ORF Start: ATG at 1ORF Stop: TAG at 1078
SEQ ID NO: 110359 aa MW at 40747.5 kD
|
NOV44a,MAAAAAQGGGGGEPRRTEGVGPGVPGEVEMVKGQPFDVGPRYTQLQYIGEGAYGMVSS
CG91643-01 Protein SequenceAYDHVRKTRVAIKKISPFEHQTYCQRTLREIQILLRFRHENVIGIRDILRASTLEAMR
DVYIVQDLMETDLYKLLKSQQLSNDHICYFLYQILRGLKYIHSANVLHRDLKPSNLLS
NTTCDLKICDFGLARIADPEHDHTGFLTEYVATRWYRAPEIMLNSKGYTKSIDIWSVG
CILAEMLSNRPIFPGKHYLDQLNHILGILGSPSQEDLNCIINMKARNYLQSLPSKTKV
AWAKLFPKSDSKALDLLDRMLTFNPNKRITVAEEPFTFAMELDDLPKERLKELIFQET
ARFQPGVLEAP
|
[0541] Further analysis of the NOV44a protein yielded the following properties shown in Table 44B.
228TABLE 44B
|
|
Protein Sequence Properties NOV44a
|
|
PSort0.4500 probability located in cytoplasm; 0.3000 probability
analysis:located in microbody (peroxisome); 0.1924 probability
located in lysosome (lumen); 0.1000 probability located in
mitochondrial matrix space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0542] A search of the NOV44a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 44C.
229TABLE 44C
|
|
Geneseq Results for NOV44a
NOV44aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAG67618Amino acid sequence of a human1..359359/379 (94%)0.0
protein - Homo sapiens, 379 aa.1..379359/379 (94%)
[WO200109316-A1, 08 Feb. 2001]
AAG67439Amino acid sequence of a human1..359359/379 (94%)0.0
polypeptide - Homo sapiens, 379 aa.1..379359/379 (94%)
[WO200109345-A1, 08 Feb. 2001]
AAY70778EGFP-Erk1 fusion protein construct -1..359359/379 (94%)0.0
Chimeric - Homo sapiens, 631 aa.253..631 359/379 (94%)
[WO200023615-A2, 27 Apr. 2000]
AAY42413Extracellular signal Regulated Kinase1..359359/379 (94%)0.0
(ERK)1 mutant - Homo sapiens, 3791..379359/379 (94%)
aa. [WO9942592-A1, 26 Aug. 1999]
AAW85006Erki-green flourescent protein fusion1..359359/379 (94%)0.0
product - Chimeric - Aequorea253..631 359/379 (94%)
victoria, 631 aa. [WO9845704-A2,
15 Oct. 1998]
|
[0543] In a BLAST search of public sequence datbases, the NOV44a protein was found to have homology to the proteins shown in the BLASTP data in Table 44D.
230TABLE 44D
|
|
Public BLASTP Results for NOV44a
NOV44aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
A48082MAP kinase 3 (EC 2.7.1.-) - human,1..359359/379 (94%)0.0
379 aa.1..379359/379 (94%)
P27361Mitogen-activated protein kinase 3 (EC1..359358/379 (94%)0.0
2.7.1.-) (Extracellular signal- regulated1..379358/379 (94%)
kinase 1) (ERK- 1) (Insulin-stimulated
MAP2 kinase) (MAP kinase 1) (MAPK
1) (p44-ERK1) (ERT2) (p44-MAPK)
(Microtubule- associated protein-2
kinase) - Homo sapiens
(Human), 379 aa.
CAB69291SEQUENCE 38 FROM PATENT1..358358/378 (94%)0.0
WO9845704 - unidentified, 630 aa253..630 358/378 (94%)
(fragment).
CAB69300SEQUENCE 56 FROM PATENT2..359346/378 (91%)0.0
WO9845704 - unidentified, 624 aa.3..380347/378 (91%)
P21708Mitogen-activated protein kinase 3 (EC2..359346/378 (91%)0.0
2.7.1.-) (Extracellular signal- regulated3..380347/378 (91%)
kinase 1) (ERK-1) (Insulin-stimulated
MAP2 kinase) (MAP kinase 1) (MAPK
1) (P44-ERK1) (ERT2) (P44-MAPK)
(Microtubule- associated protein-2
kinase) (MNK1) - Rattus norvegicus
(Rat), 380 aa.
|
[0544] PFam analysis predicts that the NOV44a protein contains the domains shown in the Table 44E.
231TABLE 44E
|
|
Domain Analysis of NOV44a
NOV44aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
pkinase: domain 1 of 142..323 94/302 (31%)2e-78
234/302 (77%)
|
Example 45
[0545] The NOV45 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 45A.
232TABLE 45A
|
|
NOV45 Sequence Analysis
|
|
SEQ ID NO:1112496 bp
NOV45a,TATGTCAGTTTCCCATGGGTCTTGAATGCAAATACAAATATCGTAAACTAAATATTTG
|
CG91911-01 DNA SequenceTGTTTTCTTTCCTAGACTCTCCAGAAAGAGCAACAGTAATGGAGTACATGAGCACTGG
|
AAGTGACAATAAAAGATTGATTTATAATTAAACATTTAAATGTGTCTGATGTA
|
ATAGACATTATGGAAAATCTTTATGCAAGTGAAGAGCCAGCAGTTTATGAACCCAGTC
|
TAATGACCATGTGTCAAGACACTAATCAAAACGATGAGCGTTCTAACTCTCTGCTGCT
|
TAGTGGCCAAAAGGTACCATGGTTGTCATCAGTCAAATACGCAACTGTGGAGGATTTG
|
CTTGCTTTTCAAAACCATATATCCAACACTGCAAACCATTTTTATGTTCAACAACCAC
|
AGGAATATGGTATTTTATTAAACATGGTAATCACTCCCCAAAATQGACGTTACCAAAT
|
AGATTCCGATCTTCTCCTGATCCCCTGGAAGCTGACTTACAGGAATATTGGTTCTGAT
|
TTTATTCCTCGGGCCGCCTTTGGAAAGGTATACTTGGCACAACATATAAAGACGAAGA
|
AAAGAATGGCGTGTAAACTGATCCCAGTAGATCAATTTAAGCCATCTCATGTCGAAAT
|
CCAGGCTTCCTTCCGGCACGAGAACATCGCAGAGCTGTATGGCGCAGTCCTGTGGGGT
|
GAAACTGTCCATCTCTTTATGGAAGCAGGCGAGGGAGGGTCTGTTCTGGAGAAACTGG
|
AGAGCTGTGGACCAATGAGAGAATTTGAAATTATTTGGGTGACAAAGCATGTTCTCAA
|
GGGACTTGATTTTCTACACTCAAAGAAAGTGATCCATCATGATATTAAACCTAGCAAC
|
ATTCTTTTCATGTCCACAAAAACTGTTTTGGTGGATTTTGGCCTAAGTGTTCAAATCA
|
CCGAAGATGTCTATTTTCCTAAGGACCTCCGAGGAACAGAGATTTACATCACCCCAGA
|
GGTCATCCTGTGCAGGGGCCATTCAACCAAAGCAGACATCTACAGCCTGGGCGCCACG
|
CTCATCCACATQCACACGGGCACCCCACCCTGGGTCAACCGCTACCCTCGCTCAGCCT
|
ATCCCTCCTACCTGTACATAATCCACAAGCAAGCACCTCCACTGGAACACATTGCACA
|
TGACTGCAGTCCAGGGATGAGAGAGCTCATAGAAGCTTCCCTGGAGAGAAACCCCAAT
|
CACCGCCCAAGAGCCGCAGACCTACTAAAACATGAGGCCCTGAACCCGCCCAGAGAGG
|
ATCAGCCACGCTCTCAGAGTCTGGACTCTGCCCTCTTGGAGCGCAAGAGGCTGCTGAG
|
TAGGAAGGAGCTGGAACTTCCTGAGAACATTCCTGATTCTTCGTGCACAGGAAGCACC
|
GAGGAATCTCAGATGCTCAAGAGGCAACGCTCTCTCTACATCGACCTCGGCGCTCTGG
|
CTGGCTACTTCAATCTTGTTCGGGAACCACCAACGCTTCAATATGGCTGAAGGATGCC
|
ATGTTTGCTCTAAATTAAGACAGCATTGATCTCCTGGAGGCTGGTTCTGCTGCCTCTA
|
CACAGGGGCCCTGTACAGTGAATGGTCCCATTTTCGAAGGAGCAGTGTGACCTCCTGT
|
GACCCGTCAATGTGCCTCCAAGCCGCCCTGTGTGTTTCACATCTCAACCTATTTGATA
|
TGCACCAGGTCTCAAGGTTCTCATTTCTCAGGTGACGTGATTCTAAGCCAGCAATTTA
|
AGAGTTCACAGAAGGATCGTGTCTGCTGACTGTTTCATTCACTGTGCACTTTGCTCAA
|
AATTTTAAAAATACCAATCACAAGGATAATAGAGTAGCCTAAAATTACTATTCTTGGT
|
TCTTATTTAAGTATGGAATATTCATTTTACTCAGAATAGCTGTTTTGTCTATATTGCT
|
GTATATTATATAACTCTTTGAGCCTTTATTGGTAAATTCTGGTATACATTGAATTCAT
|
TATAATTTGGGTGACTAGAACAACTTGAAGATTGTAGCAATAAGCTGGACTAGTGTCC
|
TAAAAATCGCTAACTGATGAATTAGAAGCCATCTGACACCAGGCCACTAGTGACAGTT
|
TCTTTTGTGTTCCTATGGAAACATTTTATACTGTACATGCTATGCTGAAGACATTCAA
|
AACGTGATGTTTTGAATGTCGATAAAACTGTGTAAACCACATAATTTTTGTACATCCC
|
AAAGGATGAGAATGTGACCTTTAAGAAAAATGAAAACTTTTGTAAATTATTGATGATT
|
TTGTAATTCTTATGACTAAATTTTCTTTTAACCATTTGTATATTAAAATAGCATACTG
|
TGTATGTTTTATATCAAATGCCTTCATGAATCTTTCATACATATATATATTTGTAACA
|
TTGTAAAGTATCTGACTAGTCTTATGTAAAGTATGTTTTTACATTATGCAAATAAAAC
|
CCAATACTTTTCTCCAATGTGCCACGTCAAATCAACTGAATAAATTCAGTATTTTGCC
|
TT
|
ORF Start: ATG at 97ORF Stop: TGA at 1498
SEQ ID NO:112467 aa MW at 53071.2 kD
NOV45a,MEYMSTGSDNKEEIDLLIKHLNVSDVTDIMENLYASEEPAVYEPSLMTMCQDSNQNDE
|
CG91911-01 Protein SequenceRSKSLLLSGQKVPWLSSVKYGTVEDLLAFENHISNTAKHFYVQRPQEYGILLNMVITP
|
QNGRYQTDSDVLLTPWKLTYRNIGSDFIPRGAFGKVYLAQDTKTKRRNACKLIPVDQF
|
KPSDVEIQACFRHENIAELYGAVLWGETVHLFMEAGECCSVLEKLESCGPMREFEIIW
|
VTKHVLKGLDFLHSKKVTHHDIKPSNIVFMSTKPNLVDFGLSVQMTEDVYFPGLRGT
|
EIYMSFEVILCRGHSTKADIYSLGATLIHMQTGTPPWVKRYPRSAYPSYLYIIHKQAP
|
PLEDIAADCSPGMRELIEASLERNPNHRRAADLLKUEALNPPREDQPRCQSLDSALL
|
ERKRLLSRKTELPENLADSSCTGSTEESEMLKRQRSLYIDLGAIAGYFNLVRGPPTL
|
EYG
|
[0546] Further analysis of the NOV45a protein yielded the following properties shown in Table 45B.
233TABLE 45B
|
|
Protein Sequence Properties NOV45a
|
|
PSort0.6500 probability located in cytoplasm; 0.1000 probability
analysis:located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen); 0.0000 probability located in
endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0547] A search of the NOV45a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 45C.
234TABLE 45C
|
|
Geneseq Results for NOV45a
NOV45aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAE10313Human Tp12 protein - Homo sapiens,1..467462/467 (98%)0.0
467 aa. [WO200166559-A1,1..467464/467 (98%)
13 Sep. 2001]
AAE05951Human cot oncoprotein encoded by1..467461/467 (98%)0.0
D14497 oncogene - Homo sapiens,1..467463/467 (98%)
467 aa. [US6265216-B1,
24 Jul. 2001]
AAY79244Human COT - Homo sapiens, 467 aa.1..467461/467 (98%)0.0
[WO200011191-A2, 02 Mar. 2000]1..467463/467 (98%)
AAE10314Rat Tp12 protein - Rattus sp, 467 aa.1..467435/467 (93%)0.0
[WO200166559-A1, 13 Sep. 2001]1..467452/467 (96%)
AAY79243Rat TPL-2 - Rattus norvegicus, 4671..467435/467 (93%)0.0
aa. [WO200011191-A2, 02 Mar. 2000]1..467452/467 (96%)
|
[0548] In a BLAST search of public sequence datbases, the NOV45a protein was found to have homology to the proteins shown in the BLASTP data in Table 45D.
235TABLE 45D
|
|
Public BLASTP Results for NOV45a
NOV45aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
A48713serine/threonine-specific protein kinase1..467462/467 (98%)0.0
cot, 58K form - human, 467 aa.1..467464/467 (98%)
P41279Mitogen-activated protein kinase kinase1..467461/467 (98%)0.0
kinase 8 (EC 2.7.1.-) (COT proto-1..467463/467 (98%)
oncogene serine/threonine-protein
kinase) (C-COT) (Cancer osaka thyroid
oncogene) - Homo sapiens
(Human), 467 aa.
Q63562Mitogen-activated protein kinase kinase1..467435/467 (93%)0.0
kinase 8 (EC 2.7.1.-) (Tumor1..467452/467 (96%)
progression locus 2) (TPL-2) - Rattus
norvegicus (Rat), 467 aa.
Q07174Mitogen-activated protein kinase kinase1..467432/467 (92%)0.0
kinase 8 (EC 2.7.1.-) (COT proto-1..467453/467 (96%)
oncogene serine/threonine-protein
kinase) (C-COT) (Cancer osaka thyroid
oncogene) - Mus musculus (Mouse),
467 aa.
A41253kinase-related transforming protein (EC1..397374/397 (94%)0.0
2.7.1.-) - human, 415 aa.1..397376/397 (94%)
|
[0549] PFam analysis predicts that the NOV45a protein contains the domains shown in the Table 45E.
236TABLE 45E
|
|
Domain Analysis of NOV45a
NOV45aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
pkinase: domain 1 of 1146..388 74/279 (27%)4.7e-54
187/279 (67%)
|
Example 46
[0550] The NOV46 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 46A.
237TABLE 46A
|
|
NOV46 Sequence Analysis
|
|
SEQ ID NO:113568 bp
NOV46aGACGAGCTGATGAGCGTGCGTCTGCGCGAGGCCCAGGCCCTCGCCCAGGGCCGCGAGC
|
CG91931-01 DNA SequenceTGCGGCAGCGCGTGGTGGAACTTGAGACGCAGGTGGACTCCGGGCAGAGGGAGGAGGAAGG
|
CCGCATCCAGGGCCAGCTGAACCACTCGGACTCATCCCAGTACATCCGCGAGCTCAAG
|
GACCAGATCGAGGAGCTGAAGGCCGAGGTGCAGGTGCGGCTGCTGAAGCGCCCGCCGC
|
CCTTCGAGGACCCGCTGGCTTTCGATGGGCTCACCCTGGCGCGCCACTTGGACGAGGA
|
CTCGCTGCCGTCGTCCGACGAGGAGCTACTTGGCGTAGGCGTGGGCGCTGCCCTGCAG
|
GACGCATTCTATCCTCTGTCCCCGCGCCATGCCCGCTTCTTCCGCCGTCTCCAGCGGC
|
CGGCCAAGGACAGCGAGGGCGCAGGACGCATTGTATCCTCTGTCCCCGCGCGATGCGC
|
GCTTCTTCCCCCGTCTGGAGCGGCCGGCCAAGCACAGCGACCGCAGCTCAGACAGCGA
|
CGCCGATGAGCTGGCCGCGCCCTACAGCCAGGGTCTGGACAACTGA
|
ORF Start: ATG at 10ORF Stop: TGA at 529
SEQ ID NO:114173 aa MW at 19187.3 kD
NOV46a,MSVRLREAQALAEGRELRQRVVELETQXIDSCQREEGRIQGQLNISDSSQYIRELKDQI
|
CG91931-01 Protein SequenceEELKAEVQVRLLKGPPPFEDPLAFDGLSLARHLDEDSLPSSDEELLGVGVGAALQDAL
|
YPLSPRDARFFRRLERPAKDSEGAGRIVSSVPARCALLPPSGAACQGQRGQLRQRRR
|
[0551] Further analysis of the NOV46a protein yielded the following properties shown in Table 46B.
238TABLE 46B
|
|
Protein Sequence Properties NOV46a
|
|
PSort0.5680 probability located in mitochondrial matrix space;
analysis:0.2832 probability located in mitochondrial inner
membrane; 0.2832 probability located in mitochondrial
intermembrane space; 0.2832 probability located in
mitochondrial outer membrane
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0552] A search of the NOV46a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 46C.
239TABLE 46C
|
|
Geneseq Results for NOV46a
NOV46aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM84422Human immune/haematopoietic 66..10224/37 (64%)4e-05
antigen SEQ ID NO:12015 - Homo 74..11024/37 (64%)
sapiens, 110 aa.
[WO200157182-A2, 09 Aug. 2001]
AAW33887Flea saliva protein PfspM(B)900 - 2..6624/67 (35%)0.13
Ctenocephalides felis, 900 aa.517..58140/67 (58%)
[WO9737676-A1, 16 Oct. 1997]
AAW82361Flea saliva protein PfspM(B)-900 - 2..6624/67 (35%)0.23
Ctenocephalides sp, 900 aa.517..58140/67 (58%)
[WO9845408-A2, 15 Oct. 1998]
AAY59273Mouse huntingtin-interacting protein 3..8026/84 (30%)0.51
(mHIP1a) - Mus sp, 1068 aa.563..64643/84 (50%)
[WO9960986-A2, 02 Dec. 1999]
AAY93405Amino acid sequence of a filament-like12..6719/61 (31%)0.67
protein 3 - Lycopersicon esculentum,312..37234/61 (55%)
582 aa. [WO200028054-A2,
18 May 2000]
|
[0553] In a BLAST search of public sequence datbases, the NOV46a protein was found to have homology to the proteins shown in the BLASTP data in Table 46D.
240TABLE 46D
|
|
Public BLASTP Results for NOV46a
NOV46aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96CN4SIMILAR TO ECOTROPIC VIRAL1..140121/140 (86%)3e-60
INTEGRATION SITE 5 - Homo644..776 127/140 (90%)
sapiens (Human), 794 aa.
Q9D9F21700084G18RIK PROTEIN - Mus1..142 99/144 (68%)1e-41
musculus (Mouse), 133 aa.1..133107/144 (73%)
O60447EVI-5 HOMOLOG - Homo sapiens1..104 55/106 (51%)5e-19
(Human), 810 aa.648..746 70/106 (65%)
P97366EVI-5 - Mus musculus (Mouse),1..104 53/106 (50%)5e-18
809 aa.648..746 70/106 (66%)
Q19101HYPOTHETICAL 53.2 KDA1..64 25/66 (37%)0.006
PROTEIN - Caenorhabditis elegans,266..331 41/66 (61%)
466 aa.
|
[0554] PFam analysis predicts that the NOV46a protein contains the domains shown in the Table 46E.
241TABLE 46E
|
|
Domain Analysis of NOV46a
NOV46aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
No Significant Matches Found
|
Example 47
[0555] The NOV47 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 47A.
242TABLE 47A
|
|
NOV47 Sequence Analysis
|
|
SEQ ID NO:1151387 bp
NOV47aCGCCACTCTGGCCACTGGCCGGCCATGCCGTCCCGGGCTGAGAACTATGAAGTGTTGT
|
CG91941-01 DNA SequenceACACCATTGGCACAGGCTCCTATGGCCGTGCCAGAAGATCCAGCGGAAAGAGTGACGG
|
CAAGATATTAGTTTGGAAAGAACTTGATTATGCCTTCATCACAGAAGCTGAGAAACAG
|
ATGCTTATTTCTGAAGTGAATTTGCAACGTCAACTGAAACATCCAAACATCGTTCATT
|
ACTATGATCGTATTATTGACCGGACCAACACAACACTGTACGTTGTAATGGAATATTG
|
TGAAGGAGGGGATCTGGCTAGTGTAATTACAAAGCGAACCATGGAAAGGCAATACTTA
|
GATCAACAGTTTGTTCTTCGAGTGATGACTCAGTTGACTCTGGCCCTGAACGAATGCC
|
ACAGACGAAGTGGTGGTGATCATACTGTATTGCGTGGGGATCTGAAACCAGCCAATGT
|
TTTCCTGGATGGCAAGCAAACGTCAAGCTTGGAGATTTGGGGGCTAGCCAGAATATTA
|
AACCACGACACGAGTTTTGCAAAAACATTTGTTGGCATACCTTATTACATGTCTCCTG
|
AACAAATGAATCGCATGTCCTACAATGAGAAACCAGATATCTGGTCATTGGGCTGCTT
|
GCTGTATGAGTTATGTGCATTAATGCCTCCATTTACAGCTTTTAGCCAGAAAGAACTC
|
GCTGGGAAAATCACAGAAGGCAAATPCACGCGAATTCTATACCCTTACTCTGATGAAT
|
TGAATGAAATTATTATGAGGATGTTAAACTTAAAGGATTACCATCAACCTTCTGTTGA
|
AGAAATTCTTGAGAACCCTTTAATAGCAGATTTGGTTGCACAAGAGCAAAGAAGAAAT
|
CTTGAGACAAGAGGGCGACAATTAGGAGACCCAGAAAAATTGCTGGATTCCAGCCCTG
|
TATTGAGTGAGCTGAAACTAAAGGAAATTCAGTTAGAGGAGCAAGACCGAGCTCTCAA
|
AGCAGGACAAGAAACATTGCAGCAGAAAGAACAGCAGCTTTGTGTTTCTGACAGACTA
|
GCAGAGCACAGACTGGCTATACCAGAAAATCTGTTCAACAATTACAGCTTGCTAAAGG
|
AACAGAAGTTCCTCTCTCTGCCAAGTAGTCCAGAACTTCTTAATCTTCCATCCTCAGT
|
AATTAAGAAGAAACTTCATTTCAGTCGGGAAAGTAAACAGAATGTCATGAGCAGTGAG
|
AATCCTCAGAGTCAGCTCACATCTAAGTCCAAGTGCAACGACCTGAAGTGCTTCATGC
|
TTCATGCTGCCCAGCTGCCGGCTCAAGCCCTCTCAGATATTAAGAAAAATTACCAACT
|
GAAAAGCAGACAGATCCTGGGCATCCGCTAGCCAGGTAGAGAGACACAGAGCT
|
ORF Start: ATG at 25ORF Stop: TAG at 1363
SEQ ID NO:116446 aa MW at 51598.9 kD
NOV47a,MPSRAENYEVLYTIGTGSYGRCQKIQRKSDCKRLVWKELDYGFMTEAEKQMLISEVNL
|
CG91941-01 Protein SequenceLRELKHPNIVHYYDRIIDRTNTTLYVVMEYCEGCDLASVITKGTMERQYLDEEFVLRV
|
MTQLTLALKECHRRSGGDHTVLRGDLKPANVFLDGKQNVKLGDLGLPRILNHDTSFAK
|
TFVGIPYYMSPEQMNRMSYNEKPDIWSLGCLLYELCALMPPFTAFSQKELACKIREGK
|
FRRILYRYSDELNEIIMRILNLKDYHQPSVREILENPLIADLVAAEQRRNLERECRQL
|
GEPEKLLDSSFVLSELKLKEIQLREQERALKACRERLEQKEQELCVCERLAEDRLAIP
|
ENLLKNYSLLKEQKFLSLASSPELLNLPSSVIKKKVHFSGESKENVMRSEEPESQLTS
|
KSKCKDLKCFMLHAAQLRAQALSDIEKNYQLKSRQILGMR
|
[0556] Further analysis of the NOV47a protein yielded the following properties shown in Table 47B.
243TABLE 47B
|
|
Protein Sequence Properties NOV47a
|
|
PSort0.6500 probability located in cytoplasm; 0.1000 probability
analysis:located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen); 0.0000 probability located in
endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0557] A search of the NOV47a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 47C.
244TABLE 47C
|
|
Geneseq Results for NOV47a
NOV47aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAU03547Human protein kinase #47 - Homo1..446421/448 (93%)0.0
sapiens, 446 aa. [W0200138503-A2,1..446427/448 (94%)
31 May 2001]
AAY92330Human N1K1 protein - Homo1..446412/446 (92%)0.0
sapiens, 445 aa. [WO200020448-A2,1..445424/446 (94%)
13 Apr. 2000]
AAY59148Human NEK2 protein fragment -1..336313/336 (93%)0.0
Homo sapiens, 336 aa.1..336321/336 (95%)
[WO9966051-A2, 23 Dec. 1999]
AAY59147E. nidulans NIMA protein fragment -3..316131/338 (38%)4e-61
Emericella nidulans, 360 aa.6..342189/338 (55%)
[WO9966051-A2, 23 Dec. 1999]
AAU03545Human protein kinase #45 - Homo6..275106/272 (38%)9e-48
sapiens, 649 aa. [WO200138503-A2,2..263158/272 (57%)
31 May 2001]
|
[0558] In a BLAST search of public sequence datbases, the NOV47a protein was found to have homology to the proteins shown in the BLASTP data in Table 47D.
245TABLE 47D
|
|
Public BLASTP Results for NOV47a
NOV47aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P51955Serine/threonine-protein kinase NEK21..446412/446 (92%)0.0
(EC 2.7.1.-) (NimA-related protein1..445424/446 (94%)
kinase 2) (NimA-like protein kinase 1)
(HSPK 21) - Homo sapiens (Human),
445 aa.
Q91Z18NIMA (NEVER IN MITOSIS GENE1..446367/446 (82%)0.0
A)-RELATED EXPRESSED KINASE1..443403/446 (90%)
2 - Mus musculus (Mouse), 443 aa.
Q921N9SIMILAR TO NIMA (NEVER IN1..446366/446 (82%)0.0
MITOSIS GENE A)-RELATED1..443402/446 (90%)
EXPRESSED KINASE 2 - Mus
musculus (Mouse), 443 aa.
O35942Serine/threonine-protein kinase NEK21..446365/446 (81%)0.0
(EC 2.7.1.-) (NimA-related protein1..443401/446 (89%)
kinase 2) - Mus musculus (Mouse),
443 aa.
Q96QN9NEK2B PROTEiN KINASE - Homo1..370341/370 (92%)0.0
sapiens (Human), 384 aa.1..370352/370 (94%)
|
[0559] PFam analysis predicts that the NOV47a protein contains the domains shown in the Table 47E.
246TABLE 47E
|
|
Domain Analysis of NOV47a
NOV47aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
pkinase: domain 1 of 18..271 90/302 (30%)2.5e-57
204/302 (68%)
|
Example 48
[0560] The NOV48 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 48A.
247TABLE 48A
|
|
NOV48 Sequence Analysis
|
|
SEQ ID NO:1171432 bp
NOV48a,CGTGACTCTGGCGACTGGCTGGCCATGCCCTCCCGGGCTCAGAACTATGAAGTGTTGG
|
CG91951-01 DNA SequenceACACCATTGGCACAGGCTCCTCTGGCCGCTGCCAGAAGATCCAGAGGAAGAGTGACGG
|
CAAGAAACTAGCTTGGAAAGAACTTGATTATGGCTTCATGACAGAAGCTGACAAACAG
|
ATGCTTATTTCTGAAGTGAATTTGCTTTGTAAACTGAAAAATCCAAACATCGTTCATT
|
ACTATCATCGTATTATTGACCGGACCAACACAACACTGTACATTCTAATGGAATATTG
|
TGAAGAAGGAGACCTGGCTAGTGTAATTACAAAGGGAACCAAGGAAAGGCAATACTTA
|
GATGAAGGAAGTTTGTCTTCGAGTGACGACTCAGTTGATCTGGCCCTGAAGGAATGCC
|
ACAGACGAAGTGGTCCTGATCATACTGTAGTCCGTCGGGGTCTGAAACCAGCCAGTGT
|
TTTCCTGGATCGCAAGCAAAACGTCAAGCTTGGAGATTTGGGGCTAGCCAGAATATTA
|
AACCACCACACGACTTTTGCAAAAACATTTGTTCGCATACCTTATTACATGTCTCCTG
|
AACAAACGAATCACATGTCCTACAATGAGAAACCAGATATCTGGTCATTGGGCTGCTT
|
GCCGTATGAGTCGCGTGCATTAATGCCTCCATTTACAGCTTTTAGCCAGAAAGAACTC
|
GCTGGGAAAATCAGACAAGGCAAATTCAGGCGAATTCTATACCATTACTCTGATGAAT
|
TGAATGAAATTATTATGAGGATGTTAAAGGATTACCATCGACCTTCTGTTGAAGAAAT
|
TCTCGAGAACCCTTTAATAGCAGATTTGGTTGCAGAAGACCAAAGAACAAATCTTGAG
|
AGAAGAGGGCGACAATTAGGAGAGCCAGAAAAATTGCCGGATTCCAGCCTTGTATTGA
|
GTGAGCTGAACTAAAGGAAATTCAGTTACAGGAGCAAGAGCGAGCTCTCAAAGCAGAG
|
ACAAGAAACGTTCGAGCACAAAGAACAGGAGCTTTGTGTTTGTGAGAGAAPAGCAGAG
|
GACAGACTGGCTATACCAGAAAATCTGTTGAACAATTACAGCTTGCTAAAGGAACAGA
|
AGTTCCTGTCTCTGGCAAGTAGTCCAGAACTTCTTAATCTTCCATCCTCAGTAATTAA
|
GAAGAAAGTTCATTTCAGTGGGGAAAGTAAAGAGAATGTCATGAGGAGTGAGAATTCT
|
GAGAGTCAGCTCACATCTAACTCCAAGTGCAACGACCTGAAGCTCCTTGCTTCATGCT
|
GTCATGCTGCCCAGCGTGCAAGGACCTGAAGGTCCTTGCTTCATGCTTCATGCTCCCCA
|
ATCCTGGGCATGCGCTAGCCGGGTAGAGAGACACAGAGCT
|
ORF Start: ATG at 25ORF Stop: TAG at 1408
SEQ ID NO:118461 aa MW at 52904.2 kD
NOV48aMPSRAENYEVLDTIGTGSSGRCQKIQRKSDGKLLAWKELDYCFMTEAEKQMLTSEVNL
|
CG91951-01 Protein SequenceLCKLKNPNIVHYYDRTIDRTNTTLYIVMEYCEEGDLASVTTKGTKBRQYLDEEFVLRV
|
TTQLTLALKECHRRSGGDHTVVRRGLKPASVPLDGKQNVKLGDLGLARTLNBDTSFAK
|
TFVGTFYYDSPEQThHNSYNEKPDIWSLGCLPYESRALMPPFTAFSQKELACKIREGK
|
FRRILYHYSDELNEIIMRMLKDYHRPSVEEILENPLIADLVAEEQRRNLERRGRQLGE
|
PEKLPDSSLVLSELKLKETQLEEQERALKAGEERLEQKEQELCVCERLAEDRLATPEN
|
LLKNYSLLKEQKFLSLASSPELLNLPSSVIKKKVHFSGESKENVNRSENSESQLTSKS
|
KCKDLKVLASCFMLPSVQGPEGPCFMLHAAQLRAQALSDFEKNYQLKSRQILGMR
|
[0561] Further analysis of the NOV48a protein yielded the following properties shown in Table 48B.
248TABLE 48B
|
|
Protein Sequence Properties NOV48a
Psort0.6500 probability located in cytoplasm; 0.1000 probability
analysis:located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen); 0.0000 probability located
in endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0562] A search of the NOV48a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 48C.
249TABLE 48C
|
|
Geneseq Results for NOV48a
NOV48aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAU03547Human protein kinase #47 - Homo1..461429/461 (93%)0.0
sapiens, 446 aa. [WO200138503-A2,1..446431/461 (93%)
31 May 2001]
AAY92330Human N1K1 protein - Homo1..461396/463 (85%)0.0
sapiens, 445 aa. [WO200020448-A2,1..445410/463 (88%)
13 Apr. 2000]
AAY59148Human NEK2 protein fragment -1..334297/336 (88%)e-166
Homo sapiens, 336 aa. [WO9966051-1..336307/336 (90%)
A2, 23 Dec 1999]
AAY59147E. nidulans NIMA protein fragment -3..314126/338 (37%)4e-55
Emericella nidulans, 360 aa.6..342185/338 (54%)
[WO9966051-A2, 23 Dec. 1999]
AAU07102Human novel human protein, NUP6..276106/282 (37%)7e-42
#2 - Homo sapiens, 1214 aa.2..274148/282 (51%)
[WO200161016-A2, 23 Aug. 2001]
|
[0563] In a BLAST search of public sequence datbases, the NOV48a protein was found to have homology to the proteins shown in the BLASTP data in Table 48D.
250TABLE 48D
|
|
Public BLASTP Results for NOV48a
NOV48aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P51955Serine/threonine-protein kinase NEK21..461396/463 (85%)0.0
(EC 2.7.1.-) (NimA-related protein1..445410/463 (88%)
kinase 2) (NimA-like protein kinase 1)
(HSPK 21) - Homo sapiens (Human),
1445aa.
Q91Z18NIMA (NEVER IN MITOSIS GENE1..461354/463 (76%)0.0
A)-RELATED EXPRESSED KINASE1..443391/463 (83%)
2 - Mus musculus (Mouse), 443 aa.
O35942Serine/threonine-protein kinase NEK21..461353/463 (76%)0.0
(EC 2.7.1-) (NimA-related protein1..443390/463 (83%)
kinase 2) - Mus musculus (Mouse), 443
aa.
Q921N9SIMILAR TO NIMA (NEVER IN1..461353/463 (76%)0.0
MITOSIS GENE A)-RELATED1..443390/463 (83%)
EXPRESSED KINASE 2 - Mus
musculus (Mouse), 443 aa.
Q96QN9NEK2B PROTEIN KINASE - Homo1..368325/370 (87%)0.0
sapiens (Human), 384 aa.1..370338/370 (90%)
|
[0564] PFam analysis predicts that the NOV48a protein contains the domains shown in the Table 48E.
251TABLE 48E
|
|
Domain Analysis of NOV48a
Identities/
NOV48aSimilarities
Matchfor theExpect
Pfam DomainRegionMatched RegionValue
|
pkinase: domain 1 of 18..269 87/302 (29%)1.5e-42
196/302(65%)
|
Example 49
[0565] The NOV49 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 49A.
252TABLE 49A
|
|
NOV49 Sequence Analysis
|
|
SEQ ID NO:1191861 bp
NOV49a,AGCCATGAACAAGAAGTTAGTGGTGCTCCCCCTGCTCGCCGTGGTCCTGGTGCTCGTC
|
CG92025-01 DNA SequenceATTCTCGCCCTCTCTCTCTGGCTGCCCTCGGCCTCCAAGGAACCTGACAACCATOTGT
|
ACACCAGGGCTGCCATGGCCGCGGATCCCAAGCAGTGCTTCGAGATTCGGAGGGGCAG
|
GGACACACTGCCGCACGGTGGCTCTGCAGTGGATGGAGCCATTGCAGCCCTGTTGTGT
|
GTGGGGCTCATGAATGCCCACAGCATGGGCATCGGGGTTGGCCTCTTCCTCACCATCT
|
ACAACACCACCACTCATGAGGTCGGTGGGCCTGCCTACCTGCTTCTTCTAGGAAAAGC
|
TGAGCTCATCAATCCCCGCGAGGTGGCCCCCAGGCTGGCCTTTGCCAGCATGTTCAAC
|
AGCTCGCAGCAGTCCCAGAAGGGAGGCCTGTCGGTGGCGGTGCCTGGGGAGATCCGAG
|
GCTATGAGCTCGCACACCACCGGCATCGCCGGCTGCCCTGGGCTCGCCTCTTCCAGCC
|
CAGCATCCAGCTCGCCCGCCAGGGCTTCCCTGTGGGCAAGGGCTTGGCGGCAGTCCTG
|
GAAAACAACCGGACTGTCATCCAQCAGCAGCCTGTCTTGCTCTGCCCCGGTGAGATGT
|
TCTGCCGGGATAGAAAGGTGCTTCGGGAGGGGGAGAGACTGACCCTGCCGCGGCTGGC
|
TGACACCTATGAGATGCTCGCCATCGAGGGTGCCCAGGCCTTCTACAACGGCAGCCTC
|
ATGCCCCACATTCTGAAGCACATCCAGGCCGCTGGTGACTCGGTAACCTCAGCAGGGG
|
GCATTGTGACAGCTGAGCACCTGAACAAGTACCGTGCTGAGCTGATCGAGCACCCGCT
|
GAACATCACCCTGGGAGACGCGGTGCTGTACATGCCCAGTGCGCGCCTCAGCCGGCCC
|
GTGCTGCCCCTCATCCTCAACATCCTCAAAGGTCGGTACAACTTCTCCCGGGAGAGCC
|
TGGAGACCCCCGAGCAGAAGGGCCTCACGTACCACCGCATCGTAGAQGCTTTCCGGTT
|
TGCCTACGCCAAGAGGACCCTGCTTCGGGACCCCAAGTTTGTGGATGTGACTGAGGTA
|
CAGGTGGTCCGCAACATGACCTCTCAGTTCTTCGCTGCCCAGCTCCGGTCCCAGATCT
|
CTGACCACACCACTCACCCGATCTCCTACTACAAGCCCGAGTTCTACACGCCGGATGA
|
CGGGGGCACTGCTCACCTGTCTGTCGTCGCAGAGGACGGCAGTGCTGTGTCCGCCACC
|
AGCACCATCAACCTCAGCTTTGGCTCCAACGTCTGCTCCCCCGTCAGTGGGATCCTGT
|
TCAATAATGAATGGACGACTTCAGCTCTCCCAGCATTCACCAATGAGTTTGGGGCACC
|
CCCCTCACCTGCCAATTTCATCCAGCCAGGTGGCAAGCAGCCGCTCTTGTCCATGTGC
|
CTGACGATCATGGTGGGCCAGGACGGCCAGGTCCGGATGGTGGTGGGAGCTGCTGGGG
|
GCACGCAGATCACCACAGACACTGCACTGGTATGTGTCGCCATCATCTACAACCTCTG
|
GTTCGGCTATGACGTGAAGAGGGCCGTGGAGGAGCCCCGGCTGCACAACAAGCTTCTG
|
CCCAACGTCACGACAGTGGAGAGAAACATTGACCAGGTGGGCCAGGGGTTCGAGAAAC
|
TGAGTCACCAGGCAGTGACTGCACCCCTGGAGACCCGGCACCATCACACCCAGATCCC
|
GTCCACCTTCATCGCTGTGGTGCAAGCCATCGTCCGCACGGCTGGTGGCTGGGCAGCT
|
GCCTCGGACTCCAGGAAAGGCGGGGAACCTGCTGGCTACTGATTGCTCCAGGCAGACA
|
AGGCT
|
ORF Start: ATG at 5ORF Stop: TGA at 1838
SEQ ID NO:120611 aa MW at 65845.9 kD
NOV49a,MKKKLVVLCLLAVVLVLVIVGLCLWLPSASKEPDNHVYTRAAMAADAKQCLETCRGRD
|
CG92025-01 Protein SequenceTLRDGGSAVDAAIAALLCVGLMNAESMGTGVGLFLTIYNSTTHEVGGPAYLLLLGKAE
|
VINAREVAPRLAFASMFNSSEQSQKGGLSVAVPGEIRGYELAHQRHGRLPWARLFQPS
|
IQLARQGFPVGKGLAAVLENKRTVIEQQPVLLCPGEVFCRDRKVLREGERLTLPRLAD
|
TYEMIAIEGAQAFYNGSLMAQIVKDTQAAGEWVTSAGCIVTAEDLNNYRAELIEHPLN
|
ISLGDAVLYMPSARLSGPVLALILNILKGGYMFSRESVETPEQKGLTYHRIVEAFRFA
|
YAKPTLLGDPKFVDVTEVQVVRNNTSEFFAAQLRSQISDHTTHPISYYKPEFYTPDDG
|
GTAHLSVVAEDCSAVSATSTINLSFGSKVCSPVSGILFNNEWTTSALPAFTNEFGAPP
|
SPANFIQPCCKQPLLSMCLTIMVGQDGQVRMVVGAAGGTQITTDTALVCVAIIYNLWF
|
GYDVKRAVEEPRTNKLLPNVTTVERNIDQVGQGLEKLSITQAVTAALETRHEETQIAS
|
TFIAVVQAIVRTAGGWAAASDSRKGCEPAGY
|
SEQ ID NO:1212089 bp
NOV49bCGGGGCAAGTGAGGTGCTGCCGTCATCCACGCTGGACAGTTCAGTGATTTGCCTGAGG
|
CG92025-02 DNA SequenceCCCACAGCAGAGTTCAACTGGAGACAGAGAAACCAGCTAGACGCAGAGGGACGTAAC
|
ACGGAGTCCCCCACAAAGGTCTGGGCTCCGCGTGCTTCAGGTAACCTCCCTTGACCTT
|
CAGGAGAACGAGAAGGCTGCCTGATCAGAGAGTCCCTGAAGAAGATTCTGTGGCTACA
|
GGCTTCAGCAGAGTGTGAGCGAGACCCCGGTTATTTCCTCAGCTATTTCCACCAAATC
|
CTCCTGTCTTTCGTGGCCAACACCCCAGGCAACCCTTGGCGCCCCCGTCTGCTCCTGG
|
ACGCAGAGCCATGAAGAAGAACTTAGTGGTCCTGGGCCTGCTGGCCGTCGTCCTGGTG
|
CTGGTCATTCTCGGCCTCTGTCTCTGGCTGCCCTCAGCCTCCAAGGAACCTGACAACC
|
ATGTGTACACCACGGCTGCCGTCGCCGCGGATGCCAAGCAGTGCTCGAAGATTGGGAG
|
GGATGCACTGCGGGACCGTGGCTCTGCGGTGGATGCAGCCATTGCAGCCCTGTTGTGT
|
GTGGGGCGCATGAATGCCCACAGCATGGGCATCGGGGGTGGCCTCTTCCTCAGCATCT
|
ACAACAGCACCACACGAAAAGCTGAGGTCATCAACGCCCGCGAGGTGGCCCCCACGCT
|
GGCCTTTGCCACCATGTTCAACAGCTCCGAGCAGTCCCAGAAGCGGGGGCTGTCGGTG
|
GCGGTGCCTGCGCAGATCCGAGGCTATCAGCTCGCACACCAGCCGCATGCGCCGCTCC
|
CCTGGGCTCGCCTCTTCCAGCCCAGCATCCAGCTGGCCCGCCAGGGCTTCCCCGTGGG
|
CAAGGGCTTGGCCGCAGCCCTGGAAAACAAGCGGACCGTCATCCAGCAGCAGCCTGTC
|
TTGTGTGAGGTGTTCTGCCGGGATAGAAACGTGCAAGCGGAGGGGGAGAGACTGACCC
|
TGCCCCAGCTGGCTGACACCTACGACACGCTGGCCATCGAGGGTGCCCAGGCCTTCTA
|
CAACGGCAGCCTCACGGCCCAGATTGTGAAGGACATCCAGGCGGCCGGGGGCATTGTG
|
ACAGCTGAGGACCTGAACAACTACCGTGCTGACCTGATCGAGCACCCGCTGAACATCA
|
GCCTGGGAGACGCGGTGCTGTACATGCCCAGTGCGCCGCTCAGCGGGCCCGTGCTGGC
|
CCTCATCCTCAACATCCTCAAAGGGTACAACTTCTCCCGGGAGAGCGTGGAGAGCCCC
|
GAGCAGAAGCGCCTGACGTACCACCGCATCGTAGAGGCTTTCCGGTTTGCCTACGCCA
|
AGAGGACCCTGCTTCGGGACCCCAACTTTGTGGATCTCACTCAGGTCGTCCGCAACAT
|
GACCTCCGACTTCTTCGCTGCCCAGCTCCGGGCCCAGATCTCTGACGACACCACTCAC
|
CCGATCTCCTACTACAAGCCCGAGTTCTACACGCCGGATGACGGGGGCACTGCTCACC
|
TGTCTGTCCTCGCAGACGACGGCAGTGCTGTGTCCGCCACCAGCACCATCAACCTCTA
|
CTTTGGCTCCAAGGTCTGCTCCCCGGTCAGTGGGATCCTGTTCAATAATGAATGGACG
|
ACTTCAGCTCTCCCAGCATTCACCAATGAGTTTGGGCCACCCCCCTCACCTGCCAATT
|
TCATCCAGCCAGGGAAGCAGCCGCTCTTGTCCATGTGCCTCACGATCATGGTGGGCCA
|
GGACGGCCAGGTCCGGATGGTGGTGCGAGCTGCTGGGGGCACGCAGATCACCACAGAC
|
ACTGCACTGGCCATCATCTACAACCTCTCGTTCGGCTATGACGTCAAGAGGGCCGTGC
|
AGGAGCCCCGGCTGCACAACAAGCTTCTGCCCAACGTCACGACAGTGGAGAGAAACAT
|
TGACCAGGCAGTGACTGCAGCCCTGGACACCCGGCACCATCACACCCAGATCCCGTCC
|
ACCTTCATCGCTGTGGTGCAAGCCATCGTCCGCATGGCTGGTGGCTGGGCAGCTGCCT
|
CGGACTCCAGGAAAGGCGGCGAACCTCCTGGCTACTGATTGCTCCAGGCAGACAAGGC
|
T
|
ORF Start: ATG at 359ORF Stop: TGA at 2066
SEQ ID NO:122569 aa MW at 61411.7 kD
NOV49b,MKKKLVVLGLLAVVLVLVIVGLCLWLPSASKEPDNHVYTRAAVAADAKQCSKIGRDAL
|
CG92025-01 Protein SequenceRDGGSAVDAAIAALLCVCLMNAIISMCIGGGLFLTIYNSTTRKAEVINAREVAPRLAFA
|
TMFNSSEQSQKGGLSVAVPGEIRGYELAHQRHGRLPWARLFQPSIQLARQGFFPVGKGL
|
AAALENKRTVIEQQPVLCEVFCRDRKVLREGERLTLPQLAETYETLAIEEGAQAFYNGS
|
LTAQIVKDIQAAGGIVTAEDLNNYRAELIEHPLNISLGDAVLYMPSAPLSGPVLALIL
|
NTLKGYNFSRESVESPEQKGLTYHRIVEAFREAYAKRTLLGDPKFVDVTEVVRNMTSE
|
FFAAQLRAQISDDTTHPISYYKPEFYTPDDGGTAHLSVVAEDGSAVSATSTINLYFGS
|
KVCSPVSGILPNNEWTTSALPAFTNEFGAPPSPANFIQPGKQPLLSMCLTIMVGQDGQ
|
VRNVVGAAGGTQITTDTALAIIYNLWFGYDVEAVEEPRLHNKLLPNVTTVERNIQDQA
|
VTAALETRHHHTQIASTFTAVVQAIVRMAGGWAAASDSRKGGEPAGY
|
[0566] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 49B.
253TABLE 49B
|
|
Comparison of NOV49a against NOV49b.
ProteinNOV49a Residues/Identities/Similarites
SequenceMatch Residuesfor the Matched Region
|
NOV49b27..611525/585 (89%)
27..569531/585 (90%)
|
[0567] Further analysis of the NOV49a protein yielded the following properties shown in Table 49C.
254TABLE 49C
|
|
Protein Sequence Properties NOV49a
|
|
PSort0.4600 probability located in plasma membrane;
analysis:0.1305 probability located in microbody (peroxisome);
0.1000 probability located in endoplasmic reticulum
(membrane); 0.1000 probability located in endoplasmic
reticulum (lumen)
SignalPCleavage site between residues 32 and 33
analysis:
|
[0568] A search of the NOV49a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 49D.
255TABLE 49D
|
|
Geneseq Results for NOV49a
NOV49aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABG30234Novel human diagnostic protein 1..611538/611 (88%)0.0
#30225 - Homo sapiens, 605 aa.37..605547/611 (89%)
[WO200175067-A2, 11 Oct. 2001]
ABG30234Novel human diagnostic protein 1..611538/611 (88%)0.0
#30225 - Homo sapiens, 605 aa.37..605547/611 (89%)
[WO200175067-A2, 11 Oct. 2001]
ABG30235Novel human diagnostic protein 1..611537/616 (87%)0.0
#30226 - Homo sapiens, 623 aa.48..623546/616 (88%)
[WO200175067-A2, 11 Oct. 2001]
ABG30235Novel human diagnostic protein 1..611537/616 (87%)0.0
#30226 - Homo sapiens, 623 aa.48..623546/616 (88%)
[WO200175067-A2, 11 Oct. 2001]
ABG30240Novel human diagnostic protein 1..611538/616 (87%)0.0
#30231 - Homo sapiens, 639 aa.64..639546/616 (88%)
[WO200175067-A2, 11 Oct. 2001]
|
[0569] In a BLAST search of public sequence datbases, the NOV49a protein was found to have homology to the proteins shown in the BLASTP data in Table 49E.
256TABLE 49E
|
|
Public BLASTP Results for NOV49a
NOV49aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P19440Gamma-glutamyltranspeptidase 11..611538/611 (88%)0.0
precursor (EC 2.3.2.2) (Gamma-1..569547/611 (89%)
glutamyltransferase 1) - Homo sapiens
(Human), 569 aa.
P20735Gamma-glutamyltranspeptidase1..611464/611 (75%)0.0
precursor (EC 2.3.2.2) (Gamma-1..568505/611 (81%)
glutamyltransferase) (GGT) - Sus
scrofa (Pig), 568 aa.
P07314Gamma-glutamyltranspeptidase1..611436/611 (71%)0.0
precursor (EC 2.3.2.2) (Gamma-1..568503/611 (81%)
glutamyltransferase) (GGT) - Rattus
norvegicus (Rat), 568 aa.
A05225gamma-glutamyltransferase (EC1..611435/611 (71%)0.0
2.3.2.2) precursor - rat, 568 aa.1..568501/611 (81%)
Q60928Gamma-glutamyltranspeptidase1..611436/611 (71%)
precursor (EC 2.3.2.2) (Gamma-1..568498/611 (81%)
glutamyltransferase) (GGT) - Mus
musculus (Mouse), 568 aa.
|
[0570] PFam analysis predicts that the NOV49a protein contains the domains shown in the Table 49F.
257TABLE 49F
|
|
Domain Analysis of NOV49a
Identities/
NOV49aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
G_glu_transpept:57..607253/618 (41%)6.2e-227
domain 1 of 1486/618 (79%)
|
Example 50
[0571] The NOV50 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 50A.
258TABLE 50A
|
|
NOV50 Sequence Analysis
|
|
SEQ ID NO:1232014 bp
NOV50a,GCAGCATGAGCCGATCACCCCTCAATCCCAGCCAACTCCGATCAGTGCGCTCCCAGGA
|
CG92078-01 DNA SequenceTGCCCTGGCCCCCTTGCCTCCACCTCCTCCCCAGAATCCCTCCACCCACTCTTCGGAC
|
CCTTTGTGTGGATCTCTGCCTTGGGGCCTCAGCTGTCTTCTGGCTCTGCAGCATGTCT
|
TGGTCATGGCTTCTCTGCTCTGTGTCTCCCACCTCCTCCTGCTTTGCAGTCTCTCCCC
|
AGGAGGACTCTCTTACTCCCCTTCTCAGCTCCTGGCCTCCAGCTTCTTTTCATGTGGT
|
ATGTCTACCATCCTGCAAACTTGGATCCGCAGCAGGAGCCTGCCTCTTCTCCAGGCTC
|
CATCCTTAGACTTCCTTATCCCTGCTCTGGTGCTGACCAGCCAGAAGCTACCCCGGGC
|
CATCCAGACACCTGGAAACGCCTCCCTCATGCTGCACCTTTGTAGGCGACCTAGCTGC
|
CATGGCCTGGGGCACTGGAACACTTCTCTCCAGGAGCTGCTGGTAGTATCTGCGCTGC
|
TGCAGGCCATGATGGGGCTGCTGGGGAGTCCCGGCCACCTGTTCCCCCACTGTGGGCC
|
CCTGGTGCTGGCTCCCAGCCTGGTTGTGGCACGGCTCTCTGCCTTTCCCCAACAGGGA
|
GTTTCCTCCTCTCACCCAGCCTGGACTGCAATGCCGATCTCGGCTCACCACTGGCTTT
|
TGGCTTCTGTCTACCCCTGCAAGGCTGGCTCAGAAGGTTCTGGGGGAGGAGTTCTTTT
|
CTCACTCTCGCCCCTCAGGTGCTGATCCCAGTGGCCTGTGTCTCGATTGTTTCTGCCT
|
TTGTGGGATTCACTGTTATCCCCCAGGAACTGTCTGCCCCCACCAAGGCACCATGGAT
|
TTGGCTGCCTCACCCAGGTGTCTGGAATTGGCCTTTGCTGACGCCCAGAGCTCTGGCT
|
GCAGGCATCTCCATGGCCTTGGCAGCCTCCACCAGTTCCCTGGGCTGCTATGCCCTGT
|
GTGGCCGGCTGCTGCATTTGCCTCCCCCACCTCCACATGCCTGCAGTCCAGGGCTGAG
|
CCTGGAGGGGCTGGGCACTGTGCTGGCCGCGCTCCTGGGAAGCCCCATGGGCACTGCA
|
TCCAGCTTCCCCAACGTGGGCAAAGTGGGTCTTATCCAGCAGGCTGGATCTCAGCAAG
|
TGGCTCACTTAGTGGGGCTACTCTGCGTGCCGCTTGGACTCTCCCCCAGGTTGGCTCA
|
GCTCCTCACCACCATCCCACTGCCTGTTGTTGGTGGTGGGCTGCTGGGGGTGACCCAG
|
GCTGTGGTTTTGTCTGCTGGATTCTCCAGCTTCTACCTGGCTGACATAGACTCTGGGC
|
GAAATATCTTCATTGTGGGCTTCTCCATCTTCATGGCCTTGCTGCTGCCAAGATGGTT
|
TCGGGAAGCCCCAGTCCTGTTCACCACAGGTCACTCACTGCTGATCGAGCCCCTTGGA
|
TGTATTACTGCACAGCCCATCTTCCTGGCTGGACTCTCAGGCTTCCTACTAGACAACA
|
CGATTCCGGGCACACAGCTTGAGCGAGGCCTAGGTCAAGGGCTACCATCTCCTTTCAC
|
TGCCCAAGAGGCTCGAATGCCTCACAACCCCACCGACAAGGCTGCTCAAGTGTACAGA
|
CTTCCTTTCCCCATCCAAAACCTCTGTCCCTGCATCCCCCAGCCTCTCCACTGCCTCT
|
GCCCACTGCCTGAAGACCCTGGGGATGAGGAAGGAGGCTCCTCTGAGCAGCAAGAGAT
|
GGCAGACTTCCTGCGTGGCTCAGGGGAGCATGCCCTGAATCTAGCAGAGAAGGGTTTA
|
GGTCCAGAAATGACCAGAACGCGTACTTCTGCCCTGGTTAATTTAGCCCTAACTCTCA
|
TCTGCTGGAGAGTCAGCTCCCAAACTGTTCTTTCGTGTAGGCAGAGGATATCTCTGTG
|
TGTATTACATCGGACTGTCTAGAGGTTCCATTTCCCAATAGG
|
ORF Start: ATG at 6ORF Stop: TGA at 1992
SEQ ID NO:124662 aa MW at 70138.5 kD
NOV50a,MSRSPLNPSQLRSVQSQDALAPLPPPAPQNPSTHSWDPLCGSLPWGLSCLLALQHVLV
|
CG92078-01 Protein SequenceMASLLCVSHLLLLCSLSPGGLSYSPSQLLASSFPSCGMSPILQTWMCSRRLPLVQAPS
|
LEFLIPALVLTSQKLPRAIQTPGNASLMLHLCRGPSCECLGHWNTSLQEVVVVSGLLQ
|
CMMGLLGSPGHVFPHCGPLVLAPSLVVACLSAFPQEGVLLLSPSLECNGMTSAHHWGE
|
ASQLCLLALECSLHPLPFGFCLPLGCWLRRFWGRSSFLSLAPQVLIPVACVWTVSAFV
|
GFSVIPQELSAPTKAPWIWLPHPGVVNWPLLTPRALAAGISMALAASTSSLGCYALCG
|
RLLHLRPPFPHACSRGLSLEGLGSVLAGLLGSPMGTASSFFNVGKVGLIQQAGSQQVA
|
HLVCIZCVGLGLSPRLAQLLTTTPLPVVGGGVLCVTQAVVLSACFSSFYLADIDSCRN
|
IFIVGFSIPMALLLPRWFREAPVLFSTGHSLLMEPLGCITAQPIFLACLSGFLLENTI
|
RGTQLERGLGQGLPSPFTAQEANPQKPREKAAQVYRLPFPIQNLCPCIPQPLHCLCP
|
LPEDPGDEEGGSSEQQEMADLLRGSGEHALNIAEKGLGPEMTRTRTSALVNLALTLIC
|
[0572] Further analysis of the NOV50a protein yielded the following properties shown in Table 50B.
259TABLE 50B
|
|
Protein Sequence Properties NOV50a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body; 0.3000
probability located in endoplasmic reticulum
(membrane); 0.3000 probability located in microbody
(peroxisome)
SignalPCleavage site between residues 17 and 18
analysis:
|
[0573] A search of the NOV50a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 50C.
260TABLE 50C
|
|
Geneseq Results for NOV50a
NOV50aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAW73924Nucleobase permease Yspl1 - Mus sp,1..617435/629 (69%)0.0
611 aa. [US5858707-A,1..608480/629 (76%)
12 Jan. 1999]
AAE06572Human protein having hydrophobic1..232215/244 (88%)e-116
domain, HP03700 - Homo sapiens,1..243219/244 (89%)
243 aa. [WO200149728-A2,
12 Jul. 2001]
AAM23509Murine EST encoded protein SEQ ID474..617 121/144 (84%)3e-64
NO: 1034 - Mus musculus, 148 aa.1..144125/144 (86%)
[WO200154477-A2, 02 Aug. 2001]
AAG54392Arabidopsis thaliana protein fragment25..569 139/567 (24%)3e-32
SEQ ID NO: 69348 - Arabidopsis10..521 239/567 (41%)
thaliana, 525 aa. [EP1033405-A2,
06 Sep. 2000]
AAG54391Arabidopsis thaliana protein fragment25..569 139/567 (24%)3e-32
SEQ ID NO: 69347 - Arabidopsis12..523 239/567 (41%)
thaliana, 527 aa. [EP1033405-A2,
06 Sep. 2000]
|
[0574] In a BLAST search of public sequence datbases, the NOV50a protein was found to have homology to the proteins shown in the BLASTP data in Table 50D.
261TABLE 50D
|
|
Public BLASTP Results for NOV50a
NOV50aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q60850YOLK SAC PERMEASE-LIKE YSPL-1..617435/629 (69%)0.0
1 FORM 1 (YOLK SAC PERMEASE-1..608480/629 (76%)
LIKE YSPL-1 FORM 4) (YOLK SAC
PERMEASE-LIKE YSPL-1 FORM 3)
(YOLK SAC PERMEASE-LIKE
YSPL- 1 FORM 2) -
Mus musculus (Mouse),
611 aa.
CAC51146SEQUENCE 22 FROM PATENT1..232215/244 (88%)e-115
WO0149728 - Homo sapiens (Human),1..243219/244 (89%)
243 aa.
Q96NA6CDNA FLJ31168 FIS,1..232184/244 (75%)2e-92
CLONE KIDNE1000152,1..212188/244 (76%)
MODERATELY SIMILAR TO
MUS MUSCULUS
YOLK SAC PERMEASE-LIKE
MOLECULE 1 (YSPL-1) MRNA -
Homo sapiens (Human), 212 aa.
Q9EPR4SODIUM-DEPENDENT VITAMIN C44..531 160/513 (31%)7e-60
TRANSPORTER TYPE 2 - Mus97..578 252/513 (48%)
musculus (Mouse), 647 aa.
Q9WTW8SODIUM-COUPLED ASCORBIC44..531 160/513 (31%)7e-60
ACID TRANSPORTER SVCT2 -42..523 252/513 (48%)
Rattus norvegicus
(Rat), 592 aa.
|
[0575] PFam analysis predicts that the NOV50a protein contains the domains shown in the Table 50E.
262TABLE 50E
|
|
Domain Analysis of NOV50a
Identities/
NOV50aSimilarities
PfamMatchfor the MatchedExpect
DomainRegionRegionValue
|
Sulfate_transp:202 . . . 483 53/352 (15%)8.8
domain 1 of 1176/352 (50%)
xan_ur_permease: 46 . . . 484117/477 (25%)5.8e−59
domain 1 of 1341/477 (71%)
|
Example 51
[0576] The NOV51 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 51A.
263TABLE 51A
|
|
NOV51 Sequence Analysis
|
|
SEQ ID NO:1251037 bp
NOV51a,TTACTGATGCCCCCTCCCGCCCCCAQCGCCTGGAACAGAATCTTCTGGCGGCAGATCC
|
CG92088-01 DNA SequenceTACTTACACTTGGCCTCTTAGGCCTGTTTCTGTATGGCCTCCCTAAATTCAGGCATCT
|
CGAAGCCCTCATCCCCATGGCCGTCTGCCCTTCGGCCACAATGTCCCAGCTCAGACAC
|
AACTTCACAGGTGCCCTGCGTCCCTCGGCCCGGCCTGAAGTTCTGACCTGTACCCCCI
|
GGGGGGCTCCCATTATTTGGGATGGCTCTTTCGACCCACATGTGGCCAAGCAAGAGGC
|
TAGACAGCAGAACCTCACCATTCGGCTGACTATCTTTGCTGTAGGCAGCTACCTGGAG
|
AAGTACCTGGACCGCTTCCTGCAGACGCCGCAGCAGCACTTCATGGCGGCCCAGAGCG
|
TGATCTACTACGTGTTCACCGAGCTTCCGGCAGCGGTGCCCCGCGTGGCGCTGCCCCC
|
GCGACGCCGGCTGCCCGTGGAGCGCGTGGCGCGCGAGCGGCGCTGGCAAGACGTGTCG
|
ATGGCGCGCATGCGCACGTTGCACGCGGCGCTGGGCCGGCTGCCGGGCCGCGAGGCGC
|
ACTTCATGTTCTGCATCGACGTGGACCAGCACTTCAGCGGCACTTTTGGGCCCGACGC
|
GCTGGCCGAGTCGGTCGCGCAGCTCCACTCCTCCCACTACCACTCGCCGTCGTGGCTC
|
CTCCCCTTCGAACGCGACGCGCATTCGCCCGCCGCGATGCCGTGGGGCCAGGGCGACT
|
TCTATAACCACCCGGCGGTGTTCGGGGGCAGCGTGGCGGCGCTGCGCGGGCTGACGGC
|
GCACTGTGCGGGGGGCCTGGACTGGCACCGCGCGCGCGGCCTGGAGGCGCGCTGGCAC
|
GACGAGAGCCACCTCAACAACTTCTTCTCCCTGCACAACCCCGCCAAGGTGCTGTCGC
|
CCGAGTTCTCCTCGAGCCCGGACATCCGCCCGCGGOCCGAGATCCGCCCCCCGCGACT
|
GCTGTGGGCGCCCAAGGGGTACCGGCTGCTGCGGAACTAGCGCCGCCGCCG
|
ORF Start: ATG at 7ORF Stop: TAG at 1024
SEQ ID NO:126339 aa MW at 38627.2 kD
NOV51a,MPPPAPRAWKRIFWRQILLTLGLLGLFLYGLPKFRULEALIPMGVCPSATMSQLRDNF
|
CG92088-01 Protein SequenceTGALRPWARPEVLTCTPWGAPIIWDGSPDPDVAKQEARQQNLTIGLTIFAVCRYLEKY
|
LERFLETPEQEFMAGQSVMYYVPTELPGAVPRVALGPGRRLPVERVARERRWQDVSMA
|
RRTLHAALGGLPGREATIFNFCMDVDQHFSCTFGPEALAESVAQLHSWHYHWPSWLLP
|
FERDAHSAAAMAWGQGDFYNHAAVFGGSVAALRGLTAHCAGGLDWDRARGLEARWHDE
|
SHLNKFFWLHRPAKVLSPEFCWSPDIGPRAEIRRPRLLWAPKGYRLLRN
|
[0577] Further analysis of the NOV51 a protein yielded the following properties shown in Table 51B.
264TABLE 51B
|
|
Protein Sequence Properties NOV51a
|
|
PSort0.8650 probability located in lysosome (lumen); 0.5565
analysis:probability located in outside; 0.3880 probability located in
microbody (peroxisome); 0.1000 probability located in
endoplasmic reticulum (membrane)
SignalPCleavage site between residues 33 and 34
analysis:
|
[0578] A search of the NOV51 a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 51C.
265TABLE 51C
|
|
Geneseq Results for NOV51a
Protein/NOV51aIdentities/
Organism/Residues/Similarities for
GeneseqLengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAW13639Murine alpha(1,3)- 64 . . . 339126/276 (45%)1e−70
galactosyltransferase - Mus sp, 394 aa.117 . . . 392175/276 (62%)
[WO9709421-A1, 13 MAR. 1997]
AAR45935A glycosyltransferase - Homo sapiens, 64 . . . 339126/276 (45%)1e−70
394 aa. [WO9402616-A, 03 FEB. 1994]117 . . . 392175/276 (62%)
AAR13750GDP-Fuc:[beta-D-Gal(1,4/1,3)]-D- 64 . . . 339126/276 (45%)1e−70
GlcNAc(/Glc)alpha(1,3/1,4) -117 . . . 392175/276 (62%)
fucosyltransferase - Mus musculus,
394 aa. [WO9112340-A, 22 AUG. 1991]
AAR80016Marmoset alpha-1,3- 53 . . . 338127/292 (43%)3e−70
galactosyltransferase - Callithrix 82 . . . 373181/292 (61%)
jacchus, 376 aa. [WO9524924-A1,
21 SEP. 1995]
AAR90573Pig alpha(1,3)-galactosyltransferase - 64 . . . 339124/276 (44%)4e−70
Sus scrofa, 359 aa. [WO9534202-A1, 82 . . . 357172/276 (61%)
21 DEC. 1995]
|
[0579] In a BLAST search of public sequence datbases, the NOV51I a protein was found to have homology to the proteins shown in the BLASTP data in Table 51D.
266TABLE 51D
|
|
Public BLASTP Results for NOV51a
NOV51aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
AAF82757IGB3 SYNTHASE - Rattus norvegicus 7 . . . 338229/332 (68%) e−137
(Rat), 339 aa. 7 . . . 338263/332 (78%)
P14769N-acetyllactosaminide alpha-1,3- 64 . . . 339125/276 (45%)2e−71
galactosyltransferase (EC 2.4.1.151) 91 . . . 366177/276 (63%)
(Galactosyltransferase) (UDP-
galactose:beta-D-galactosyl-1,4-N-acetyl-
D-glucosaminide alpha-1,3-
galactosyltransferase) - Bos taurus
(Bovine), 368 aa.
Q91V22ALPHA-1,3- 64 . . . 339126/276 (45%)3e−70
GALACTOSYLTRANSFERASE - Mus musculus 94 . . . 369175/276 (62%)
(Mouse), 371 aa.
Q91W00UNKNOWN (PROTEIN FOR 64 . . . 339126/276 (45%)3e−70
MGC:11545) - Mus musculus (Mouse), 95 . . . 370175/276 (62%)
372 aa.
Q9DBU1GLYCOPROTEIN 64 . . . 339126/276 (45%)3e−70
GALACTOSYLTRANSFERASE129 . . . 404175/276 (62%)
ALPHA 1,3 - Mus musculus (Mouse),
406 aa.
|
[0580] PFam analysis predicts that the NOV51a protein contains the domains shown in the Table 51E.
267TABLE 51E
|
|
Domain Analysis of NOV51a
Identities/
NOV51aSimilaritiesExpect
Pfam DomainMatch Regionfor the Matched RegionValue
|
No Significant Matches Found
|
Example 52.
[0581] The NOV52 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 52A.
268TABLE 52A
|
|
NOV52 Sequence Analysis
|
|
SEQ ID NO:1272684 bp
NOV52a,ATCAGAATTTTGAGTTCTAGTATTTACTCTCTCGATTCCTTGTTAATTTAAATGGTAC
|
CG92142-01 DNA SequenceCTATTTTTTATAGCACATGATTTGGGAATTACACTTTGTGACATGGATGAATCTGCAC
|
TGACCCTTGGTACAATAGATGTTTCTTATCTGCCACATTCATCAGAATACAGTGTTGC
|
TCGATGTAAGCACACAAGTGAGGAATGGGTAGAGTGTGGCTTTAGACCCACCATCTTC
|
ACATCTGCAACTTTAAAATGGAAAGAAACCCTAATGAGTCGGAAAAGGCCATTTGTTG
|
GAAGATGTTGTTACTCCTGCACTCCCCAGAGCTCCGACAAATTTTTCAACCCCAGTAT
|
CCCGTCTTTGGGTTTGCGGAATGTTATTTATATCAATCAAACTCACACAAGACACCCC
|
GGATGGCTTGCAAGACGCCTTTCTTACGTTCTTTTTATTCAAGAGCGAGATGTGCATA
|
AGGGCATGTTTGCCACCAATGTCACTGAAAATCTGCTGAACAGCACTACAGTACAAGA
|
CGCAATTGCAGAAGTGGCTGCTGAATTAAACCCTGATGGTTCTCCCCAGCAGCAATCA
|
AAACCCGTTAACAAAGTCAAAAAGAAAGCTAAAACGATTCTTCAACAAATCGTTGCCA
|
CTCTCTCACCGGCAATGATCAGACTGACTGGGTGGGTGCTGCTAAAACTCTTCAACAG
|
CTTCTTTTCCAACATTCAAATTCACAAACGTCAACTTGACATGCTTAAAGCTGCAACT
|
GAGACGAATTTGCCGCTTCTGTTTCTACCAGTTCATAGATCCCATATTGACTATCTGC
|
TGCTCACTTTCATTCTCTTCTOCCATAACATCAAACCACCATACATTGCTTCAGGCAA
|
TAATCTCAACATCCCAATCTTCAGTACCTTGATCCATAACCTTGGGCGCTTCTTCATA
|
CGACGAACGCTCGATGAAACACCAGATGGACGGAAAGATGTTCTCTATACAGCTTTGC
|
TCCATGGGCATATAGTTGAATTACTTCGACAGCAGCAATTCTTCGAGATCTTCCTGGA
|
AGGCACACGTTCTAGGAGTGGAAAAACCTCTTGTGCTCGGGCACCACTTTTGTCAGTT
|
GTGGTAGATACTCTGTCTACCAATGTCATCCCAGACATCTTGATAATACCTGTTGGAA
|
TCTCCTATGATCGCATTATCGAAGGTCACTACAATGGTCAACAACTGGGCAAACCTAA
|
GAAGAATCACAGCCTGTGGAGTGTAGCAAGACGTGTTATTAGAATGTTACGAAAAAAC
|
TATGGTTGTGTCCGAGTGGATTTTCCACAGCCATTTTCCTTAAAGGAATATTTAGAAA
|
GCCAAAGTCAGAAACCGGTGTCTGCTCTACTTTCCCTGCACCAACCGTTGTTACCAGC
|
TATACTTCCTTCAAGACCCACTGATGCTGCTGATCAAGGTAGAGACACCTCCATTAAT
|
CAGTCCAGAAATGCAACACATGAATCCCTACCAAGGAGGTTGATTGCAAATCTGGCTG
|
AGCATATTCTATTCACTGCTAGCAAGTCCTGTGCCATTATGTCCACACACATTGTCGC
|
TTGCCTCCTCCTCTACAGACACACCCACGGAATTGATCTCTCCACATTGGTCGAACAC
|
TTCTTTGTGATGAAAGAGGAAGTCCTGGCTCGTGATTTTGACCTCOGCTTCTCAGGAA
|
ATTCAGAAGATGTAGTAATGCATGCCATACAGCTGCTCGCAAATTCTCTCACAATCAC
|
CCACACTAGCAGGAACGATGAGTTTTTTATCACCCCCAGCACAACTGTCCCATCACTC
|
TTCGAACTCAACTTCTACAGCAATGCGGTACTTCATGTCTTTATCATCGAGGCCATCA
|
TAGCTTGCAGCCTTTATGCAGTTCTGAACAAGAGGGGACTGGGGGGTCCCACTACCAC
|
CCCACCTAACCTGATCAGCCAGGAGCAGCTGGTGCGGAAGGCGGCCAGCCTCTGCTAC
|
CTTCTCTCCAATGAAGGCACCATCTCACTGCCTTGCCAGACATTTTACCAAGTCTGCC
|
ATGAAACAGTAGGAAAGTTTATCCAGTATGGCATTCTTACAGTGGCAGAGCACGATGA
|
CCAGGAAGATATCAGTCCTAGTCTTGCTGAGCAGCAGTGGGACAAGAAGCTTCCTGAA
|
CCTTTGTCTTGGAUAAGTGATGAACAAGATCAAGACAGTGACTTTGGGCAGGAACAGC
|
GAGATTCCTACCTGAAGGTGAGCCAATCCAAGGAGCACCAGCAGTTTATCACCTTCTT
|
ACAGACACTCCTTGGGCCTTTGCTGGAGGCCTACAGCTCTGCTGCCATCTTTGTTCAC
|
AACTTCAGTGGTCCTGTTCCAGAACCTGAGTATCTGCAAAAGTTGCACAAATACCTAA
|
TAACCAGAACAGAAAGAAATGTTGCAGTATATGCTGAGAGTGCCACATATTGTCTTGT
|
GAAGAATGCTGTGAAAATGTTTAAGCATATTGGGGTTTTCAAGGAGACCAAACAAAAG
|
AGAGTGTCTGTTTTAGAACTGAGCAGCACTTTTCTACCTCAATGCAACCGACAAAAAC
|
TTCTAGAATATATTCTGAGTrTTGTGGTGCTGTAGGTAACGTGTGGCACTGCTGGCAA
|
ATGAAGGTCATGAGATGAGTTCCTTGTAGGTACCAGCTTCTGGCTCAAGAGTTGAAGG
|
TGCCGTCGCAGGGTCA
|
ORF Start: ATG at 101ORF Stop: TAG at 2585
SEQ ID NO:128828 aa MW at 93835.7 kD
NOV52a,MDESALTLGTIDVSYLPHSSEYSVGRCKHTSEEWVECGFRPTIFRSATLKWKESLMSR
|
CG92142-01 Protein SequenceKRPFVGRCCYSCTPQSWDKFFNPSIPSLGLRNVIYINETHTRERGWLARRLSYVLFIQ
|
ERDVHKGMFATNVTENVLNSSRVQEAIAEVAAELNPDGSAQQQSKAVNKVKKKAKRIL
|
QEMVATVSPAMIRLTCNVLLKLPNSFFWNIQIEKGQLEMVKAATETNLPLLFLPVHRS
|
HIDYLLLTFILFCHNIKAPYIASGNNLNIPIFSTLIHKLGGFFIRRRLDETPDGRKDV
|
LYRALLHGHIVELLRQQQFLEIFLEGTRSRSGKTSCARAGLLSVVVDTLSTNVIPDIL
|
IIPVGISYDRIIEGHYNGEQLGKPKKNESLWSVARGVIRMLRKNYGCVRVDFAQPFSL
|
KEYLESQSQKPVSALLSLEQALLPAILPSRRSDAADEGRDTSINESRNATDESLRRRL
|
IANLAEEILFTASKSCAIMSTHIVACLLLYRHRQGIDLSTLVEDFFVMKEEVLARDFD
|
LGFSGNSEDVVMHAIQLLGNCVTITHTSRNDEFFITPSTTVPSVFELNFYSNGVLHVF
|
IMEAIIACSLYAVLHKRGLGGPTSTPPNLISQEQLVRKAASLCYLLSNEGTTSLPCQT
|
FYQVCHETVGKFIQYGILTVAEHDDQEDISPSLAEQQWDKKLPEPLSWRSDEEDEDSD
|
FGEEQRDCYLKVSQSKEHQQFITFLQRLLGPLLEAYSSAAIFVHNFSCPVPEPEYLQK
|
LHKYLTTRTERNVAVYAESATYCLVKNAVKMFKDIGVFKETKQKRVSVLELSSTFLPQ
|
CNRQKLLEYILSFVVL
|
[0582] Further analysis of the NOV52a protein yielded the following properties shown in Table 52B.
269TABLE 52B
|
|
Protein Sequence Properties NOV52a
|
|
PSort0.8500 probability located in endoplasmic reticulum
analysis:(membrane); 0.4400 probability located in plasma membrane;
0.3000 probability located in nucleus; 0.1000 probability
located in mitochondrial inner membrane
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0583] A search of the NOV52a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 52C.
270TABLE 52C
|
|
Geneseq Results for NOV52a
Protein/NOV52aIdentities/
Organism/Residues/Similarities for
GeneseqLengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABB11171Human sn-glycerol-3-P 89 . . . 233145/145 (100%)8e−78
acyltransferase homologue, SEQ ID 12 . . . 156145/145 (100%)
NO:1541 - Homo sapiens, 156 aa.
[WO200157188-A2, 09 AUG. 2001]
ABG23092Novel human diagnostic protein178 . . . 822209/678 (30%)8e−72
#23083 - Homo sapiens, 807 aa.142 . . . 799324/678 (46%)
[WO200175067-A2, 11 OCT. 2001]
ABG23092Novel human diagnostic protein178 . . . 822209/678 (30%)8e−72
#23083 - Homo sapiens, 807 aa.142 . . . 799324/678 (46%)
[WO200175067-A2, 11 OCT. 2001]
AAY72134E. coli glycerol-3-phosphate131 . . . 410 89/281 (31%)1e−30
acyltransferase with ER retention234 . . . 499146/281 (51%)
sequence - Escherichia coli, 827 aa.
[WO200078974-A2, 28 DEC. 2000]
AAY72133Escherichia coli glycerol-3-phosphate131 . . . 410 89/281 (31%)1e−30
acyltransferase (GPAT) - Escherichia234 . . . 499146/281 (51%)
coli, 827 aa. [WO200078974-A2,
28 DEC. 2000]
|
[0584] In a BLAST search of public sequence datbases, the NOV52a protein was found to have homology to the proteins shown in the BLASTP data in Table 52D.
271TABLE 52D
|
|
Public BLASTP Results for NOV52a
NOV52aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9HCL2Glycerol-3-phosphate 1 . . . 828828/828 (100%)0.0
acyltransferase, mitochondrial 1 . . . 828828/828 (100%)
precursor (EC 2.3.1.15) (GPAT) -
Homo sapiens (Human), 828 aa.
AAH19201GLYCEROL-3-PHOSPHATE 1 . . . 828769/828 (92%)0.0
ACYLTRANSFERASE, 1 . . . 827799/828 (95%)
MITOCHONDRIAL -
Mus musculus (Mouse), 827 aa.
Q61586Glycerol-3-phosphate 1 . . . 828767/828 (92%)0.00
acyltransferase, mitochondrial 1 . . . 827799/828 (95%)
precursor (EC 2.3.1.15) (GPAT)
(P90) - Mus musculus (Mouse),
827 aa.
P97564Glycerol-3-phosphate 1 . . . 828760/828 (91%)0.0
acyltransferase, mitochondrial 1 . . . 828794/828 (95%)
precursor (EC 2.3.1.15) (GPAT) -
Rattus norvegicus (Rat), 828 aa.
Q9Y137BCDNA:GH07066 PROTEIN -163 . . . 809196/654 (29%)1e−81
Drosophila melanogaster (Fruit fly),194 . . . 820353/654 (53%)
850 aa.
|
[0585] PFam analysis predicts that the NOV52a protein contains the domains shown in the Table 52E.
272TABLE 52E
|
|
Domain Analysis of NOV52a
Identities/
Similarities
PfamNOVA52a Matchfor the Matched
DomainRegionRegionExpect Value
|
Acyltransferase:215 . . . 412 47/207 (23%)6.4e−34
domain 1 of 1151/207 (73%)
|
Example 53
[0586] The NOV53 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 53A.
273TABLE 53A
|
|
NOV53 Sequence Analysis
|
|
SEQ ID NO:129906 bp
NOV53a,AGTAACCTGGAACATTATGAATGAACTTACATGCAAGAAAAGTACTGAGAAATGTGGC
|
CG92152-01 DNA SequenceATCCCTCCATTTAGTCCCCAGTGGCTTTCCAGAAGAATCGCAGGAGGGGAAGAAGCCT
|
GCCCCCACTGTTCGCCATGGCAGGTCGGTCTCAGCTTTCTAGGCGATTACCAATGTCG
|
AGGTGCCATCATCAACCCAGTGTGGATTCTGACCGCAGCCCACTGTGTGCAAAAGAAT
|
AATCCACTCTCCTGGACTATTATTCCTGGGGACCATGACAGAAACCTGAAGGAATCAA
|
CAGAGCAGGTGAAAAAATGTTTTTACATAATAGTGCATGAAGACTTTAACACACTAAG
|
TTATGACTCTCACATTGCCCTAATACAACTAAGCTCTCCTCTGGAGTACAACTCCGTG
|
GTGAGGCCAGTATGTCTCCCACACAGCGCACAGCCTCTATTTTCCTCGGAGATCTGTG
|
CTGTGACCQGATGGGGAAGCATCAGTGATGGTGGCCTAGCAACTCGCCTACAGCAGAT
|
TCAAGTGCATGTGTTAGAAAGACACGTCTGTGAACACACTTACTATTCTGCCCATCCA
|
GGACGGATCACAGAGAAGATGATCTGTGCTGGCTTTGCAGCATCTGGAGAGAAAGATT
|
TCTGCCAGGGAGACTCTGGTGGGCCACTAGTATGTAGACATGAAAATGGTCCCTTTGT
|
CCTCTATGGCATTGTCACCTCGGGACCTGGCTCTCTCCACCCATGGAAGCCGGCTGTA
|
TTTGCCAGAGTGATGATCTTCTTGGACTGGATCCAATCAAAAATCAATGGTAAATTGT
|
TTTCAAATGTTATTAAAACAATAACCTCTTTCTTTAGAGTGGGTTTGCGAACAGTGAG
|
TGTGGTATAAATTAGCATGAAAGAGAAAACCATAGA
|
ORF Start: ATG at 17ORF Stop: TAA at 878
SEQ ID NO:130287 aa MW at 31625.0 kD
NOV53a,MNELTCKKSTEKCGIPPFSPQWLSRRTAGGEEACPHCWPWQVCLRPLGDYQCGGAIIN
|
CG92152-01 Protein SequencePVWILTAAHCVQKNNPLSWTIIAGDHDRNLKESTEQVKKCFYIIVHEDFNTLSYDSDI
|
ALIQLSSPLEYNSVVRPVCLPHSAEPLBSSEICAVTGWGSISDGGLASRLQQIQVHVL
|
EREVCEHTYYSAHPGGITEKNICAGFAASGEKDFCQGDSGGPLVCRHENGPFVLYGIV
|
SWGAGCVQPWKPGVFARVMIFLDWIQSKINCKLFSNVIKTITSFFRVCLGTVSVV
|
[0587] Further analysis of the NOV53a protein yielded the following properties shown in Table 53B.
274TABLE 53B
|
|
Protein Sequence Properties NOV53a
|
|
PSort0.6564 probability located in microbody (peroxisome); 0.1000
analysis:probability located in mitochondrial matrix space; 0.1000
probability located in lysosome (lumen); 0.0000 probability
located in endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0588] A search of the NOV53a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 53C.
275TABLE 53C
|
|
Geneseq Results for NOV53a
Protein/NOV53aIdentities/
Organism/Residues/Similarities for
GeneseqLengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABG24246Novel human diagnostic protein 13 . . . 268196/257 (76%)e−110
#24237 - Homo sapiens, 913 aa. 506 . . . 743201/257 (77%)
[WO200175067-A2, 11 OCT. 2001]
ABG24246Novel human diagnostic protein 13 . . . 268196/257 (76%)e−110
#24237 - Homo sapiens, 913 aa. 506 . . . 743201/257 (77%)
[WO200175067-A2, 11 OCT. 2001]
ABG19887Novel human diagnostic protein 13 . . . 262194/251 (77%)e−109
#19878 - Homo sapiens, 1576 aa.1345 . . . 1576199/251 (78%)
[WO200175067-A2, 11 OCT. 2001]
ABG14588Novel human diagnostic protein 13 . . . 262194/251 (77%)e−109
#14579 - Homo sapiens, 1576 aa.1345 . . . 1576199/251 (78%)
[WO200175067-A2, 11 OCT. 2001]
ABG10218Novel human diagnostic protein 13 . . . 262194/251 (77%)e−109
#10209 - Homo sapiens, 1576 aa.1345 . . . 1576199/251 (78%)
[WO200175067-A2, 11 OCT. 2001]
|
[0589] In a BLAST search of public sequence datbases, the NOV53a protein was found to have homology to the proteins shown in the BLASTP data in Table 53D.
276TABLE 53D
|
|
Public BLASTP Results for NOV53a
NOV53aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q91674POLYPROTEIN - Xenopus laevis 13 . . . 259130/248 (52%)2e−76
(African clawed frog), 1524 aa.570 . . . 814175/248 (70%)
Q9BK47SEA STAR REGENERATION- 13 . . . 261108/255 (42%)8e−52
ASSOCIATED PROTEASE SRAP - 18 . . . 264152/255 (59%)
Luidia foliolata, 267 aa.
O96899PLASMINOGEN ACTIVATOR 12 . . . 273115/269 (42%)1e−51
SPA - Scolopendra subspinipes, 19 . . . 276163/269 (59%)
277 aa.
Q90WD8OVIDUCTIN - Bufo japonicus 11 . . . 261 99/253 (39%)2e−49
(Japanese toad), 974 aa.574 . . . 818150/253 (59%)
CAC17064SEQUENCE 3 FROM PATENT 26 . . . 263107/245 (43%)3e−49
WO0065067 - Homo sapiens255 . . . 492148/245 (59%)
(Human), 492 aa.
|
[0590] PFam analysis predicts that the NOV53a protein contains the domains shown in the Table 53E.
277TABLE 53E
|
|
Domain Analysis of NOV53a
Identities/
NOV53aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
trypsin:27 . . . 257108/263 (41%)2.8e−79
domain 1 of 1180/263 (68%)
|
Example 54
[0591] The NOV54 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 54A.
278TABLE 54A
|
|
NOV54 Sequence Analysis
|
|
SEQ ID NO:1311155 bp
NOV54a,TTATTCTCCTGCCTCAGCCTCCTGAGTAGCTGAGATTATAGGCATGTGCCACCACGCC
|
CG92228-01 DNA SequenceTGGCTAATTTTTGTATTTTTAGTAGAGACTGGGTTTCACCATGTTGGTCACGCTGGTC
|
TCAAACTTGTGACCTTGCCAGAAGCCCAGCTGGCTTCACCTCTCAATGGCCCTTGTGG
|
GCGGCCCACAGTCTCATCTGGTATTGCCTCAGGCTGGGGGGCTAGTGTGGGGCAGTGG
|
CCCTGGCAGCTCAGCATCCGCCAGGGCTTGATTCACGTCTGCTCAGATACCCTCATCT
|
CAGAGGAGTGGGTGCTGACAGTGGCGATCTGCTTCCCGTCCTCCTGCCTCAGCCTCCC
|
GAGTAGCTGGGGACTTCAGGTGCACATCACTACACCAGGCGCAGGCCTCCTTTCTTTT
|
CCCCATAGGACCTGGACTACTTTTTCATATACTCTGTGGCTAGGATCGATTACAGTAG
|
GTGACTCAACGAAACGTGTGAAGTACTACGTGTCCAAAATCGTCATCCATCCCAAGTA
|
CCAAGATACAACGGCAGACGTCGCCTTGTTGAAACTGTCCTCTCAAGTCACCTTCACT
|
TCTGCCATCCTGCCTATTTGCTTGCCCAGTGTCACAAACCAGTTCGCAATTCCACCCT
|
TTTGTTGGGTGACCCCATGGCGAAAAGTTAAGGAAAGTTCACTTCACCAACCAGAAGT
|
ACCCATTATTGACCGCCAGGCTTGTGAACAGCTCTACGGTCATTCTGGAGCGCCTCTG
|
TCGTGTCACATTGATGGTGTATGGATCCAGACACGACTAGTAAGCTGGGGATTAGAAT
|
GTGGTAAATCTCTTCCTGGAGTCTACACCAATGTAATCTACTACCAAAAATGGATTAA
|
TGCCACTATTTCAACAGCCAACAATCTAGACTTCTCTGACTTCTTGTTCCCTATTGTC
|
CTACTCTCTCTGGCTCTCCTGCGTCCCTCCTGTGCCTTTGCACCTAACACTATACACA
|
GAGTAGGCACTGTAGCTGAAGCTGTTGCTTGCATACAGGGCTGCGAACAGAATGCATG
|
GAGATTTAGTCCCAGGGGCAGATAACTCACAGGAGAGCCACTGCTAACCCTGGGTGAC
|
TTTATTTACAATTTGAAATGATTTTGTTTTTAAGGTTTTTGATTTTGGAAGTT
|
ORF Start: ATG at 44ORF Stop: TAA at 1067
SEQ ID NO:132341 aa MW at 37339.7 kD
NOV54a,MCHHAWLIFVELVETGFHHVGQAGLKLVTLPEAQLASPLNGPCGRPTVSSGIASGWGA
|
CG92228-01 Protein SequenceSVGQWPWQVSIRQGLIHVCSDTLISEEWVLTVAICFPSSCLSLPSSWGLQVHITTPGA
|
GLLSFPHRTWTTFSYTVWLGSITVGDSRKRVKYYVSKIVIHPKYQDTTADVALLKLSS
|
QVTPTSAILPICLPSVTKQLAIPPFCWVTGWGKVKESSLQEAEVPIIDRQACEQLYGD
|
SGGPLSCIDGVWIQTGVVSWGLECGKSLPGVYTNVIYYQKWINATISRPANNLDFSDF
|
LFPIVLLSLALLRPSCAFGPNTIHRVGTVAEAVACIQGWEENAWRFSPRGR
|
SEQ ID NO:1331022 bp
NOV54b,TCTGAGGACAGAGACATGGGCCCTGCTGGCTGTGCCTTCACGCTGCTCCTTCTGCTGG
|
CG92228-02 DNA SequenceGGATCTCAGTGTGTGGGCAACCTGTATACTCCAGCCGCGTTGTAGGTGGCCAGGATGC
|
TGCTGCAGGGCGCTCGCCTTGGCAGGTCAGCCTACACTTTGACCACAACTTTATCTGT
|
GGAGGTTCCCTCGTCAGTGAGAGGTTGATACTGACAGCAGCTCACTGCATAGCAACCT
|
ATACTGTGTGGCTAGGATCGATTACAGTAGCTGACTCAACGAAACGTGTGAAGTACTA
|
CGTGTCCAAAATCGTCATCCATCCCAAGTACCAAGATACAACCGCACACGTCGCCTTG
|
TTGAAACTGTCCTCTCAAGTCACCTTCACTTCTGCCATCCTGCCTATTTGCTTGCCCA
|
GTGTCACAAAGCAGTTCGCAATTCCACCCTTTTGTTGGGTGACCGGATCGCGAAAAGT
|
TAAGGAAAGTTCAGATAGAGATTACCATTCTGCCCTTCAGGAAGCAGAAGTACCCATT
|
ATTGACCGCCAGGCTTGTGAACAGCTCTACAATCCCATCGGTATCTTCTTGCCAGCAC
|
TGGAGCCAGTCATCAAGGAACACAAGATTTGTGCTCCTGATACTCAAAACATGAACGA
|
TAGTTGCAAGGGTGATTCTGGAGGGCCTCTGTCGTGTCACATTGATGGTGTATGGATC
|
CAGACAGGAGTACTAAGCTGGGGATTAGAATGTGGTAAATCTCTTCCTGGAGTCTACA
|
CCAATGTAATCTACTACCAAAAATGGATTAATGCCACTATTTCAAGAGCCAACAATCT
|
AGACTTCTCTGACTTCTTCTTCCCTATTGTCCTACTCTCTCTGGCTCTCCTCCGTCCC
|
TCCTGTGCCTTTGGACCTAACACTATACACAGACTAGGCACTGTAGCTGAACCTGTTG
|
CTTGCATACAGGGCTGGCAAGAGAATGCATGGAGATTTAGTCCCACGGGCACATAACT
|
CACAGGAGAGCCACTGCTAACCCTCGGTGACTTTAT
|
ORF Start: ATG at 16ORF Stop: TAA at 982
SEQ ID NO:134322 aa MW at 35193.2 kD
NOV54b,MGPAGCAFTLLLLLGISVCGQPVYSSRVVGGQDAAAGRWPWQVSLHFDHNFICGGSLV
|
CG92228-02 Protein SequenceSERLILTAAHCIATYTVWLGSITVGDSRKRVKYYVSKIVIHPKYQDTTADVALLKLSS
|
QVTFTSAILPICLPSVTKQLAIPPFCWVTGWGKVKESSDRDYHSALQEAEVPLIDRQA
|
CEQLYNPIGIFLPALEPVIKEDKICAGDTQNMKDSCKGDSGGPLSCHIDGVWIQTGVVC~QLYNPIGI FLPALEPVI KEDKICAGDTQNMKDSCKGDSGGPLSCHIDCVNIQTCVV
|
SWGLECGKSLPGVYTNVIYYQKWINATISRANNLDFSDFLFPIVLLSLALLRPSCAFG
|
PNTIHRVGTVAEAVACIQGWEENAWRFSPRGR
|
[0592] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 54B.
279TABLE 54B
|
|
Comparison of against NOV54b.
Identities/
NOV54a Residues/Similarities for
Protein SequenceMatch Residuesthe Matched Region
|
NOV54b43 . . . 341233/338 (68%)
19 . . . 322243/338 (70%)
|
[0593] Further analysis of the NOV54a protein yielded the following properties shown in Table 54C.
280TABLE 54C
|
|
Protein Sequence Properties NOV54a
|
|
PSort0.7300 probability located in plasma membrane; 0.6400
analysis:probability located in endoplasmic reticulum (membrane);
0.3200 probability located in microbody (peroxisome); 0.1000
probability located in endoplasmic reticulum (lumen)
SignalPCleavage site between residues 24 and 25
analysis:
|
[0594] A search of the NOV54a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 54D.
281TABLE 54D
|
|
Geneseq Results for NOV54a
Protein/NOV54aIdentities/
Organism/Residues/Similarities for
GeneseqLengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAW77297Amino acid sequence of long isoform20 . . . 304117/322 (36%)3e−48
of HELA2 - Homo sapiens, 314 aa.13 . . . 309170/322 (52%)
[WO9836054-A1, 20 AUG. 1998]
AAB80256Human PR0303 protein - Homo20 . . . 304116/322 (36%)1e−47
sapiens, 314 aa. [WO200104311-A1,13 . . . 309170/322 (52%)
18 JAN. 2001]
AAU01569Human secreted protein immunogenic20 . . . 304116/322 (36%)1e−47
epitope encoded by gene #9 - Homo13 . . . 309170/322 (52%)
sapiens, 315 aa. [WO200123547-A1,
05 APR. 2001]
AAU02223Human extracellular serine protease20 . . . 304116/322 (36%)1e−47
TADG-16 - Homo sapiens, 314 aa.13 . . . 309170/322 (52%)
[WO200127257-A1, 19 APR. 2001]
AAY91871Human cancer-specific gene protein,20 . . . 304116/322 (36%)1e−47
Pro104 - Homo sapiens, 327 aa.26 . . . 322170/322 (52%)
[WO200016805-A1, 30 MAR. 2000]
|
[0595] In a BLAST search of public sequence datbases, the NOV54a protein was found to have homology to the proteins shown in the BLASTP data in Table 54E.
282TABLE 54E
|
|
Public BLASTP Results for NOV54a
NOV54aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9Y6M0Testisin precursor (EC 3.4.21.-)20 . . . 304116/322 (36%)3e−47
(Eosinophil serine protease 1) (ESP-1) -13 . . . 309170/322 (52%)
Homo sapiens (Human), 314 aa.
Q9JHJ7Testisin precursor (EC 3.4.21.-)31 . . . 305115/315 (36%)1e−43
(Tryptase 4) - Mus musculus34 . . . 324157/315 (49%)
(Mouse), 324 aa.
Q920S2TESTIS SERINE PROTEASE-1 -39 . . . 305101/308 (32%)6e−38
Mus musculus (Mouse), 322 aa.39 . . . 322149/308 (47%)
Q9D4I34931440B09RIK PROTEIN -42 . . . 305100/305 (32%)1e−37
Mus musculus (Mouse), 282 aa. 2 . . . 282147/305 (47%)
Q9QUL7Tryptase gamma precursor (EC43 . . . 305103/302 (34%)1e−35
3.4.21.-) (Transmembrane tryptase) -18 . . . 285128/302 (42%)
Mus musculus (Mouse), 311 aa.
|
[0596] PFam analysis predicts that the NOV54a protein contains the domains shown in the Table 54F.
283TABLE 54F
|
|
Domain Analysis of NOV54a
Identities/
NOV54aSimilarities forExpect
Pfam DomainMatch Regionfor the Matched RegionValue
|
trypsin:52 . . . 275 89/284 (31%)1.2e−48
domain 1 of 1163/284 (57%)
|
Example 55
[0597] The NOV55 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 55A.
284TABLE 55A
|
|
NOV55 Sequence Analysis
|
|
SEQ ID NO:135991 bp
NOV55a,TCCCTCTTGGCCATCACATTCCCCTTGCCCTATGGCGGCCCTCACAGACCTCTCATTT
|
CG92425-01 DNA SequenceATGTATCGCTGGTTCAAGAACTGCAATCTGGTTGCCAACCTCTCAGAGAAGTACGTCT
|
TCATCACAGGCTGTGACTCTGCCTTCGCGAACCTCCTGGCCAAACAGCTGGTTGATCG
|
GGGCATGCAGGTGCTGGCTGCTTGCTTCACTGAGGAGGGATCCCAGAAACTTCAGCGG
|
GATACCTCCTATCGGCTGCAGACCACCCTACTGGATGTCACCAAGAGCGAAAGCATCA
|
AGGCGGCGGCCCAGTGGGTCAGCGACAAAGTGGGCGAACAAGGTCTCTCGGCCCTGGT
|
GAACAATGCTGGTGTGGGCCTGCCCAGTCGTCCCAACGAATGGCTGACCAACGATGAC
|
TTTGTGAAGGTGATTAATGTGAACCTGGTGCCACPGATCGAAGTGACCCTTCACATGC
|
TGCCCATGGTCAAGAGAGCCCGGGGCAGGGTTGTCAACATGTCCAGCTCTGGTGGTCG
|
TGTCGCTCTCATTGGTCGTCGCTACTGCGTCTCCAACTTTGGCCTTGACGCCTTCTCT
|
GACAGCATAAGGCGTGAGCTCTACTACTTTGGGGTGAAAGTCTGCATCATTGAGCCAG
|
GGAACTATCGGACAGCCATTCTCGGCAAGGAGAACCTGGAGTCACGCATGCGAAAGCT
|
TTGGGAGAGGCTGCCTCAGGACACCCGGGACACCTACGGAGAGCATTATATGCCTCTT
|
ATCCTAAAGGCATTAGAGAGCCTTCTAAATGGCTCTGAPATGAAGGATCAGATTGTCA
|
TCAACAGCATCGAGCATGCTATTGTTTCCCGGAGCCCTCCCATCCGCTACAACCCTGG
|
CCTGGATGCCAAACTCCTCTACATCCCTCTGGCTAAGTTGCCCACCCCTGTGACAGAT
|
TTCATCCTAAGCCGCTACCTTCCAAGGCCAGCCGACAGTGTCTAAACTCGGGAGGATC
|
AATGG
|
ORF Start: ATG at 32ORF Stop: TAA at 971
SEQ ID NO:136313 aa MW AT 34973.1 kD
NOV55a,MAALTDLSFMYRWFKNCNLVGNLSEKYVFITGCDSGFGNLLAKQLVDRGMQVLAACFT
|
CG92425-01 Protein SequenceEEGSQKLQRDTSYRLQTTLLDVTKSESIKAAAQWVRDKVGEQGLWALVNNAGVGLPSG
|
PNEWLTKDDFVKVINVNLVGLIEVTLHMLPMVKRARGRVVNMSSSGGRVAVICGGYCV
|
SKFGVEAFSDSIRRELYYFGVKVCIIEPGNYRTAILGKENLESRMRKLWERLPQETRD
|
SYGEDYMALILKALESLVMGSEMKDQIVINSMEHAIVSRSPRIRYNPGLDAKLLYIPL
|
AKLPTPVTDFILSRYLPRPADSV
|
SEQ ID NO:137991 bp
NOV55b,TCCCTCTTGGCCATCACATTCCCCTTGCCCTATGGCGGCCCTCACAGACCTCTCATTT
|
CG92425-02 DNA SequenceATGTATCGCTGGTTCAAGAACTGCAATCTGGTTGGCAACCTCTCAGAGAAGTACGTCT
|
TCATCACAGGCTGTGACTCTGGCTTCGGGAACCTGCTGGCCAAACAGCTGGTTGATCG
|
GGGCATGCAGGTGCTGGCTGCTTGCTTCACTGAGGAGGGATCCCAGAAACTTCAGCGG
|
GATACCTCCTATCGGCTGCAGACCACCCTACTGGATGTCACCAAGAGCGAAAGCATCA
|
AGGCGGCGGCCCAGTGGGTGAGGGACAAAGTGGGCGAACAAGGCCTCTGGGCCCTGGT
|
GAACAATGCTGGTGTGGGCCTGCCCAGTGGTCCCAACGAATCGCTGACCAAGGATGAC
|
TTTGTGAAGGTGATTAATGTGAACCTGGTGGGACTGATCGAAGTGACCCTTCACATGC
|
TGCCCATGGTCAAGAGAGCCCGGGGCAGGGTTGTCAACATGTCCAGCTCTGGTGGTCG
|
TGTGGCTGTCATTGGTGGTGGCTACTGCGTCTCCAAGTTTGGCGTTGAGGCCTTCTCT
|
GACAGCATAAGGCGTGAGCTCTACTACTTTGGGGTGAAAGTCTGCATCATTGAGCCAG
|
GGAACTATCGGACAGCCATTCTCGGCAAGGAGAACCTGGAGTCACGCATGCGAAAGCT
|
TTGGGAGAGGCTGCCTCAGGAGACCCGGGACAGCTACGGAGAGGATTATTTCCGCATC
|
TATACTGACAAGTTAAAAAACATAATGCAGGTGGCAGAGCCCAGAGTCAGAGATGTCA
|
TCAACAGCATGGAGCATGCTATTGTTTCCCGGAGCCCTCGCATCCGCTACAACCCTGG
|
CCTGGATGCCAAACTCCTCTACATCCCTCTGGCTAAGTTCCCCACCCCTGTCACAGAT
|
TTCATCCTAAGCCGGTACCTTCCAAGGCCAGCGGACAGTGTCTAAACTGGGGAGGATC
|
AATGG
|
ORF Start: ATG at 32ORF Stop: TAA at 971
SEQ ID NO:138313 aa MW AT 25262.4 kD
NOV55b,MAALTDLSFMYRWFKNCNLVGNLSEKYVFITGCDSGFGNLLAKQLVDRGMQVLAACFT
|
CG9425-02 Protein SequenceEEGSQKLQRDTSYRLQTTLLDVTKSESIKAAAQWVRDKVGEQGLWALVNNAGVGPLSG
|
PNEWLTKDDFVKVINVNLVGLIEVTLHMLPMVKRARGRVVNMSSSGGRVAVIGGGYCV
|
SKFGVEAFSDSIRRELYYFGVKVCIIEPGNYRTAILGKENLESRMRKLWERLPQETRD
|
SYGEDYFRIYTDKLKNIMQVAEPRVRDVINSMEHAIVSRSPRIRYNPGLDAKLLYIPL
|
AKLPTPVTDFILSRYLPRPADSV
|
[0598] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 55B.
285TABLE 55B
|
|
Comparison of NOV55a against NOV55b.
Identities/
ProteinNOV55a Residues/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV55b1 . . . 313294/313 (93%)
1 . . . 313302/313 (95%)
|
[0599] Further analysis of the NOV55a protein yielded the following properties shown in Table 55C.
286TABLE 55C
|
|
Protein Sequence Properties NOV55a
|
|
PSort0.5813 probability located in mitochondrial matrix space;
analysis:0.3000 probability located in microbody (peroxisome);
0.2927 probability located in mitochondrial inner
membrane; 0.2927 probability located in mitochondrial
intermembrane space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0600] A search of the NOV55a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 55D.
287TABLE 55D
|
|
Geneseq Results for NOV55a
NOV55aIdentities/
ResiduesSimilarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM41630 Human polypeptide SEQ ID NO1..313294/313 (93%)e−169
6561 - Homo sapiens, 360 aa.48..360 302/313 (95%)
[WO200153312-A1, 26 Jul. 2001]
AAM39844Human polypeptide SEQ ID NO1..313294/313 (93%)e−169
2989 - Homo sapiens, 313 aa.1..313302/313 (95%)
[WO200153312-A1, 26 Jul. 2001]
AAY02003 A retinol dehydrogenase nrotein of1..309170/310 (54%)9e−94
the invention - Mus sp, 318 aa.5..314227/310 (72%)
[WO9916788-A1, 08 Apr. 1999]
AAW18335Murine liver p32 11-cis-retinol1..302169/302 (55%)9e−94
dehydrogenase - Mus musculus, 3175..306219/302 (71%)
aa. [WO9719167-A1, 29 May 1997]
AAY02002A retinol dehydrogenase protein of1..302168/302 (55%)2e−93
the invention - Mus sp, 317 aa.5..306219/302 (71%)
[WO9916788-A1, 08 Apr. 1999]
|
[0601] In a BLAST search of public sequence datbases, the NOV55a protein was found to have homology to the proteins shown in the BLASTP data in Table 55E.
288TABLE 55E
|
|
Public BLASTP Results for NOV55a
NOV55aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P50169Retinol dehydrogenase type I (EC1..310170/310 (54%)4e−99
1.1.1.105) (RODH I) - Rattus5..314236/310 (75%)
norvegicus (Rat), 317 aa.
P55006Retinol dehydrogenase type III (EC1..310169/310 (54%)9e−98
1.1.1.105) (RODHIII) - Rattus5..314235/310 (75%)
norvegicus (Rat), 317 aa.
AAK30001RETINOL DEHYDROGENASE1..302166/302 (54%)3e−96
TYPE 1 - Mus musculus (Mouse),5..306224/302 (73%)
317 aa.
AAH18263SIMILAR TO RETINOL1..309173/310 (55%)5e−94
DEHYDROGENASE TYPE 6 -5..314226/310 (72%)
Mus musculus (Mouse), 318 aa.
O75452STEROL/RETINOL1..313162/313 (51%)6e−94
DEHYDROGENASE - Homo5..317231/313 (73%)
sapiens (Human), 317 aa.
|
[0602] PFam analysis predicts that the NOV55a protein contains the domains shown in the Table 55F.
289TABLE 55F
|
|
Domain Analysis of NOV55a
NOV55aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
adh_short:24..265 78/275 (28%)5.4e−42
domain 1 of 1178/275 (65%)
|
Example 56
[0603] The NOV56 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 56A.
290TABLE 56A
|
|
NOV56 Sequence Analysis
|
|
SEQ ID NO:1391553 bp
NOV56A,TTTTAATGTGGTTCGTGACTGAGCCAATTCTCTGGGCATTTTTCCTATTCTTAATGCT
|
CG92477-01 DNA SequenceAATGACTATCCACTGCTGGTCCTCTTATCTGTTTACTCAAAAACACAAACTGACTCTC
|
ATCCTTGTGTTTCAGGATGTCATCAGTATAGCTGACAATATCCTTAATTCAGCCTCAG
|
TAACCAACTGGACAGTCTTACTGCGGGAAGAAAAGTATGCCAGCTCACCGTTACTAGA
|
GACATTAGAAAACATCAGCACTCTCGTGCCTCCGACAGCTCTTCCTCTGAATTTTTCT
|
CGGAAATTCATTGACTGGAAAGGGATTCCAGTGAACAAAAGCCAACTCAAAAGGGGTT
|
ACAGCTATCAGATTAAAATGTGTCCCCAAAATACATCTATTCCCATCAGACGCCGTGT
|
GTTAATTGGGTCAGACCAATTCCAGAGATCCCTTCCAGAAACTATTATCAGCATGGCC
|
TCGTTGACTCTGGCGAACATTCTACCCCTTTCCAAAAATGCAAATGCTCAGGTCAATG
|
GACCTGTGATATCCACGGTTATTCAAAACTATTCCATAAATGAAGTTTTCCTATTTTT
|
TTCCAAGATAGAGTCAAACCTGAGCCAGCCTCATTGTGTGTTTTGGGATTTCAGTCAT
|
TTGCAGPCCAACGATGCAGGCTCCCACCTAGTGAATGAAACTCAAGACATCGTGACGT
|
GCCAATGTACTCACTTCACCTCCTTCTCCATGTTGATGTCACCTTTTGTCCCCTCTAC
|
AATCTTCCCCGTTGTAAAATGGATCACCTATGTGGGACTGGGTATCTCCATTGGAAGT
|
CTCATTTTATGCCTGATCATCGAGGCTTTGTTTTGGAAGCAGATTAAAAAAACCCAAA
|
CCTCTCACACACGTCGTATTTGCATGGTGAACATAGCCCTGTCCCTCTTGATTGCTGA
|
TGTCTGGTTTATTGTTGGTGCCACAGTGGACACCACGGTGAACCCTTCTGGAGTCTGC
|
ACAGCTGCTGTGTTCTTTACACACTTCTTCTACCTCTCTTTGTTCTTCTGGATGCTCA
|
TGCTTGCCATCCTGCTCCCTTACCGGATCATCCTCGTGTTCCATCACATGGCCCAGCA
|
TTTGATGATGGCTGTTGCATTTTGCCTCCGTTATGGGTGCCCTCTCATTATATCTGTC
|
ATTACCATTGCTGTCACGCAACCTAGCAATACCTACAAAAGGAAAGATGTGTGTTGGC
|
TTAACTGGTCCAATCGAAGCAAACCACTCCTGGCTTTTGTTGTCCCTGCACTGCCTAT
|
TGTGGCTGTGAACTTCGTTGTCGTGCTGCTAGTTCTCACAAAGCTCTGGAGGCCGACT
|
GTTGGGGAAAGACTGAGTCGGGATGACAAGGCCACCATCGTCCCCGTGCGGAAGAGCC
|
TCCTCATTCTGACCCCTCTGCTACGGCTCACCTGGGCCTTTGGAATAGGAACAATAGT
|
GGACAGCCAGAATCTGGCTTGGCATGTTATTTTTGCTTTACTCAATCCATTCCAGCTG
|
AGAACAGTAACAATAACCTATTGTATTGTCAAGTGATTGGAATAA
|
ORF Start: ATG at 6ORF Stop: TGA at 1542
SEQ ID NO:140512 aa MW at 57687.7 kD
NOV56a,MWFVTEPILWAFFLFLMLMTIHCWSSYLFTQKHKTLTLVFQDVISIADNILNTSASVT
|
CG92477-01 Protein SequenceNWTVLLREEKYASSRLLETLENISTLVPPTALPLNFSRKFIDWKGIPVNKSQLKRGYS
|
YQIKMCPQNTSIPIRGRVLIGSDQBQRSLPETIISMASLTLGNILPVSKNGNAQVNGP
|
VISTVIQNYSINEVFLFFSKIESNLSQPHCVFWDFSHLQWNDAGCELVNETQDIVTCQ
|
CTHLTSFSMLMSPFVPSTIPPVVKWITYVGLGTSIGSLILCLIIEALFWKQIKKSQTS
|
HTRRICMVNIALSLLIADVWFIVGATVDTTVNPSGVCTAAVFFTHFFYLSLFFWMLML
|
GILLAYRIILVFHHMAQHLMMAVGFCLCYCCPLIISVITIAVTQPSNTYKRKDVCWLN
|
WSNGSKPLLAFVVPALAIVAVNIVVVLLVLTKLWRPTVGERLSRDDKATIVRVGKSLL
|
ILTPLLGLTWGFGIGTIVDSQNLAWHVIFALLNAFQVRTVTITYCIVK
|
[0604] Further analysis of the NOV56a protein yielded the following properties shown in Table 56B.
291TABLE 56B
|
|
Protein Sequence Properties NOV56a
|
|
PSort0.6850 probability located in endoplasmic reticulum
analysis:(membrane); 0.6400 probability located in plasma
membrane; 0.4600 probability located in Golgi body;
0.1000 probability located in endoplasmic reticulum
(lumen)
SignalPCleavage site between residues 24 and 25
analysis:
|
[0605] A search of the NOV56a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 56C.
292TABLE 56C
|
|
Geneseq Results for NOV56a
NOV56aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAG68126Human 7TM-GPCR protein sequence42..502459/461 (99%)0.0
SEQ ID NO:6 - Homo sapiens, 928 aa.195..655 461/461 (99%)
[WO200168701-A2, 20 Sep. 2001]
AAE12023Human G-protein coupled receptor,42..500456/459 (99%)0.0
GCREC-2 - Homo sapiens, 910 aa.372..830 459/459 (99%)
[WO200172836-A2, 04 Oct. 2001]
AAB61616Human protein HP10678 - Homo21..500196/491 (39%)3e−95
sapiens, 542 aa. [WO200102563-A2,10..491297/491 (59%)
11 Jan. 2001]
AAY57288Human GPCR protein (HGPRP)44..510187/480 (38%)8e−94
sequence (clone ID 3036563) - Homo256..729 283/480 (58%)
sapiens, 807 aa. [WO200015793-A2,
23 Mar. 2000]
AAU04581Human G-protein coupled receptor44..510187/480 (38%)1e−93
like protein, GPCR #12 - Homo795..1268282/480 (57%)
sapiens, 1346 aa. [WO200153454-A2,
26 Jul/ 2001]
|
[0606] In a BLAST search of public sequence datbases, the NOV56a protein was found to have homology to the proteins shown in the BLASTP data in Table 56D.
293TABLE 56D
|
|
Public BLASTP Results for NOV56a
NOV56aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
AAH19217HYPOTHIETICAL 101.5 KDA39..500349/462 (75%)0.0
PROTEIN - Mus musculus368..828 410/462 (88%)
(Mouse), 908 aa.
Q96DQ1CDNA FLJ30646 FIS, CLONE42..283240/242 (99%)e−138
CTONG2004716, WEAKLY60..301242/242 (99%)
SIMILAR TO RATTUS
NORVEGICUS SEVEN
TRANSMEMBRANE RECEPTOR -
Homo sapiens (Human), 302 aa.
Q9WVT0SEVEN TRANSMEMBRANE40..510191/487 (39%)9e−98
RECEPTOR - Rattus norvegicus791..1271294/487 (60%)
(Rat), 1349 aa.
CAC27252SEQUENCE 29 FROM PATENT21..500196/491 (39%)8e−95
WO0102563 - Homo sapiens10..491297/491 (59%)
(Human), 542 aa.
Q9UIZ3DJ365O12.1 (KIAA0758 PROTEIN) -44..510187/480 (38%)2e−93
Homo sapiens (Human), 1346 aa.795..1268283/480 (58%)
|
[0607] PFam analysis predicts that the NOV56a protein contains the domains shown in the Table 56E.
294TABLE 56E
|
|
Domain Analysis of NOV56a
NOV56aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
GPS: domain 1 of 1200..25024/56 (43%)3e−12
40/56 (71%)
7tm_2:252..511 75/290 (26%)9.4e−20
domain 1 of 1193/290 (67%)
|
Example 57
[0608] The NOV57 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 57A.
295TABLE 57A
|
|
NOV57 Sequence Analysis
|
|
SEQ ID NO:1412320 bp
NOV57a,TTCAGAAGGAAATATCATCAGCAGAAGCTCCTAAAAAATAATGAGTCCTTGGATGAAG
|
CG92499-01 DNA SequenceGCTTGAGGCTACACACAGTGAATCTGAGACAACTGGGTCATTGTCTTGCCATGGAGGA
|
ACCCAAAGGCTACTACTGGCCATCTATCCAACCTTCTGAATACGTTCTTCCTTGTCCA
|
GACAAGCCTGGGTTTTCTGCTTCTCGGATATGTTTTTACAATGCTACCAACCCATTGG
|
TAACCTACTGGGGACCTGTTGATATCTCCAACTGTTTAAAAGAAGCAAATGAACTTGC
|
TAACCAGATTTTAAATTTAACTGCTGATGGCCAGAACTTAACCTCAGCCAATATTACC
|
AACATTGTGGAACAGGTCAAAACAATTGTGAATAAAGAAGAAAACATTGATATAACAC
|
TTGGCTCAACTCTAATGAATATATTTTCTAATATCTTAAGCAGTTCAGACAGTGACTT
|
GCTTGAGTCATCTTCTGAAGCTTTAAAAACAATTGATGAATTGGCCTTCAACATAGAC
|
CTAAATAGCACATCACATGTGAATATTACAACTCGGAACTTGGCTCTCAGCGTATCAT
|
CCCTGTTACCAGGGACAAATGCAATTTCAAATTTTAGCATTGGTCTTCCAAGCAATAA
|
TGAATCGTATTTCCACATGGATTTTGAGAGTCGACAAGTGGATCCACTGGCATCTGTA
|
ATTTTGCCTCCAAACTTACTTGAGAATTTAAGTCCAGAAGATTCTGTATTAGTTAGAA
|
GAGCACAGTTTACTTTCTTCAACAAAACTGGACTTTTCCAGGATGTAGGACCCCAAAG
|
AAAAACTTTAGTGAGTTATGTGATGGCGTGCAGTATTGCAAACATTACTATCCAGAAT
|
CTGAAGGATCCTGTTCAAATAAAAATCAAACATACAAGAACTCAGGAAGTGCATCATC
|
CCATCTGTGCCTTCTGCGATCTGAACAAAAACAAAAGTTTTCCAGGATGCAACACGTC
|
AGGATGTGTTGCACACAGAGATTCAGATGCAAGTGACACAGTCTGCCTGTGTAACCAC
|
TTCACACACTTTCGAGTTCTGATGGACCTTCCAAGAAGTCCCTCACAGTTACATGCAA
|
GAAACACTAAAGTCCTCACTTTCATCAGCTATATTGGGTGTCGAATATCTGCTATTTT
|
TTCAGCAGCAACTCTCCTGACATATGTTGCTTTTGAGAAATTGCGAAGGGATTATCCC
|
TCCAAAATCTTGATCAACCTGAGCACAGCCCTGCTGTTCCTGAATCTCCTCTTCCTCC
|
TAGATGGCTGGATCACCTCCTTCAATQTCCATGGACTTTGCATTGCTGTTGCAGTCCT
|
GTTGCATTTCTTCCTTCTGGCAACCTTTACCTGGATGCGGCTAGAAGCAATTCACATG
|
TACATTGCTCTAGTTAAAGTATTTAACACTTACATTCGCCGATACATTCTAAAATTCT
|
GCATCATTGGCTGCGGTTTGCCTGCCTTAGTGGTGTCAGTTGTTCTAGCGAGCAGAAA
|
CAACAATCAAGTCTATGGAAAAGAAAGTTATCGCAAAGAAAAAGGTGATGAATTCTGT
|
TGGATTCAAGATCCAGTCATATTTTATGTGACCTGTGCTGGGTATTTTGGAGTCATGT
|
TTTTTCTGAACATTGCCATGTTCATTGTGGTAATGGTGCAGATCTGTGGCAGCAATGG
|
CAAGAGAAGCAACCGGACCCTGAGAGAAGAAGTGTTAAGGAACCTGCGCAGTGTGGTT
|
AGCTTGACCTTTCTGTTGCGCATGACATGGGGTTTTGCATTCTTTGCCTGGGGACCCT
|
TAAATATCCCCTTCATGTACCTCTTCTCCATCTTCAATTCATTACAAGGCTTATTTAT
|
ATTCATCTTCCACTGTCCTATGAACGAGAATGTTCAGAAACAGTGGCGGCGGCATCTC
|
TGCTGTGGTAGATTTCGGTTAGCACATAACTCAGATTCGAGTAAGACAGCTACCAATA
|
TCATCAAGAAAAGTTCTCATAATCTAGGAAAATCTTTGTCTTCAAGCTCCATTGGTTC
|
CAACTCAACCTATCTTACATCCAAATCTAAATCCAGCTCTAACCACCTATTTCAAAAG
|
GAATAGCCACACAGATTATGTCTCCTATAACTTTCCTCCCCAAAGTGGTTCTCCGACG
|
GGTTCCTTGAAAGTCTTTTAAAATGCCCTTTATGGAAAAAATCAAACCCCTCCCCGGT
|
TTTTTGGGGGGTTTGGGGTCTCCCCTTTTTTATTTTTTAAATTGGGGGGGTTTTTTAA
|
AAAAAATTTATTTTTGGGGGGTTTTTTTTTATTAAAAAAAAAAAAAAACAATTTTTTA
|
ORF Start: ATG at 109ORF Stop: TAG at 2092
SEQ ID NO:142661 aa MW at 74148.5 kD
NOV57a,MEEPKGYYWPSIQPSEYVLPCPDKPGFSASRICFYNATNPLVTYWGPVDISNCLKEANMEEPKGYYWPS I QPSEYVLPCPDKPGFSASRI CPYNATNPLVrYWGPVD ISNCLKEAN
|
CG92499-01 Protein SequenceEVANQILNLTADGQNLTSANITNTVEQVKRIVNKBENIDITLGSTLMNIFSNILSSSD
|
SDLLESSSEALKTIDELAFKIDLNSTSHVNITTRNLALSVSSLLPGTNAISNFSIGLP
|
SNNESYFQMDFESGQVDPLASVILPPNLLENLSPEDSVLVRRAQFTFFNKTGLFQDVG
|
PQRKTLVSYVMACSIGNITTQNLKDPVQIKIKHTRTQEVHHPICAFWDLNKNKSFGGW
|
NTSGCVAHRDSDASETVCLCNHFTHFGVLMDLPRSASQLDARNTKVLTFISYIGCGIS
|
AIFSAATLLTYVAFSKLRRDYPSKILMNLSTALLFLNLLFLLDGWITSFNVDGLCIAV
|
AVLLHFFLLATFTWMGLEAIHMYIALVKVFNTYIRRYILKFCIIGWGLPALVVSVVLA
|
SRNNNEVYGKESYGKEKGDEFCWIQDPVXFYVTCAGYFGVMFFLNIAMFIVVMVQICG
|
RNGKRSNRTLRBEVLRNLRSVVSLTFLLGMTWGFAFFAWGPLNIPFMYLFSIFNSLQG
|
LFIFIFHCAMKENVQKQWRRHLCCGRFRLADNSDWSKTATNITKKSSDNLGKSLSSSS
|
IGSNSTYLTSKSKSSSNHLFQKE
|
SEQ ID NO:1432428 bp
NOV57b,GGATGAGCCAAGCTTGGTGCTTTCGCCCCTTCTAGTTTACAATGCTACCAACAATACT
|
CG92499-02 DNA SequenceAATTTAGAAGGAAAAATCATTCAGCAGAACCTCCTAAAAAATAATGAGTCCTTGGATG
|
AAGGCTTGAGGCTACATACAGTGAATGTCACACAACTGGGTCATTGTCTTGCCATGCA
|
GGAACCCAAAGGCTACTACTGGCCATCTATCCAACCTTCTGAATACGTTCTTCCTTGT
|
CCAGACAAGCCTGGCTTTTCTGCTTCTCGGATATGTTTTTACAATGCTACCAACCCAT
|
TGGTAACCTACTGGCGACCTGTTGATATCTCCAACTGTTTAAAACAAGCAAATGAAGT
|
TGCTAACCAGATTPTAAATTTAACTGCPGATGGGCAGAACTTAACCTCAGCCAATATT
|
ACCAACATTGTGGAACAGGTCAAAAGAATTGTGAATAAAGAAGAAAACATTGATATAA
|
CACTTGGCTCAACTCTAATGAATATATTTTCTAATATCTTAAGCAGTTCAGACAGTCA
|
CTTGCTTGAGTCATCTTCTGAAGCTTTAAAAACAATTQATGAATTGGCCTTCAAGATA
|
GACCTAAATACCACATCACATGTGAATATTACAACTCCGAACTTGGCTCTCAGCGTAT
|
CATCCCTCTTACCAGGGACAAATGCAATTTCAAATTTTAGCATTGGTCTTCCAAGCAA
|
TAATGAATCGTATTTCCAGGTACACATGCATTTTGAGAGTGGACAACTCGATCCACTG
|
GCATCTGTAATTTTCCCTCCAAACTTACTTGACAATTTAAGTCCAGAACATTCTGTAT
|
TAGTTAGAAGAGCACAGTTTACTTTCTTCAACAAAACTGGACTTTTCCAGGATGTAGG
|
ACCCCAAAGAAAAACTTTAGTGAGTTATGTGATGGCGTGCAGTATTGGAAACATTACT
|
ATCCAGAATCTGAAGGATCCTGTTCAAATAAAAATCAAACATTATAGAGAAGAAACTG
|
ACCTGCTGTTTTCACACTGTTTGTTGATTCCTTCAACAGAAAGTTTTCGAGGATGGAA
|
CACGTCAGGATGTGTTGCACACAGAGATTCAGATGCAACTGAGACAGTCTGCCTGTGT
|
AACCACTTCACACACTTTGGAGTTCTGATGGACCTTCCAAGAAGTGCCTCACAGTTAG
|
ATGCAAGAAACACTAAAGTCCTCACTTTCATCAGCTATATTGGGTGTGGAATATCTGC
|
TATTTTTTCAGCAGCAACTCTCCTGACATATGTTGCTTTTGAGAAATTGCGAAGGGAT
|
TATCCCTCCAAAATCTTGATGAACCTGAGCACAGCCCTGCTGTTCCTCAATCTCCTCT
|
TCCTCCTAGATGGCTGGATCACCTCCTTCAATGTGGATGGACTTTGCATTGCTGTTGC
|
AGTCCTGTTCCATTTCTTCCTTCTCGCAACCTTTACCTGGATGCGGCTAGAAGCAATT
|
CACATCTACATTGCTCTAGTTAAAGTATTTAACACTTACATTCCCCGATACATTCTAA
|
AATTCTGCATCATTGGCTGGGGTTTGCCTGCCTTAGTGGTGTCAGTTGTTCTAGCGAG
|
CAGAAACAACAATCAAGTCTATGGAAAAGAAAGTTATGGGAAAGAAAAAGGTGATGAA
|
TGTTGGATTCAAGATCCAGTCATATTTTATGTGACCTGTGCTGGGTATTTTGGAGTCA
|
TGTTTTTTCTGAACATTGCCATGTTCATTGTGGTAATGGTGCAGATCTCTGGGAGGAA
|
TGGCAAGAGAAGCAACCGGACCCTGAGAGAACAAGTCTTAAGGAACCTGCGCACTGTG
|
GTTAGCTTGACCTTTCTGTTGGGCATGACATGGGGTTTTGCATTCTTTCCCTGGGGAC
|
CCTTAAATATCCCCTTCATCTACCTCTTCTCCATCTTCAATTCATTACAAGGTTTATT
|
TATATTCATCTTCCACTCTGCTATCAAGGAGAATGTTCACAAACAGTCGCGGCAGCAT
|
CTCTGCTGTGGTAGATTTCGGTTAGCACATAACTCAGATTGGAGTAAGACAGCTACCA
|
ATATCATCAAGAAAAGTTCTGATAATCTAGGAAAATCTTTGTCTTCAAGCTCCATTGG
|
TTCCAACTCAACCTATCTTACATCCAAATCTAAATCCACCTCTACCACCTATTTCAAA
|
AGGAATAGCCACACAGACAGTGCTTCCATGGACAAGTCCTTGTCAAAACTGGCCCATG
|
CTGATGGAGATCAAACATCAATCATCCCTGTCCATCAGGTCATTGATAAGGTCAAGGG
|
TTATTCCAATGCTCATTCACACAACTTCTATAAAAATATTATCATGTCAGACACCTTC
|
AGCCACAGCACAAAGTTTTAATGTCTTTAAGAAAAAGAAATCAATCTGCAGAAATGTG
|
AAGATTTGCAAGCAGTGTAAACTGCAACTAGTGATGTAAATGTGCTATTA
|
ORF Start: ATG at 170ORF Stop: TAA at 2339
SEQ ID NO:144723 aa MW at 80895.8 kD
NOV57b,MEEPKGYYWPSIQPSEYVLPCPDKPGFSASRICFYNATNPLVTYWCPVDISNCLKEAN
|
CG92499-02 Protein SequenceEVANQILNLTANCQNLTSANTTNIVEQVKRIVNKEENIDITLCSTLMNIFSNILSSSD
|
SDLLESSSEALKTIDELAFKIDLNSTSHVNITTRNLALSVSSLLPGTNAISNFSIGLP
|
SNNESYFQVQMDFESGQVDPLASVILPPNLLENLSPEDSVLVRRAQPTFFNKTGLFQD
|
VGPQRKTLVSYVMACSIGNITIQNLKDPVQIKIKHYREETDLLFSHCLLIPSTESFGG
|
WNTSGCVAHRDSDASETVCLCNHFTHFGVLNDLPRSASQLDARNTKVLTPISYIGCGI
|
SAIFSAATLLTYVAFEKLRRDYPSKILMNLSTALLFLNLLFLLDGWITSFNVDGLCIA
|
VAVLLHFFLLATFTWNCLEAIHMYIALVKVFNTYIRRYILKFCIICWGLPALVVSVVL
|
ASRNNNEVYGKESYGKEKGDECWIQDPVIFYVTCAGYFGVMFFLNIAMFIVVMVQICG
|
RNGKRSNRTLREEVLRNLRSVVSLTFLLGMTWCFAFFAWGPLNIPFMYLFSTFNSLQG
|
LFIFIFHCAKKENVQKQWRQHLCCGRFRLADNSDWSKTATNIIKKSSDNLCKSLSSSS
|
IGSNSTYLTSKSKSSSTTYFKRNSHTDSASMDKSLSKLAHADGDQTSIIPVHQVIDKV
|
KGYCNAHSDNFYKNIIMSDTFSESTKF
|
[0609] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 57B.
296TABLE 57B
|
|
Comparison of NOV57a against NOV57b.
ProteinNOV57a Residues/Identitites/Similarities
SequenceMatch Residuesfor the Matched Region
|
NOV57b1..624536/627 (85%)
1..624542/627 (85%)
|
[0610] Further analysis of the NOV57a protein yielded the following properties shown in Table 57C.
297TABLE 57C
|
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Protein Sequence Properties NOV57a
|
|
PSort0.6000 probability located in plasma membrane; 0.4000
analysis:probability located in Golgi body; 0.3000 probability
located in endoplasmic reticulum (membrane);
0.0300 probability located in mitochondrial inner
membrane
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0611] A search of the NOV57a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 57D.
298TABLE 57D
|
|
Geneseq Results for NOV57a
NOV57aIdentities
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
AAU00719Human novel G-protein coupled 1..660655/660 (99%)0.0
receptor, NGPCR#29 - Homo sapiens, 499..1158657/660 (99%)
1192 aa. [WO200118207-A1,
15 Mar. 2001]
AAU00210Human novel G-protein coupled 1..660655/660 (99%)0.0
recentor NGPCR#21 - Homo sapiens, 499..1158657/660 (99%)
1221 aa. [WO200118207-A1,
15 Mar. 2001]
AAU00207Human novel G-protein coupled 1..660655/660 (99%)0.0
receptor, NGPCR#18 - Homo sapiens, 500..1159657/660 (99%)
1193 aa. [WO200118207-A1,
15 Mar. 2001]
AAU00206Human novel G-protein coupled 1..660655/660 (99%)0.0
receptor, NGPCR#17 - Homo sapiens, 500.. 1159657/660 (99%)
1222 aa. [WO200118207-A1,
15 Mar. 2001]
AAU00199Human novel G-protein coupled 1..660655/660 (99%)0.0
receptor, NGPCR#10 - Homo sapiens, 528..1187657/660 (99%)
1221 aa. [WO200118207-A1,
15 Mar. 2001]
|
[0612] In a BLAST search of public sequence datbases, the NOV57a protein was found to have homology to the proteins shown in the BLASTP data in Table 57E.
299TABLE 57E
|
|
Public BLASTP Results for NOV57a
NOV57aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe Matched ValueExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96JW0CDNA FLJ14937 FIS, CLONE183..660473/478 (98%)0.0
PLACE1010231, WEAKLY SIMILAR 1..478475/478 (98%)
TO CELL SURFACE
GLYCOPROTEIN EMR1
PRECURSOR - Homo sapiens
(Human), 512 aa.
Q9Y3K0DJ287G14.2 (PUTATTVE NOVEL183..660472/478 (98%)0.0
SEVEN TRANSMEMBRANE 1..478475/478 (98%)
DOMAIN PROTEIN) - Homo sapiens
(Human), 541 aa (fragment).
CAB69577SEQUENCE 1 FROM PATENT 78..655278/588 (47%)e-155
EP0805204 - unidentified, 1038 aa391..968396/588 (67%)
(fragment).
O00406G-PROTEIN-COUPLED RECEPTOR 78..655278/588 (47%)e-155
HE6 PRECURSOR - Homo sapiens367..944396/588 (67%)
(Human), 1014 aa.
CAC43514SEQUENCE 1 FROM PATENT221..591135/385 (35%)1e-57
WO0144281 - Homo sapiens (Human),135..507210/385 (54%)
528 aa.
|
[0613] PFam analysis predicts that the NOV57a protein contains the domains shown in the Table 57F.
300TABLE 57F
|
|
Domain Analysis of NOV57a
NOV57aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionfor the MatchedValue
|
HRM: domain 1 of 1 3..5818/79 (23%)9.6
38/79 (48%)
GPS: domain 1 of 1272.32525/55 (45%)2.1e-16
44/55 (80%)
LHC: domain 1 of 1404..422 6/21 (29%)5.7
15/21 (71%)
DUF32: domain 1 of 1486..542 14/163 (9%)1.1
42/163 (26%)
7tm_2: domain 1 of 1334..594 74/281 (26%)1.3e-59
200/281 (71%)
|
Example 58
[0614] The NOV58 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 58A.
301TABLE 58A
|
|
NOV58 Sequence Analysis
|
|
SEQ ID NO:1454483 bp
NOV58a,CCCCTGACAGAGCGTGGCAGCCCCAGGCTCTTTGCATAATCCTGTGGCTTCGCTGTCT
|
CG92541-01 DNA SequenceTCACCCAGCACCACCGGACAGGGAAGGGCAGAGAAGGCCACCATGGCGACACTCCTCT
|
CCCATCCGCAGCAGCGCCCTCCCTTCTTGCGCCAOCCCATCAACATAAGGCGCCGCAG
|
AGTCAGAGATCTACAGGATCCCCGCCCCAAATGGCCCCGAAGGATCCAGCCTCCATCC
|
CACCACTTCTCCCCCGAGCAGCCGGCCCTGCTCTACGAGGACGCACTCTACACTGTCT
|
TGCACCCCCTGGGTCATCCTGAGCCCAACCATGTGACGGACGCCTCTGACCTGCTGCG
|
ATACCTGCAGGAGGCCTTCCACGTGGACCCCGAGGAGCACCAGCAGACACTGCAGCGG
|
GTCAGGGAGCTTGAGAAGCCAATATTTTGTCTGAAGGCAACAGTGAAACAGGCCAAGG
|
GCATTCTGGGCAAAGATGTCAGTGGGTTCAGCGACCCCTACTGCCTGCTGGGCATTGA
|
GCACGCGGTAGGTGTGCCAGGCCCCAGCCCCCGGTCCCGQCATCGGCAGAAGGCTGTG
|
GTGAGGCACACCATCCCCGACGAGGAGACCCACCGCACGCAGGTCATCACCCAGACAC
|
TCAACCCCGTCTGCGACGAGACCTTCATCCTGGAGTTTCAGGACATCACCAATGCGAG
|
CTTTCATCTGCACATGTCGGACCTGGACACTGTGGAGTCTGTCCQACAGAAGCTTGGG
|
GAGCTCACGGATCTGCATGGGCTTCGCACGATCTTTAAAGAGGCCCGGAACGACAAAG
|
GCCAGGACGACTTTCTGCGGAACGTGGTTCTGAGGCTGCAGGACCTGCCCTGCCGAGA
|
CGACCAGTGGTACCCCCTGCAACCCCGCACTGAGACCTACCCAGACCGAGGCCAGTGC
|
CACCTCCAGTTCCAACTCATCCATAAGCGGCAGAGAGCCACTTCGGCCAGCCGCTCGC
|
AGCCGAGCTACACCGTGCACCTCCACCTCCTGCAGCAGCTTGTGTCCCACGAGGTCAC
|
CCAGCACCAGGCGGGAAGCACCTCCTGGGACGGGTCGCTGAGTCCCCAGGCTGCCACC
|
GTCCTCTTTCTGCACGCCACACAGAACGACCTATCCGACTTCCACCAGTCCATCGCGC
|
AGTGGCTGGCCTACAGCCGCCTCTACCACAGCCTGGAGTTCCCCAGCACCTGCCTCCT
|
GCACCCCATCACCAGCATCGACTACCAQTGGATCCAGGGTCGGCTCAACGCAGAACAG
|
CAGGAGGAGCTGGCCGCCTCATTCAACTCCCTGATGACCTACGGCCTCTCCCTCATCC
|
GGACGTTCCGCTCTCTCATCCACCTCTCTGTCTCGGACTCCCCAGCCAGACTCCAGTC
|
TCTTCTAAGGGTCCTGCTACAGATGTGCAAGATGAAGGCCTTTCCAGAACTGTGCCCC
|
AACACCGCCCCATTGCCCCACCTGGTGACTGAGGCCCTGCAGACTGGCACCACTGAAT
|
GGTTCCACCTGAAGCACCAGCACCATCAACCCATGGTGCAGGGCATCCCCGACGCAGG
|
CAAGGCCTTCCTGCGCCTOGTACAGGATGTCATTGGCGACCTGCACCAGTGCCAGCGC
|
ACATGGGACAAGATCTTCCACAGTACCCTCAAGATCCACCTCTTCTCCATGGCTTTCC
|
GGGAGCTGCAGTGCCTGGTGCCCAAGCGGGTCCAGGACCACACCACGGTTGTGGCTGA
|
TGTAGTGTCCCCAGACATGGGCGAGAGTCTGTTCCAGCTCTACATCAGCCTCAAGGAG
|
CTCTGCCAGCTGCCCATCAGCTCCTCAGAGAGGGATGGAGTCCTCGCCCTGGATAATT
|
TCCACCGCTGCTTCCAGCCCGCCATCCCCTCCTGGCTGCAGAAGACGTACAACGAGGC
|
CCTGGCGCGCGTGCAGCGCGCTGTGCACATCGATGAGCTCGTGCCCCTGCCTGAACTG
|
ACCAAGCACAGCACATCAGCGGTGGATCTATCCAACTGCTTTGCCCAGATCAGCCACA
|
CTGCCCGGCAGCTGGACTGGCCAGACCCAAGAGAGGCCTTCATGATTACCGTCAAGTT
|
TGTGGAGGACACCTGTCGCCTGGCCCTGGTGTACTCCAGCCTTATAAAGGCCCGGGCC
|
CGCGAGCTCTCTTCAGGCCAGAAGGACCAAGGCCAGGCAGCCAACATGCTGTGTGTGG
|
TGGTGAATGACATGGAGCAGCTGCGGCTGGTGATCGGCAAGTTGCTCGCCCAGCTGGC
|
ATGGGAGGCCCTGGAGCAGCGGGTAGGGGCCGTGCTGGAGCAGGGGCAGCTGCACAAC
|
ACGCTGCATGCCCAGCTGCAGAGCGCGCTGGCCGGGCTGGGCCATGAGATCCGCACTG
|
GCGTCCGCACCCTGGCCGAGCAGTTGGAGGTGGGCATCGCCAAGCACATCCAGAAACT
|
GGTGGGCGTCAGGGAGTCTGTCCTGCCTGACGATGCCATTCTGCCCCTGATGAAGTTC
|
CTGGAGGTGGAGCTTTGCTACATGAACACCAACTTGGTGCAGGAGAACTTCAGCAGCC
|
TCCTGACCCTCCTCTGGACCCACACACTCACAGTGCTGGTGGAGGCGGCCCCCTCCCA
|
GCGCAGCTCATCCCTGGCTTCCAACAGGCTGAAGATTCCCCTGCAGAACCTCGAGATC
|
TGCTTCCACGCTGAGGGCTGTGGCCTGCCACCCAAGGCCCTGCACACTGCCACCTTCC
|
AGGCTCTGCAGAGGGACCTGGAGCTGCAGGCGACCTCCAGCGGGGAACTCATCCGGAA
|
GTACTTCTGCAGCCCAATCCAGCAGCAGCCAGAAACCACCTCTGACGAGCTGGGGGCT
|
GTGACAGTCAAGGCCTCCTACCGCGCCTCTCACCAGAAGCTGCCTGTCGAGCTGCTCA
|
GCGCCTCCAGCCTGCTGCCCCTGGACTCCAATGGTTCCAGCGACCCCTTTGTCCAGCT
|
GACCTTGGAGCCCAGGCATGAGTTCCCTGACCTGGCCGCCCGGGAGACCCAGAAGCAC
|
AAGAAGGACCTTCACCCATTGTTTGATGAGACCTTTGAATTCCTGGTGCCTCCTGAGC
|
CGTGCCGCAACGCTCGGGCATGCCTCCTGCTCACCGTGCTGGACTACGACACGCTGGG
|
GGCCGACGACCTGGAAGGCGAGGCCTTCCTGCCGCTGCGTGAGGTGCCCGGGCTGACT
|
GGCTCTGAGGAGCCTGGTGAGGTGCCTCAGACCCGCCTGCCCCTCACGTACCCCGCAC
|
CCAACGGGGACCCAATCCTCCAGCTGCTGGAGGGCCGGAAGGGTGACCCAGAAGCCCA
|
GGTCTTTGTGAGGCTGCGGCGGCACCGGGCCAAGCAGCCCTCCCAGCATGCCTTGCGG
|
CCGGCACCGTAGCCGTAGAGGTTTGCGGTGGGGCTCCGTCCCCGGTGGGGACTTGCAA
|
GGGCCTTCCTGTAGGGTCTGGGGCTTCCCCGCCACATCGCGGCCCTCCAGCCTGGCCT
|
AACACTTGGCGAGCCCCAGCATGCGGAGTGCCCAGAGTGCAGACCTCCCCTGCCTCCC
|
ATGGTGATGGCGCCTCAGCAGCGACATCTCTACTCCCGCCTCCCTGCCTCCAGCCCTG
|
GCTGCAATGTCTCTACCACATCCCAGCACCAGGGGGAGCAAACCCTGCCCCTGCCCGC
|
CTCTCAGAAAAGCTGCTGTGGTGGGCAGCGGATTGGGCCATCTGTCTCCTGGCCCTGG
|
CCCATCTGCCTCCTGGCCTTCCTGTTCCAGCCACTGGGGTGGGGGCCAGGTTCACTGG
|
GACCAGGGCTACACGCACAGAGTCTCCTGGAAAAGGGACAGGGGACCCTGCCAAAGAT
|
GAGGCTCCAGCTCCCCTGGCGGGAGGGTGGTGGCCATTACTAGAGCGGGCCTGGGTCC
|
TCTCCCCAGGGGCTGCCAGCATCCAGGCCAGCAAGCCTGGAGCCAAGAACCTTCTGGC
|
TCTGAGGGAGCAAGAGCTGCCAGGCGGCAGGGCTGGCACAGACAGACGGAAGCAGAAA
|
GGACAGTTTGGCTGCTGTGTCTGCTGCGCACGCCCCCTCCCCGGACAGCACCTGCCAC
|
CTAGAAACTTTCTTAGCAAAAAAATTAATAAAAACAAATCCATTGTCCTCTTAAAATA
|
TCCTTTGGCCTACAGTCGCGCCTGGAATGCGAGCCAGGCCGGCTAGCTTCCTCCCCAG
|
CCCTCAGGGGACTTTGACGTACCGCCACCTTGGGGTAGCTACAAACCAGGGGGGTAGG
|
TGTGGAAATAACTCAGGCAGAGCCACGGCTAGGGTCATTTPTGGCCGTGCGCTTTGAA
|
TAAATTGCTTTACCACGCATACCACTTCCTGTGGTCACACCCAGGACAGCGACCCGTT
|
CCTCGGGGGACCACAGTGAGCACGCGCCTCCCCAGGGTGCAGGTTGAGGCCTGAGGGC
|
TGCTCTTGAGACAGTAGGGCGTAGAGCAACTGGGTCCTTCCCCTCCCTGGGGGGTCAA
|
AACCTGAGCCTGGGCTG
|
ORF Start: ATG at 101ORF Stop: TAG at 3374
SEQ ID NO:1461091 aa MW at 123545.0 kD
NOV58a,MATLLSHPQQRPPFLRQAIKIRRRRVRDLQDPPPQMAPRIQPPSHHFSPEQRALLYED
|
CG92541-01 Protein SequenceALYTVLHRLGHPEPNHVTEASELLRYLQEAFHVEPEEEQQTLQRVRELEKPIFCLKAT
|
VKQAKGILGKDVSGFSDPYCLLGIEQGVGVFGGSPGSRHRQKAVVRHTTPEEETHRTQ
|
VITQTLNPVWDETFTLEFEDITNASPHLDMWDLDTVESVRQKLGELTDLHGLRRIFKE
|
ARKDKGQDDPLRNVVLRLQDLRCREDQWYPLEPRTETYPDRGQCHLQFQLIHKRQRAT
|
SASRSQPSYTVHLHLLQQLVSHEVTQHQACSTSWDGSLSPQAATVLFLHATQKDLSDF
|
HQSMAQWLAYSRLYQSLEEPSSCLLHPITSIEYQWIQGRLKAEQQEELAASFNSLMTY
|
GLSLIRRFRSVIELSVSDSPARLQSLLRVLVQMCKMKAFGELCPNTAPLPQLVTEALQ
|
TGTTEWFHLKQQEHQPMVQGIPEAGKALLGLVQDVIGDLHQCQRTWDKIFHSTLKIHL
|
FSMAFRELQWLVAKRVQDHTTVVGDVVSPEMGESLFQLYISLKELCQLRNSSSERDGV
|
LALDNFHRWFQPAIPSWLQKTYNBALARVQRAVQMDELVPLGELTKHSTSAVDLSNCF
|
AQISHTARQLDWPDPREAFMITVKFVEDTCRLALVYCSLIKARARELSSGQKDQGQAA
|
NMLCVVVNDMEQLRLVTGKLLAQLAWEALEQRVGAVLEQGQLQNTLHAQLQSALAGLG
|
HEIRTGVRTLAEQLEVGIAKHIQKLVOVRESVLPEDAILPLMKFLEVELCYMNTNLVQ
|
ENFSSLLTLLWTHTLTVLVEAAASQRSSSLASNRLKIALQNLEICFHAEGCGLPPKAL
|
HTATFQALQRDLELQATSSGELIRKYFCSRIQQQAETTSEELGAVTVKASYRASEQKL
|
RVELLSASSLLPLDSNGSSDPFVQLTLEPRHEFPELAARETQKHKKDLHPLFDSTFEF
|
LVPAEPCRKAGACLLLTVLDYDTLGADDLEGEAFLPLREVPGLSGSEEPGEVPQTRLP
|
LTYPAPNCDPILQLLEGRKGDREAQVFVRLRRHRAKQASQHALRPAP
|
[0615] Further analysis of the NOV58a protein yielded the following properties shown in Table 58B.
302TABLE 58B
|
|
Protein Sequence Properties NOV58a
|
|
PSort0.9600 probability located in nucleus: 0.3092 probability
analysis:located in microbody (peroxisome); 0.1776 probability
located in lysosome (lumen); 0.1000 probability located
in mitochondrial matrix space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0616] A search of the NOV58a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 58C.
303TABLE 58C
|
|
Geneseq Results for NOV58a
NOV58aIdentities
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB41695Human ORFX ORF1459 polypeptide 587..1091498/505 (98%)0.0
sequence SEQ ID NO:2918 - Homo 5..509498/505 (98%)
sapiens, 509 aa.
[WO200058473-A2, 05 Oct. 2000]
ABG08373Novel human diagnostic protein 48..1049356/1048 (33%)e-155
#8364 - Homo sapiens, 1433 aa. 132..1168547/1048 (51%)
[WO200175067-A2, 11 Oct. 2001]
ABG08373Novel human diagnostic protein 48..1049356/1048 (33%)e-155
#8364 - Homo sapiens, 1433 aa. 132..1168547/1048 (51%)
[WO200175067-A2, 11 Oct. 2001]
AAU19712Human novel extracellular matrix 1..158152/158 (96%)5e-86
protein, Seq ID No 362 - Homo 32..189154/158 (97%)
sapiens, 191 aa.
[WO200155368-A1, 02 Aug. 2001]
AAM83868Human immune/haematopoietic 1..8887/88 (98%)3e-45
antigen SEQ ID NO:11461 - Homo 36..12387/88 (98%)
sapiens, 140 aa.
[WO200157182-A2, 09 Aug. 2001]
|
[0617] In a BLAST search of public sequence datbases, the NOV58a protein was found to have homology to the proteins shown in the BLASTP data in Table 58D.
304TABLE 58D
|
|
Public BLASTP Results for NOV58a
NOV58aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9R189MUNC13-4 PROTEIN - Rattus 1..1091941/1091 (86%)0.0
norvegicus (Rat), 1088 aa. 1..1088992/1091 (90%)
Q9H7K5FLJ00067 PROTEIN - Homo318..816 491/499 (98%)0.0
sapiens (Human), 574 aa8..506 494/499 (98%)
(fragment).
Q96RZ3BAI ASSOCIATED PROTEIN 3 -48..1073359/1055 (34%)e-160
Homo sapiens (Human), 1187 aa.132..1174 556/1055 (52%)
Q9UJK1C316G12.1 (KIAA0734 (C248..1073359/1055 (34%)e-160
DOMAIN PROTEIN)) - Homo97..1139556/1055 (52%)
sapiens (Human), 1152 aa.
O94839KIAA0734 PROTEIN - Homo48..1073359/1055 (34%)e-160
sapiens (Human), 1186 aa131..1173 556/1055 (52%)
(fragment).
|
[0618] PFam analysis predicts that the NOV58a protein contains the domains shown in the Table 58E.
305TABLE 58E
|
|
Domain Analysis of NOV58a
NOV58aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Match RegionValue
|
C2: domain 1 of 2113..21428/116 (24%)0.075
74/116 (64%)
C2: domain 2 of 2 928..102031/101 (31%)2e-07
66/101 (65%)
|
Example 59
[0619] The NOV59 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 59A.
306TABLE 59A
|
|
NOV59 Sequence Analysis
|
|
SEQ ID NO:1471006 bp
NOV 59a,CTGTGGGTGGTGATGGATCTCAACCAAAGCCATTCCGTGACGCTGAATGATGGACATG
|
CG92662-01 DNA SequenceTCATGCCGATGCTGGGATTTCGCACTTATGCTCCTGATCATGTACCCAAGAGCAAGGC
|
TGGTCAAGCCACCCAACTGGCTATTGATGCAGCCTTCCCTCACATTGATGCGGCGTTC
|
TTCTACCAAAACGAGGAGGAGGTCGGAAAGGCCATTCCACAGAAGATTCCTGATGGCC
|
CTGTGAACAGAGAGGACATTTTCTACACCACTGAGCTTTGGACAACTTTCTTTAGACC
|
AGAATTAGTTCGCCCAGCCCTGGAAGGCTCACTGAAGAAACTTCAACTGGACTATGTC
|
GATCTCTTCATTATCCACAATCCATTGGCTATGAAGCCTGGGGAGGAATTGCTGCCTA
|
AGGATGCCAGTGGAAACATTATTTTTGATACTGTGCATCTTCCTGACACATGGGAGCT
|
ACTGGAGAAGTGCAAAGAAGCAGGTTTAACCAAGTCCATCGGGGTGTCCAATTTCAAT
|
CACAAACTGCTGGAACTCATCCTCAACAAGCCAGGGCTCAAGTACAAGCCCACCTGCA
|
ACCAGGTCCAATGTCACCCTTACCTCAACCAGACCAAACTCCTGGAGTTCTGCAACTC
|
CAAGGACATTGTTCTAGTTGCCTACAGGGCCCTGGCATCTCACACAGACCCAAACTGG
|
ATGGACCCAGATAGCCCATATCTCTTAGAGCAGCCAACCTTGAAATCCATTGCCAAGA
|
AATACAATAGAAGCCCAGGCCAGGTTGCCCTGTGCTATGAGCTGCACCGGGGGGTGGT
|
GGTCCTGGCCAAGAGCTTCTCTGAGAAGAGAATCAAAGAGAACTTCCAGCAGGTTTTT
|
GACTTTCAGTTGACTCCAGAGGACATGAAAGCCACTGATGGCCTCAACAGAAATTTCC
|
GATATGAAACTCTAGACTTTTATCTCAGATATCAAGAGGAAAACCACATGAATAAATA
|
CCTGTTTCTTTAAAGCTGAT
|
ORF Start: ATG at 13ORF Stop: TAA at 997
SEQ ID NO:148328 aa MW at 37820.9 kD
NOV59a,MDLKQSHSVRLNDGHVMPMLGFGTYAPDHVPKSKAGEATEVAIDAGFRHIDAAFFYQN
|
CG92662-01 Protein SequenceEEEVGKAIREKIADGPVKREDTFYTTELWTTFFRPELVRPALEGSLKKLQLDYVDLFI
|
IHNPLAMKPGEELLPKDASGNIIFDTVDLRDTWEVLEKCKEAGLTKSIGVSNFNHKLL
|
ELILNKPGLKYKPTCNQVECHPYLNQSKLLEFCKSKDIVLVAYRALASHRDFNWMDPD
|
SPYLLEEPTLKSIAKKYNRSPGQVALCYELQRGVSVLAKSFSEKRIKENFQQVFDFEL
|
TPEDMKATDGLNRNPRYETLDFYLRYQEENHMNKYLFL
|
[0620] Further analysis of the NOV59a protein yielded the following properties shown in Table 59B.
307TABLE 59B
|
|
Protein Sequence Properties NOV59a
|
|
PSort0.4500 probability located in cytoplasm; 0.3726 probability
analysis:located in microbody (peroxisome); 0.1000 probability
located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0621] A search of the NOV59a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 59C.
308TABLE 59C
|
|
Geneseq Results for NOV59a
NOV59aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM79455Human protein SEQ ID NO 3101-1..325218/325 (67%)e-127
Homo sapiens, 325 aa.3..325265/325 (81%)
[WO200157190-A2, 09 Aug. 2001]
AAM78471Human protein SEQ ID NO 1133 -1..325218/325 (67%)e-127
Homo sapiens, 323 aa.1..323265/325 (81%)
[WO200157190-A2, 09 Aug. 2001]
AAW14799Type 5 17-beta-hydroxysteroid1..325218/325 (67%)e-127
dehydrogenase - Homo sapiens,1..323265/325 (81%)
323 aa. [WO9711162-A1,
27 Mar. 1997]
AAB43444Human cancer associated protein9..313215/305 (70%)e-126
sequence SEQ ID NO:889 - Homo21..324 257/305 (83%)
sapiens, 336 aa.
[WO200055350-A1, 21 Sep. 2000]
AAB76865Human lung tumour protein related30..313 201/284 (70%)3-117
protein sequence SEQ ID NO:783 -70..352 242/284 (84%)
Homo sapiens, 364 aa.
[WO200100828-A2, 04 Jan. 2001]
|
[0622] In a BLAST search of public sequence datbases, the NOV59a protein was found to have homology to the proteins shown in the BLASTP data in Table 59D.
309TABLE 59D
|
|
Public BLASTP Results for NOV59a
NOV59aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/Length ResiduesRegionValue
|
AAH20744ALDO-KETO REDUCTASE9..325214/317 (67%)e-128
FAMILY 1, MEMBER C48..323267/317 (83%)
(CHLORDECONE REDUCTASE,
3-ALPHA
HYDROXYSTEROID
DEHYDROGENASE, TYPE I,
DIHYDRODIOL DEHYDROGENASE
4) - Homo sapiens (Human), 323 aa.
P80508Prostaglandin-E2 9-reductase (EC9..325218/317 (68%)e-128
1.1.1.189) (20-alpha-hydroxysteroid8..323260/317 (81%)
dehydrogenase) (EC 1.1.1.149)
(20-alpha-HSD) -
Oryctolagus cuniculus
(Rabbit), 323 aa.
Q96A71ALDO-KETO REDUCTASE FAMILY9..313217/305 (71%)e-128
1, MEMBER C2 (DIHYDRODIOL8..311260/305 (85%)
DEHYDROGENASE 2, BILE ACID
BINDING PROTEIN, 3-ALPHA
HYDROXYSTEROID
DEHYDROGENASE, TYPE III)
(DD2/BILE ACID-BINDING
PROTEIN/AKR1C2/3ALPHA-
HYDROXYSTEROID
DEHYDROGENASE TYPE 3) - Homo
sapiens (Human), 323 aa.
P17516Chlordecone reductase (EC 1.1.1.225)9..325214/317 (67%)e-127
(CDR) (3-alpha-hydroxysteroid8..323266/317 (83%)
dehydrogenase) (EC 1.1.1.50) (3-alpha-
HSD) (Dihydrodiol dehydrogenase 4)
(DD4) (HAKRA) - Homo sapiens
(Human), 323 aa.
Q9NS543ALPHA-HYDROXYSTEROID9..325214/317 (67%)e-127
DEHYDROGENASE VARIANT (EC8..323266/317 (83%)
1.1.1.213) - Homo sapiens
(Human), 323 aa.
|
[0623] PFam analysis predicts that the NOV59a protein contains the domains shown in the Table 59E.
310TABLE 59E
|
|
Domain Analysis of NOV59a
NOV59aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
aldo_ket_red:11..305160/368 (43%)9.7e−140
domain 1 of 1216/368 (71%)
|
Example 60
[0624] The NOV60 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 60A.
311TABLE 60A
|
|
NOV60 Sequence Analysis
|
|
SEQ ID NO:1491922 bp
NOV60a,CTGTGGCTTGACCCTGAGCTTGCCCATATTCCCTGGCTGGATGGAGTGGCTAAGCCCT
|
CG92683-01 DNA SequenceGATATCGCTCTGCCCAGAAGAGATGAGTCGACTCAAAcTTCTCCAGCCAGGAAGAGGA
|
TCACGCATGCCAAAGTCCACGGTGCAGGTCAGCTCAGGCTGTCCATTGATGCCCAGGA
|
CCGGGTTCTGCTGCTTCACGTTATACAAGGTAAAGGCCTGATCAGCAAACAQCCTGGC
|
ACCTGTGATCCGTATGTGAAGGTATCTTTGATCCCTGAAGATAGTAGACTACGCCACC
|
AGAAGACGCAGACCGTTCCAGACTGCAGAGACCCCGCTTTCCACGAGCACTTCTTCTT
|
TCCTGTCCAAGAGCAGGATGATCAGAAGCGTCTCTTGGTTACTGTCTGGAACAGGGCC
|
AGCCAGTCCAGGAGACAGAGTCTCATTGGCTGCATGAGCTTTGGGGTGAAGTCTCTCC
|
TGACTCCAGACAAGCACATCAGTGGTTGGTACTACCTCCTAGGGGACCACCTGGCCCC
|
GACCAAGCACTTGAACCTGGCCAGGCGGCGACTGCCCCCGCTGAGAGACCCGCTGCTG
|
AGAATGCCAGGAGGTGGGGACACTGAGAATGGGAAGAAACTAAAGATCACCATCCCGA
|
CGCGAAAGGACGGCTTTCGCTTCACCATCTGCTGCCACTCTCCAGTTCGAGTCCAGGC
|
CGTGGATTCCGGTGGTCCGGCGGAACGGGCAGGGCTGCAGCAGCTGGACACGGTGCTG
|
CAGCTCAATGAGACGCCTGTGOAGCACTCGAAATGTGTGGACCTGGCCCACGAGATCC
|
GGAGCTGCCCCAGTGAGATCATCCTACTCGTGTGGCGCATGGTCCCCCAGGTCAAGCC
|
AGGACCAGATGGCCGGGTCCTCCGGCCCCCCTCCTGCAAGTCCACACATCACCTCCAC
|
TCACCCCCCAACAAACGGGACAAGAACTGCACCCATCCGGTCCAGGCACGGCCTGAGC
|
AGCGCCACACCTGCCACCTGGTATCTGACAGCTCTGATGGGCTGCTGCTCGGCGGCTG
|
GCACCGCTACACCGAGGTGGCCAAGCGCGGGGGCCAGCACACCCTGCCTGCACTGTCC
|
CGTGCCACTGCCCCCACCGACCCCAACTACATCATCCTCGCCCCGCTGAATCCTGGGA
|
GCCAGCTGCTCCCGCCTGTGTACCAGGACGATACCATCCCCCAACAATCAGGGAGTCC
|
CAGTAAAGGGAAGTCCEACACAGGCCTCGGGAAGAAGTCCCGGCTCATGAAGACAGTG
|
CAGACCATGAAGGGCCACGCCAACTACCAAAACTGCCCGGTTGTGAGGCCGCATGCCA
|
CGCACTCAAGCTATGGCACCTACGTCACCCTCGCCCCCAAAGTCCTGGTGTTCCCTGT
|
CTTTGTTCAGCCTCTAGATCTCTGTAATCCTGCCCGGACCCTCCTGCTGTCAGAGGAG
|
CTGCTGCTGTATGAAGGGAGCAACAAGTCAACCCAGGTGACACTGTTTGCCTATTCGG
|
ACCTGCTGCTCTTCACCAAGGAGGACGAGCCTGGCCGCTGCGACCTCCTGAGGAACCC
|
CCTCTACCTCCAGAGTGTGAAGCTGCAGGAAGGTTCTTCAGAAGACCTGAAATTCTGC
|
GTGCTCTATCTAGCACAGAAGGCAGAGTGCTTATTCACTTTGGAAGCGCACTCGCAGG
|
AGCAGAAGAAGAGAGTGTGCTGGTGCCTGTCGGAGAACATCCCCAAGCAGCAACAGCT
|
GGCAGCACCCCCCGACAGCAAGCACAAACTCCACCCTTTCGGCTCTCTCCAGCAGGAG
|
ATGGGGCCGGTCAACTCAACCAATGCCACCCAGGATAGAAGCTTTACCTCACCAGGAC
|
AGACTCTGATTGGCTGAGCAAACCCAAGGGCGGGACTCTGCTTCTGGCAACTTAACCC
|
TTTCTTGG
|
ORF Start: ATG at 41ORF Stop: TGAat 1871
SEQ ID NO:150610 aa MW at 68306.6 kD
NOV60a,MEWLSPDIALPRRDEWTQTSPARKRITEAKVQGAGQLRLSIDAQDRVLLLHVTEGKGL
|
CG92683-01 Protein SequenceISKQPGTCDPYVKVSLIPEDSRLRHQKTQTVPDCRDPAFHEHFFFPVQEEDDQKRLLV
|
TVWNRASQSRRQSLICCMSFCVKSLLTPDKEISGWYYLLGEHLGRTKHLKVARRRLRP
|
LRDPLLRMPGGGDTENGKKLKITIPRGKDGFGFTICCDSPVRVQAVDSGGPAERAGLQ
|
QLDTVLQLNERPVEHWKCVELAHEIRSCPSEIILLVWPNVPQVKPGPDGGVLRRASCK
|
STHDLQSPPNKREKNCTHGVQARPEQRESCHLVCDSSDGLLLGGWERYTEVAKRGGQH
|
TLPALSRATAPTDPNYIILAPLNPGSQLLRPVYQEDTIPEESGSPSKCKSYTGLGKKS
|
RLMKTVQTMKGEGNYQNCPVVRPHATHSSYGTYVTLAPKVLVPPVPVQPLDLCNPART
|
LLLSEELLLYEGRNKSTQVTLFAYSDLLLPTKEDEPGRCDVLRNPLYLQSVKLQEGSS
|
EDLKFCVLYLAQKAECLFTLEAESQEQKKRVCWCLSENIAKQQQLAAPPDSKQKLHPF
|
GSLQQEMGPVNSTNATQDRSFTSPGQTLIG
|
SEQ ID NO:1512874 bp
NOV60b,GCAGGACCCGCAGCCATGAACCCCTTCAATGGGCTCTGCAACGTQTGCTCGGAGCGCC
|
CG92683-02 DNA SequenceGCTACCGCCAGATCACCATCCCCAGCGGAAAGGACGGCTTTCGCTTCACCATCTGCTG
|
CGACTCTCCAGTTCCAGTCCAGGCCGTGCATTCCCGTGGTCCGGCGGAACCGGCAGGG
|
CTCCAGCACCTCCACACGGTGCTGCACCTGAATGAGAGGCCTGTGCACCACTCGAAAP
|
CTGTCCACCTGGCCCACGACATCCCCAGCTGCCCCAGTGACATCATCCTACTCGTCTG
|
GCGCATCGTCCCCCAGGTCAAGCCAGGACCAGATGGCGGGGTCCTGCGGCCGGCCTCC
|
TCCAAGTCGACACATGACCTCCAGTCACCCCCCAACAAACCGGAGAAGAACTGCACCC
|
ATGGGGTCCACCCACGGCCTGAGCAGCGCCACAGCTGCCACCTGGTATGTGACAGCTC
|
TGATGGGCTGCTGCTCGGCGGCTGGGAGCGCTACACCGAGGTGGCCAAGCGCGGGGGC
|
CAGCACACCCTCCCTGCACTGTCCCGTGCCACTGCCCCCACCGACCCCAACTACATCA
|
TCCTGGCCCCGCTGAATCCTGCGAGCCAGGTACCTGTCTTTCCCTTGCAGCTGCTCCG
|
GCCTCTGTACCAGGAGCATACCATCCCCGAAGAATCACOGAGTCCCAGTAAAGGGAAG
|
TCCTACACAGGCCTGGGGAAGAAGTCCCCGCTCATGAAGACAGTGCAGACCATGAAGG
|
GCCACGGGAACTACCAAAACTGCCCGGTTCTGAGGCCGCATGCCACGCACTCAAGCTA
|
TGGCACCTACGTCACCCTCGCCCCCAAAGTCCTGGTCTTCCCTGTCTTTGTTCAGCCT
|
CTAGATCTCTGTAATCCTGCCCGGACCCTCCTGCTGTCAGAGGAGCTGCTGCTGTATG
|
AAGGGAGGAACAACTCAACCCAGGTGACACTGTTTGCCTATTCGGACCTCCTCCTCTT
|
CACCAAGCAGGACGAGCCTCGCCGCTGCGACGTCCTGAGGAACCCCCTCTACCTCCAG
|
AGTGTGAAGCTGCAGGAACGTTCTTCAGAAGACCTCAAATTCTGCGTGCTCTATCTAG
|
CAGAGAAGGCACACTCCTTATTCACTTTCGAAGCGCACTCGCAGGAGCACAAGAAGAG
|
AGTGTGCTGGTGCCTGTCGGAGAACATCCCCAAGCAGCAACAGCTCGCACCATCACCC
|
CCGGACAGCAAGATCTTTGAGACCGACCCAGATGACAAGAGGGAGATCGCCTTGGAGG
|
AAGGGAAGGGGCCTGGTGCCGAGGATTCCCCACCCAGCAAGGAGCCCTCTCCTGGCCA
|
GGAGCTTCCTCCACGACAAGACCTTCCACCCAACAAGGACTCCCCTTCTGCGCAGGAA
|
CCCGCTCCCAGCCAAGAACCACTGTCCAGCAAAGACTCAGCTACCTCTGAAGGATCCC
|
CTCCAGGCCCAGATGCTCCGCCCACCAACGATGTGCCACCATGCCAGGAACCCCCTCC
|
AGCCCAAGACCTCTCACCCTCCCAGGACCTACCTGCTGGTCAAGAACCCCTGCCTCAC
|
CAGGACCCTCTACTCACCAAAGACCTCCCTGCCATCCAGGAATCCCCCACCCGGCACC
|
TTCCACCCTGTCAAGATCTGCCTCCTAGCCAGGTCTCCCTGCCAGCCAAGGCCCTTAC
|
TGAGGACACCATGAGCTCCCGCGACCTACTAGCAGCTACTCGGGACCCACCTGCGGCC
|
CCCAGGCCAGCCTTCGTGATCCCTGAGGTCCGGCTGGATAGCACCTACAGCCAGAAGG
|
CAGCGGCAGAGCACGCCTGCTCGGCACATCACGAGGATGCAGAAGACGCCCAGGAGGT
|
GGAGGAGGGGGAGGAAGGCCAGGAGGACGAGGATGAGGACACCAGCGATGACAACTAC
|
GGAGAGCGCAGTGAGGCCAAGCGCAGCAGCATGATCGACACGGGCCAGGGGGCTGAGG
|
GTGGCCTCTCACTGCGTGTGCAGAACTCGCTGCGGCGCCGGACGCACAGCGAGGGCAG
|
CCTGCTGCAGGAGCCCCGAGGGCCCTGCTTTGCCTCCGACACCACCTTGCACTGCTCA
|
GACGGTGAGGCCGCCGCCTCCACCTGGCGCATGCCTTCGCCCACCACCCTCAAGAAAG
|
AGCTGGGCCGCAATGGTGGCTCCATGCACCACCTTTCCCTCTTCTTCACAGGACACAG
|
GAAGATGAGCGGGGCTGACACCGTTGGGGATGATGACGAAGCCTCCCGGAAGAGAAAG
|
AGCAAAAACCTGGCCAAGGACATGAAGAACAAGCTGGGGATCTTCACACGGCGGAATG
|
AGTCCCCTGGAGCCCCTCCCGCGGGCAAGGCAGACAAAATGATGAACTCATTCAAGCC
|
CACCTCAGAGGAAGCCCTCAAGTGGGGCGAGTCCTTCGAGAAGCTGCTGGTTCACAAA
|
GGGTTAGCAGTGTTCCAAGCCTTCCTTCGCACTGAGTTCAGTGAGGAGAATCTGGAGT
|
TCTGGTTGGCTTGTGAGGACTTCAAGAAGGTCAAGTCACAGTCCAAGATGGCATCCAA
|
GGCCAAGAAGATCTTTGCTCAATACATCGCGATCCAGGCATGCAAGGAGGTAAACCTG
|
GACPCCTACACGCGGGAGCACACCAAGGACAACCTGCAGAGCGTCACGCGGGGCTGCT
|
TCGACCTGGCACAGAAGCGCATCTTCGGGCTCATGGAAAAGGACTCGTACCCTCGCTT
|
TCTCCGTTCTGACCTCTACCTGGACCTEATTAACCAGAAGAAGATGAGTCCCCCGCTT
|
TAGGGGCCACTGGAGTCGAGCTCACCGTTCACACCAGGCGGGCTGGGTCCCCTGCCCA
|
CCTGCCTCCCTGCCCCCTGTGACGGAGCGGGC
|
ORF Start: ATG at 16ORF Stop: TAG at 2785
SEQ ID NO:152923 aa MW at 101609.2 kD
NOV60b,MNRFNGLCKVCSERRYRQITIPRGKDGFGFTICCDSPVRVQAVDSGGPAERAGLQQLD
|
CG92683-02 Protein SequenceTVLQLNERPVEEWKCVELAUETRSCPSEIILLVWRNVPQVKPGPDGGVLRRASCKSTH
|
DLQSPPNKREKNCTHGVQARFEQRHSCHLVCDSSDGLLLGGWERYTEVAKRGGQETLP
|
ALSRATAPTDPNYTILAPLNPGSQVPVFPLQLLRPVYQEDTTPEESGSPSKGKSYTGL
|
GKKSRLMKTVQTMKGHGNYQNCPVVRPHATHSSYGTYVTLAPKVLVFPVFVQPLDLCN
|
PARTLLLSEELLLYEGRNKSTQVTLPAYSDLLLFTKEDEPGRCDVLRNPLYLQSVKLQ
|
EGSSEDLKPCVLYLAEKAECLFTLEAHSQEQKKRVCWCLSENIAKQQQLAASPPDSKM
|
FETEADEKREMALEEGKGPGAEDSPPSKEPSPGQELPPCQDLPPNKDSPSGQEPAPSQ
|
EPLSSKDSATSEGSPPGPDAPPSKDVPPCQEPPPAQDLSPCQDLPAGQEPLPHQDPLL
|
TKDLPAIQESPTRDLPPCQDLPPSQVSLPAKALTEDTMSSGDLLAATGDPPAAPRPAF
|
VIPEVRLDSTYSQKAGAEQGCSCDEEDAEEAEEVEEGEEGEEDEDEDTSDDNYGERSE
|
AKRSSMIETGQGAEGGLSLRVQNSLRRRTHSEGSLLQEPRGPCFASDTTLHCSDGEGA
|
ASTWGMPSPSTLKKELGRNGGSMHHLSLFFTGHRKMSGADTVGDDDEASRKRKSKNLA
|
KDMKNKLGIPRRRNESPGAPPAGKADKMMKSFKPTSEEALKWGESLEKLLVHKGLAVF
|
QAFLRTEFSEENLEFWLACEDPKKVKSQSKMASKAKKIFAEYIAIQACKEVNLDSYTR
|
EHTKDNLQSVTRGCEDIAQKRIFGLMEKDSYPRFLRSDLYLDLINQKKMSPPL
|
[0625] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 60B.
312TABLE 60B
|
|
Comparison of NOV60a against NOV60b.
NOV60a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV60b195..574367/388 (94%)
18..405369/388 (94%)
|
[0626] Further analysis of the NOV60a protein yielded the following properties shown in Table 60C.
313TABLE 60C
|
|
Protein Sequence Properties NOV60a
|
|
PSort0.3000 probability located in microbody (peroxisome);
analysis:0.3000 probability located in nucleus; 0.1000
probability located in mitochondrial matrix space;
0.1000 probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0627] A search of the NOV60a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 60D.
314TABLE 60D
|
|
Geneseq Results for NOV60a
NOV60aIdentities/
Residues/Similarities
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]Residues RegionValue
|
AAM93446Human polypeptide, SEQ ID NO:35..610560/576 (97%)0.0
3092 - Homo sapiens, 602 aa.27..602567/576 (98%)
[EP1130094-A2, 05 Sep. 2001]
AAU17386Novel signal transduction pathway195..363 148/170 (87%)2e−80
protein, Seq ID 951 - Homo sapiens,45..212149/170 (87%)
212 aa. [WO200154733-A1,
02 Aug. 2001]
ABG01916Novel human diagnostic protein453..601 123/203 (60%)2e−57
#1907 - Homo sapiens, 336 aa.14..216134/203 (65%)
[WO200175067-A2, 11 Oct. 2001]
ABG01916Novel human diagnostic protein453..601 123/203 (60%)2e−57
#1907 - Homo sapiens, 336 aa.14..216134/203 (65%)
[WO200175067-A2, 11 Oct. 2001]
ABG01914Novel human diagnostic protein455..519 61/65 (93%)8e−29
#1905 - Homo sapiens, 66 aa.1..6564/65 (97%)
[WO200175067-A2, 11 Oct. 2001]
|
[0628] In a BLAST search of public sequence datbases, the NOV60a protein was found to have homology to the proteins shown in the BLASTP data in Table 60E.
315TABLE 60E
|
|
Public BLASTP Results for NOV60a
NOV60aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9QXA2C2PA PROTEIN - Mus musculus1..610530/610 (86%)0.0
(Mouse), 610 aa.1..610564/610 (91%)
AAL37173ENDOTHELIAL PDZ PROTEIN-195..610 381/416 (91%)0.0
1 - Mus musculus18..433396/416 (94%)
(Mouse), 433 aa.
AAL68829PDZ-RGS3 - Homo sapiens195..574 374/381 (98%)0.0
(Human), 917 aa.18..398378/381 (99%)
Q925G9PDZ-RGS3 PROTEIN - Mus195..608 359/418 (85%)0.0
musculus (Mouse), 930 aa.18..434378/418 (89%)
Q920Q9SRB-RGS - Rattus norvegicus195..575 349/381 (91%)0.0
(Rat), 967 aa.18..398362/381 (94%)
|
[0629] PFam analysis predicts that the NOV60a protein contains the domains shown in the Table 60F.
316TABLE 60F
|
|
Domain Analysis of NOV60a
NOV60aIdentities/
MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegioValue
|
C2: domain 1 of 1 48..13529/97 (30%)7.8e−13
68/97 (70%)
PDZ: domain 1 of 1195..27124/86 (28%)1.5e−07
56/86 (65%)
PH: domain 1 of 1465..55212/88 (14%)3.4
61/88 (69%)
|
Example 61
[0630] The NOV61 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 61 A.
317TABLE 61A
|
|
NOV61 Sequence Analysis
|
|
SEQ ID NO: 1531482 bp
NOV61a,TCAGAGTAGGAGTCCATGCGGACATCGGCTAAGATTCCAGAATGACTACCGTCTTGAC
CG92694-01 DNA Sequence
TTACACTTTTAAAAAGTGCACTTCTGATGCACTTCTCACTGCATCAGAATGGTCCCTC
|
AGATTTTCCATAAGACCTCTGAGCTATTCCTCCCAGTTGCGACCTGCCCCAGCCATCC
|
AGACCGAAACCACCAAGACATTACCCAAACCCAATATAACGAATATTGTGGTGGTGGA
|
TGGTGTTCCCACTCCATTTTTGCTGTCAGGCACTTCGTATAAAGACCTGATACCACAT
|
GATTTCCCTAGAGCAACACTTACCCGTTTGTTGCATCAGCCCAGTGTCCCCAAGGAAG
|
TAGTTGATTATATCATCTTTGGCACAGTCATTCACGAACTGAAAACAAGCCATGTGGC
|
TGGAGAGGCTGCCCTTGGAACTGCCTTCTCTGACAAGACTCCTCCTCACACTGTCACC
|
ATGGCTTGTATCTCTGCCAACCAAGCCATGACCACAGGTGTTOGTTTCATTGCTTCTG
|
GCCAGTCTGATGTGATCATGGCAGGTCGTGTCGAGTTGATGTTCCATGTCCCTATTCG
|
TCAGTCAAGGAAAATGAGAAAACTGATGCTTGATCTAAGTAAGGCCAAATTTGTGGGC
|
CAGAGACTGTCTTCAATCTCTAAATGCCAATTGAATTTCCCAGCACCTGAGCTCCCTG
|
AGGTTTCTGAGTTCTCCACCAGTGAGACCACGGGCCACTCTGCAGACCAATTGGCTGT
|
TTCTCGACTGGAACAGGATGAATATGCACTGCGCTGTCCCAGTCTCQCCAAGAAGGCA
|
CAGGATGAAGGACTCCTTTCTGATGTCQTACCCTTCAAAGTACCAGGAAAAGATACAG
|
TTACCAAACATAATGGCATCCGTCCTTCCTCACTGGAGCAGATGGCTAAACTAAAACC
|
TGCATTCATCAAGCCCTACGGCACACCGACAGCTGCAAATTCTTCTTTCTTGACTGAT
|
GGCGCATCTGAGGCAGAGGAAAAGGCTCTGGCAATGGGTTATAAGCCGAAGGAGTGTT
|
TGAGGGATTTTATGTACGTGTCTCACAATCTCAAAGATCAACTATTATTCACACCAAC
|
ATATGCGACTCCAAAAATTCTAGAAAAGGCAGCATTAACCATGAGTGATATTGATGCT
|
TTTGAATTTCACAAAGCTTTCTCAGGTAAGATTTTAGCTAATTTTAAAGCCATGGATT
|
CTGATTGGTTTGCACAAAATTACATGGGTAGAAAAACCAACGTTACATTGCTTCCTTC
|
GGAGAAGTTTAATAACTGGGGTGGATCTCTGTCCCTGGAACACCCATTTGGAGCTACT
|
GGCTGCAGGTTGGTCGTCGCAGCTGCCAACACATTACGGAAGGAAGCACCCCAGTATG
|
CCTTAGCCGCTGCCTGTGCAGCTGGAGGGCAGGGCCATGGTATGATAGTGGAAGCTTA
|
CCCAAAATAATAGATCCAGAAGAAATGACCAG
|
ORF Start: ATG at 42ORF Stop: TAA at 1458
SEQ ID NO: 154472 aaMW at 51283.5kD
NOV61a,MTTVLTYTFKKCTSDALLTASEWSLRFSIRPLSYSSQLRAAPAIQTETRKTLAKPNIR
CG92694-01 Protein Sequence
NIVVVDGVRTPFLLSGTSYKDLIPHDLARATLTGLLHQPSVPKEVVDYITFGTVTQEV
|
KTSHVAGEAALGTAFSDKTPAHTVTMACISANQAMTTGVGLIASGQCDVIMAGGVELM
|
FHVPIRQSRKMRKLMLDLSKAKFVGQRLSSISKCQLNFPAPELPEVSEFSTSETTGHS
|
ADQLAVSRLEQDEYALRCPSLAKKAQDEGLLSDVVPFKVPGKDTVTKHNGIRPSSLEQ
|
MAKLKPAFIKPYGTATAANSSFLTDGASEAEEKALAMGYKPKECLRDFMYVSHNLKDQ
|
LLFRPTYATPKTLEKAGLPMSDIDAFEFHKAFSGKILAIFKANDSDWFAQNYMGRKSK
|
VTLLPSEKFNNWCGSLSLEHPFCATGCRLVVAAANRLRKEGGQYALAAACAACGQGHG
|
MIVEAYPK
|
[0631] Further analysis of the NOV61 a protein yielded the following properties shown in Table 61B.
318TABLE 61B
|
|
Protein Sequence Properties NOV61a
|
|
PSort0.6079 probability located in mitochondrial matrix space;
analysis:0.3717 probability located in microbody (peroxisome);
0.3122 probability located in mitochondrial inner membrane
0.3122 probability located in mitochondrial intermembrane
space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0632] A search of the NOV61 a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 61 C.
319TABLE 61C
|
|
Geneseq Results for NOV61a
NOV61aIdentities
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
ABG11653Novel human diagnostic protein125..472305/356 (85%)e−168
#11644 - Homo sapiens, 417 aa. 62..417318/356 (88%)
[WO200175067-A2, 11 Oct. 2001]
ABG11653Novel human diagnostic protein125..472305/356 (85%)e−168
#11644 - Homo sapiens, 417 aa. 62..417318/356 (88%)
[WO200175067-A2, 11 Oct. 2001]
ABG11657Novel human diagnostic protein125..472294/361 (81%)e−156
#11648 - Homo sapiens, 470 aa.110..470308/361 (84%)
[WO200175067-A2, 11 Oct. 2001]
ABG11657Novel human diagnostic protein125..472294/361 (81%)e−156
#11648 - Homo sapiens, 470 aa.110..470308/361 (84%)
[WO200175067-A2, 11 Oct. 2001]
AAU30648Novel human secreted protein #1139 - 90..472295/403 (73%)e−144
Homo sapiens, 478 aa. 76..478318/403 (78%)
[WO200179449-A2, 25 Oct. 2001]
|
[0633] In a BLAST search of public sequence datbases, the NOV61 a protein was found to have homology to the proteins shown in the BLASTP data in Table 61D.
320TABLE 61D
|
|
Public BLASTP Results for NOV61a
NOV61aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe Matched ValueExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9H3F5MSTP029 - Homo sapiens (Human),1..472407/480 (84%)0.0
475 aa.1..475430/480 (88%)
Q96C77HYPOTHETICAL 51.4 KDA1..472406/480 (84%)0.0
PROTEIN - Homo sapiens1..475429/480 (88%)
(Human), 475 aa.
P55084Trifunctional enzyme beta subunit,3..472405/478 (84%)0.0
mitochondrial precursor (TP-beta)2..474428/478 (88%)
[Includes: 3-ketoacyl-CoA thiolase
(EC 2.3.1.16) (Acetyl-CoA
acyltransferase)
(Beta-ketothiolase)] -
Homo sapiens (Human), 474 aa.
O46629Trifunctional enzyme beta subunit,1..472385/480 (80%)0.0
mitochondrial precursor (TP-beta)1..475420/480 (87%)
[Includes: 3-ketoacyl-CoA thiolase
(EC 2.3.1.16) (Acetyl-CoA
acyltransferase)
(Beta-ketothiolase)] - Bos taurus
(Bovine), 475 aa.
Q99JY0SIMILAR TO HYDROXYACYL-1..472378/480 (78%)0.0
COENZYME A DEHYDROGENASE/1..475423/480 (87%)
3-KETOACYL-COENZYME A
THIOLASE/ENOYL-COENZYME A
HYDRATASE (TRIFUNCTIONAL
PROTEIN), BETA SUBUNIT - Mus
musculus (Mouse), 475 aa.
|
[0634] PFam analysis predicts that the NOV61 a protein contains the domains shown in the Table 61 E.
321TABLE 61E
|
|
Domain Analysis of NOV61a
Identities/
Similarities
NOV61afor the MatchedExpect
Pfam DomainMatch RegionRegionValue
|
thiolase: domain 1 of 152..323 99/294 (34%)1.6e-83
229/294 (78%)
thiolase_C: domain 1 of 1327..469 59/144 (41%)1.4e-58
121/144 (84%)
|
Example 62
[0635] The NOV62 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 62A.
322TABLE 62A
|
|
NOV62 Sequence Analysis
|
|
SEQ ID NO: 1551280 bp
NOV62a,ACTATGGCCTCCTCCTCGGTCCCACCAGCCACGGTATCCGCGGCGACAGCACGCCCCG
CG92896-01 DNA Sequence
GCCCAGGTTTCGGCTTCGCCTCCAAGACCAAGAAGAAGCATTTCGTGCAGCAGAAGGT
|
GAAGGTGTTCCGGGCGGCCCACCCGCTGGTGCGTGTGTTCCTGTGGGCCGTAGCCCAC
|
TCGATCAATGAGCTCAGCCAGGTGCCTCCCCCGGTGATGCTCCTGCCAGATGACTTTA
|
AGGCCAGCTCCAAGATCAAGGTCAACAATCACCTTTTCCACAGGGAAAATCTGCCCAG
|
TCATTTCAAGTTCAAGGACTATTGTCCCCAGGTCTTCAGGAACCTCCGTGATCGATTT
|
GGCATTGATGACCAAGATTACTTGGTGTCCCTTACCCGAAACCCCCCCAGCGAAAGTG
|
AAGGCAGTGATGGTCGCTTCCTTATCTCCTACGATCGGACTCTGGTCATCAAAGAAGT
|
ATCCAGTGAGGACATTGCTGACATGCATAGCAACCTCTCCAACTATCACCAGTACATT
|
GTGAACTGCCATGGCAACACCCTTCTGCCCCAGTTCCTGCGGATGTACCGAGTCAGTG
|
TGGACAACGAAGACAGCTACATCCTTGTGATGCGCAATATGTTTAGCCACCGTCTTCC
|
TGTGCACAGGAAGTATGACCTCAAGGGTTCCCTAGTGTCCCGGGAAGCCAGCGATAAG
|
GAAAAGGTGAAAGAATTGCCCACCCTTAACGATATGGACTTTCTCAACAAGAACCAGA
|
AAGTATATATTCGTGAAGAGGAGAACAAAATATTTCTGCAGAAGCTGAAGAGAGATGT
|
GGAGTTTCTAGTGCAGCTGAAGATCATGGACTACAGCCTTCTGCTAGGCATCCACGAC
|
ATCATTCGGGGCTCTGAACCAGAGGAGGAAGCGCCCGTGCGGGAGGATGAGTCAGAGG
|
TGGATGGGGACTGCAGCCTGACTGGACCTCCTGCTCTTGTGGGCTCCTATGGCACCTC
|
CCCAGAGGGTATCGGAGGCTACATCCATTCCCATCCGCCCCTGGCCCCACGACAGTTT
|
GAGTCCTTCATTGATGTCTATGCCATCCGGAGTGCTGAAGCTGCCCCCCAGAAGGAGG
|
TCTACTTCATGGGCCTCATTGATATCCTTACACAGTATGATGCTAAGAAGAAAGCAGC
|
TCATGCAGCCAAAACTGTCAAGCATGGGGCTGGGGCAGAGATCTCTACTGTCCATCCG
|
GAGCAGTATGCTAAGCGATTCCTGGATTTTATTACCAACATCTTTGCCTAAGAGACTG
|
CCTG
|
ORF Start: ATG at 4ORF Stop: TAA at 1267
SEQ ID NO: 156421 aaMW at 47299.4kD
NOV62a,MASSSVPPATVSAATAGPGPGFGFASKTKKKHFVQQKKVKVRAADPLVGVFLWGVAHS
CG92896-01 Protein Sequence
INELSQVPPPVMLLPDDFKASSKIKVNNHLFHRENLPSHFKFKEYCPQVFRNLRDRFG
|
IDDQDYLVSLTRNPPSESEGSDGRFLISYDRTLVIKEVSSEDIADMHSNLSNYHQYIV
|
KCHGNTLLPQFLGMYRVSVDNEDSYMLVMRNMFSHRLPVHRKYDLKGSLVSREASDKE
|
KVKELPTLKDMDFLNKNQKVYIGEEEKKIFLEKLKRDVEFLVQLKIMDYSLLLGIHDI
|
IRGSEPEEEAPVREDESEVDGDCSLTGPPALVGSYGTSPEGIGCYIHSURPLGPGEFE
|
SFIDVYAIRSAEGAPQKEVYPMGLIDILTQYDAKKKAAHAAKTVKHCAGAETSTVHPE
|
QYAKRFLDFITNIFA
|
SEQ ID NO: 1571280 bp
NOV62b,ACTATGGCGTCCTCCTCGGTCCCACCAGCCACGGTATCGGCGGCGACAGCAGGCCCCG
CG92896-02 DNA Sequence
GCCCACGTTTCCGCTTCGCCTCCAAGACCAACAAGAAGCATTTCGTGCAGCAGAACCT
|
GAAGGTGTTCCGGGCGGCCCACCCCCTGGTGGGTGTGTTCCTGTGGGCCGTAGCCCAC
|
TCGATCAATCAGCTCAGCCAGGTGCCTCCCCCGGTGATGCTGCTGCCAGATGACTTTA
|
AGGCCAGCTCCAAGATCAACGTCAACAATCACCTTTTCCACAGGCAAAATCTCCCCAG
|
TCATTTCAAGTTCAAGCACTATTCTCCCCAGGTCTTCAGGAACCTCCGTGATCGATTT
|
GGCATTGATGACCAAGATTACTTGGTGTCCCTTACCCGAAACCCCCCCAGCGAAAGTG
|
AAGGCAGTGATGGTCGCTTCCTTATCTCCTACGATCGGACTCTGGTCATCAAAGAAGT
|
ATCCAGTGAGGACATTGCTGACATGCATAGCAACCTCTCCAACTATCACCAGTACATT
|
GTGAAGTGCCATGCCAACACGCTTCTGCCCCAGTTCCTGGGGATCTACCCAGTCAGTG
|
TGGACAACGAAGACAGCTACATGCTTGTGATGCGCAATATGTTTAGCCACCGTCTTCC
|
TGTGCACAGGAAGTATGACCTCAAGGGTTCCCTAGTGTCCCGGGAAGCCAGCGATAAG
|
GAAAAGGTTAAAGAATTGCCCACCCTTAAGGATATGGACTTTCTCAACAAGAACCAGA
|
AAGTATATATTGGTGAAGAGGAGAAGAAAATATTTCTGGAGAAGCTGAAGAGAGATGT
|
GGAGTTTCTAGTGCAGCTGAAGATCATGGACTACAGCCTTCTGCTAGGCATCCACGAC
|
ATCATTCGGGGCTCTGAACCAGACGAGGAAGCGCCCGTGCCGGAGGATGAGTCAGAGG
|
TGGATGGGGACTGCAGCCTGACTCCACCTCCTGCTCTGGTGGGCTCCTATGCCACCTC
|
CCCAGAGGGTATCCGAGGCTACATCCATTCCCATCGGCCCCTGGGCCCAGGAGAGTTT
|
GAGTCCTTCATTGATCTCTATGCCATCCGGAGTGCTGAAGGAGCCCCCCAGAAGGAGG
|
TCTACTTCATGGGCCTCATTGATATCCTTACACAGTATGATGCCAAGAAGAAAGCAGC
|
TCATGCAGCCAAAACTGTCAAGCATGGGGCTGGGGCAGAGATCTCTACTGTCCATCCG
|
GAGCAGTATCCTAAGCGATTCCTGGATTTTATTACCAACATCTTTGCCTAAGAGACTG
|
CCTG
|
ORF Start: ATG at 4ORF Stop: TAA at 1267
SEQ ID NO: 158421 aaMW at 47299.4kD
NOV62b,MASSSVPPATVSAATAGPCPGPGFASKTKKKHFVQQKVKVFRAADPLVGVFLWGVAHS
CG92896-02 Protein Sequence
INELSQVPPPVMLLPDDFKASSKIKVNNHLFHRENLPSHFKFKEYCPQVFRNLRDRFG
|
IDDQDYLVSLTRNPPSESEGSDGRFLISYDRTLVIKEVSSEDIADMHSNLSNYHOVIV
|
KCHGNTLLPQFLGMYRVSVDNEDSYMLVMRNMFSERLPVHRKYDLKGSLVSREASDKE
|
KVKELPTLKDMDFLNKNQKVYIGEEEKKIFLEKLKRDVEELVQLKIMDYSLLLGIHDI
|
IRGSEPEEEAPVREDESEVDGDCSLTGPPALVGSYGTSPEGIGGYIHSHRPLGPGEFE
|
SFIDVYAIRSAEGAPQKEVYFMGLIDILTQYDAKKKAAHAAKTVKHGAGAEISTVHPE
|
QYAKRFLDFITNIFA
|
[0636] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 62B.
323TABLE 62B
|
|
Comparison of NOV62a against NOV62b.
NOV62a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV62b42..421334/380 (87%)
42..421334/380 (87%)
|
[0637] Further analysis of the NOV62a protein yielded the following properties shown in Table 62C.
324TABLE 62C
|
|
Protein Sequence Properties NOV62a
|
|
PSort0.3000 probability located in nucleus; 0.2053 probability
analysis:located in microbody (peroxisome); 0.1000 probability located
in mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPCleavage site between residues 59 and 60
analysis:
|
[0638] A search of the NOV62a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 62D.
325TABLE 62D
|
|
Geneseq Results for NOV62a
NOV62aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAG63222Amino acid sequence of a human1..421421/421 (100%)0.0
lipid metabolism enzyme—Homo1..421421/421 (100%)
sapiens, 421 aa. [WO200153468-A2,
26-JUL-2001]
AAB88462Human membrane or secretory1..262234/262 (89%)e-135
protein clone PSEC0253—Homo1..262245/262 (93%)
sapiens, 265 aa. [EP1067182-A2, 10-
JAN-2001]
ABB12247Human PI 5-phosphate 4-kinase1..171171/171 (100%)1e-96
gamma homologue, SEQ ID15..185171/171 (100%)
NO:2617—Homo sapiens, 223 aa.
[WO200157188-A2, 09-AUG-2001]
ABG09799Novel human diagnostic protein47..418125/381 (32%)6e-40
#9790—Homo sapiens, 605 aa.128..490182/381 (46%)
[WO200175067-A2, 11-OCT-2001]
ABG09796Novel human diagnostic protein47..418125/381 (32%)6e-40
#9787—Homo sapiens, 556 aa.128..490182/381 (46%)
[WO200175067-A2, 11-OCT-2001]
|
[0639] In a BLAST search of public sequence datbases, the NOV62a protein was found to have homology to the proteins shown in the BLASTP data in Table 62E.
326TABLE 62E
|
|
Public BLASTP Results for NOV62a
NOV62aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q91XU3PHOSPHATIDYL INOSITOL1 . . . 421404/421 (95%)0.0
PHOSPHATE KINASE TYPE II1 . . . 421410/421 (96%)
GAMMA-Mus musculus (Mouse),
421 aa.
O88370PHOSPHATIDYLINOSITOL 5-1 . . . 421401/421 (95%)0.0
PHOSPHATE 4-KINASE GAMMA-1 . . . 420409/421 (96%)
Ratttus norvegicus (Rat), 420 aa.
O88377PHOSPHATIDYLINOSITOL 5-26 . . . 419270/397 (68%)e-154
PHOSPHATE 4-KINASE BETA-21 . . . 414323/397 (81%)
Rattus norvegicus (Rat), 416 aa.
P78356PHOSPHATIDYLINOSITOL-4-26 . . . 419267/397 (67%)e-153
PHOSPHATE 5-KINASE TYPE II21 . . . 414324/397 (81%)
BETA (EC 2.7.1.68)-Homo sapiens
(Human), 416 aa.
AAL18245PHOSPHATIDYLINOSITOL-4-30 . . . 419267/393 (67%)e-152
PHOSPHATE 5-KINASE TYPE II1 . . . 390322/393 (80%)
BETA-Mus musculus (Mouse), 392
aa (fragment).
|
[0640] PFam analysis predicts that the NOV62a protein contains the domains shown in the Table 62F.
327TABLE 62F
|
|
Domain Analysis of NOV62a
Identities/
Similarities
NOV62athe for
Pfam DomainMatch RegionMatched RegionValue
|
PIP5K: domain 1 of 1124 . . . 420139/359 (39%)1.5e − 1 55
279/359 (78%)
|
Example 63
[0641] The NOV63 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 63A.
328TABLE 63A
|
|
NOV63 Sequence Analysis
|
|
SEQ ID NO: 1592056 bp
NOV63a,ATGGCCAAACCAGCACAGGGTGCCAAGTACCGGGGCTCCATCCATGACTTCCCAGGCT
CG92987-O1 DNA Sequence
TTGACCCCAACCAGGATGCCGAGGCTCTGTACACTGCCATGAAGGGCTTTGGCAGTGA
|
CAAGGAGGCCATATTGGACATAATCACCTCACGGAGCAACAGGCAGAGGCAGGAGGTC
|
TGCCAGAGCTACAAGTCCCTCTACCCCAACCACCTCATTGCTGATTTAAAGTATGAAT
|
TGACGGGCAAGTTTGAACGGTTGATTGTGGGCCTGATGAGGCCACCTGCCTATTGTGA
|
TGCCAAAGAAATTAAAGATGCCATCTCGGGCATTGGCACTGATGAGAAGTCCCTCATT
|
GAGATCTTCGCTTCCCGGACCAATGAGCAGATGCACCAGCTGGTGGCAGCATACAAAG
|
ATGCCTACGACCGGGACCTGGAGGCTGACATCATCGGCGACACCTCTGGCCACTTCCA
|
GAAGATCCTTGTGGTCCTGCTCCAGGGAACCAGGGAGGAGGATGACGTAGTGAGCGAG
|
GACCTCCTACAACACGATCTCCAGGACCTATACCAGGCACGCGAACTGAAATGCGGAA
|
CAGATGAAGCCCAGTTCATTTACATCTTGGGAAATCGCAGCAAGCAGCATCTTCGGTT
|
GGTGTTCGATGAGTATCTGAACACCACAGGGAAGCCGATTGAAGCCAGCATCCCAGGG
|
GAGCTGTCTGGGGACTTTGAGAAGCTAATGCTGGCCGTAGTGAAGTGTATCCGGAGCA
|
CCCCGGAATATTTTGCTGAAAGGCTCTTCAAGGCTATGAAGGCCCTGGGGACTCGGGA
|
CAACACCCTGATCCGCATCATGGTCTCCCGTAGTGAGTTGGACATGCTCGACATTCGG
|
GAGATCTTCCGGACCAAGTATGAGAAGTCCCTCTACACCATGATCAAGAATGACACCT
|
CTGGCGAGTACAAGAAGACTCTGCTGAAGCTGTCTGGCGGAGATGATGATGCTGCTGG
|
CCAGTTCTTCCCGCAGGCACCGCAGCTGGCCTATCAGATGTCGGAACTTAGTGCAGTC
|
GCCCGAGTAGAGCTGAAGGCAGTGGCCCGAGTAGAGCTGAAGGGAACTGTGCGCCCAG
|
CCAATGACTTCAACCCTGACGCAGATGCCAAAGCCCTGCGGAAAGCCAPGAAGGGACT
|
CCCGACTGACGAAGACACAATCATCGATATCATCACGCACCGCAGCAATGTCCAGCGG
|
CAGCAGATCCGGCAGACCTTCAACTCTCACTTTCGCCGGGACTTAATGACTGACCTGA
|
AGTCTGAGATCTCTGGAGACCTGGCAAGGCTCATTCTGGCGCTCATCATCCCACCCGC
|
CCATTACGATGCCAAGCAGTTGAAGAAGGCCATGGAGGGAGCCGGCACAGATGAAAAG
|
GCTCTTATTCAAATCCTGGCCACTCGGACCAATGCTGAAATCCGGGCCATCAATCAGG
|
CCTATAACGAGGACTATCACAAGTCCCTGGAGGATGCTCTGAGCTCAGACACAPCTGG
|
CCACTTCAGGAGGATCCTCATTTCTCTGCCCACGGCQCATCGTGAGGAGGGAGGAGAA
|
AACCTGGACCAGGCACGGGAAGATGCCCAGGAAATAGCAGACACACCCAGTGGAGACA
|
AAACTTCCTTGGAGACACGTTTCATGACGATCCTGTGTACCCGGAGCTATCCGCACCT
|
CCGCAGAGTCTTCCAGGAGTTCATCAAGATGACCAACTATGACGTGGAGCACACCATC
|
AAGAACGACATGTCTGGGGATGTCACGCATGCATTTGTGCCCATTGTTCAAAGTGTCA
|
AGAACAAGCCTCTCTTCTTTGCCGACAAACTTTACAAATCCATCAACGGTGCTGGCAC
|
AGATGAGAAGACTCTGACCAGCATCATCGTATCCCACAGTGAGATTGACCTGCTCAAC
|
ATCCGGAGGGAATTCATTGAGAAATAPGACAAGTCTCTCCACCAAGCCATTCAGGGTG
|
ACACCTCCGGAGACTTCCTGAAGGCCTTGCTGGCTCTCTGTGGTGGTGAGGACTAGGG
|
CCACTGCTTTCAGGTGTGATATCTAT
|
ORF Start: ATG at 1ORF Stop: TAG at 2026
SEQ ID NO: 160675 aaMW at 76123.6kD
NOV63a,MAKPAQGAKYRGSIHDFPGFDPNQDAEALYTANKGFGSDKEAILDIITSRSNRQRQEV
CG92987-01 Protein Sequence
CQSYKSLYGKDLIADLKYELTGKFERLIVGLMRPPAYCKAKEIKDAISGIGTDEKCLI
|
EILASRTNEQMHQLVAAYKDAYERDLEADIIGDTSGHFQKMLVVLLQGTREEDDVVSE
|
DLVQQDVQDLYEAGELKWCTDEAQFIYILGNRSKQHLRLVFDEYLKTTGKPIEASIRG
|
ELSCDFEKLMLAVVKCIRSTPEYFAERLFKAMKGLGTRDNTLIRIMVSRSELDMLDIR
|
EIFRTKYEKSLYSMIKNDTSGEYKKTLLKLSGGDDDAAGQFFPEAAQVAYQMWELSAV
|
ARVELKAVARVELKGTVRPANDFNPDADAKALRKANKGLCTOEDTIIDIITERSNVQR
|
QQIRQTFKSHFGRDLMTDLKSEISGDLARLILGLMMPFAHYDAKQLKKAEEGAGTDEK
|
ALIEILATRTNAEIRAINEAYKEDYHKSLEDALSSDTSGHFRRILISLATGHREEGGE
|
NLDQAREDAQEIADTPSGDKTSLETRFMTILCTRSYPHLRRVFQEFIKMTNYDVEHTI
|
KKEMSGDVRDAFVAIVQSVKNKPLFFADKLYKSMKGAGTDEKTLTRIMVSHSEIDLLN
|
IRREFIEKYDKSLHQAIEGDTSGDFLKALLALCGGED
|
[0642] Further analysis of the NOV63a protein yielded the following properties shown in Table 63B.
329TABLE 63B
|
|
Protein Sequence Properties NOV63a
|
|
PSort0.4500 probability located in cytoplasm; 0.3000 probability
analysis:located in microbody (peroxisome); 0.1000 probability
located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0643] A search of the NOV63a protein against the Gene seq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 63C.
330TABLE 63C
|
|
Geneseq Results for NOV63a
NOV63aIdentities/
Residues/Similarities
GeneseqProtein/Organism/LengthMatchfor theExpect
Identifier[Patent #, Date]ResiduesMatched RegionValue
|
AAR03726Human placenta-derived coagulation1 . . . 675665/681 (97%)0.0
inhibitor protein-Homo sapiens, 7861 . . . 673665/681 (97%)
aa. [EP351826-A, JAN. 24, 1990]
AAR03725Human placenta-derived coagulation2 . . . 675662/680 (97%)0.0
inhibitor-Homo sapiens, 672 aa.1 . . . 672664/680 (97%)
[EP351826-A, JAN. 24, 1990]
ABG19948Novel human diagnostic protein1 . . . 673633/685 (92%)0.0
#19939-Homo sapiens, 736 aa.60 . . . 736641/685 (93%)
[WO200175067-A2, OCT. 11, 2001]
ABG19948Novel human diagnostic protein1 . . . 673633/685 (92%)0.0
#19939-Homo sapiens, 736 aa.60 . . . 736641/685 (93%)
[WO200175067-A2, OCT. 11, 2001]
ABB57067Mouse ischaemic condition related11 . . . 326180/316 (56%)e-101
protein sequence SEQ ID NO: 136-4 . . . 319237/316 (74%)
Mus musculus, 319 aa.
[WO200188188-A2, NOV. 22, 2001]
|
[0644] In a BLAST search of public sequence datbases, the NOV63a protein was found to have homology to the proteins shown in the BLASTP data in Table 63D.
331TABLE 63D
|
|
Public BLASTP Results for NOV63a
NOV63aIdentities/
ProteinResidues/Similarities
AccessionMatchfor theExpect
NumberProtein/Organisn/LengthResiduesMatched PortionValue
|
AAH17046ANNEXIN A6-Homo sapiens1 . . . 675666/681 (97%)0.0
(Human), 673 aa.1 . . . 673666/681 (97%)
P08133Annexin VI (Lipocortin VI) (P68)12 . . . 675665/680 (97%)0.0
(P70) (Protein III) (Chromobindin 20)1 . . . 672665/680 (97%)
(67 kDa calelectrin) (Calphobindin-II)
(CPB-II)-Homo sapiens (Human),
672 aa.
Q99JX6SIMILAR TO ANNEXIN A6-Mus1 . . . 675629/675 (93%)0.0
musculus (Mouse), 667 aa.1 . . . 667646/675 (95%)
S01786annexin VI-mouse, 673 aa.1 . . . 675629/681 (92%)0.0
1 . . . 673646/681 (94%)
S52844annexin VI-rat, 673 aa.1 . . . 675629/681 (92%)0.0
1 . . . 673647/681 (94%)
|
[0645] PFam analysis predicts that the NOV63a protein contains the domains shown in the Table 63E.
332TABLE 63E
|
|
Domain Analysis of NOV63a
Identities/
NOV63aSimilarities
Matchfor theExpect
Pfam DomainRegionMatched RegionValue
|
annexin: domain 1 of 822 . . . 8928/68 (41%) 1e−7
46/68 (68%)
annexin: domain 2 of 894 . . . 16126/68 (38%)8.9e−18
46/68 (68%)
annexin: domain 3 of 8178 . . . 24523/68 (34%)1.2e−14
43/68 (63%)
annexin: domain 4 of 8253 . . . 32027/68 (40%)5.1e−19
46/68 (68%)
annexin: domain 5 of 8373 . . . 44031/68 (46%)2.7e−22
49/68 (72%)
annexin: domain 6 of 8445 . . . 51229/68 (43%)1.7e−24
55/68 (81%)
annexin: domain 7 of 8527 . . . 59519/69 (28%)5.3e−06
41/69 (59%)
annexin: domain 8 of 8603 . . . 67030/68 (44%) 5e−20
50/68 (74%)
|
Example 64
[0646] The NOV64 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 64A.
333TABLE 64A
|
|
NOV64 Sequence Analysis
|
|
SEQ ID NO: 161987 bp
NOV64a,TCTGAAGCAATGCTCAATACAACCTCAGTCACTGAATTTCTCCTTTTCGCAGTGACAG
CG93042-01 DNA Sequence
ACATTCAAGAACTGCAGCCTTTTCTCTTCGTTCTTTTCCTTACCATCTACTTCATCAG
|
TGTGGCTGGGAATGGAGCCATTCTGATGATTCTCATCTCPGATCATAGAGTCCATTCC
|
CCTATGTATTTCTTCCTGCGAAACCTGTCCTGCCTGGACATCTGCTACTCCAGCGTAA
|
CACTGCCAAAAATGCTGCAGAACTTCCTCTCTCCACACAAAGCAATTTCTTTCTTGGG
|
ATGCATAAGCCAACTCCATTTCTTCCACTTCCTGGGCACCACAGAGGCCATGTTGTTG
|
GCCGTGATGGCATTTGACCGCTTTGTCGCTATTTGCAAGCCACTTCGCTACACTGTCA
|
TTATGAACCCTCAGCTCTGTACCCACATGGCCATCACAATCTGGATGATTGGTTTTTT
|
CCATGCCCTGCTGCACTCCCTAATGACCTCTCGCTTGAACTTCTGTGGTTCTAACCGT
|
ATCTATCACTTCTTCTGTGATGTGAAGCCATTGCTAAAGCTGGCCTGTGGGAACACTG
|
AGCTTAATCAGTGGCTGCTCAGTACTGTCACACGGACAAPCGCCATGGGCCCCTTCTT
|
TCTCACATTACTCTCCTATTTCTACATTATCACCCATCTCTTCPTCAAGACTCATTCT
|
TTTAGCATGCTCCGCAAACCACTGTCCACTTGTOCCTCCCACTTCATGGTAGTTATTC
|
TTTTGTATGCACCTGTTCTCTTCACCTATATTCATCATGCCTCAGCCACCTCCATGGA
|
CCAGGACCGGATCACTGCCATCATGTATACTGTGGTCACTCCAGTACTAAACCCACTG
|
ATCTACACTTTGAAGCGAACCAAAGGAAAGTGAAAAGGGGCCTTTATATATGAGCCAA
|
TGAAAAAGGTGCCTTTCGCCTAAAGAAATCTTGAAGAACTCTTTTGAACCATAAATAA
|
A
ORF Start: ATG at 10ORF Stop: TGA at 901
SEQ ID NO: 162297 aaMW at 33589.7dK
NOV64a,MLNTTSVTEFLLLGVTDIQELQPFLFVVFLTIYFISVAGNGAILMIVISDHRLHSPMY
CG93042-01 Protein Sequence
FFLGNLSCLDICYSSVTLPKMLQNFLSAHKAISFLGCISQLHFFHFLGSTEAMLLAVM
|
AFDRFVAICKPLRYTVTMNPQLCTQMAPTIWMICFFHALLHSLMTSRLNFCOSNRIYH
|
FFCDVKPLLKLACGNTELNQWLLSTVTGTIAMGPFFLTLLSYFYIITHLFFKTHSFSM
|
LRKALSTCASEFMVVILLYAPVLFTYIHHASGTSMDQDRITAIMYTVVTPVLNPLTYT
|
LKGTKGK
|
[0647] Further analysis of the NOV64a protein yielded the following properties shown in Table 64B.
334TABLE 64B
|
|
Protein Sequence Properties NOV64a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body; 0.3000
probability located in endoplasmic reticulum (membrane);
0.3000 probability located in microbody (peroxisome)
SignalPCleavage site between residues 39 and 40
analysis:
|
[0648] A search of the NOV64a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 64C.
335TABLE 64C
|
|
Geneseq Results for NOV64a
NOV64aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAG72212Human olfactory receptor polypeptide,1 . . . 297281/297 (94%)e−159
SEQ ID NO: 1893 - Homo sapiens,1 . . . 292286/297 (95%)
309 aa. [WO200127158-A2, 19-APR-2001]
AAG71661Human olfactory receptor polypeptide,1 . . . 297281/297 (94%)e−159
SEQ ID NO: 1342 - Homo sapiens,1 . . . 292286/297 (95%)
309 aa. [WO200127158-A2, 19-APR-2001]
AAG72870Human olfactory receptor data1 . . . 297265/297 (89%)e−151
exploratorium sequence, SEQ ID NO:7 . . . 303276/297 (92%)
2552 - Homo sapiens, 315 aa.
[WO200127158-A2, 19-APR-2001]
AAG72203Human olfactory receptor polypeptide,1 . . . 297265/297 (89%)e−151
SEQ ID NO: 1884 - Homo sapiens,1 . . . 297276/297 (92%)
307 aa. [WO200127158-A2, 19-APR-2001]
AAG72658Murine OR-like polypeptide query1 . . . 292235/292 (80%)e−138
sequence, SEQ ID NO: 2340 - Mus7 . . . 298260/292 (88%)
musculus, 325 aa. [WO200127158-A2,
19-APR-2001]
|
[0649] In a BLAST search of public sequence datbases, the NOV64a protein was found to have homology to the proteins shown in the BLASTP data in Table 64D.
336TABLE 64D
|
|
Public BLASTP Results for NOV64a
NOV64aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P58182Olfactory receptor 12D2 (Hs6M1-20) -1 . . . 297265/297 (89%)e−151
Homo sapiens (Human), 307 aa.1 . . . 297276/297 (92%)
Q920Y9BM332P19.2 (NOVEL 71 . . . 292235/292 (80%)e−138
TRANSMEMBRANE RECEPTOR1 . . . 292260/292 (88%)
(RHODOPSIN FAMILY)
(OLFACTORY RECEPTOR LIKE)
PROTEIN (MM17M1-13),
ORTHOLOG OF HUMAN DJ994E9.8
(HS6M1-20)) - Mus musculus (Mouse),
308 aa.
Q920Y8BM332P19.3 (NOVEL 71 . . . 292228/292 (78%)e−133
TRANSMEMBRANE RECEPTOR1 . . . 292257/292 (87%)
(RHODOPSIN FAMILY)
(OLFACTORY RECEPTOR LIKE)
PROTEIN (MM17M1-14)) - Mus
musculus (Mouse), 313 aa.
Q920Z0BM332P19.1 (NOVEL 71 . . . 292228/292 (78%)e−133
TRANSMEMBRANE RECEPTOR1 . . . 292253/292 (86%)
(RHODOPSIN FAMILY)
(OLFACTORY RECEPTOR LIKE)
PROTEIN (MM17M1-12)) - Mus
musculus (Mouse), 308 aa.
AAL60921OLFACTORY RECEPTOR MOR250-1 -1 . . . 292210/294 (71%)e−120
Mus musculus (Mouse), 314 aa.1 . . . 294243/294 (82%)
|
[0650] PFam analysis predicts that the NOV64a protein contains the domains shown in the Table 64E.
337TABLE 64E
|
|
Domain Analysis of NOV64a
Identities/
NOV64aSimilarities for theExpect
Pfam DomainMatch RegionMatched RegionValue
|
7tm_1: domain 1 of 139 . . . 289 46/269 (17%)4.9e−33
169/269 (63%)
|
Example 65
[0651] The NOV65 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 65A.
338TABLE 65A
|
|
NOV65 Sequence Analysis
|
|
SEQ ID NO: 1631137 bp
NOV65a,GGGCAGGAGCTGGGGCCACGCTGGTCTGGGATAGTTGGGCAGGGAGGCTGTCTACCTG
CG93265-01 DNA Sequence
GTCTCCAGAATGGACGGCCCTGTGGCAGAGCATGCCAAGCAGGAGGCCTTTCACGTGG
|
TCACACCTCTGTTGGAGAGCTGGGCGCTGTCCCAGGTGGCGGGCATGCCTGTCTTCCT
|
CAAGTGTGAGAATGTGCAGCCCAGCGGCTCCTTCAAGATTCGGGGCATTGGGCATTTC
|
TGCCAGGAGATGGCCAAGAAGGGATGCAGACACCTGGTGTGCTCCTCAGGGGGTAATG
|
CGGGCATCGCTGCTGCCTATGCTGCTAGGAAGCTGGGCATTCCTGCCACCATCGTGCT
|
CCCCGAGAGCACCTCCCTGCAGGTGGTGCAGAGGCTGCAGGGGGAGGGGGCCGAGGTT
|
CAGCTGACTGGAAAGGTCTGGGACGAGGCCAATCTGAGGGCCCAAGAGTTGGCCAAGA
|
GGGACGGCTGGGAGAATGTCCCCCCGTTTGACCACCCCCTAATATGGAAAGGCCACGC
|
CAGCCTGGTGCAGGAGCTGAAAGCAGTGCTGAGGACCCCACCAGGTGCCCTGGTGCTG
|
GCAGTTGGGGGTGGGGGTCTCCTGGCCGGGGTGGTGGCTGGCCTGCTGGAGGTGGGCT
|
GGCAGCATGTACCCATCATTGCCATGGAGACCCATGGGGCACACTGCTTCAATGCGGC
|
CATCACAGCCGGCAAGCTGGTCACACTTCCAGACATCACCAGTGTGGCCAAGAGCCTG
|
GGTGCCAAGACGGTGGCCGCTCGGGCCCTTAGTTCGATGCAGGTGTGCAAGATTCACT
|
CTGAAGTGGTGGAGGACACCGAGGCTGTGAGCGCTGTGCAGCAGCTCCTGGATGATGA
|
GCGTATGCTGGTGGAGCCTGCCTGTGGGGCAGCCTTAGCAGCCATCTACTCAGGCCTC
|
CTGCGGAGGCTCCAGGCCGAGGGCTGCCTGCCCCCTTCCCTGACTTCAGTTGTGGTAA
|
TCGTGTGTGGAGGCAACAACATCAACAGCCGAGAGCTGCAGGCTTTGAAAACCCACCT
|
GGGCCAGGTCTGAGGGGTCCCATCCTGGCCCCAAAGACCCCTGAGAGGCCCATGGACA
|
GTCCTGTGTCTGGATGAGGAGGACTCAGTGCTGGC
|
ORF Start ATG at 68ORF Stop: TGA at 1055
|
SEQ ID NO: 164329 aaMW at 34615.8kD
NOV65a,NDGPVAEHAKQEPFHVVTPLLESWALSQVAGMPVFLKCENVQPSGSFKIRGIGHFCQE
CG93265-01 Protein Sequence
MAKKGCRHLVCSSGGNAGIAAAYAARKLGIPATIVLPESTSLQVVQRLQGEGAEVQLT
|
GKVWDEANLRAQELAKRDGWENVPPFDHPLIWKGHASLVQELKAVLRTPPGALVLAVG
|
GGGLLAGVVAGLLEVGWQHVPIIANETHGAHCFNAAITAGKLVTLPDITSVAKSLGAK
|
TVAARALSSMQVCKIHSEVVEDTEAVSAVQQLLDDERMLVEPACGAALAAIYSGLLRR
|
LQAEGCLPPSLTSVVVIVCGGNNINSRELQALKTHLGQV
|
SEQ ID NO: 1651137 bp
NOV65b,GGGCAGGAGCTGGGGCCACGCTGGTCTGGGATAGTTGGGCAGGGAGGCTGTCTACCTG
CG93265-02 DNA Sequence
GTCTCCAGAATGGACGGCCCTGGGCCAGAGCATGCCAAGCAGGAGCCCTTCACGTGG
|
TCACACCTCTGTTGGAGAGCTGGGCGCTGTCCCAGGTGGCGGGCATGCCTGTCTTCCT
|
CAAGTGTGAGAATGTGCAGCCCAGCGGCTCCTTCAAGATTCGGGGCATTGGGCATTTC
|
TGCCAGGAGATGGCCAAGAAGGGATGCAGACACCTGGTGTGCTCCTCAGGGGGTAATG
|
CGGGCATCGCTGCTGCCTATGCTGCTAGGAAGCTGGGCATTCCTGCCACCATCGTGCT
|
CCCCGAGAGCACCTCCCTGCAGGTGGTGCAGAGGCTGCAGGGGGAGGGGGCCGAGGTT
|
CAGCTGACTGGAAAGGTCTGGGACGAGGCCAATCTGAGGGCGCAAGAGTTGGCCAAGA
|
GGGACGGCTGGGAGAATGTCCCCCCGTTTGACCACCCCCTAATATGGAAAGGCCACGC
|
CAGCCTGGTGCAGGAGCTGAAAGCAGTGCTGAGGACCCCACCAGGTGCCCTGGTGCTG
|
GCAGTTGGGGGTGGGGGTCTCCTGGCCGGGGTGGTGGCTGGCCTGCTGGAGGTGGGCT
|
GGCAGCATGTACCCATCATTGCCATGGAGACCCATGGGGCACACTGCTTCAATGCGGC
|
CATCACAGCCGGCAAGCTGGTCACACTTCCAGACATCACCAGTGTGGCCAAGAGCCTG
|
GGTGCCAAGACGGTGGCCGCTCGGGCCCTTAGTTCGATGCAGGTGTGCAAGATTCACT
|
CTGAAGTGGTGGAGGACACCGAGGCTGTGAGCGCTGTGCAGCAGCTCCTGGATGATGA
|
GCGTATGCTGGTGGAGCCTGCCTGTGGGGCAGCCTTAGCAGCCATCTACTCAGGCCTC
|
CTGCGGAGGCTCCAGGCCGACGGCTGCCTGCCCCCTTCCCTGACTTCAGTTGTGGTAA
|
TCGTGTGTGGAGGCAACAACATCAACAGCCGAGAGCTGCAGGCTTTGAAAACCCACCT
|
GGGCCAGGTCTGAGGGGTCCCATCCTGGCCCCAAAGACCCCTGAGAGGCCCATGGACA
|
GTCCTGTGTCTGGATGAGGAGGACTCAGTGCTGGC
|
ORF Start: ATG at 68ORF Stop: TGA at 1055
SEQ ID NO: 166329 aaMW at 346 15.8kD
NOV65b,MDGPVAEHAKQEPFHVVTPLLESWALSQVAGMPVFLKCENVQPSGSFKIRGIGHFCQE
CG93265-02 Protein Sequence
MAKKGCRHLVCSSGGNAGIAAAYAARKLGIPATIVLPESTSLQVVQRLQGEGAEVQLT
|
GKVWDEANLRAQELAKRDGWENVPPFDHPLIWKGHASLVQELKAVLRTPPGALVLAVG
|
GGGLLAGVVAGLLEVGWQHVPIIAMETHGAHCFNAAITAGKLVTLPDITSVAKSLGAK
|
TVAARALSSMQVCKIHSEVVEDTEAVSAVQQLLDDERMLVEPACGAALAAIYSGLLRR
|
LQAEGCLPPSLTSVVVIVCGGNNINSRELQALKTHLGQV
|
[0652] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 65B.
339TABLE 65B
|
|
Comparison of NOV65a against NOV65b.
NOV65a Residues/Identities/Similarities for
Protein SequenceMatch Residuesthe Matched Region
|
NOV65b1 . . . 329284/329 (86%)
1 . . . 329284/329 (86%)
|
[0653] Further analysis of the NOV65a protein yielded the following properties shown in Table 65C.
340TABLE 65C
|
|
Protein Sequence Properties NOV65a
|
|
PSort0.8500 probability located in endoplasmic
analysis:reticulum (membrane);
0.4400 probability located in plasma membrane;
0.1000 probability located in mitochondrial inner membrane;
0.1000 probability located in Golgi body
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0654] A search of the NOV65a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 65D.
341TABLE 65D
|
|
Geneseq Results for NOV65a
NOV65aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAM40622Human polypeptide SEQ ID NO:1 . . . 329327/329 (99%)0.0
5553 - Homo sapiens, 340 aa.12 . . . 340 327/329 (99%)
[WO200153312-A1, 26-JUL-2001]
AAM38836Human polypeptide SEQ ID NO:1 . . . 329327/329 (99%)0.0
1981 - Homo sapiens, 329 aa.1 . . . 329327/329 (99%)
[WO200153312-A1, 26-JUL-2001]
AAU23238Novel human enzyme polypeptide1 . . . 329327/329 (99%)0.0
#324 - Homo sapiens, 340 aa.12 . . . 340 327/329 (99%)
[WO200155301-A2, 02-AUG-2001]
AAM52238Human DHY SEQ ID NO: 2 - Homo1 . . . 329326/329 (99%)0.0
sapiens, 329 aa. [WO200181559-A2,1 . . . 329326/329 (99%)
01-NOV-2001]
AAU23764Novel human enzyme polypeptide1 . . . 329326/329 (99%)0.0
#850 - Homo sapiens, 340 aa.12 . . . 340 326/329 (99%)
[WO200155301-A2, 02-AUG-2001]
|
[0655] In a BLAST search of public sequence datbases, the NOV65a protein was found to have homology to the proteins shown in the BLASTP data in Table 65E.
342TABLE 65E
|
|
Public BLASTP Results for NOV65a
NOV65aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96GA7UNKNOWN (PROTEIN FOR1 . . . 329327/329 (99%)0.0
MGC: 15400) - Homo sapiens1 . . . 329327/329 (99%)
(Human), 329 aa.
AAH22601RIKEN CDNA 4432411H131 . . . 327263/327 (80%)e−149
GENE - Mus musculus (Mouse),1 . . . 327290/327 (88%)
329 aa.
AAL56988SERINE DEHYDRATASE (EC1 . . . 324262/324 (80%)e−148
4.2.1.13) - Mus musculus (Mouse),1 . . . 324289/324 (88%)
359 aa.
DWRTTL-serine dehydratase (EC 4.2.1.13) -9 . . . 327197/320 (61%)e−107
rat, 327 aa.2 . . . 321243/320 (75%)
AAH21605SIMILAR TO SERINE9 . . . 327199/320 (62%)e−106
DEHYDRATASE - Mus musculus2 . . . 321242/320 (75%)
(Mouse), 327 aa.
|
[0656] PFam analysis predicts that the NOV65a protein contains the domains shown in the Table 65F.
343TABLE 65F
|
|
Domain Analysis of NOV65a
Identities/
Similarities
NOV65a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Adaptin_N: domain 1 of238 . . . 272 10/35(29%)8.9
128/35(80%)
PALP: domain 1 of 111 . . . 31196/385(25%)1.4e−64
226/385(59%)
|
Example 66
[0657] The NOV66 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 66A.
344TABLE 66A
|
|
NOV66 Sequence Analysis
|
|
SEQ ID NO: 1672786 bp
NOV66a,CTTGATACACACAGCCACATATGCACACAGATGTATAAAACGGAAAAGTTTCACAAAG
CG93464-01 DNA Sequence
CAACATCCTGTGAACAATATGTTAATATTACAGGAAACAGCTATGACCATGATTACGA
|
ATTCGAGCTCGGTACCCATAAAGCTGTCCTGGAAGACCCAATGTTGAAGTTCTCAGGA
|
ACTATATCAAGAGACATGCTCTGATCTTTATGTTACTTGTCAAGTTTTTGCAGAAGGGA
|
AGCCTTTGGCCTTGCCAGTTAGAACATCCTCCCCAGCATTTAGTACAAGATGGAGCTG
|
GAATGAATGGCTGAAACTACCAGTAAAATACCCTGACCTGCCCAGGAATGCCCAAGTG
|
GCCCTCACCATATGGGATGTGTATGGTCCCGGAAAAGCAGTGCCTGTAGGAGGAACAA
|
CGGTTTCGCTCTTTGGAAAATACGGGATGTTTCGCCAAGGGATGCATGACTTGAAAGT
|
CTGGCCTAATGTAGAAGCAGATGGATCAGAACCCACAAAAACTCCTGGCAGAACAAGT
|
AGCACTCTCTCAGAAGATCAGATGAGCCGTCTTGCCAAGCTCACCAAAGCTCATCGAC
|
AAGGACACATGGTGAAAGTAGATTGGCTGGATAGATTGACATTTAGAGAAATAGAAAT
|
GATAAATGAGAGTGAAAAACGAAGTTCTAATTTCATGTACCTGATGGTTGAATTTCGA
|
TGTGTCAAGTGTGATGATAAGGAATATGGTATTGTTTATTATGAAAAGGACGGTGATG
|
AATCATCTCCAATTTTAACAAGTTTTGAATTAGTGAAAGTTCCTGACCCCCAGATGTC
|
TATGGAGAATTTAGTTGAGAGCAAACACCACAAGCTTGCCCGGAGTTTAAGAAGTGGA
|
CCTTCTGACCACGATCTGAAACCCAATGCTGCCACGAGAGATCAGTTAAATATTATTG
|
TGAGTTATCCACCAACCAAGCAACTTACATATGAAGAACAAGATCTTGTTTGGAAGTT
|
TAGATATTATCTTACGAATCAAGAAAAAGCCTTGACAAAATTCTTGAAATGTGTTAAT
|
TGGGATCTACCTCAAGAGGCCAAACAGGCCTTGGAACTTCTGGGAAAATGGAAGCCGA
|
TGGATGTAGAGGACTCCTTGGAGCTGTTATCCTCTCATTACACCAACCCAACTGTGAG
|
GCGTTATGCTGTTGCCCGGTTGCGACAGGCCGATGATGAGGATTTGTTGATGTACCTA
|
TTACAATTGGTCCAGGCTCTCAAATATGAAAATTTTGATGATATAAAGAATGGATTGG
|
AACCTACCAAGAAGGATAGTCAGAGTTCAGTGTCAGAAAATGTGTCAAATTCTGGAAT
|
AAATTCTGCAGAAATAGATAGCTCCCAAATTATAACCAGCCCCCTTCCTTCAGTCTCT
|
TCACCTCCTCCTGCATCAAAAACAAAAGAAGTTCCAGATGGCGAAAATCTGGAACAAG
|
ATCTCTGTACCTTCTTGATATCGAGAGCCTGCAAAAACTCAACACTGGCTAATTATTT
|
ATACTGGTATGTGATAGTGGAATGTGAAGATCAAGATACTCAGCAGAGAGATCCAAAG
|
ACCCATGAGATGTACTTGAACGTAATGAGAAGATTCAGCCAAGCATTGTTGAAGGGTG
|
ATAAGTCTGTCAGAGTTATGCGTTCTTTGCTGGCTGCACAACAGACATTTGTAGATCG
|
GTTGGTGCATCTAATGAAGGCAGTACAACGCGAAAGTGGAAATCGTAAGAAAAAGAAT
|
GAGAGACTACAGGCATTGCTTGGAGATAATGAAAAGATGAATTTGTCAGATGTGGAAC
|
TTATCCCGTTGCCTTTAGAACCCCAAGTGAAAATTAGAGGAATAATTCCGGAAACAGC
|
TACACTGTTTAAAAGTGCCCTTATGCCTGCACAGTTGTTTTTTAAGACGGAAGATGGA
|
GGCAAATATCCAGTTATATTTAAGCATGGAGATGATTTACGTCAAGATCAACTTATTC
|
TTCAAATCATTTCACTCATGGACAAGCTGTTACGGAAAGAAAATCTGGACTTGAAATT
|
GACACCTTATAAGGTGTTAGCCACCAGTACAAAACATGGTTTCATGCAGTTTATCCAG
|
TCAGTTCCTGTGGCTGAAGTTCTTGATACAGAGGGAAGCATTCAGAACTTTTTTAGAA
|
AATATGCACCAAGTGAGAATGGGCCAAATGGGATTAGTGCTGAGGTCATGGACACTTA
|
CGTTAAAAGCTGTGCTGGATATTGCGTGATCACCTATATACTTGGAGTTGGAGACAGG
|
CACCTGGATAACCTTTTGCTAACAAAAACAGGTAAACTCTTCCACATAGACTTTGGAT
|
ATATTTTGGGTCGGGATCCAAAGCCTCTTCCTCCACCAATGAAGCTGAATAAAGAAAT
|
GGTAGAAGGAATGGGGGGCACACAGAGTGAGCAGTACCAAGAGTTCCGTAAACAGTGT
|
TACACGGCTTTCCTCCACCTGCGAAGGTATTCTAATCTGATTTTGAACTTGTTTTCCT
|
TGATGGTTGATGCAAACATTCCAGATATTGCACTTGAACCAGATAAAACTGTGAAAAA
|
GGTTCAGGATAAATTCCGCTTAGACCTGTCGGATGAAGAGGCTGTGCATTACATGCAG
|
AGTCTGATTGATGAGAGTGTCCATGCTCTTTTTGCTGCAGTGGTGGAACAGATTCACA
|
AGTTTGCCCAGGTAAGTTCCCTGAGTGCAGTGCATTTTTCTCACCTTCTCCGTCTATA
|
CTGCCCCAGAGTCCTTGGAGAGCCATGCATTTCTCCTGCCAGTTGACATCTTTTCTGT
|
TT
|
ORF Start: ATG at 31ORE Stop: TGA at 2770
SEQ ID NO: 168913 aaMW at 104082.1 kD
NOV66a,MYKTEKFHKATSCEQYVNITGNSYDHDYEFELGTHKAVLEDPMLKFSGLYQETCSDLY
CG93464-01 Protein Sequence
VTCQVFAEGKPLALPVRTSSPAFSTRWSWNEWLKLPVKYPDLPRNAQVALTIWDVYGP
|
GKAVPKGGTTVSLFGKYGMFRQGMHDLKVWPNVEADGSEPTKTPGRTSSTLSEDQMSR
|
LAKLTKAHRQGHMVKVDWLDRLTFREIEMINESEKRSSNFMYLMVEFRCVKCDDKEYG
|
IVYYEKDGDESSPILTSFELVKVPDPQMSMENLVESKHHKLARSLRSGPSDHDLKPNA
|
ATRDQLNIIVSYPPTKQLTYEEQDLVWKFRYYLTNQEKALTKFLKCVNWDLPQEAKQA
|
LELLGKWKPMDVEDSLELLSSHYTNPTVRRYAVARLRQADDEDLLMYLLQLVQALKYE
|
NFDDIKNGLEPTKKDSQSSVSENVSNSGINSAEIDSSQIITSPLPSVSSPPPASKTKE
|
VPDGENLEQDLCTFLISRACKNSTLANYLYWYVIVECEDQDTQQRDPKTHEMYLNVMR
|
RFSQALLKGDKSVRVMRSLLAAQQTFVDRLVHLMKAVQRESGNRKKKNERLQALLGGN
|
EKMNLSDVELIPLPLEPQVKIRGIIPETATLFKSALMPAQLFFKTEDGGKYPVIFKHG
|
DDLRQDQLILQIISLMDKLLRKENLDLKLTPYKVLATSTKHGFMQFIQSVPVAEVLDT
|
EGSIQNFFRKYAPSENGPNGISAEVMDTYVKSCAGYCVITYILGVGDRHLDNLLLTKT
|
GKLFHIDFGYILGRDPKPLPPPMKLNKEMVEGMGGTQSEQYQEFRKQCYTAFLHLRRY
|
SNLILNLFSLMVDANIPDIALEPDKTVKKVQDKFRLDLSDEEAVHYNQQSLIDESVHAL
|
FAAVVEQIHKFAQVSSLSAVHFSHLLRVYCPRVLGEPCISPAS
|
SEQ ID NO: 1693040 bp
NOV66b,TGTAGGTGGTACCTTTGCAGACGGTGCGATGGGGGAAGCAGAGAAGTTTCACTACATC
CG93464-02 DNA Sequence
TATAGTTGTGACCTGGATATCAACGTCCAGCTTAAGATAGGAAGCTTGGAAGGGAAGA
|
GAGAACAAAAGAGTTATAAAGCTGTCCTGGAAGACCCAATGTTGAAGTTCTCAGGACT
|
ATATCAAGAGACATGCTCTGATCTTTAGGGTACTTGTCAAGTTTTTGCAGAAGGGAAG
|
CCTTTGGCCTTGCCAGTTAGAACATCCTCCCCAGCATTTAGTACAAGATGGAGCTGGA
|
ATGAATGGCTGAAACTACCAGTAAAATATCCTGACCTGCCCAGGAATGCCCAAGTGGC
|
CCTCACCATATGGGATGTGTATGGTCCCGGAAAAGCAGTGCCTGTAGGAGGAACAACG
|
GTTTCGCTCTTTGGAAAATACGGGATGTTTCGCCAAGGGATGCATGACTTGAAAGTCT
|
GGCCTAATGTAGAAGCAGATGGATCAGAACCCACAAAAACTCCTGGCAGAACAAGTAG
|
CACTCTCTCAGAAGATCAGATGAGCCGTCTTGCCAAGCTCACCAAAGCTCATCGACAA
|
TAAATGAGAGTGAAAAACGAAGTTCTAATTTCATGTACCTGATGGTTGAATTTCGATG
|
TGTCAAGTGTGATGATAAGGAATATGGTATTGTTTATTATGAAAAGGACGGTGATGAA
|
TCATCTCCAATTTTAACAAGTTTTGAATTAGTGAAAGTTCCTGACCCCCAGATGTCTA
|
TGGAGAATTTAGTTGAGAGCAAACACCACAAGCTTGCCCGGAGTTTAAGAAGTCGACC
|
TTCTGACCACGATCTGAAACCCAATGCTGCCACGAGAGATCAGTTAAATATTATTGTG
|
AGTTATCCACCAACCAAGCAACTTACATATGAAGAACAAGATCTTGTTTGGAAGTTTA
|
GATATTATCTTACGAATCAAGAAAAAGCCTTGACAAAATTCTTGAAATGTGTTAATTG
|
GGATCTACCTCAAGAGGCCAAACAGGCCTTGGAACTTCTGGGAAAATGGAAGCCGATG
|
GATGTAGAGGACTCCTTGGAGCTGTTATCCTCTCATTACACCAACCCAACTGTGAGGC
|
GTTATGCTGTTGCCCGGTTGCGACAGGCCGATGATGAGGATTTGTTGATGTACCTATT
|
ACAATTGGTCCAGGCTCTCAAATATGAAAATTTTGATGATATAAAGAATGGATTGGAA
|
CCTACCAAGAAGGATAGTCAGAGTTCAGTGTCAGAAAATGTGTCAAATTCTGGAATAA
|
ATTCTGCAGAAATAGATAGCTCCCAAATTATAACCAGCCCCCTTCCTTCAGTCTCTTC
|
ACCTCCTCCTGCATCAAAAACAAAAGAAGTTCCAGATGGCGAAAATCTGGAACAAGAT
|
CTCTGTACCTTCTTGATATCGAGAGCCTGCAAAAACTCAACACTGGCTAATTATTTAT
|
ACTGGTATGTGATAGTGGAATGTGAAGATCAAGATACTCAGCAGAGAGATCCAAAGAC
|
CCATGAGATGTACTTGAACGTAATGAGAAGATTCAGCCAAGCATTGTTGAAGGGTGAT
|
AAGTCTGTCAGAGTTATGCGTTCTTTGCTGGCTGCACAACAGACATTTGTAGATCGGT
|
TGGTGCATCTAATGAAGGCAGTACAACGCGAAAGTGGAAATCGTAAGAAAAAGAATGA
|
GAGACTACAGGCATTGCTTGGAGATAATGAAAAGATGAATTTGTCAGATGTGGAACTT
|
ATCCCGTTGCCTTTAGAACCCCAAGTGAAAATTAGAGGAATAATTCCGGAAACAGCTA
|
CACTGTTTAAAAGTGCCCTTATGCCTGCACAGTTGTTTTTTAAGACGGAAGATGGAGG
|
CAAATATCCAGTTATATTTAAGCATGGAGATGATTTACGTCAAGATCAACTTATTCTT
|
CAAATCATTTCACTCATGGACAAGCTGTTACGGAAAGAAAATCTGGACTTGAAATTGA
|
CACCTTATAAGGTGTTAGCCACCAGTACAAAACATGGCTTCATGCAGTTTATCCAGTC
|
AGTTCCTGTGGCTGAAGTTCTTGATACAGAGGGAAGCATTCAGAACTTTTTTAGAAAA
|
TATGCACCAAGTGAGAATGGGCCAAATGGGATTAGTGCTGAGGTCATGGACACTTACG
|
TTAAAAGCTGTGCTGGATATTGCGTGATCACCTATATACTTGGAGTTGGAGACAGGCA
|
CCTGGATAACCTTTTGCTAACAAAAACAGGCAAACTCTTCCACATAGACTTTGGATAT
|
ATTTTGGGTCGGGATCCAAAGCCTCTTCCTCCACCAATGAAGCTGAATAAAGAAATGG
|
TAGAAGGAATGGGGGGCACACAGAGTGAGCAGTACCAAGAGTTCCGTAAACAGTGTTA
|
CACGGCTTTCCTCCACCTGCGAAGGTATTCTAATCTGATTTTGAACTTGTTTTCCTTG
|
ATGGTTGATGCAAACATTCCAGATATTGCACTTGAACCAGATAAAACTGTGAAAAAGG
|
TTCAGGATAAATTCCGCTTAGACCTGTCGGATGAAGAGGCTGTGCATTACATGCAGAG
|
TCTGATTGATGAGAGTGTCCATGCTCTTTTTGCTGCAGTGGTGGAACAGATTCACAAG
|
TTTGCCCAGTACTGGAGAAAATGAAACTGGGATTGACCCATCAAGATGCTTGGCTCAA
|
TAAGAAAACCACGTTAGGAGCAACCTTTGTATATTGGAGACTTCAGAGTAACCAGCAA
|
GGAAGAGAAATCTTAATCTTCAAGTTACCATATTTTCCAAATATTACATGGTACCTGA
|
GTTCTGCTTCCTTGGATGTCATTGCTTAAATATAGTCTTGAAGGGCTTGTTTTGAAAT
|
ATTGTATATATTTTTTCAAATGTATACATTGTTAATAAATTAAGAAATGAGAAACATT
|
CTTATTTATGTACATGTTATGAGAGTTCTGGAGGAAGTAATATGTTGAAATAGTAAAC
|
CCTTTTAGTTTTTGAGTTTAAAAA
|
ORF Start: ATG at 29ORF Stop: TGA at 2690
SEQ ID NO: 170887 aaMW at 101414.2kD
NOV66b,MGEAEKFHYIYSCDLDINVQLKIGSLEGKREQKSYKAVLEDPMLKFSGLYQETCSDLY
CG93464-02 Protein Sequence
VTCQVFAEGKPLALPVRTSSPAFSTRWSWNEWLKLPVKYPDLPRNAQVALTIWDVYGP
|
GKAVPVGGTTVSLFGKYGMFRQGMHDLKVWPNVEADGSEPTKTPGRTSSTLSEDQMSR
|
LAKLTKAHRQGHMVKVDWLDRLTFREIEMINESEKRSSNFMYLMVEFRCVKCDDKEYG
|
IVYYEKDGDESSPILTSFELVKVPDPQMSMENLVESKHHKLARSLRSGSPDHDLKPNA
|
ATRDQLNIIVSYPPTKQLTYEEQDLVWKFRYYLTNQEKALTKFLKCVNWDLPQEAKQA
|
LELLGKWKPMDVEDSLELLSSHYTNPTVRRYAVARLRQADDEDLLMYLLQLVQALKYE
|
NFDDIKNGLEPTKKDSQSSVSENVSNSGINSAEIDSSQIITSPLPSVSSPPPASKTKE
|
VPDGENLEQDLCTFLISRACKNSTLANYLYWYVIVECEDQDTQQRDPKTHEMYLNVMR
|
RFSQALLKGDKSVRVMRSLLAAQQTFVDRLVHLMKAVQRESGNRKKKNERLQALLGDN
|
EKMNLSDVELIPLPLEPQVKIRGIIPETATLFKSALMPAQLFFKTEDGGKYPVIFKHG
|
DDLRQDQLILQIISLMDKLLRKENLDLKLTPYKVLATSTKHGFMQFIQSVPVAEVLDT
|
EGSIQNFFRKYAPSENGPNGISAEVMDTYVKSCAGYCVITYILGVGDRHLDNLLLTKT
|
GKLFHIDFGYILGRDPKPLPPPMKLNKEMVEGMGGTQSEQYQEFRKQCYTAFLHLRRY
|
SNLILNLFSLMVDANIPDIALEPDKTVKKVQDKFRLDLSDEEAVHYMQSLIDESVHAL
|
FAAVVEQIHKFAQYWRK
|
SEQ ID NO: 1712654 bp
NOV66c,TGTAGGTGGTACCTTTGCAGACGGTGCGATGGGGGAAGCAGAGAAGTTTCACTACATC
CG93464-03 DNA Sequence
TATAGTTGTGACCTGGATATCAACGTCCAGCTTAAGATAGGAAGCTTGGAAGGGAAGA
|
GAGAACAAAAGAGTTATAAAGCTGTCCTGGAAGACCCAATGTTGAAGTTCTCAGGACT
|
ATATCAAGAGACATGCTCTGATCTTTATGTTACTTGTCAAGTTTTTGCAGAAGGGAAG
|
CCTTTGGCCTTGCCAGTTAGAACATCCTCCCCAGCATTTAGTACAAGATGGAGCTGGA
|
ATGAATGGCTGAAACTACCAGTAAAATACCCTGACCTGCCCAGGAATGCCCAAGTGGC
|
CCTCACCATATGGGATGTGTATGGTCCCGGAAAAGCAGTGCCTGTAGGAGGAACAACG
|
GTTTCGCTCTTTGGAAAATACGGGATGTTTCGCCAAGGGATGCATGACTTGAAAGTCT
|
GGCCTAATGTAGAAGCAGATGGATCAGAACCCACAAAAACTCCTGGCAGAACAAGTAG
|
CACTCTCTCAGAAGATCAGATGAGCCGTCTTGCCAAGCTCACCAAAGCTCATCGACAA
|
GGACACATGGTGAAAGTAGATTGGCTGGATAGATTGACATTTAGAGAAATAGAAATGA
|
TAAATGAGAGTGAAAAACGAAGTTCTAATTTCATGTACCTGATGGTTGAATTTCGATG
|
TGTCAAGTGTGATGATAAGGAATATGGTATTGTTTATTATGAAAAGGACGGTGATGAA
|
TCATCTCCAATTTTAACAAGTTTTGAATTAGTGAAAGTTCCTGACCCCCAGATGTCTA
|
TGGAGAATTTAGTTGAGAGCAAACACCACAAGCTTGCCCGGAGTTTAAGAAGTGGACC
|
TTCTGACCACGATCTGAAACCCAATGCTGCCACGAGAGATCAGTTAAATATTATTGTG
|
AGTTATCCACCAACCAAGCAACTTACATATGAAGAACAAGATCTTGTTTGGAAGTTTA
|
GATATTATCTTACGAATCAAGAAAAAGCCTTGACAAAATTCTTGAAATGTGTTAATTG
|
GGATCTACCTCAAGAGGCCAAACAGGCCTTGGAACTTCTGGGAAAATGGAAGCCGATG
|
GTTATGCTGTTGCCCGGTTGCGACAGGCCGATGATGAGGATTTGTTGATGTACCTATT
|
ACAATTGGTCCAGGCTCTCAAATATGAAAATTTTGATGATATAAAGAATGGATTGGAA
|
CCTACCAAGAAGGATAGTCAGAGTTCAGTGTCAGAAAATGTGTCAAATTCTGGAATAA
|
ATTCTGCAGAAATAGATAGCTCCCAAATTATAACCAGCCCCCTTCCTTCAGTCTCTTC
|
ACCTCCTCCTGCATCAAAAACAAAAGAAGTTACAGATGGCGAAAATCTGGAACAAGAT
|
CTCTGTACCTTCTTGATATCGAGAGCCTGCAAAAACTCAACACTGGCTAATTATTTAT
|
ACTGGTATGTGATAGTGGAATGTGAAGATCAAGATACTCAGCAGAGAGATCCAAAGAC
|
CCATGAGATGTACTTGAACGTAATGAGAAGATTCAGCCAAGCATTGTTGAAGGGTGAT
|
AAGTCTGTCAGAGTTATGCGTTCTTTGCTGGCTGCACAACAGACATTTGTAGATCGGT
|
TGGTGCATCTAATGAAGGCAGTACAACGCGAAAGTGGAAATCGTAAGAAAAAGAATGA
|
GAGACTACAGGCATTGCTTGGAGATAATGAAAAGATGAATTTGTCAGATGTGGAACTT
|
ATCCCGTTGCCTTTAGAACCCCAAGTGAAAATTAGAGGAATAATTCCGGAAACAGCTA
|
CACTGTTTAAAAGTGCCCTTATGCCTGCACAGTTGTTTTTTAAGACGGAAGATGGAGG
|
CAAATATCCAGTTATATTTAAGCATGGAGATGATTTACGTCAAGATCAACTTATTCTT
|
CAAATCATTTCACTCATGGACAAGCTGTTACGGAAAGAAAATCTGGACTTGAAATTGA
|
CACCTTATAAGGTGTTAGCCACCAGTACAAAACATGGCTTCATGCAGTTTATCCAGTC
|
AGTTCCTGTGGCTGAAGTTCTTGATACAGAGGGAAGCATTCAGAACTTTTTTAGAAAA
|
TATGCACCAAGTGAGAATGGGCCAAATGGGATTAGTGCTGAGGTCATGGACACTTACG
|
TTAAAAGCTGTGCTGGATATTGCGTGATCACCTATATACTTGGAGTTGGAGACAGGCA
|
CCTGGATAACCTTTTGCTAACAAAAACAGTACTGGAGAAAATGAAACTGGGATTGACC
|
CATCAAGATGCTTGGCTCAATAAGAAAACCACGTTAGGAGCAACCTTTGTATATTGGA
|
GACTTCAGAGTAACCAGCAAGGAAGAGAAATCTTAATCTTCAAGTTACCATATTTTCC
|
AAATATTACATGGTACCTGAGTTCTGCTTCCTTGGATGTCATTGCTTAAATATAGTCT
|
TGAAGGGCTTGTTTTGAAATATTGTATATATTTTTTCAAATGTATACATTGTTAATAA
|
ATTAAGAAATGAGAAACATTCTTATTTATGTACATGTTATGAGAGTTCTGGAGGAAGT
|
AATATGTTGAAATAGTAAACCCTTTTAGTTTTTGAGTTTAAAAA
|
ORF Start: ATG at 29ORF Stop: TAA at 2483
SEQ ID NO: 172818 aaMW at 93341.0kD
NOV66c,MGEAEKFHYIYSCDLDINVQLKIGSLEGKREQKSYKAVLEDPMLKFSGLYQETCSDLY
CG93464-03 Protein Sequence
VTCQVFAEGKPLALPVRTSSPAFSTRWSWNEWLKLPVKYPDLPRNAQVALTIWDVYGP
|
GKAVPVGGTTVSLFGKYGMFRQGMHDLKVWPNVEADGSEPTKTPORTSSTLSEDQMSR
|
LAKLTKAHRQGHMVKVDWLDRLTFREIEMINESEKRSSNFMYLMVEFRCVKCDDKEYG
|
IVYYEKDGDESSPILTSFELVKVPDPQMSMENLVESKHHKLARSLRSGPSDHDLKPNA
|
ATRDQLNIIVSYPPTKQLTYEEQDLVWKFRYYLTNQEKALTKFLKCVNWDLPQEAKQA
|
LELLGKWKPMDVEDSLELLSSHYTNPTVRRYAVARLRQADDEDLLMYLLQLVQALKYE
|
NFDDIKNGLEPTKKDSQSSVSENVSNSGINSAEIDSSQIITSPLPSVSSPPPASKTKE
|
VPDGENLEQDLCTFLISRACKNSTLANYLYWYVIVECEDQDTQQRDPKTHEMYLNVMR
|
RFSQALLKGDKSVRVMRSLLAAQQTFVDRLVHLMKAVQRESGNRKKKNERLQALLGDN
|
EKMNLSDVELIPLPLEPQVKIRGIIPETATLFKSALMPAQLFFKTEDGGKYPVIFKHG
|
DDLRQDQLILQIISLMDKLLRKENLDLKLTPYKVLATSTKHGFMQFIQSVPVAEVLDT
|
EGSIQNFFRKYAPSENGPNGISAEVMDTYVKSCAGYCVITYILGVGDRHLDNLLLTKT
|
VLEKMKLGLTHQDAWLNKKTTLGATFVYWRLQSNQQGREILIFKLPYFPNITWYLSSA
|
SLDVIA
|
[0658] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 66B.
345TABLE 66B
|
|
Comparison of NOV66a against NOV66b and NOV66c.
NOV66a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV66b1 . . . 883806/883 (91%)
1 . . . 883814/883 (91%)
NOV66c1 . . . 754687/754 (91%)
1 . . . 754695/754 (92%)
|
[0659] Further analysis of the NOV66a protein yielded the following properties shown in Table 66C.
346TABLE 66C
|
|
Protein Sequence Properties NOV66a
|
|
PSort0.4600 probability located in nucleus; 0.3000 probability
analysis:located in microbody (peroxisome); 0.1000 probability
located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0660] A search of the NOV66a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 66D.
347TABLE 66D
|
|
Geneseq Results for NOV66a
NOV66aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAM39431Human polypeptide SEQ ID NO1 . . . 883844/883 (95%)0.0
2576 - Homo sapiens, 887 aa.1 . . . 883855/883 (96%)
[WO200153312-A1, 26 Jul. 2001]
AAM41217Human polypeptide SEQ ID NO1 . . . 883822/885 (92%)0.0
6148 - Homo sapiens, 901 aa.13 . . . 897 838/885 (93%)
[WO200153312-A1, 26 Jul. 2001]
AAW97880Maize phosphatidylinositol-3-kinase49 . . . 883 328/839 (39%)e−163
- Zea mays, 803 aa. [WO9905298-32 . . . 800 475/839 (56%)
A1, 04 Feb. 1999]
AAG00176Human secreted protein, SEQ ID358 . . . 506 149/149 (100%)1e−80
NO: 4257 - Homo sapiens, 149 aa.1 . . . 149149/149 (100%)
[EP1033401-A2, 06 Sep. 2000]
AAR46552Human PITR-c, 127 aa.638 . . . 764 126/127 (99%)1e−67
[WO9321328-A, 28 Oct. 1993]1 . . . 127127/127 (99%)
|
[0661] In a BLAST search of public sequence datbases, the NOV66a protein was found to have homology to the proteins shown in the BLASTP data in Table 66E.
348TABLE 66E
|
|
Public BLASTP Results for NOV66a
NOV66aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
O88763PHOSPHATIDYLINOSITOL 3-1 . . . 883837/883 (94%)0.0
KINASE - Rattus norvegicus (Rat),1 . . . 883858/883 (96%)
887 aa.
Q15134PHOSPHATIDYLINOSITOL 3-1 . . . 883820/883 (92%)0.0
KINASE - Homo sapiens (Human),1 . . . 883836/883 (93%)
887 aa.
S57219phosphatidylinositol 3-kinase -1 . . . 883819/883 (92%)0.0
human, 887 aa.1 . . . 883834/883 (93%)
AAH24675SIMILAR TO1 . . . 842792/842 (94%)0.0
PHOSPHATIDYLINOSITOL 3-1 . . . 841812/842 (96%)
KINASE - Mus musculus (Mouse),
847 aa.
Q9TXI7HYPOTHETICAL 103.1 KDA23 . . . 883 349/915 (38%)e−154
PROTEIN - Caenorhabditis elegans,12 . . . 898 520/915 (56%)
901 aa.
|
[0662] PFam analysis predicts that the NOV66a protein contains the domains shown in the Table 66F.
349TABLE 66F
|
|
Domain Analysis of NOV66a
Identities/
Similarities for
NOV66a Matchthe Matched
Pfam DomainRegionRegionValue
|
PI3K_C2: domain 1 of 1 34 . . . 202 68/197 (35%)6.4e−64
153/197 (78%)
PI3Ka: domain 1 of 1283 . . . 530 88/273 (32%)1.6e−69
192/273 (70%)
PI3_PI4_kinase:630 . . . 833103/286 (36%) 8.4e−105
domain 1 of 1191/286 (67%)
|
Example 67
[0663] The NOV67 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 67A.
350TABLE 67A
|
|
NOV67 Sequence Analysis
|
|
SEQ ID NO: 1735873 bp
NOV67aCACGTGCATGTGTAGCATGCCTTGGTTTTTCCTTTGGCATCTGAAAAAGGCACAACCT
CG93495-01 DNA Sequence
GAAAGACCTAGAACCCAGTGTCGGTCCCCAGGCCCTTTGGGACAGGAAGAGAAGAGCC
|
GTGTGGCCGCGGGGAGGATGTCCTGCGGCGGGGCTGTCCTCGCGGACTGACTGGACTC
|
CATCTCCCAGCGGGCGCCGCGGCGCGGCCACGCCCCCCCACTCCCCGCGCGCGCCCGG
|
TGGAGACTTCGATTTTCAGAATTCCTCCTGGGAATGCTGACTCCTTGCTTGGTGCCCT
|
GATGCTTCTCTGAGATAAACTGATGAATTGGAACCATGGTGCAAAAGAAGAAGTTCTG
|
TCCTCGGTTACTTGACTATCTAGTGATCGTAGGGGCCAGGCACCCGAGCAGTGATAGC
|
GTGGCCCAGACTCCTGAATTGCTACGGCGATACCCCTTGGAGGATCACACTGAGTTTC
|
CCCTGCCCCCAGATGTAGTGTTCTTCTGCCAGCCCGAGGGCTGCCTGAGCGTGCGGCA
|
GCGGCGCATGAGCCTTCGGGATGATACCTCTTTTGTCTTCACCCTCACTGACAAGGAC
|
ACTGGAGTCACGCGATATGGCATCTGTGTTAACTTCTACCGCTCCTTCCAAAAGCGAA
|
TCTCTAAGGAGAAGGGGGAAGGTGGGGCAGGGTCCCGTGGGAAGGAAGGAACCCATGC
|
CACCTGTGCCTCAGAAGAGGGTGGCACTGAGAGCTCAGAGAGTGGCTCATCCCTGCAG
|
CCTCTCAGTGCTGACTCTACCCCTGATGTGAACCAGTCTCCTCGGGGCAAACGCCGGG
|
CCAAGGCGGGGAGCCGCTCCCGCAACAGTACTCTCACGTCCCTGTGCGTGCTCAGCCA
|
CTACCCTTTCTTCTCCACCTTCCGAGAGTGTTTGTATACTCTCAAGCGCCTGGTGGAC
|
TGCTGTAGTGAGCGCCTTCTGGGCAAGAAACTGGGCATCCCTCGAGGCGTACAAAGGG
|
ACACCATGTGGCGGATCTTTACTGGATCGCTGCTGGTAGAGGAGAAGTCAAGTGCCCT
|
TCTGCATGACCTTCGAGAGATTGAGGCCTGGATCTATCGATTGCTGCGCTCCCCAGTA
|
CCCGTCTCTGGGCAGAAGCGAGTAGACATCGAGGTCCTACCCCAAGAGCTCCAGCCAG
|
CTCTGACCTTTGCTCTTCCAGACCCATCTCGATTCACCCTAGTGGATTTCCCACTGCA
|
CCTTCCCTTGGAACTTCTAGGTGTGGACGCCTGTCTCCAGGTGCTAACCTGCATTCTG
|
TTAGAGCACAAGGTGGTGCTACAGTCCCGAGACTACAATGCACTCTCCATGTCTGTGA
|
TGGCATTCGTGGCAATGATCTACCCACTGGAATATATGTTTCCTGTCATCCCGCTGCT
|
ACCCACCTGCATGGCATCAGCAGAGCAGCTGCTGTTGGCTCCAACCCCGTACATCATT
|
GGGGTTCCTGCCAGCTTCTTCCTCTACAAACTGGACTTCAAAATGCCTGATGATGTAT
|
GGCTAGTGGATCTGGACAGCAATAGGGTGATTGCCCCCACCAATGCAGAAGTGCTGCC
|
TATCCTGCCAGAACCAGAATCACTAGAGCTGAAAAAGCATTTAAAGCAGGCCTTGGCC
|
AGCATGAGTCTCAACACCCAGCCCATCCTCAATCTGGAGAAATTTCATGAGGGCCAGG
|
AGATCCCCCTTCTCTTGGGAAGGCCTTCTAATGACCTGCAGTCCACACCGTCCACTGA
|
ATTCAACCCACTCATCTATGGCAATGATGTGGATTCTGTGGATGTTGCAACCAGGGTT
|
GCCATGGTACGGTTCTTCAATTCCGCCAACGTGCTGCAGGGATTTCAGATGCACACGC
|
GTACCCTGCGCCTCTTTCCTCGGCCTGTGGTAGCTTTTCAAGCTGGCTCCTTTCTAGC
|
CTCACGTCCCCGGCAGACTCCTTTTGCCGAGAAATTGGCCAGGACTCAGGCTGTGGAG
|
TACTTTGGGGAATGGATCCTTAACCCCACCAACTATGCCTTTCAGCGAATTCACAACA
|
ATATGTTTGATCCAGCCCTGATTGGTGACAAGCCAAAGTGGTATGCTCATCAGCTGCA
|
GCCTATCCACTATCGCGTCTATGACAGCAATTCCCAGCTGGCTGAGGCCCTGAGTGTA
|
CCACCAGAGCGGGACTCTGACTCCGAACCTACTGATGATAGTGGCAGTGATAGTATGG
|
ATTATGACGATTCAAGCTCTTCTTACTCCTCCCTTGGTGACTTTGTCAGTGAAATGAT
|
GAAATGTGACATTAATGGTGATACTCCCAATGTGGACCCTCTGACACATGCAGCACTG
|
GGGGATGCCAGCGAGGTGGAGATTGACGAGCTGCAGAATCAGAAGGAAGCAGAAGAGC
|
CTGGCCCAGACAGTGAGAACTCTCAGGAAAACCCCCCACTGCGCTCCAGCTCTAGCAC
|
CACAGCCAGCAGCAGCCCCAGCACTGTCATCCACGGAGCCAACTCTGAACCTGCTGAC
|
TCTACGGAGATGGATGATAAGGCAGCAGTAGGCGTCTCCAAGCCCCTCCCTTCCGTGC
|
CTCCCAGCATTGGCAAATCGGACGTGGACAGACGTCAGGCAGAAATTGGAGAGGGGGC
|
TCAAAAGCTGCTGCGGCCCAACAGCTTGAGACTGGCAAGTGACTGAGATGCAGAGTCA
|
GACTCTCGGGCAAGCTCTCCCAACTCCACCGTCTCCAACACCAGCACCGAGGGCTTCG
|
GGGGCATCATGTCTTTTGCCAGCAGCCTCTATCGGAACCACAGTACCAGCTTCAGTCT
|
TTCAAACCTCACACTGCCCACCAAAGGTGCCCGAGAGAAGGCCACGCCCTTCCCCAGT
|
CTGAAAGGAAACAGGAGGGCGTTAGTGGATCAGAAGTCATCTGTCATTAAACACAGCC
|
CAACAGTGAAAAGAGAACCTCCATCACCCCAGGGTCGATCCAGCAATTCTAGTGAGAA
|
CCAGCAGTTCCTGAAGGAGGTGGTGCACAGCGTGCTGGACGGCCAGGGAGTTGGCTGG
|
CTCAACATGAAAAAGGTGCGCCGGCTGCTGGAGAGCGAGCAGCTGCGAGTTCTTGTCC
|
TGAGCAAGCTGAACCGCATGGTGCAGTCAGAGGACGATGCCCGGCAGGACATCATCCC
|
GGATGTGGAGATCAGTCGGAAGGTGTACAAGGGAATGTTAGACCTCCTCAAGTGTACA
|
GTCCTCAGCTTGGAGCAGTCCTATGCCCACGCGGGTCTGGGTGGCATGGCCAGCATCT
|
TTGGGCTTTTGGAGATTGCCCAGACCCACTACTATAGTAAAGAACCAGACAAGCGGAA
|
GAGAAGTCCAACAGAAAGTGTAAATACCCCAGTTGGCAAGGATCCTGGCCTAGCTGGG
|
CGGGGGGACCCAAAGGCTATGGCACAACTGAGAGTTCCACAACTGGGACCTCGGGCAC
|
CAAGTGCCACAGGAAAGGGTCCTAAGGAACTGGACACCAGAAGTTTAAAGGAAGAAAA
|
TTTTATAGCATCTATTGGGCCTGAAGTAATCAAACCTGTCTTTGACCTTGGTGAGACA
|
GAGGAGAAAAAGTCCCAGATCAGCGCAGACAGTGGTGTGAGCCTGACGTCTAGTTCCC
|
AGAGGACTGATCAAGACTCTGTCATCGGCGTGAGTCCAGCTGTTATGATCCGCAGCTC
|
AAGTCAGGATTCTGAAGTTAGCACCGTGGTGAGTAATAGCTCTGGAGAGACCCTTGGA
|
GCTGACAGTGACTTGAGCAGCAATGCAGGTGATGGACCAGGTGGCGAGGGCAGTGTTC
|
ACCTGGCAAGCTCTCGGGGCACTTTGTCTGATAGTGAAATTGAGACCAACTCTGCCAC
|
AAGCACCATCTTTGGTAAAGCCCACAGCTTGAAGCCAAGCATAAAGGAGAAGCTGGCA
|
GGCAGCCCCATTCGTACTTCTGAAGATGTGAGCCAGCGAGTCTATCTCTATGAGGGAC
|
TCCTAGGCAAAGAGCGTTCTACTTTATGGGACCAAATGCAATTCTGGGAAGATGCCTT
|
CTTAGATGCTGTGATGTTGGAGAGAGAAGGGATGGGTATGGACCAGGGTCCCCAGGAA
|
ATGATCGACAGGTACCTGTCCCTTGGAGAACATGACCGGAAGCGCCTGGAAGATGATG
|
AAGATCGCTTGCTGGCCACACTTCTGCACAACCTCATCTCCTACATGCTGCTGATGAA
|
GGTAAATAAGAATGACATCCGCAAGAAGGTGAGGCGCCTAATGGGAAAGTCGCACATT
|
GGGCTTGTGTACAGCCAGCAAATCAATGAGGTGCTTGATCAGCTGGCGAACCTGAATG
|
GACGCGATCTCTCTATCTGGTCCAGTGGCAGCCGGCACATGAAGAAGCAGACATTTGT
|
GGTACATGCAGGGACAGATACAAACGGAGATATCTTTTTCATGGAGGTGTGCGATGAC
|
TGTGTGGTGTTGCGTAGTAACATCGGAACAGTGTATGAGCGCTGGTGGTACGAGAAGC
|
TCATCAACATGACCTACTGTCCCAAGACGAAGGTGTTGTGCTTGTGGCGTAGAAATGG
|
CTCTGAGACCCAGCTCAACAAGTTCTATACTAAAAAGTGTCGGGAGCTGTACTACTGT
|
GTGAAGGACAGCATGGAGCGCGCTGCCGCCCGACAGCAAAGCATCAAACCCGGACCTG
|
AATTGGGTGGCGAGTTCCCTGTGCAGGACCTGAAGACTGGTGAGGGTGGCCTGCTGCA
|
GGTGACCCTGGAAGGGATCAACCTCAAATTCATGCACAATCACGTTTTCATAGAGCTG
|
AATCACATTAAAAAGTGCAATACAGTTCGACGCGTCTTTGTCCTGGAGGAATTTGTTC
|
CTGAAATTAAAGAAGTGGTGAGCCACAAGTACAAGACACCAATGGCCCACGAAATCTG
|
CTACTCCGTATTATGTCTCTTCTCGTACGTGGCTGCAGTTCATAGCAGTGAGGAAGAT
|
CTCAGAACCCCGCCCCGGCCTGTCTCTAGCTGATGGAGAGGGGCTACGCAGCTGCCCC
|
AGCCCAGGGCACGCCCCTGGCCCCTTGCTGTTCCCAAGTGCACGATGCTGCTGTGACT
|
GAGGAGTGGATGATGCTCGTGTGTCCTCTGCAAGCCCCCTGCTGTGGCTTGGTTGGTT
|
ACCGGTTATGTGTCCCTCTGAGTCTGTCTTGAGCGTGTCCACCTTCTCCCTCTCCACT
|
CCCAGAAGACCAAACTGCCTTCCCCTCAGGGCTCAAGAATGTGTACAGTCTGTGGGGC
|
CGGTGTGAACCCACTATTTTGTGTCCTTGAGACATTTGTGTTGTGGTTCCTTGTCCTT
|
GTCCCTGGCGTTATAACTGTCCACTGCAAGAGTCTGGCTCTCCCTTCTCTGTGACCCG
|
GCATGACTGGGCGCCTGGAGCAGTTTCACTCTGTGAGGAGTGAGGGAACCCTGGGGCT
|
CACCCTCTCAGAGGAAGGGCACAGAGAGGAAGGGAAGAATTGGGGGGCAGCCGGAGTG
|
AGTGGCAGCCTCCCTGCTTCCTTGTGCATTCCCAAGCCGGCAGCTACTGCCCACGGCC
|
CGCAGTGTTGGCTGCTGCCTGCCACAGCCTCTGTGACTGCAGTGGAGCGGCGAATTCC
|
CTGTGGCCTGCCACGCCTTCGGCATCAGAGGATGGAGTGGTCGAGGCTAGTGGAGTCC
|
CAGGGACCGCTGGCTGCTCTGCCTGAGCATCAGGGAGGGGGCAGGAAAGACCAAGCTG
|
GGTTTGCACATCTGTCTGCAGGCTGTCTCTCCAGGCACCGGGTGTCACGAGGGAGAGA
|
CAGCCTGGGTATGGGCAAGAAATGACTGTAAATATTTCAGCCCCACATTATTTATAGA
|
AAATGTACAGTTGTGTGAATGTGAAATAAATGTCCTCAACTCCCAAAAAAAAAAAAAA
|
AAAAAAAAAAAAAAA
|
ORF Start: ATG at 326ORF Stop: TGA at 4961
SEQ ID NO: 1741545 aaMW at 171357.1kD
NOV67a,MVQKKKFCPRLLDYLVIVGARHPSSDSVAQTPELLRRYPLEDHTEFPLPPDVVFFCQP
CG93495-01 Protein Sequence
EGCLSVRQRRMSLRDDTSFVFTLTDKDTGVTRYGICVNFYRSFQKRISKEKGEGGAGS
|
RGKEGTHATCASEEGGTESSESGSSLQPLSADSTPDVNQSPRGKRRAKAGSRSRNSTL
|
TSLCVLSHYPFFSTFRECLYTLKRLVDCCSERLLGKKLGIPRGVQRDTMWRIFTGSLL
|
VEEKSSALLHDLREIEAWIYRLLRSPVPVSGQKRVDIEVLPQELQPALTFALPDPSRF
|
TLVDFPLHLPLELLGVDACLQVLTCILLEHKVVLQSRDYNALSMSVMAFVAMIYPLEY
|
MFPVIPLLPTCMASAEQLLLAPTPYIIGVPASFFLYKLDFKMPDDVWLVDLDSNRVIA
|
PTNAEVLPILPEPESLELKKHLKQALASMSLNTQPILNLEKFHEGQBIPLLLGRPSND
|
LQSTPSTEFNPLIYGNDVDSVDVATRVAMVRFFNSANVLQGFQMHTRTLRLFPRPVVA
|
FQAGSFLASRPRQTPFAEKLARTQAVEYFGEWILNPTNYAFQRIHNNMFDPALIGDKP
|
KWYAHQLQPIHYRVYDSNSQLAEALSVPPERDSDSEPTDDSGSDSMDYDDSSSSYSSL
|
GDFVSEMMKCDINGDTPNVDPLTHAALGDASEVEIDELQNQKEAEEPGPDSENSQENP
|
PLRSSSSTTASSSPSTVIHGANSEPADSTEMDDKAAVGVSKPLPSVPPSIGKSDVDRR
|
QAEIGEGAQKLLRPNSLRLASDSDAESDSRASSPNSTVSNTSTEGFGGIMSFASSLYR
|
NHSTSFSLSNLTLPTKGAREKATPFPSLKGNRRALVDQKSSVIKHSPTVKREPPSPQG
|
RSSNSSENQQFLKEVVHSVLDGQGVGWLNMKKVRRLLESEQLRVLVLSKLNRMVQSED
|
DARQDIIPDVEISRKVYKGMLDLLKCTVLSLEQSYAHAGLGGMASIFGLLEIAQTHYY
|
SKEPDKRKRSPTESVNTPVGKDPGLAGRGDPKAMAQLRVPQLGPRAPSATGKGPKELD
|
TRSLKEENFIASAGPEVIKPVFDLGETEEKKSQISADSGVSLTSSSQRTDQDSVIGVS
|
PAVMIRSSSQDSEVSTVVSNSSGETLGADSDLSSNAGDGPGGEGSVHLASSRGTLSDS
|
EIETNSATSTIFGKAHSLKPSIKEKLAGSPIRTSEDVSQRVYLYEGLLGKERSTLWDQ
|
MQFWEDAFLDAVMLEREGMGMDQGPQEMIDRYLSLGEHDRKRLEDDEDRLLATLLHNL
|
ISYMLLMKVNKNDIRKKVRRLMGKSHIGLVYSQQINEVLDQLANLNGRDLSIWSSGSR
|
HMKKQTFVVHAGTDTNGDIFFMEVCDDCVVLRSNIGTVYERWWYEKLINMTYCPKTKV
|
LCLWRRNGSETQLNKFYTKKCRELYYCVKDSMERAAARQQSIKPGPELGGEFPVQDLK
|
TGEGGLLQVTLEGINLKFMHNQVFIELNHIKKCNTVRGVFVLEEFVPEIKEVVSHKYK
|
TPMAHEICYSVLCLFSYVAAVHSSEEDLRTPPRPVSS
|
[0664] Further analysis of the NOV67a protein yielded the following properties shown in Table 67B.
351TABLE 67B
|
|
Protein Sequence Properties NOV67a
|
|
PSort0.8200 probability located in endoplasmic reticulum (mem-
analysis:brane); 0.1900 probability located in plasma membrane;
0.1000 probability located in endoplasmic reticulum (lumen);
0.1000 probability located in outside
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0665] A search of the NOV67a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 67C.
352TABLE 67C
|
|
Geneseq Results for NOV67a
NOV67aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAW35576TNF-R1-DD ligand protein clone1 . . . 15451542/1588(97%)0.0
57TU4A - Homo sapiens, 1588 aa.1 . . . 15881544/1588(97%)
[WO9730084-A1, 21 Aug. 1997]
AAW64453Rat brain Rab3 GEP protein - Rattus1 . . . 15451477/1609(91%)0.0
sp, 1602 aa. [EP856583-A2, 051 . . . 16021501/1609(92%)
Aug. 1998]
AAB37805Human TNF receptor death domain939 . . . 1545 607/607(100%)0.0
ligand protein #4 - Homo sapiens,1 . . . 607 607/607(100%)
607 aa. [WO200064479-A1, 02
Nov. 2000]
AAW35574TNF-R1-DD ligand protein clone939 . . . 1545 607/607(100%)0.0
27TU - Homo sapiens, 607 aa.1 . . . 607 607/607(100%)
[WO9730084-A1, 21 Aug. 1997]
AAR95331Tumor necrosis factor receptor 1939 . . . 1545 607/607(100%)0.0
death domain ligand (clone 27TU) -1 . . . 607 607/607(100%)
Homo sapiens, 607 aa.
[WO9612735-A1, 02 May 1996]
|
[0666] In a BLAST search of public sequence datbases, the NOV67a protein was found to have homology to the proteins shown in the BLASTP data in Table 67D.
353TABLE 67D
|
|
Public BLASTP Results for NOV67a
NOV67aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
AAL40268INSULINOMA-1 . . . 15451538/1545 (99%)0.0
GLUCAGONOMA PROTEIN 201 . . . 15451540/1545 (99%)
SPLICE VARIANT 3 - Homo
sapiens (Human), 1545 aa.
AAL40267INSULINOMA-1 . . . 15451538/1565 (98%)0.0
GLUCAGONOMA PROTEIN 201 . . . 15651540/1565 (98%)
SPLICE VARIANT 2 - Homo
sapiens (Human), 1565 aa.
O15293MAP KINASE-ACTIVATING1 . . . 15451542/1588 (97%)0.0
DEATH DOMAIN PROTEIN -1 . . . 15881544/1588 (97%)
Homo sapiens (Human), 1588 aa.
AAL35261INSULINOMA-1 . . . 14771467/1477 (99%)0.0
GLUCAGONOMA PROTEIN 201 . . . 14761471/1477 (99%)
SPLICE VARIANT 4 - Homo
sapiens (Human), 1479 aa.
O08873RAB3 GDP/GTP EXCHANGE1 . . . 15451477/1609 (91%)0.0
PROTEIN - Rattus norvegicus1 . . . 16021501/1609 (92%)
(Rat), 1602 aa.
|
[0667] PFam analysis predicts that the NOV67a protein contains the domains shown in the Table 67E.
354TABLE 67E
|
|
Domain Analysis of NOV67a
Identities/
Similarities
NOV67a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
DENN: domain 1 of 1254 . . . 40283/154(54%)7e−86
147/154(95%)
Latrophilin: domain 1 of 759 . . . 104883/419(20%)4.4
1142/419(34%)
Ribosomal_L14: domain1046 . . . 10629/19(47%)5
1 of 115/19(79%)
Ribosomal_L30: domain1460 . . . 14786/19(32%)6.4
1 of 117/19(89%)
|
Example 68
[0668] The NOV68 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 68A.
355TABLE 68A
|
|
NOV68 Sequence Analysis
|
|
SEQ ID NO: 1756327 bp
NOV68a,ATGGCCTTCTGGACACAGCTGATGCTGCTGCTCTGGAAGAATTTCATGTATCGCCGGA
CG93529-01 DNA Sequence
GGACAGCCGGTCCAGCTCCTGGTCGAATTGCTGTGGCCTCTCTTCCTCTTCTTCATCCT
|
GGTGGCTGTTCGCCACTCCCACCCGCCCCTGGAGCACCATGAATGCCACTTCCCAAAC
|
GAAGCCACTGCCATCGGCGGGCACCGTGCCCTGGCTCCAGGGTCTCATCTGTAATGTGA
|
GACAACACCTGCTTTCCGCAGCTGACACCGGGCGAGGAGCCCGGGCGCCTGAGCAACTT
|
GCAACGACTCCCTGGTCTCCCGGCTGCTAGCCGATGCCCGCACTGTGCTGGGAGGGGCC
|
GAGTGCCCACAGGACGCTGGCTGGCCTAGGGAAGCTGATCGCCACGCTGAGGGCTGCAC
|
GGCAGCACGGCCCAGCCTCAACCAACCAAGCAGTCTCCACTGGAACCACCCATGCTGGA
|
GTGTCGCGGAGCTGCTGACGTCACTGCTGCGCACGGAATCCCTGGGGTTGGCACTGGGC
|
GCAAGCCCAGGAGCCCTTGCACAGCTTGTTGGAGGCCGCTGAGGACCTGGCCCAGGAGC
|
GTCCTGGCGCTGCGCAGCCTGGTGGAGCTTCGGGCACTGCTGCAGAGACCCCGAGGGAC
|
GCAGCGGCCCCCTGGAGTTGCTGTCAGAGGCCCTCTGCAGTGTCAGGGGACCTAGCAGC
|
GACAGTGGGCCCCTCCCTCAACTGGTACGAGGCTAGTGACCTGATGGAGCTGGTGGGGC
|
GAGGAGCCAGAATCCGCCCTGCCAGACAGCAGCCTGAGCCCCGCCTGCTCGGAGCTGAT
|
GTGGAGCCCTGGACAGCCACCCGCTGTCCCGCCTGCTCTGGAGACGCCTGAAGCCTCTG
|
GATCCTCGGGAAGCTACTCTTTGCACCAGATACACCTTTTACCCGGAAGCTCATGGCCC
|
GAGGTCAACCGGACCTTCGAGGAGCTCACCCTGCTGAGGGATGTCCGGGAGGTGTGGGA
|
GGATGCTGGGACCCCGGATCTTCACCTTCATGAACGACAGTTCCAATGTGGCCATGCTG
|
GCAGCGGCTCCTGCAGATGCAGGATGAAGGAAGAAGGCAGCCCAGACCTGGAGGCCGGG
|
GACCACATGGAGGCCCTGCGATCCTTTCTGGACCCTGGGAGCGGTGGCTACAGCTGGCA
|
GGGACGCACACGCTGATGTGGGGCACCTGGTGGGCACGCTGGGCCGAGTGACGGAGTGC
|
GCTGTCCTTGGACAAGCTGGAGGCGGCACCCTCAGAGGCAGCCCTGGTGTCGCGGGCCC
|
GTGCAACTGCTCGCGGAACATCGATTCTGGGCCGGCGTCGTCTTCTTGGGACCTGAGGA
|
GCTCTTCAGACCCCACAGAGCACCCAACCCCAGACCTGGGCCCCGGCCACGTGCGCATC
|
GAAAATCCGCATGGACATTGACGTGGTCACGAGGACCAATAAGATCAGGGACAGGTTTT
|
GGGGACCCTGGCCCAGCCGCGGACCCCCTGACCGACCTGCGCTACGTGTGGGGCGGCTT
|
GCGTGTACCTGCAAGACCTGGTGGAGCGTGCAGCCGTCCGCGTGCTCAGCGGCGCCAAC
|
GCCCCGGGCCGGCCTCTACCTGCAGCAGATGCCCTATCCGTGCTATGTGGACGACGTGT
|
GTCCTGCGTGTGCTGAGCCGGTCGCTGCCGCTCTTCCTGACGCTGGCCTGGATCTACTC
|
GCGTGACACTGACAGTGAAGGCCGTGGTGCGGGAGAAGGAGACGCGGCTGCGGGACACC
|
GATGCGCGCCATGGGGCTCAGCCGCGCGGTGCTCTGGCTAGGCTGGTTCCTCAGCTGCC
|
GTCGGGCCCTTCCTGCTCAGCGCCGCACTGCTGGTTCTGGTGCTCAAGCTGGGAGACAT
|
GCCTCCCCTACAGCCACCCGGGCGTGGTCTTCCTGTTCTTGGCAGCCTTCGCGGTGGCC
|
GACGGTGACCCAGAGCTTCCTGCTCAGCGCCTTCTTCTCCCGCGCCAACCTGGCTGCGG
|
GCCTGCGGCGGCCTGGCCTACTTCTCCCTCTACCTGCCCTACGTGCTGTGTGTGGCTTG
|
GGCGGGACCGGCTGCCCGCGGGTGGCCGCGTGGCCGCGAGCCTGCTGTCGCCCGTGGCC
|
GTTCGGCTTCGGCTGCGAGAGCCTGGCTCTGCTGGAGGAGCAGGGCGAGGGCGCGCAGT
|
GGGCACAACGTGGGCACCCGGCCTACGGCAGACGTCTTCAGCCTGGCCCAGGTCTCTGG
|
GCCTTCTGCTGCTGGACGCGGCGCTCTACGGCCTCGCCACCTGGTACCTGGAAGCTGTG
|
GTGCCCAGGCCAGTACGGGATCCCTGAACCATGGAATTTTCCTTTTCGGAGGAGCTACT
|
GGGTGCGGACCTCGGCCCCCCAAGAGTCCAGCCCCTTGCCCCACCCCGCTGGACCCAAA
|
GGGTGCTGGTAGAAGAGGCACCGCCCGGCCTGAGTCCTGGCGTCTCCGTTCGCAGCCTG
|
GGAGAAGCGCTTTCCTGGAAGCCCGCAGCCAGCCCTGCGGGGGCTCAGCCTGGACTTCT
|
GACCAGGGCCACATCACCGCCTTCCTGGGCCACAACGGGGCCGGCAAGACCACCACCCT
|
GGTCCATCTTGAGTGGCCTCTTCCCACCCAGTGGTGGCTCTGCCTTCATCCTGGGCCAC
|
GGACGTCCGCTCCAGCATGGCCGCCATCCGGCCCCACCTGGGCGTCTGTCCTCAGTACA
|
GACGTGCTGTTTGACATGCTGACCGTGGACGAGCACGTCTGGTTCTATGGGCGGCTGAA
|
GGGGTCTGAGTGCCGCTGTAGTGGGCCCCGAGCAGGACCGTCTGCTGCAGGATGTGGGG
|
GCTGGTCTCCAAGCACAGTGTGCAGACTCGCCACCTCTCTGGTGGGATGCAACGGAAGC
|
GTGTCCGTGGCCATTGCCTTTGTGGGCGGCTCCCAAGTTGTTATCCTGGACGAGCCTAC
|
GGGCTGGCGTGGATCCTGCTTCCCGCCGCGGTATTTGGGAGCTGCTGCTCAAATACCGA
|
GGAAGGTCGCACGCTGATCCTCTCCACCCACCACCTGGATGAGGCAGAGCTGCTGGGAG
|
GACCGTGTGGCTGTGGTGGCAGGTGGCCGCTTGTGCTGCTGTGGCTCCCCACTCTTCCT
|
GGCGCCGTCACCTGGGCTCCGGCTACTACCTGACGCTGGTGAAGGCCCGCCTGCCCCTG
|
GACCACCAATGAGAAGGCTGACACTGACATGGAGGGCAGTGTGGACACCAGGCAGGAAA
|
GAGAAGAATGGCAGCCAGGGCAGCAGAGTCGGCACTCCTCAGCTGCTGGCCCTGGTACA
|
GGCACTGGGTGCCCGGGGCACGGCTGGTGGAGGAGCTGCCACACGAGCTGGTGCTGGTG
|
GCTGCCCTACACGGGTGCCCATGACGGCAGCTTCGCCACACTCTTCCGAGAGCTAGACA
|
GCGCGGCTGGCGGAGCTGAGGCTCACTGGCTACGGGATCTCCGACACCAGCCTCGAGGA
|
GGATCTTCCTGAAGGTGGTGGAGGAGTGTGCTGCGGACACAGATATGGAGGATGGCAGC
|
GTGCGGGCAGCACCTATGCACAGGCATTGCTGGCCTAGACGTAACCCTGCGGCTCAAGA
|
GTGCCGCCACAGGAGACAGCGCTGGAGAACGGGGAACCAGCTGGGTCAGCCCCAGAGAC
|
GTGACCAGGGCTCTGGGCCAGACGCCGTGGGCCCGGTACAGGGCTGGGCACTGACCCGC
|
GCAGCAGCTCCAGGCCCTGCTTCTCAAGCGCTTTCTGCTTGCCCGCCGCAGCCGCCGCG
|
GGCCTGTTCGCCCAGATCGTGCTGCCTGCCCTCTTTGTGGGCCTGGCCCTCGTGTTCAG
|
GCCTCATCGTGCCTCCTTTCGGGCACTACCCGGCTCTGCGGCTCAGTCCCACCATGTAC
|
GGGTGCTCAGGTGTCCTTCTTCAGTGAGGACGCCCCAGGGGACCCTGGACGTGCCCGGC
|
GTGCTCGAGGCGCTGCTGCAGGAGGCAGGACTGGAGGAGCCCCCAGTGCAGCATAGCTC
|
GCCACAGGTTCTCGGCACCAGAAGTTCCTGCTGAAGTGGCCAAGGTCTTGGCCAGTGGC
|
GAACTGGACCCCAGAGTCTCCATCCCCAGCCTGCCAGTGTAGCCAGCCCGGTGCCCGGC
|
GGCCTGCTGCCCGACTGCCCGGCTGCAGCTGGTGGTCCCCCTCCGCCCCAGGCAGTGAC
|
GCGGCTCTGGGGAAGTGGTTCAGAACCTGACAGGCCGGAACCTGTCTGACTTCCTGGTC
|
GAAGACCTACCCGCGCCTGGTGCGCCAGGGCCTGAAGACTAAGAAGTGGGTGAATGAGG
|
GTCAGGTACGGAGGCTTCTCGCTGGGGGGCCGAGACCCAGGCCTGCCCTCTGGCCAAGA
|
GGTTGGGCCGCTCAGTGGAGGAGTTGTGGGCGCTGCTGAGTCCCCTGCCTGGCGGGGCC
|
GCTCGACCGTGTCCTGAAAAACCTCACAGCCTGGGCTCACAGCCTGGATGCTCAGGACA
|
GGTCTCAAGATCTGGTTCAACAACAAAGGCTGGCACTCCATGGTGACCTTTGTCAACCG
|
GAGCCAGCAACGCAATCCTCCGTGCTCACCTGCCCCCAGGCCCGGCCCGCCACGCCCAC
|
GAGCATCACCACACTCAACCACCCCTTGAACCTCACCAAGGAGCAGCTGTCTGAGGCTG
|
GCACTGATGGCCTCCTCGGTGGACGTCCTCGTCTCCATCTGTGTGGTCTTTGCCATGTC
|
GCTTTGTCCCGGCCAGCTTCACTCTTGTCCTCATTGAGGAGCGAGTCACCCGGGCCAAG
|
GCACCTGCAGCTCATGGGGGGCCTGTCCCCCACCCTCTACTGGCTTGGCAACTTTCTCT
|
GGGGACATGAAGCTGCAGGAGGTGAGCCGGATCTTGAAACAGGTCTTCCTTATCTTCCC
|
GCCACTTCTGCTTGGGCCGGGGGCTCATTGACATGGTGCGGAACCAGGCCATGGCTGAT
|
GGCCTTTGAGCGCTTGGGAGACAGGCAGTTCCAGTCACCCCTGCGCTGGGAGGTGGTCG
|
GGCAAGAACCTCTTGGCCATGGTGATACAGGGGCCCCTCTTCCTTCTCTTCACACTACT
|
GGCTGCAGCACGAAGCCAACTTCCTGCCACAGCCCAGGGTGAGGTCTCTGCCACTCCTG
|
GGGAGAGGAGGACGAGGATGTAGCCCGTGAACGGGAGCGGGTGGTCCAAGGAGCCACCC
|
GAGGGGGATGTGTTGGTGCTGAGGAACTTGACCAAGGTATACCGTGGGCAGAGGATGCC
|
GACCTGTTGACCGCTTGTGCCTGGGGATTCCCCCTGGTGAGTGTTTTGGGCTGCTGGGT
|
GGTGAATGGAGCAGGGAAGACGTCCACGTTTCGCATGGTGACGGGGGACACATTGGCCA
|
GGCAGGGGCGAGGCTGTGCTGGCAGGCCACAGCGTGGCCCGGGAACCCAGTGCTGCGCA
|
GCCTCAGCATGGGATACTGCCCTCAATCCGATGCCATCTTTGAGCTGCTGACGGGCCGC
|
GGAGCACCTGGAGCTGCTTGCGCGCCTGCGCGGTGTCCCGGAGGCCCAGGTTGCCCAGA
|
GCCGCTGGCTCGGGCCTGGCGCGTCTGGGACTCTCATGGTACGCAGACCGGCCTGCAGG
|
GCACCTACAGCGGAGGCAACAAACGCAAGCTGGCGACGGCCCTGGCGCTGGTTGGGGAC
|
GCCAGCCGTGGTGTTTCTGGACGAGCCGACCACAGGCATGGACCCCAGCGCGCGGCGCT
|
GTCCTTTGGAACAGCCTTTTGGCCGTGGTGCGGGAGGGCCGTTCAGTGATGCTCACCTC
|
GCCATAGCATGGAGGAGTGTGAAGCGCTCTGCTCGCGCCTAGCCATCATGGTGAATGGG
|
GCGGTTCCGCTGCCTGGGCAGCCCGCAACATCTCAAGGGCAGATTCGCGGCGGGTCACA
|
GCACTGACCCTGCGGGTGCCCGCCGCAAGGTCCCAGCCGGCAGCGGCCTTCGTGGCGGC
|
GCGAGTTCCCTGGGTCGGAGCTGCGCGAGGCACATGGAGGCCGCCTGCGCTTCCAGCTG
|
GCCGCCGGGAGGGCGCTGCGCCCTGGCGCGCGTCTTTGGAGAGCTGGCGGTGCACGGCG
|
GCAGAGCACGGCGTGGAGGACTTTTCCGTGAGCCAGACGATGCTGGAGGAGGTATTCTT
|
GGTACTTCTCCAAGGACCAGGGGAAGGACGAGGACACCGAAGAGCAGAAGGAGGCAGGA
|
GGTGGGAGTGGACCCCGCGCCAGGCCTGCAGCACCCCAAACGCGTCAGCCAGTTCCTCG
|
GATGACCCTAGCACTGCCGAGACTGTGCTCTGAGCCTCCCTCCCCTGCGGGGCCGCGGG
|
GGAGGCCCTGGGAATGGCAAGGGCAAGGTAGAGTGCCTAGGAGCCCTGGACTCAGGCTG
|
GGCAGAGGGGCTGGTGCCCTGGAGAAAATAAAGAGAAGGCTGGAGAGAAGCCGTGCTTG
|
GGTGAA
|
ORF Start: ATG at 1ORF Stop: TGA at 6178
SEQ ID NO: 1762059 aaMW at 224783.2kD
NOV68a,MAFWTQLMLLLWKNFMYRRRQPVQLLVELLWPLFLFFILVAVRHSHPPLEHHECHFPN
CG93529-01 Protein Sequence
KPLPSAGTVPWLQGLICNVNNTCFPQLTPGEEPGRLSNFNDSLVSRLLADARTVLGGA
|
SAHRTLAGLGKLIATLRAARSTAQPQPTKQSPLEPPMLDVAELLTSLLRTESLGLALG
|
QAQEPLHSLLEAAEDLAQELLALRSLVELRALLQRPRGTSGPLELLSEALCSVRGPSS
|
TVGPSLNWYEASDLMELVGQEPESALPDSSLSPACSELIGALDSHPLSRLLWRRLKPL
|
ILGKLLFAPDTPFTRKLMAQVNRTFEELTLLRDVREVWEMLGPRIFTFMNDSSNVAML
|
QRLLQNQDEGRRQPRPGGRDHMEALRSFLDPGSGGYSWQDAHADVGHLVGTLGRVTEC
|
LSLDKLEAAPSEAALVSRALQLLAEHRFWAGVVFLGPEDSSDPTEHPTPDLGPGHVRI
|
KIRMDIDVVTRTNKIRDRFWDPGPAADPLTDLRYVWGGFVYLQDLVERAAVRVLSGAN
|
PRAGLYLQQMPYPCYVDDVFLRVLSRSLPLFLTLAWIYSVTLTVKAVVREKETRLRDT
|
MRAMGLSRAVLWLGWFLSCLGPFLLSAALLVLVLKLGDILPYSHPGVVFLFLAAFAVA
|
TVTQSFLLSAFFSRANLAAACGGLAYFSLYLPYVLCVAWRDRLPAGGRVAASLLSPVA
|
FGFGCESLALLEEQGEGAQWHNVGTRPTADVFSLAQVSGLLLLDAALYGLATWYLEAV
|
CPGQYGIPEPWNFPFRRSYWCGPRPPKSPAPCPTPLDPKVLVEEAPPGLSPGVSVRSL
|
EKRFPGSPQPALRGLSLDFYQGHITAFLGHNGAGKTTTLSILSGLFPPSGGSAFILGH
|
DVRSSMAAIRPHLGVCPQYNVLFDMLTVDEHVWFYGRLKGLSAAVVGPEQDRLLQDVG
|
LVSKQSVQTRHLSGGMQRKLSVAIAFVGGSQVVILDEPTAGVDPASRRGIWELLLKYR
|
EGRTLILSTHHLDEAELLGDRVAVVAGGRLCCCGSPLFLRRHLGSGYYLTLVKARLPL
|
TTNEKADTDMEGSVDTRQEKKNGSQGSRVGTPQLLALVQHWVPGARLVEELPHELVLV
|
LPYTGAHDGSFATLFRELDTRLAELRLTGYGISDTSLEEIFLKVVEECAADTDMEDGS
|
CGQHLCTGIAGLDVTLRLKMPPQETALENGEPAGSAPETDQGSGPDAVGRVQGWALTR
|
QQLQALLLKRFLLARRSRRGLFAQIVLPALFVGLALVFSLIVPPFGHYPALRLSPTMY
|
NWTPESPSPACQCSQPGARRLLPDCPAAAGGPPPPQAVTGSGEVVQNLTGRNLSDFLV
|
KTYPRLVRQGLKTKKWVNEVRYGGFSLGGRDPGLPSGQELGRSVEELWALLSPLPGGA
|
LDRVLKNLTAWAHSLDAQDSLKIWFNNKGWHSMVTFVNRASNAILRAHLPPGPARHAH
|
SITTLNHPLNLTKEQLSEAALMASSVDVLVSICVVFAMSFVPASFTLVLIEERVTRAK
|
HLQLMGGLSPTLYWLGNFLWDMKLQEVSRILKQVFLIFPHFCLGRGLIDMVRNQAMAD
|
AFERLGDRQFQSPLRWEVVGKNLLAMVIQGPLFLLFTLLLQHRSQLLPQPRVRSLPLL
|
GEEDEDVARERERVVQGATQGDVLVLRNLTKVYRGQRMPAVDRLCLGIPPGECFGLLG
|
VNGAGKTSTFRNVTGDTLASRGEAVLAGHSVAREPSAAHLSMGYCPQSDAIFELLTGR
|
EHLELLARLRGVPEAQVAQTAGSGLARLGLSWYADRPAGTYSGGNKRKLATALALVGD
|
PAVVFLDEPTTGMDPSARRFLWNSLLAVVREGRSVMLTSHSMEECEALCSRLAIMVNG
|
RFRCLGSPQHLKGRFAAGHTLTLRVPAARSQPAAAFVAAEFPGSELREAHGGRLRFQL
|
PPGGRCALARVFGELAVHGAEHGVEDFSVSQTMLEEVFLYFSKDQGKDEDTEEQKEAG
|
VGVDPAPGLQHPKRVSQFLDDPSTAETVL
|
[0669] Further analysis of the NOV68a protein yielded the following properties shown in Table 68B.
356TABLE 68B
|
|
Protein Sequence Properties NOV68a
|
|
PSort0.8000 probability located in plasma membrane; 0.7548
analysis:probability located in mitochondrial inner membrane; 0.6258
probability located in mitochondrial intermembrane space;
0.5172 probability located in mitochondrial matrix space
SignalPCleavage site between residues 46 and 47
analysis:
|
[0670] A search of the NOV68a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 68C.
357TABLE 68C
|
|
Geneseq Results for NOV68a
NOV68aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAU04484Human PD-ATP-binding cassette 1 . . . 15881585/1588 (99%)0.0
(PD-ABC) protein form #2 - Homo 1 . . . 15881586/1588 (99%)
sapiens, 1873 aa. [WO200153490-
A1, 26-JUL-2001]
AAU04483Human PD-ATP-binding cassette 1 . . . 15881585/1588 (99%)0.0
(PD-ABC) protein form #1 - Homo 1 . . . 15881586/1588 (99%)
sapiens, 2146 aa. [WO200153490-
A1, 26-JUL-2001]
AAB38114Human ABC1 cholesterol transporter158 . . . 1588 732/1482 (49%)0.0
mutant, E1172D - Homo sapiens,234 . . . 1707 982/1482 (65%)
2261 aa. [WO200055318-A2, 21-SEP-2000]
AAB38111Human ABC1 cholesterol transporter158 . . . 1588 732/1482 (49%)0.0
mutant, V771M - Homo sapiens,234 . . . 1707 982/1482 (65%)
2261 aa. [WO200055318-A2, 21-SEP-2000]
AAB31365Amino acid sequence of a human158 . . . 1588 732/1482 (49%)0.0
ABC1 polypeptide - Homo sapiens,234 . . . 1707 982/1482 (65%)
2261 aa. [WO200078971-A2, 28-DEC-2000]
|
[0671] In a BLAST search of public sequence datbases, the NOV68a protein was found to have homology to the proteins shown in the BLASTP data in Table 68D.
358TABLE 68D
|
|
Public BLASTP Results for NOV68a
NOV68aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9BZC4ABC TRANSPORTER MEMBER 1 . . . 15881585/1588 (99%)0.0
7 - Homo sapiens (Human), 2146 1 . . . 15881586/1588 (99%)
aa.
Q9NR73MACROPHAGE ABC 1 . . . 15881585/1588 (99%)0.0
TRANSPORTER - Homo sapiens 1 . . . 15881585/1588 (99%)
(Human), 2146 aa.
Q96S58ABCA-SSN - Homo sapiens167 . . . 15881419/1422 (99%)0.0
(Human), 2008 aa. 29 . . . 14501419/1422 (99%)
Q91V24ATP-BINDING CASSETTE 1 . . . 15881200/1625 (73%)0.0
TRANSPORTER SUB-FAMILY A 1 . . . 16021334/1625 (81%)
MEMBER 7 - Mus musculus
(Mouse), 2159 aa.
AAL56247ATP-BINDTNG CASSETTE136 . . . 1588 740/1508 (49%)0.0
TRANSPORTER 1 - Gallus gallus210 . . . 1706 995/1508 (65%)
(Chicken), 2260 aa.
|
[0672] PFam analysis predicts that the NOV68a protein contains the domains shown in the Table 68E.
359TABLE 68E
|
|
Domain Analysis of NOV68a
Identities/
Similarities
NOV68a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Ca_channel_B:224 . . . 247 12/25 (48%)1.3
domain 1 of 1 15/25 (60%)
photoRC: domain 1 of 1591 . . . 608 9/20 (45%)0.31
15/20 (75%)
SRP54: domain 1 of 2832 . . . 851 7/20 (35%)0.28
15/20 (75%)
ABC_tran: 834 . . . 1014 73/199 (37%)2.3e−58
domain 1 of 2139/199 (70%)
Pterin_4a: domain 1 of 11440 . . . 1539 25/111 (23%)1.1
46/111 (41%)
SRP54: domain 2 of 21738 . . . 1749 7/12 (58%)13
11/12 (92%)
ABC_tran:1733 . . . 1914 61/199 (31%)2.4e−39
domain 2 of 2132/199 (66%)
|
Example 69
[0673] The NOV69 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 69A.
360TABLE 69A
|
|
NOV69 Sequence Analysis
|
|
SEQ ID NO: 1771702 bp
NOV69a,TAATCTGGCTCAATTTCTGACACAAGAACAATATGCAGCTGAGATGAGTAAAGCTATT
CG93594-01 DNA Sequence
GCTTTTGAGATCATTCAGAAATACGAGCCTATCGAAGAAGTTAGGAAAGCACACCAAA
|
TGTCATTAGAAGGTTTTACAAGATACATGGATTCACGTGAATGTCTACTGTTTAAAAA
|
TGAATGTAGAAAAGTTTATCAAGATATGACTCATCCATTAAATGATTATTTTATTTCA
|
TCTTCACATAACACATATTTGGTATCTGATCAATTATTGGGACCAAGTGACCTTTGGG
|
GATATGTAAGTGCCCTTGTGAAAGGATGCCGTTGTTTGGAGATTGACTGCTGGGATGG
|
AGCACAAAATGAACCTGTTGTATATCATGGCTACACACTCACAAGCAAACTTCTGTTT
|
AAAACTGTTATCCAAGCTATACACAAGTATGCATTCATGACATCTGACTACCCAGTGG
|
TGCTCTCTTTAGAAAATCACTGCTCCACTGCCCAACAAGAAGTAATGGCAGACAATTT
|
GCAGGCTACTTTTGGAGAGTCCTTGCTTTCTGATATGCTTGATGATTTTCCTGATACT
|
CTACCATCACCAGAGGCACTAAAATTCAAAATATTAGTTAAAAATAAGAAAATAGGAA
|
CCTTAAAGGAAACCCATGAAAGAAAAGGTTCTGATAAGCGTGGTAAGGTGGAGGAATG
|
GGAAGAAGAAGTGGCAGATGGAGAGGAGGAGGAGGAGGAGGAGGAGGAGGAGGAGGAG
|
GAGGAGGAGGATAAATTCAAAGAATCAGAAGTATTGGAATCTGTTTTAGGAGACAATC
|
AAGACAAGGAAACAGGGGTAAAAAGGAAGCTAAAAATTGCTCTGTCCTTATCTGATCT
|
TGTCATTTATACGAAAGCTGAGAAATTCAAAAGCTTTCAACATTCAAGATTATATCAG
|
CAATTTAATGAAAATAATTCTATTGGGGAGACACAAGCCCGAAAACTTTCAAAATTGC
|
GAGTCCATGAGTTTATTTTTCACACCAGGAAGTTCATTACCAGAATATATCCCAAAGC
|
AACAAGAGCAGACTCTTCTAATTTTAATCCCCAAGAATTTTGGAATATAGGTTGTCAA
|
ATGGTGGCTTTAAATTTCCAGACCCCTGGTCTGCCCATGGATCTGCAAAATGGGAAAT
|
TTTTGGATAATGGTGGTTCTGGATATATTTTGAAACCACATTTCTTAAGAGAGAGTAA
|
ATCATACTTTAACCCAAGTAACATAAAAGAGGGTATGCCAATTACACTTACAATAAGG
|
CTCATCAGTGGTATCCAGTTGCCTCTTACTCATTCATCATCTAACAAAGGTGATTCAT
|
TAGTAATTATAGAAGTTTTTGGTGTTCCAAATGATCAAATGAAGCAGCAGACTCGTGT
|
AATTAAAAAAAATGCTTTTAGTCCAAGATGGAATGAAACATTCACATTTATTATTCAT
|
GTCCCAGAATTGGCATTGATACGTTTTGTTGTTGAAGGTCAAGGTTTAATAGCAGGAA
|
ATGAATTTCTTGGGCAATATACTTTGCCACTTCTATGCATGAACAAAGGTTATCGTCG
|
TATTCCTCTGTTTTCCAGAATGGGTGAGAGCCTTGAGCCTGCTTCACTGTTTGTTTAT
|
GTTTGGTACGTCAGATAACAGCTAATGATAAATGACATATCATTAGCTATGCATCGCA
|
ATAAAACAGCCAAAATGAAT
|
ORE Start: ATG at 44JRF Stop: TAA at 1640
SEQ ID NO: 178532 aaMW at 61203.1kD
NOV69a,MSKAIAFEIIQKYEPIEEVRKAHQMSLEGFTRYMDSRECLLFKNECRKVYQDMTHPLN
CG93594-01 Protein Sequence
DYFISSSHNTYLVSDQLLGPSDLWGYVSALVKGCRCLEIDCWDGAQNEPVVYHGYTLT
|
SKLLFKTVIQAIHKYAFMTSDYPVVLSLENHCSTAQQEVMADNLQATFGESLLSDMLD
|
DFPDTLPSPEALKFKILVKNKKIGTLKETHERKGSDKRGKVEEWEEEVADGEEEEEEE
|
EEEEEEEEDKFKESEVLESVLGDNQDKETGVKRKLKIALALSDLVIYTKAEKFKSFQH
|
SRLYQQFNENNSIGETQARKLSKLRVHEFIFHTRKFITRIYPKATRADSSNFNPQEFW
|
NIGCQMVALNFQTPGLPMDLQNGKFLDNGGSGYILKPHFLRESKSYFNPSNIKEGMPI
|
TLTIRLISGIQLPLTHSSSNKGDSLVIIEVFGVPNDQMKQQTRVIKKNAFSPRWNETF
|
TFIIHVPELALIRFVVEGQGLIAGNEFLGQYTLPLLCMNKGYRRIPLFSRMGESLEPA
|
SLFVYVWYVR
|
[0674] Further analysis of the NOV69a protein yielded the following properties shown in
361TABLE 69B
|
|
Protein Sequence Properties NOV69a
|
|
PSort0.4500 probability located in cytoplasm; 0.4223
analysis:probability located in microbody (peroxisome); 0.1000
probability located in mitochondrial matrix space;
0.1000 probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0675] A search of the NOV69a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 69C.
362TABLE 69C
|
|
Geneseq Results for NOV69a
NOV69aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAM95867Human reproductive system related279 . . . 532253/254 (99%)e−147
antigen SEQ ID NO: 4525 - Homo 1 . . . 254253/254 (99%)
sapiens, 254 aa. [WO200155320-A2,
02-AUG-2001]
AAU22938Novel human enzyme polypeptide279 . . . 532253/254 (99%)e−147
#24 - Homo sapiens, 254 aa. 1 . . . 254253/254 (99%)
[WO200155301-A2, 02-AUG-2001]
AAE14268Human phospholipase C delta 5 5 . . . 528249/528 (47%)e−133
(PLCD5) protein #1 - Homo sapiens,244 . . . 751335/528 (63%)
762 aa. [WO200183771-A2, 08-NOV-2001]
AAE10440Novel human phospholipase protein 5 . . . 528249/528 (47%)e−133
#7 - Homo sapiens, 762 aa.244 . . . 751335/528 (63%)
[WO200168871-A2, 20-SEP-2001]
AAE14270Human phospholipase C delta 5 5 . . . 528247/529 (46%)e−129
(PLCD5) protein #3 - Homo sapiens,241 . . . 748333/529 (62%)
759 aa. [WO200183771-A2, 08-NOV-2001]
|
[0676] In a BLAST search of public sequence datbases, the NOV69a protein was found to have homology to the proteins shown in the BLASTP data in Table 69D.
363TABLE 69D
|
|
Public BLASTP Results for NOV69a
NOV69aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q95JS1HYPOTHETICAL 74.6 KDA 1 . . . 532503/545 (92%)0.0
PROTEIN - Macaca fascicularis (Crab105 . . . 641513/545 (93%)
eating macaque) (Cynomolgus
monkey), 641 aa.
Q95JS0HYPOTHETICAL 74.4 KDA 1 . . . 532501/545 (91%)0.0
PROTEIN - Macaca fascicularis (Crab105 . . . 640512/545 (93%)
eating macaque) (Cynomolgus
monkey), 640 aa.
Q96J70TESTIS-DEVELOPMENT RELATED 1 . . . 532489/545 (89%)0.0
NYD-SP27 - Homo sapiens (Human), 1 . . . 504490/545 (89%)
504 aa.
Q9D9N4ADULT MALE TESTIS CDNA, 8 . . . 532388/533 (72%)0.0
RIKEN FULL-LENGTH ENRICHED 10 . . . 536452/533 (84%)
LIBRARY, CLONE: 1700041H07,
FULL INSERT SEQUENCE - Mus
musculus (Mouse), 537 aa.
Q9BRC7HYPOTHETICAL 87.6 KDA 5 . . . 528249/528 (47%)e−133
PROTEIN - Homo sapiens (Human),244 . . . 751335/528 (63%)
762 aa.
|
[0677] PFam analysis predicts that the NOV69a protein contains the domains shown in the Table 69E.
364TABLE 69E
|
|
Domain Analysis of NOV69a
Identities/
Similarities
NOV69a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
PI-PLC-X: domain 1 of 1 52 . . . 19678/153 (51%) 3.9e−64
115/153 (75%)
UvrD-helicase:187 . . . 24011/64 (17%)9.5
domain 1 of 140/64 (62%)
PI-PLC-Y: domain 1 of 1272 . . . 38963/128 (49%) 1.6e−50
89/128 (70%)
C2: domain 1 of 1408 . . . 49633/97 (34%)4.9e−20
73/97 (75%)
|
Example 70
[0678] The NOV70 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 70A.
365TABLE 70A
|
|
NOV70 Sequence Analysis
|
|
SEQ ID NO:1792257 bp
NOV70a,CCGCAAGTCCCTCGCCGCCTTGGGGTCTGGGCGCGCGGTCCGTGGGGGTCAGCAGGGC
CG93669-01 DNA Sequence
GAGCGGCTTTTCCAGGAGAAAGGGCCCTCACGGGTGAGCGGGGCGACTGGGCTCCCCC
|
GCGGTGCAGTTGCCCCGCGCGACCGGCCCCGGCTTCAACGGATTCTTCTCGCTCGCTG
|
CCCGGAAAGAACCATTTGGGAGAGCCCATGGTGACTGCGTGAGTGGAGCCCAGCTGTG
|
TGGATGCCCCAGCATGGATGACTACATGGTCCTGAGAATGATTGGGGAGGGCTCCTTC
|
GGCAGAGCTCTTTTGGTTCAGCATGAAAGCAGTAATCAGATGTTTGCCATGAAAGAAA
|
TAAGGCTTCCCAAGGTCACTACTAATACACAGAATTCTAGGAAGGAGGCTGTTCTTTT
|
AGCCAAAATGAAACACCCTAATATTGTTGCCTTCAAAGAATCATTTGAAGCTGAAGGA
|
CACTTCTATATTGTGATGGAATACTGTGATGGAGGGGATCTAATGCAAAAGATTAAAC
|
AGCAGAAAGGAAAGTTATTTCCTGAAGACCAGATACTTAATTGGTTTACCCAAATGTG
|
CCTTGGAGTAAATCACATTCACAAGAAACGTGTGCTACACAGAGATATCAAGTCCCAG
|
AATATCTTCCTCACTCAGAATGGAAAAGTGAAATTGGGAGACTTTGGATCTGCCCGTC
|
TTCTCTCCAATCCGATGGCATTTGCTTGTACCTATGTGGGAACTCCTTATTATGTGCC
|
TCCAGAAATTTGGGAAAACCTGCCTTATAACAATAAAAGTGACATCTGGTCCTTGGGT
|
TGCATCCTGTATGAACTCTGTACCCTTAAGCATCCATTTCAGGCAAATAGTTGGAAAA
|
ATCTTATCCTCAAAGTATGTCAAGGGTGCATCAGTCCACTGCCGTCTCATTACTCCTA
|
TGAACTTCAGTTCCTAGTCAAGCAGATGTTTAAAAGGAATCCCTCACATCGCCCCTCG
|
GCTACAACGCTTCTCTCTCGAGGCATCGTAGCTCGGCTTGTCCAGAAGTGCTTACCCC
|
CCGAGATCATCATGGAATATGGTGAGGAAGTATTAGAAGAAATAAAAAATTCGAAGCA
|
TAACACACCAAGAAAAAAAACAAACCCCAGCAGAATCAGGATAGCTTTGGGAAATGAA
|
GCAAGCACAGTGCAAGAGGAAGAACAAGATAGAAAGGGTAGCCATACTGATTTGGAAA
|
GCATTAATGAAAATTTAGTTGAAAGTGCATTGAGAAGAGTAAACAGAGAAGAAAAAGG
|
TAATAAGTCAGTCCATCTGAGGAAAGCCAGTTCACCAAATCTTCATAGACGACAGTGG
|
GAGAAAAATGTACCCAATACAGCTCTTACAGCTTTGGAAAATGCATCCATACTCACCT
|
CCAGTTTAACAGCAGAGGACGATAGAGGTGGTTCTGTAATAAAGTACAGCAAAAATAC
|
TACTCGTAAGCAGTGGCTCAAAGAGACCCCTGACACTTTGTTGAACATCCTTAAGAAT
|
GCTGATCTCAGCTTGGCTTTTCAAACATACACAATATATAGACCAGGTTCAGAAGGGT
|
TCTTGAAAGGCCCCCTGTCTGAAGAAACAGAAGCATCGGACAGTGTTGATGGAGGTCA
|
CGATTCTGTCATTTTGGATCCAGAGCGACTTGAGCCTGGGCTAGATGAGGAGGACACG
|
GACTTTGAGGAGGAAGATGACAACCCCGACTGGGTGTCAGAGCTGAAGAAGCGAGCTG
|
GATGGCAAGGCCTGTGCGACAGATAATGCCTGAGGAAATGTTCCTGAGTCACGCTGAG
|
GAGAGCCTTCACTCAGGAGTTCATGCTGAGATGATCATGAGTTCATGCGACGTATATT
|
TTCCTTTGGAAACAGAATGAAGCAGAGGAAACTCTTAATACTTAAAATCGTTCTTGAT
|
TAGTATCGTGAGTTTGAAAAGTCTAGAACTCCTGTAAGTTTTTGAACTCAAGGGAGAA
|
GGTATAGTGGAATGAGTGTGAGCATCGGGCTTTGCAGTCCCATAGAACAGAAATGGGA
|
TGCTAGCGTGCCACTACCTACTTGTGTGATTGTGGGAAATTACTTAACCTCTTCAAGC
|
CCCAATTTCCTCAACCATAAAATGAAGATAATAATGCCTACCTCAGAGGGATGCTGAC
|
CACAGACCTTTATAGCAGCCCGTATGATATTATTCACATTATGATATGTGTTTATTAT
|
TATGTGACTCTTTTTACATTTCCTAAAGGTTTGAGAATTAAATATATTTAATT
|
ORF Start: ATG at 246ORF Stop: TAA at 1764
SEQ ID NO:180506 aaMW at 57681.0 kD
NOV70a,MDDYMVLRMIGEGSFGRALLVQHESSNQMFAMKEIRLPKVTTNTQNSRKEAVLLAKMK
CG93669-01 Protein Sequence
HPNIVAFKESFEAEGHLYIVMEYCDGGDLMQKIKQQKGKLFPEDQILNWFTQMCLGVN
|
HIHKKRVLHRDIKSQNIFLTQNGKVKLGDFGSARLLSNPMAFACTYVGTPYYVPPEIW
|
ENLPYNNKSDIWSLGCILYELCTLKHPFQANSWKNLILKVCQGCISPLPSHYSYELQF
|
LVKQMFKRNPSHRPSATTLLSRGIVARLVQKCLPPEIIMEYGEEVLEEIKNSKHNTPR
|
KKTNPSRIRIALGNEASTVQEEEQDRKGSHTDLESINENLVESALRRVNREEKGNKSV
|
HLRKASSPNLHRRQWEKNVPNTALTALENASILTSSLTAEDDRGGSVIKYSKNTTRKQ
|
WLKETPDTLLNILKNADLSLAFQTYTIYRPGSEGFLKGPLSEETEASDSVDGGHDSVI
|
LDPERLEPGLDEEDTDFEEEDDNPDWVSELKKRAGWQGLCDR
|
SEQ ID NO:1811781 bp
NOV70b,CCGCAAGTCCCTCGCCGCCTTGGGGTCTGGGCGCGCGGTCCGTGGGGGTCAGCAGGGC
CG93669-02 DNA Sequence
GAGCGGCTTTTCCAGGAGAAAGGGCCCTCACGGGTGAGCGGGGCGACTGGGCTCCCCC
|
GCGGTGCAGTTGCCCCGCGCGACCGGCCCCGGCTTCAACGGATTCTTCTCGCTCGCTG
|
CCCGGAAAGAACCATTTGGGAGAGCCCATGGTGACTGCGTGAGTGGAGCCCAGCTGTG
|
TGGATGCCCCAGCATGGATGACTACATGGTCCTGAGAATGATTGGGGAGGGCTCCTTC
|
GGCAGAGCTCTTTTGGTTCAGCATGAAAGCAGTAATCAGATGTTTGCCATGAAAGAAA
|
TAAGGCTTCCCAAGGTCACTACTAATACACAGAATTCTAGGAAGGAGGCTGTTCTTTT
|
AGCCAAAATGAAACACCCTAATATTGTTGCCTTCAAAGAATCATTTGAAGCTGAAGGA
|
CACTTGTATATTGTGATGGAATACTGTGATGGAGGGGATCTAATGCAAAAGATTAAAC
|
AGCAGAAAGGAAAGTTATTTCCTGAAGACCAGATACTTAATTGGTTTACCCAAATGTG
|
CCTTGGAGTAAATCACATTCACAAGAAACGTGTGCTACACAGAGATATCAAGTCCCAG
|
AATATCTTCCTCACTCAGAATGGAAAAGTGAAATTGGGAGACTTTGGATCTGCCCGTC
|
TTCTCTCCAATCCGATGGCATTTGCTTGTACCTATGTGGGAACTCCTTATTATGTGCC
|
TCCAGAAATTTGGGAAAACCTGCCTTATAACAATAAAAGTGACATCTGGTCCTTGGGT
|
TGCATCCTGTATGAACTCTGTACCCTTAAGCATCCATTTCAGGCAAATAGTTGGAAAA
|
ATCTTATCCTCAAAGTATGTCAAGGGTGCATCAGTCCACTGCCGTCTCATTACTCCTA
|
TGAACTTCAGTTCCTAGTCAAGCAGATGTTTAAAAGGAATCCCTCACATCGCCCCTCG
|
GCTACAACGCTTCTCTCTCGAGGCATCGTAGCTCGGCTTGTCCAGAAGTGCTTACCCC
|
CCGAGATCATCATGGAATATGGTGAGGAAGTATTAGAAGAAATAAAAAATTCGAAGCA
|
TAACACACCAAGAAAAAAACAAGAGGAAGAACAAGATAGAAAGGGTAGCCATACTGAT
|
TTGGAAAGCATTAATGAAAATTTAGTTGAAAGTGCATTGAGAAGAGTAAACAGAGAAG
|
AAAAAGGTAATAAGTCAGTCCATCTGAGGAAAGCCAGTTCACCAAATCTTCATAGACG
|
ACAGTGGGAGAAAAATGTACCCAATACAGCTCTTACAGCTTTGGAAAATGCATCCATA
|
CTCACCTCCAGTTTAACAGCAGAGGACGATAGAGGTGGTTCTGTAATAAAGTACAGCA
|
AAAATACTACTCGTAAGCAGTGGCTCAAAGAGACCCCTGACACTTTGTTGAACATCCT
|
TAAGAATGCTGATCTCAGCTTGGCTTTTCAAACATACACAATATATAGACCAGGTTCA
|
GAAGGGTTCTTGAAAGGCCCCCTGTCTGAAGAAACAGAAGCATCGGACAGTGTTGATG
|
GAGGTCACGATTCTGTCATTTTGGATCCAGAGCGACTTGAGCCTGGGCTAGATGAGGA
|
GGACACGGACTTTGAGGAGGAAGATGACAACCCCGACTGGGTGTCAGAGCTGAAGAAG
|
CGAGCTGGATGGCAAGGCCTGTGCGACAGATAATGCCTGAGGAAATGTTCCTGAGTCA
|
CGCTGAGGAGAGGCTTCACTCTAGGAGTTCATGCTGAGATG
|
ORF Start: ATG at 246ORF Stop: TAA at 1713
SEQ ID NO:182489 aaMW at 55900.0 kD
NOV70b,MDDYMVLRMIGEGSFGRALLVQHESSNQMFAMKEIRLPKVTTNTQNSRKEAVLLAKMK
CG93669-02 Protein Sequence
HPNIVAFKESFEAEGHLYIVMEYCDGGDLMQKIKQQKGKLFPEDQILNWFTQMCLGVN
|
HIHKKRVLHRDIKSQNIFLTQNGKVKLGDFGSARLLSNPMAFACTYVGTPYYVPPEIW
|
ENLPYNNKSDIWSLGCILYELCTLKHPFQANSWKNLILKVCQGCISPLPSHYSYELQF
|
LVKQMFKRNPSHRPSATTLLSRGIVARLVQKCLPPEIIMEYGEEVLEEIKNSKHNTPR
|
KKQEEEQDRKGSHTDLESINENLVESALRRVNREEKGNKSVHLRKASSPNLHRRQWEK
|
NVPNTALTALENASILTSSLTAEDDRGGSVIKYSKNTTRKQWLKETPDTLLNILKNAD
|
LSLAFQTYTIYRPGSEGFLKGPLSEETEASDSVDGGHDSVILDPERLEPGLDEEDTDF
|
EEEDDNPDWVSELKKRAGWQGLCDR
|
SEQ ID NO:1831588 bp
NOV70c,CCGCAAGTCCCTCGCCGCCTTGGGGTCTGGGCGCGCGGTCCGTGGGGGTCAGCAGGGC
CG93669-03 DNA Sequence
GAGCGGCTTTTCCAGGAGAAAGGGCCCTCACGGGTGAGCGGGGCGACTGGGCTCCCCC
|
GCGGTGCAGTTGCCCCGCGCGACCGGCCCCGGCTTCAACGGATTCTTCTCGCTCGCTG
|
CCCGGAAAGAACCATTTGGGAGAGCCCATGGTGACTGCGTGAGTGGAGCCCAGCTGTG
|
TGGATGCCCCAGCATGGATGACTACATGGTCCTGAGAATGATTGGGGAGGGCTCCTTC
|
GGCAGAGCTCTTTTGGTTCAGCATGAAAGCAGTAATCAGATGTTTGCCATGAAAGAAA
|
TAAGGCTTCCCAAGGTCACTACTAATACACAGAATTCTAGGAAGGAGGCTGTTCTTTT
|
AGCCAAAATGAAACACCCTAATATTGTTGCCTTCAAAGAATCATTTGAAGCTGAAGGA
|
CACTTGTATATTGTGATGGAATACTGTGATGGAGGGGATCTAATGCAAAAGATTAAAC
|
AGCAGAAAGGAAAGTTATTTCCTGAAGACCAGATACTTAATTGGTTTACCCAAATGTG
|
CCTTGGAGTAAATCACATTCACAAGAAACGTGTGCTACACAGAGATATCAAGTCCCAG
|
AATATCTTCCTCACTCAGAATGGAAAAGTGAAATTGGGAGACTTTGGATCTGCCCGTC
|
TCCTCTCCAATCCGATGGCATTTGCTTGTACCTATGTGGGAACTCCTTATTATGTGCC
|
TCCAGAAATTTGGGAAAACCTGCCTTATAACAATAAAAGTGACATCTGGTCCTTGGGT
|
TGCATCCTGTATGAACTCTGTACCCTTAAGCATCCATTTCAGGCAAATAGTTGGAAAA
|
ATCTTATCCTCAAAGTATGTCAAGGGTGCATCAGTCCACTGCCGTCTCATTACTCCTA
|
TGAACTTCAGTTCCTAGTCAAGCAGATGTTTAAAAGGAATCCCTCACATCGCCCCTCG
|
GCTACAGCGCTTCTCTCTCGAGGCATCGTAGCTCGGCTTGTCCAGAAGTGCTTACCCC
|
CCGAGATCATCATGGAATATGGTGAGGAAGTATTAGAAGAAATAAAAAATTCGAAGCA
|
TAACACACCAAGAAAAAAACAAGAGGAAGAACAAGATAGAAAGGGTAGCCATACTGAT
|
TTGGAAAGCATTAATGAAAATTTAGTTGAAAGTGCATTGAGAAGAGTAAACAGAGAAG
|
AAAAAGGTAATAAGTCAGTCCATCTGAGGAAAGCCAGTTCACCAAATCTTCATAGACG
|
ACAGTGGGAGAAAAATGTACCCAATACAGCTCTTACAGCTTTGGAAAATGCATCCATA
|
CTCACCTCCAGTTTAACAGCAGAGGACGATAGAGGTTCAGAAGGGTTCTTGAAAGGCC
|
CCCTGTCTGAAGAAACAGAAGCATCGGACAGTGTTGAGGAGGACACGGACTTTGAGGA
|
GGAAGATGACAACCCCGACTGGGTGTCAGAGCTGAAGAAGCGAGCTGGATGGCAAGGC
|
CTGTGCGACAGATAATGCCTGAGGAAATGTACCTGAGTCACGCTGAGGAGAGGCTTCA
|
CTCAGGAGTTCATGCTGAGATG
|
ORF Start: ATG at 246ORF Stop: TAA at 1521
SEQ ID NO:184425 aaMW at 48684.0 kD
NOV70c,MDDYMVLRMIGEGSFGRALLVQHESSNQMFAMKEIRLPKVTTNTQNSRKEAVLLAKMK
CG93669-03 Protein Sequence
HPNIVAFKESFEAEGHLYIVMEYCDGGDLMQKIKQQKGKLFPEDQILNWFTQMCLGVN
|
HIHKKRVLHRDIKSQNIFLTQNGKVKLGDFGSARLLSNPMAFACTYVGTPYYVPPEIW
|
ENLPYNNKSDIWSLGCILYELCTLKHPFQANSWKNLILKVCQGCISPLPSHYSYELQF
|
LVKQMFKRNPSHRPSATALLSRGIVARLVQKCLPPEIIMEYGEEVLEEIKNSKHNTPR
|
KKQEEEQDRKGSHTDLESINENLVESALRRVNREEKGNKSVHLRKASSPNLHRRQWEK
|
NVPNTALTALENASILTSSLTAEDDRGSEGFLKGPLSEETEASDSVEEDTDFEEEDDN
|
PDWVSELKKRAGWQGLCDR
|
[0679] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 70B.
366TABLE 70B
|
|
Comparison of NOV70a against NOV70b and NOV70c.
NOV70a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV70b1 . . . 506477/506 (94%)
1 . . . 489477/506 (94%)
NOV70c1 . . . 506410/506 (81%)
1 . . . 425410/506 (81%)
|
[0680] Further analysis of the NOV70a protein yielded the following properties shown in
367TABLE 70C
|
|
Protein Sequence Properties NOV70a
|
|
PSort0.6000 probability located in nucleus;
0.3000 probability located in microbody
analysis:(peroxisome); 0.1000 probability located in
mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0681] A search of the NOV70a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 70D.
368TABLE 70D
|
|
Geneseq Results for NOV70a
NOV70aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM78344Human protein SEQ ID NO 1006 -1 . . . 506501/506 (99%)0.0
Homo sapiens, 506 aa.1 . . . 506503/506 (99%)
[WO200157190-A2, 09 AUG. 2001]
AAM79328Human protein SEQ ID NO 2974 -1 . . . 506500/506 (98%)0.0
Homo sapiens, 527 aa.22 . . . 527 502/506 (98%)
[WO200157190-A2, 09 AUG. 2001]
AAY68778Amino acid sequence of a human42 . . . 506 462/465 (99%)0.0
phosphorylation effector PHSP-10 -46 . . . 510 464/465 (99%)
Homo sapiens, 510 aa.
[WO200006728-A2, 10 FEB. 2000]
AAU07102Human novel human protein, NHP #21 . . . 272152/273 (55%)3e−92
Homo sapiens, 1214 aa.1 . . . 273209/273 (75%)
[WO200161016-A2, 23 AUG. 2001]
AAM39211Human polypeptide SEQ ID NO 2356 -1 . . . 272152/273 (55%)3e−92
Homo sapiens, 1214 aa.1 . . . 273209/273 (75%)
[WO200153312-A1, 26 JUL. 2001]
|
[0682] In a BLAST search of public sequence datbases, the NOV70a protein was found to have homology to the proteins shown in the BLASTP data in Table 70E.
369TABLE 70E
|
|
Public BLASTP Results for NOV70a
NOV70aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P51956Serine/threonine-protein kinase NEK348 . . . 506 456/459 (99%)0.0
(EC 2.7.1.-) (NimA-related protein1 . . . 459457/459 (99%)
kinase 3) (HSPK 36) - Homo sapiens
(Human), 459 aa (fragment).
AAH19916HYPOTHETICAL 51.3 KDA57 . . . 506 448/450 (99%)0.0
PROTEIN - Homo sapiens (Human),1 . . . 450449/450 (99%)
450 aa.
Q99K72SIMILAR TO NIMA (NEVER IN1 . . . 500369/503 (73%)0.0
MITOSIS GENE A)-RELATED1 . . . 496423/503 (83%)
EXPRESSED KINASE 3 - Mus
musculus (Mouse), 509 aa.
Q9R0A5Serine/threonine-protein kinase NEK31 . . . 500370/505 (73%)0.0
(EC 2.7.1.-) (NimA-related protein1 . . . 498423/505 (83%)
kinase 3) - Mus musculus (Mouse),
511 aa.
Q96PY6KIAA1901 PROTEIN - Homo sapiens1 . . . 272152/273 (55%)8e−92
(Human), 1265 aa (fragment).8 . . . 280209/273 (75%)
|
[0683] PFam analysis predicts that the NOV70a protein contains the domains shown in the Table 70F.
370TABLE 70F
|
|
Domain Analysis of NOV70a
Identities/
NOV70a MatchSimilarities for
Pfam DomainRegionthe Matched RegionExpect Value
|
RIO1: domain 1 of 15 . . . 18334/234(15%)6.5
86/234(37%)
pkinase: domain 1 of 14 . . . 25795/293(32%)1.4e−88
212/293(72%)
Vmethyltransf: domain 1 of 1248 . . . 263 7/16(44%)6.8
15/16(94%)
|
Example 71
[0684] The NOV71 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 71A.
371TABLE 71A
|
|
NOV71 Sequence Analysis
|
|
SEQ ID NO: 1851453 bp
NOV71 a,ATGGATAAGTACGATGTGATTAAGGCCATCGGGCAAGGTGCCTTCGGGAAAGCATACT
CG93896-01 DNA Sequence
TAGCTAAAGGGAAATCAGATAGCAAGCACTGTGTCATAAAAGAGATCAATTTTGAAAA
|
GCAAGAAAAAGAAGCTTCAAAGAAAGAAGTGATTCTTCTGGAAAAGATGAAACATCCC
|
AACATTGTAGCCTTCTTCAATTCATTTCAAGAGAATGGCAGGCTGTTTATTGTAATGG
|
AATATTGTGATGGAGGGGATCTCATGAAAAGGATCAATAGACAACGGGGTGTGTTATT
|
TAGTGAAGATCAGATCCTCGGTTGGTTTGTACAGATTTCTCTAGGACTAAAACATATT
|
CATGACAGGAAGATATTACACAGGGACATAAAAGCTCAGAACATTTTTCTTAGCAAGA
|
ACGGAATGGTGGCAAAGCTTGGGGACTTTGGTATAGCAAGAGTCCTGAATAATTCCAT
|
GGAACTTGCTCGAACTTGTATTGGAACACCTTACTACCTGTCCCCAGAGATCTGTCAG
|
AATAAACCCTACAACAATAAAACGGATATTTGGTCTCTTGGCTGTGTCTTATATGAGC
|
TCTGCACACTTAAACATCCTTTTGAGGGTAACAACTTACAGCAGCTGGTTCTGAAGAT
|
TTGTCAAGCACATTTTGCCCCAATATCTCCGGGGTTTTCTCGTGAGCTCCATTCCTTG
|
ATATCTCAGCTCTTTCAAGTATCTCCTCGAGACCGACCATCCATAAATTCCATTTTGA
|
AAAGGCCCTTTTTAGAGAATCTTATTCCCAAATATTTGACTCCTGAGGTCATTCAGGA
|
AGAATTCAGTCACATGCTTATATGCAGAGCAGGAGCGCCAGCTTCTCGACATGCTGGG
|
AAGGTGGTCCAGAAGTGTAAAATACAAAAAGTGAGATTCCAGGGAAAGTGCCCACCAA
|
GATCAAGGATATCTGTGCCAATTAAAAGGAATGCTATATTGCATAGAAATGAATGGAG
|
ACCACCAGCTGGAGCCCAGAAGGCCAGATCTATAAAAATGATAGAAAGACCCAAAATT
|
GCTGCTGTCTGTGGACATTATGATTATTATTATGCTCAACTTGATATGCTGAGGAGGA
|
GAGCCCACAAACCAAGTTATCACCCCATTCCTCAAGAAAATACTGGAGTTGAGGATTA
|
CGGTCAGGAAACGAGGCATGGTCCATCCCCAAGTCAATGATTCTGTAACTGTGAACTA
|
CTTCTTGAACTTGGAACTTCAAGCCACTGGTGAATTGTGAATCTCATTACTAAACTGA
|
AAATTACTCGTCAAATTGGTGCCTAAGATTCGTTCAAGTTTCTACTTAAGCTGAACAT
|
TCTTATTTTCTAAGGCCTGCTGAGTACCTTCAGAGAAAATTTGAAGCTCAACAATATA
|
AGTTGAAAGTGGAGAAGCAATTGGGTCTTCGTCCATCTTCTGCCGAGCCAAATTACAA
|
CCA
|
ORE Start: ATG at 1ORF Stop: TGA at 1198
SEQ ID NO: 186399 aaMW at 45662.6kD
NOV71a,MDKYDVIKAIGQGAFGKAYLAKGKSDSKHCVIKEINFEKQEKEASKKEVILLEKMKHP
CG93896-01 Protein Sequence
NIVAFFNSFQENGRLFIVMEYCDGGDLMKRINRQRGVLFSEDQILGWFVQISLGLKHI
|
HDRKILHRDIKAQNIFLSKNGMVAKLGDFGIARVLNNSMELARTCIGTPYYLSPEICQ
|
NKPYNNKTDIWSLGCVLYELCTLKHPFEGNNLQQLVLKICQAHFAPISPGFSRELHSL
|
ISQLFQVSPRDRPSINSILKRPFLENLIPKYLTPEVIQEEFSHMLICRAGAPASRHAG
|
KVVQKCKIQKVRFQGKCPPRSRISVPIKRNAILHRNEWRPPAGAQKARSIKMIERPKI
|
AAVCGHYDYYYAQLDMLRRRAHKPSYHPIPQENTGVEDYGQETRHGPSPSQ
|
SEQ ID NO: 1871587 bp
NOV71b,TGGACACTGACATGGACTGAAGGAGTAGAAAATCCTTCCGGGACGCTTCGTTGGCCCC
CG93896-02 DNA Sequence
GCGGAGCCGGCGGAGCAGGAAACTCAGCCCATTGGAGACCATGGATAAGTACGATGTG
|
ATTAAGGCCATCGGGCAAGGTGCCTTCGGGAAAGCATACTTAGCTAAAGGGAAATCAG
|
ATAGCAAGCACTGTGTCATAAAAGAGATCAATTTTCAAAAGATCCCCATACAAGAAAA
|
AGAAGCTTCAAAGAAAGAAGTGATTCTTCTGGAAAAGATGAAACATCCCAACATTGTA
|
GCCTTCTTCAATTCATTTCAAGAGAATGGCAGGCTGTTTATTGTAATGGAATATTGTG
|
ATGGAGGGGATCTCATGAAAAGGATCAATAGACAACGGGGTGTGTTATTTAGTGAAGA
|
TCAGATCCTCGGTTGGTTTGTACAGATTTCTCTAGGACTAAAACATATTCATGACAGG
|
AAGATATTACACAGGGACATAAAAGCTCAGAACATTTTTCTTAGCAAGAACGGAATGG
|
TGGCAAAGCTTGGGGACTTTGGTATAGCAAGAGTCCTGAATAGTTCCATGGAACTTGC
|
TCGAACTTGTATTGGAACACCTTACTACCTGTCCCCAGAGATCTGTCAGAATAAACCC
|
TACAACAATAAAACGGATATTTGGTCTCTTGGCTGTGTCTTATATGAGCTCTGCACAC
|
TTAAACATCCTTTTGAGGGTAACAACTTACAGCAGCTGGTTCTGAAGATTTGTCAAGC
|
ACATTTTGCCCCAATATCTCCGGGGTTTTCTCGTGAGCTCCATTCCTTGATATCTCAG
|
CTCTTTCAAGTATCTCCTCGAGACCGACCATCCATAAATTCCATTTTGAAAAGGCCCT
|
TTTTAGAGAATCTTATTCCCAAATATTTGACTCCTGAGGTCAGTTTTGAGGAAGAATT
|
CAGTCACATGCTTATATGCAGAGCAGGAGCGCCAGCTTCTCGACATGCTGGGAAGGTG
|
GTCCAGAAGTGTAAAATACAAAAAGTGAGATTCCAGGGAAAGTGCCCACCAAGATCAA
|
GGATATCTGTGCCAATTAAAAGGAATGCTATATTGCATAGAAATGAATGGAGACCACC
|
AGCTGGAGCCCAGAAGGCCAGATCTATAAAAATGATAGAAAGACCCAAAATTGCTGCT
|
GTCTGTGGACATTATGATTATTATTATGCTCAACTTGATATGCTGAGGAGGAGAGCCC
|
ACAAACCAAGTTATCACCCCATTCCTCAAGAAAATACTGGAGTTGAGGATTACGGTCA
|
GGAAACGAGGCATGGTCCATCCCCAAGTCAATGATTCTGTAACTGTGAACTACTTCTT
|
GAACTTGGAACTTCAAGCCACTGGTGAATTGTGAATCTCATTACTAAACTGAAAATTA
|
CTCGTCAAATTGGTGCCTAAGATTCGTTCAAGTTTCTACTTAAGCTGAACATTCTTAT
|
TTTCTAAGGCCTGCTGAGTACCTTCAGAGAAAATTTGAAGCTCAACAATATAAGTTGA
|
AAGTGGAGAAGCAATTGGGTCTTCGTCCATCTTCTGCCGAGCCAAATTACAACCAAAA
|
AAAAAAAAAAAAAAAAAACGG
|
ORF Start: ATG at 99ORF Stop: TGA at 1308
SEQ ID NO: 188403 aaMW at 46099.1kD
NOV71b,MDKYDVIKAIGQGAFGKAYLAKGKSDSKHCVIKEINFEKMPIQEKEASKKEVILLEKM
CG93896-02 Protein Sequence
KHPNIVAFFNSFQENGRLFIVMEYCDGGDLMKRINRQRGVLFSEDQILGWFVQISLGL
|
KHIHDRKILHRDIKAQNIFLSKNGMVAKLGDFGIARVLNSSMELARTCIGTPYYLSPE
|
ICQNKPYNNKTDIWSLGCVLYELCTLKHPFEGNNLQQLVLKICQAHFAPISPGFSREL
|
HSLISQLFQVSPRDRPSINSILKRPFLENLIPKYLTPEVSFEEEFSHMLICRAGAPAS
|
RHAGKVVQKCKIQKVRFQCKCPPRSRISVPIKRNAILHRNEWRPPAGAQKARSIKMIE
|
RPKIAAVCGHYDYYYAQLDMLRRRAHKPSYHPIPQENTGVEDYGQETRHGPSPSQ
|
[0685] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 71B.
372TABLE 71B
|
|
Comparison of NOV71a against NOV71b.
Identities/
NOV71a Residues/Similarities for
Protein SequenceMatch Residuesthe Matched Region
|
NOV71b1 . . . 399396/403 (98%)
1 . . . 403398/403 (98%)
|
[0686] Further analysis of the NOV71a protein yielded the following properties shown in Table 71C.
373TABLE 71C
|
|
Protein Sequence Properties NOV71a
|
|
PSort0.4500 probability located in cytoplasm;
0.3000 probability located in microbody
analysis:(peroxisome); 0.1000 probability located in
mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0687] A search of the NOV71 a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 71D.
374TABLE 71D
|
|
Geneseq Results for NOV71a
NOV71aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAU03545Human protein kinase #45 - Homo1 . . . 399338/402 (84%)0.0
sapiens, 649 aa. [WO200138503-A2,1 . . . 377344/402 (85%)
31 MAY 2001]
AAU07102Human novel human protein, NHP1 . . . 289183/295 (62%)e−103
#2 - Homo sapiens, 1214 aa.1 . . . 294231/295 (78%)
[WO200161016-A2, 23 AUG. 2001]
AAM39211Human polypeptide SEQ ID NO1 . . . 289183/295 (62%)e−103
2356 - Homo sapiens, 1214 aa.1 . . . 294231/295 (78%)
[WO200153312-A1, 26 JUL. 2001]
AAM39210Human polypeptide SEQ ID NO1 . . . 289183/295 (62%)e−103
2355 - Homo sapiens, 1242 aa.1 . . . 294231/295 (78%)
[WO2001533 12-A1, 26 JUL. 2001]
AAM78344Human protein SEQ ID NO 1006 -1 . . . 271148/273 (54%)3e−85
Homo sapiens, 506 aa.1 . . . 272200/273 (73%)
[WO200157190-A2, 09 AUG. 2001]
|
[0688] In a BLAST search of public sequence datbases, the NOV71 a protein was found to have homology to the proteins shown in the BLASTP data in Table 71E.
375TABLE 71E
|
|
Public BLASTP Results for NOV71a
NOV71aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P51954Serine/threonine-protein kinase NEK11 . . . 347197/379 (51%)e−105
(EC 2.7.1.-) (NimA-related protein1 . . . 375261/379 (67%)
kinase 1) - Mus musculus (Mouse),
774 aa.
Q96PY6KIAA1901 PROTEIN - Homo sapiens1 . . . 289183/295 (62%)e−103
(Human), 1265 aa (fragment).8 . . . 301231/295 (78%)
Q9R0A5Serine/threonine-protein kinase NEK31 . . . 271149/271 (54%)2e−85
(EC 2.7.1.-) (NimA-related protein1 . . . 270199/271 (72%)
kinase 3) - Mus musculus (Mouse),
511 aa.
Q99K72SIMILAR TO NIMA (NEVER IN1 . . . 271148/271 (54%)9e−85
MITOSIS GENE A)-RELATED1 . . . 270199/271 (72%)
EXPRESSED KINASE 3 - Mus
musculus (Mouse), 509 aa.
P51956Serine/threonine-protein kinase NEK346 . . . 271 130/226 (57%)1e−75
(EC 2.7.1.-) (NimA-related protein1 . . . 225171/226 (75%)
kinase 3) (HSPK 36) - Homo sapiens
(Human), 459 aa (fragment).
|
[0689] PFam analysis predicts that the NOV71 a protein contains the domains shown in the
376TABLE 71F
|
|
Domain Analysis of NOV71a
Identities/
Similarities for
Pfam DomainNOV71a Match Regionthe Matched RegionExpect Value
|
NadA: domain 1 of 131 . . . 213 47/369 (13%)7.5
112/369 (30%)
pkinase: domain 1 of 14 . . . 256 89/292 (30%)7.1e−88
205/292 (70%)
|
Example 72
[0690] The NOV72 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 72A.
377TABLE 72A
|
|
NOV72 Sequence Analysis
|
|
SEQ ID NO: 1892290 bp
NOV72a,GTGGCATCAATGCCGAAGAACAGCAAAGTGACCCAGCGTGAGCACAGCAGTGAGCATG
CG93939-01 DNA Sequence
TCACTGAGTCCGTGGCCGACCTGCTGGCCCTCCACGAGCCTGTGGACTATAAGCAGAG
|
TGTACTGAATCTGGCTGGTGAGGCAGCCGGCAAGCACAAGGCGGTGGAGGACGACCTG
|
GATGCAGAGGACCGGCCGGCCTGGAACAGTAAGCTGCAGTACATCCTGGCCCACATTG
|
GCTTCTCTGTGGGCCTCGGCAACATCTGGAGGTTCCCCTACCTGTGCCAGAAAAATGG
|
AGGAGGTGCTTACCTGGTGCCCTACCTGGTGCTGCTGATCATCATCGGGATCCCCCTC
|
TTCTTCCTGGAGCTGGCTGTGGGTCAGAGGATCCGCCGCGGCAGCATCGGTGTGTGGC
|
ACTATATATGTCCCCGCCTGGGGGGCATCGGCTTCTCCAGCTGCAPAGTCTGTCTCTT
|
TGTGGGGCTGTATTATAATGTGATCATCGGGTGGAGCATCTTCTATTTCTTCAAGTCC
|
TTCCAGTACCCGCTGCCCTGGAGTGAATGTCCTGTCGTCAGGAATGGCAGCGTCGCAG
|
TGGTGGAGGCAGAGTGTGAAAAGAGCTCAGCCACTACCTACTTCTGGTACCGAGAGGC
|
CTTGGACATCTCTCACTCCATCTCGGAGAGTCGGGGCCTCAACTGGAAGATGACCCTG
|
TGCCTCCTCGTGGCCTGGAGCATCGTCGGGATGGCTGTCGTTAAGGGCATCCAGTCCT
|
CGGGGAAGGTGATGTATTTCAGCTCCCTCTTCCCCTACGTGGTGCTGGCCTGCTTCCT
|
GGTCCGGGGGCTGTTGCTGCGAGGGGCAGTTGATGGCATCCTACACATGTTCACTCCC
|
AAGCTGGACAAGATGCTGGACCCCCAGGTGTGGCGGGAGGCAGCTACCCAGGTCTTCT
|
TTGCCTTGGGCCTCGGCTTTGCTGGTGTCATTGCCTTCTCCAGCTACAATAAGCAGGA
|
CAACAACTGCCACTTCGATGCCGCCCTGGTGTCCTTCATCAACTTCTTCACGTCAGTG
|
TTGGCCACCCTCGTGGTGTTTGCTGTGCTGCGCTTCAAGGCCAACATCATGAATGAGA
|
AGTGTGTGGTCGAGAATGCTGAGAAAATCCTAGGGTACCTTAACACCAACGTCCTGAG
|
CCGGGACCTCATCCCACCCCACGTCAACTTCTCCCACCTCACCACAAAGGACTACATG
|
GAGATGTACAATGTCATCATGACCGTGAAGGAGGACCAGTTCTCAGCCCTGGGTCTTG
|
ACCCCTGCCTTCTGGAGGACGAGCTCGACAAGTCCGTGCAGGGCACACGCCTGGCCTT
|
CATCGCCTTCACTGAGGCCATGACGCACTTCCCCGCCTCCCCGTTCTGGTCCGTCATC
|
TTCTTCTTGATGCTTATCAACCTGGGCCTGGGCAGCATGATCGGGACCATGGCAGGCA
|
TCACCACGCCCATCATCGACACCTTCAACGTGCCCAAGGAGATGTTCACACTGGCCTG
|
CTGTGTCTTTGCATTCCTCGTGGGGCTGTTGTTCGTCCAGCGCTCCGGAAACTACTTT
|
GTCACCATGTTCGATGACTACTCAGCCACCCTGCCACTCACTCTCATCGTCATCCTTG
|
AGAACATCGCTGTGGCCTGGATTTATGGAACCAAGAAGTTCATGCAGGAGCTGACGGA
|
GATGCTGGGCTTCCGCCCCTACCGCTTCTATTTCTACATGTGGAAGTTCGTGTCTCCA
|
CTATGCATGGCTGTGCTCACCACAGCCAGCATCATCCAGCTGGGGGTCACGCCCCCGG
|
GCTACAGCGCCTGGATCAAGGAGGAGGCTGCCGAGCGCTACCTGTATTTCCCCAACTG
|
CGCCATGGCACTCCTGATCACCCTCATCGTCGTGGCGACGCTGCCCATCCCTGTCGTG
|
TTCGTCCTGCGGCACTTCCACCTGCTCTGTGATGGCTCCAACACCCTCTCCGTGTCCT
|
ACAAGAAGCGCCGCATGATGAAGGACATCTCCAACCTGGAGGAGAACCATGAGACCCG
|
CTTCATCCTCAGCAAGGTGCCCAGTGAGGCACCTTCCCCCATGCCCACTCACCGTTCC
|
TATCTGGGGCCCGGCAGCACATCACCCCTGGAGACCAGCGGTAACCCCAATGGACGCT
|
ATGGGAGCGGCTACCTGCTGGCCAGCACCCCTGAGTCGGAGCTGTGACCACTGCCCAA
|
GCCCT
|
ORF Start: ATG at 10ORF Stop: TGA at 2191
SEQ ID NO: 190727 aaMW at 81000.5kD
NOV72a, MPKNSKVTQREHSSEHVTESVADLLALEEPVDYKQSVLNVAGEAGGKQKAVEEELDAE
CG93939-01 Protein Sequence
DRPAWNSKLQYILAQIGFSVGLGNIWRFPYLCQKNGGGAYLVPYLVLLIIIGIPLFFL
|
ELAVGQRIRRGSIGVWHYICPRLGGIGFSSCIVCLFVGLYYNVIIGWSIFYFFKSFQY
|
PLPWSECPVVRNGSVAVVEAECEKSSATTYFWYREALDISDSISESGGLNWKMTLCLL
|
VAWSIVGMAVVKGIQSSGKVMYFSSLFPYVVLACFLVRGLLLRGAVDGILHMFTPKLD
|
KMLDPQVWREAATQVFFALGLGFGGVIAFSSYNKQDNNCHFDAALVSFINFFTSVLAT
|
LVVFAVLGFKANIMNEKCVVENAEKILGYLNTNVLSRDLIPPHVNFSHLTTKDYMEMY
|
NVIMTVKEDQFSALGLDPCLLEDELDSKSQGTGLAFIAFTEAMTHFPASPFWSVMFFL
|
MLINLGLGSMIGTMAGITTPIIDTFKVPKEMFTVGCCVFAFLVGLLFVQRSGNYFVTM
|
FDDYSATLPLTLIVILENIAVAWIYGTKKFMQELTEMLGFRPYRFYFYMWKFVSPLCM
|
AVLTTASIIQLGVTPPGYSAWIKEEAAERYLYFPNWAMALLITLIVVATLPIPVVFVL
|
RHFHLLSDGSNTLSVSYKKGRMMKDISNLEENDETRFILSKVPSEAPSPMPTHRSYLG
|
PGSTSPLETSGNPNGRYGSGYLLASTPESEL
|
[0691] Further analysis of the NOV72a protein yielded the following properties shown in Table 72B.
378TABLE 72B
|
|
Protein Sequence Properties NOV72a
|
|
PSort0.8000 probability located in plasma membrane;
0.4000 probability located in
analysis:Golgi body; 0.3000 probability located in
endoplasmic reticulum (membrane);
0.3000 probability located in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0692] A search of the NOV72a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 72C.
379TABLE 72C
|
|
Geneseq Results for NOV72a
NOV72aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAR88390Human neurotransmitter transporter1 . . . 727706/727 (97%)0.0
protein - Homo sapiens, 727 aa.1 . . . 727706/727 (97%)
[WO9531539-A1, 23 NOV. 1995]
AAY72908Human NTT7 protein - Homo sapiens,1 . . . 727487/735 (66%)0.0
730 aa. [WO200119854-A2,1 . . . 730603/735 (81%)
22 MAR. 2001]
AAG67159Amino acid sequence of a human 5791 . . . 727487/735 (66%)0.0
transporter polypeptide - Homo1 . . . 730603/735 (81%)
sapiens, 730 aa. [WO200164875-A2,
07 SEPT. 2001]
AAE05100Drosophila melanogasterdmNTT459 . . . 684 280/629 (44%)e−168
protein - Drosophila melanogaster,54 . . . 662 407/629 (64%)
675 aa. [WO200149848-A2, 12 JUL.
2001]
AAG64193Human nerve mass-transferring25 . . . 634 262/617 (42%)e−147
protein - Homo sapiens, 616 aa.3 . . . 596390/617 (62%)
[CN1287170-A, 14 MAR. 2001]
|
[0693] In a BLAST search of public sequence datbases, the NOV72a protein was found to have homology to the proteins shown in the BLASTP data in Table 72D.
380TABLE 72D
|
|
Public BLASTP Results for NOV72a
NOV72aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P31662Orphan sodium- and chloride-1 . . . 727705/727 (96%)0.0
dependent neurotransmitter transporter1 . . . 727718/727 (97%)
NTT4 - Rattus norvegicus (Rat), 727
aa.
I56506Na+/Cl(−)-dependent neurotransmitter1 . . . 727703/727 (96%)0.0
transporter, brain - rat, 727 aa.1 . . . 727716/727 (97%)
Q9H2J7Orphan sodium- and chloride-1 . . . 727487/735 (66%)0.0
dependent neurotransmitter transporter1 . . . 730603/735 (81%)
NTT73 (Orphan transporter v7-3) -
Homo sapiens (Human), 730 aa.
Q9XS59Orphan sodium- and chloride-1 . . . 724480/727 (66%)0.0
dependent neurotransmitter transporter1 . . . 723593/727 (81%)
NTT73 (Orphan transporter v7-3) -
Bos taurus (Bovine), 729 aa.
Q08469Orphan sodium- and chloride-1 . . . 724484/734 (65%)0.0
dependent neurotransmitter transporter1 . . . 723595/734 (80%)
NTT73 (Orphan transporter v7-3) -
Rattus norvegicus (Rat), 729 aa.
|
[0694] PFam analysis predicts that the NOV72a protein contains the domains shown in the
381TABLE 72E
|
|
Domain Analysis of NOV72a
Identities/
NOV72a MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
SNF: domain 1 of 160 . . . 657310/638 (49%)0
572/638 (90%)
|
Example 73
[0695] The NOV73 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 73A.
382TABLE 73A
|
|
NOV73 Sequence Analysis
|
|
SEQ ID NO: 1911547 bp
NOV73a,GGAATTATATTAAATCTGTAAATCAATTGAAAAATAATTGTCAAAACTGTGGCGCCAG
CG94245-01 DNA Sequence
ATACCGCGGTTGAGGTGGATGGAAGGGACCTGCTGCAGGGAGATGAGGAGGCTGCAGG
|
GCAGTGACTGCAGCTCCACAGCTCCCGCAGTGGCTCTGCCCAGGTACTTCTGCCCCAG
|
AGAGGACAAATATGAATACCCAGACAACAGTCTCATCTGTCTTTTTGCTATATGTGAG
|
TCAAGGGCAACAGGAAAACACTGTGCGGCATCGAGCTGTGGTCGGTGCAACGGCTTCT
|
TCAGATGCAGCATATGCAAGAGTAACGTTTATTCTTGCAGGTTCCGTCGGCAATGTTT
|
TGTTGACAAGGCCAAAAGAAATGAATGTACATACAGTCAATTAAGAAAGTGTTTTAGA
|
GCAGGAATGGAAAAAGAAGTTGTGCATAATGAATGCAATAGAACAAGCACCAGAAGAA
|
GCACATGTGATGGCAGCAATATCCCCTCCATTAACACACTGGCACAAGCTGAAGTTCT
|
GTCTTGCCAGATCTCAGTCTCAAGCCTTGGGGCAAGTACTGATATACATGTTAAAAAA
|
AATAAAAGTGTTGGTGATGTCTGTCAATTTCTGAAACAGCAGCTCTTAGTCTTGGTGG
|
AATGGGCTAAATATATTCCTGCCTTCTGTCAGTTACCATTGGATGACAAGGTCGCGCT
|
GTTGAGATATCATGCAGGAGAACACTTACTGCCTGGAGCTATAAAGAGATCCATGATG
|
TATAAAGATATTTTGCTTTTGGGAAACAACTATATTATTCACTGCAACAGCTGTGAAT
|
TTGAGATTAGCAGTGTAATCAGTCGGGTTCTAGTTGAGCTGGTTCAACCATTTCAAGA
|
AATCCAAAAGGCAATTGTATTTTTTGGCCCAGATGTGAAAGGGCTACACGATCCAATA
|
AACATTAAGAACATGCAATTCCAAGGGCGGATGGGTTTGCAGAACTCCGTCAATGATT
|
CCGCACGCAGTATGGCTCCCTGGGGGAGGTTTGGACAGTTGCTTCTGCTGCTGCCCAC
|
ACTGCTGCGCATCATTTGGCAAATGATTGAGCAAATAAGTTTGTTTAAAAAACTTTTG
|
CGGTTGACTAAAATTGGCCACCTACTTCAGGAAATGTTAAATGGGGCTTCCAATGATA
|
GTAGTCATCTCCATCATCCAATACATCCACATTCATCTCAAGATCCATTAACTGGACA
|
AACTGTACTTTTACGTCCCATGTACACACTGGTACACACTGGTTCTTATGAAGACCAC
|
ATCAGAACTCCTGAAACCCCACTCCCTTCCCCACCACAAGGCTTTGCACAAGAAGATT
|
ACAGAACAGCTACAAATCAAGCTTCAGTCATTTCACCAGCCTCTCTTCAAACAAAAAC
|
AATTGTTTCAAACAAAAACAATTGTGAAAATGTGTTTATTTCTGAACAGCACTGCATA
|
AATGTGAAAAGCTGTTTGTCTTGAAACATCTCAAGATAGTACTTTTGGCAAACTCTGA
|
TCCAAGGCTTCTTCATGGAACTGTTATAAGACAGTATCC
|
ORF Start: ATG at 77ORF Stop: TGA at 1472
SEQ ID NO: 192465 aaMW AT 52258.7kD
NOV73a, MEGTCCREMRRLQGSDCSSRAPGVALRRYFCPREDKYEYPDNSLICLFAICESRATGK
CG94245-01 Protein Sequence
HCGASSCGGCKGFFRCSICKSNVYSCRFRRQCFVDKAKRNQCRYSQLRKCFRAGMEKE
|
VVHNECNRTSTRRSTCDGSNIPSINTLAQAEVLSCQISVSSLGASTDIHVKKNKSVGD
|
VCEFLKQQLLVLVEWAKYIPAFCQLPLDDKVALLRYHAGEHLLPGAIKRSMMYKDILL
|
LGNNYIIHCNSCEFEISSVISRVLVELVQPFQEIQKAIVFFGPDVKGLHDPINIKNMQ
|
FQGRMGLENSVNDSARSMAPWGRFGELLLLLPTLLRIIWQMIEQISLFKKLLRLTKIG
|
HLLQEMLNGASNDSSHLHHPIHPHSSQDPLTGQTVLLGPMYTLVHTGSYEDHIRTPET
|
PLPSPPQGFAQEDYRTATNQASVISPASLQTKTIVSNKNNCENVFISEQHCINVKSCL
|
S
|
[0696] Further analysis of the NOV73a protein yielded the following properties shown in
383TABLE 73B
|
|
Protein Sequence Properties NOV73a
|
|
PSort0.4721 probability located in
mitochondrial matrix space; 0.3000 probability
analysis:located in microbody (peroxisome);
0.1752 probability located in mitochondrial
inner membrane; 0.1752 probability located
in mitochondrial intermembrane space
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0697] A search of the NOV73a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 73C.
384TABLE 73C
|
|
Geneseq Results for NOV73a
NOV73aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAM79298Human protein SEQ ID NO 1960 -47 . . . 434230/407 (56%)e−116
Homo sapiens, 474 aa.59 . . . 461279/407 (68%)
[WO200157190-A2, 09 AUG. 2001]
AAW71574Human native hepatocyte nuclear47 . . . 434230/407 (56%)e−116
factor 4 alpha - Homo sapiens, 46550 . . . 452279/407 (68%)
aa. [WO9811254-A1, 19 MAR.
1998]
AAW71587Human hepatocyte nuclear factor 447 . . . 434230/407 (56%)e−115
alpha - Homo sapiens, 516 aa.101 . . . 503 279/407 (68%)
[WO9811254-A1, 19 MAR. 1998]
AAM80282Human protein SEQ ID NO 3928 -47 . . . 434228/407 (56%)e−115
Homo sapiens, 505 aa.90 . . . 492277/407 (68%)
[WO200157190-A2, 09 AUG. 2001]
AAY77496Rat hepatocyte nuclear factor 447 . . . 444226/412 (54%)e−115
(HNF-4) - Rattus sp, 455 aa.50 . . . 453281/412 (67%)
[US6025196-A, 15 FEB. 2000]
|
[0698] In a BLAST search of public sequence datbases, the NOV73a protein was found to have homology to the proteins shown in the BLASTP data in Table 73D.
385TABLE 73D
|
|
Public BLASTP Results for NOV73a
NOV73aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
JC6095hepatocyte nuclear factor 4 gamma41 . . . 431293/401 (73%)e−158
chain - human, 774 aa.371 . . . 764 320/401 (79%)
Q14541Hepatocyte nuclear factor 4-gamma41 . . . 431292/401 (72%)e−158
(HNF-4-gamma) - Homo sapiens 5 . . . 398320/401 (78%)
(Human), 408 aa.
Q9WUU6Hepatocyte nuclear factor 4-gamma41 . . . 435288/405 (71%)e−156
(HNF-4-gamma) - Mus musculus 5 . . . 402319/405 (78%)
(Mouse), 408 aa.
Q91766Hepatocyte nuclear factor 4-alpha47 . . . 444235/411 (57%)e−118
(HNF-4-alpha) - Xenopus laevis50 . . . 453283/411 (68%)
(African clawed frog), 455 aa.
JC4937hepatocyte nuclear factor 4, splice47 . . . 434231/407 (56%)e−116
form B - human, 465 aa.50 . . . 452280/407 (68%)
|
[0699] PFam analysis predicts that the NOV73 a protein contains the domains shown in the
386TABLE 73E
|
|
Domain Analysis of NOV73a
Identities/
NOV73a MatchSimilarities forExpect
Pfam DomainRegionthe Matched RegionValue
|
zf-C4: domain 1 of 1 49 . . . 12135/74(47%)9.9e−25
60/74(81%)
hormone_rec:180 . . . 35559/205(29%)6.3e−27
domain 1 of 1128/205(62%)
|
Example 74
[0700] The NOV74 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 74A.
387TABLE 74A
|
|
NOV74 Sequence Analysis
|
|
SEQ ID NO: 1937386 bp
NOV74a,TCAAAAAAGCCAAGTTTGATGGTGCCCAAGAGAAATTCAACACGTACGTGACCCTGAA
CG94302-01 DNA Sequence
AGTGCAGAATGTCAAGAGCACGACCATCGCGGTGCGGGGCAGCCAGCCCAGCTGGGAG
|
CAGGATTTCATGTTCGAGATTAACCGTCTGGATTTGGGACTGACGGTGGAGGTGTGGA
|
ATAAGCGPCTCATCTGGGACACAATGGTGGGCACTGTGTGCATCCCACTGAGGACCAT
|
CCGCCAGTCCAATGAGGAGGGCCCTGGAGAGTGGCTCACGCTGGACTCCCACGTCATC
|
ATGGCAGACAGTGAGATCTGTGGCACAAAGGACCCCACCTTCCACCGCATCCTCCTGG
|
ACACGCGCTTTGAGCTACCCTTAGACATTCCTGAAGAGGACGCTCCCTACTGGCCCAA
|
GAAGCTGGAGCAGCTCAATGCTATGCGGGACCAGGATGAATATTCGTTCCAAGATGAG
|
CAAGACAAGCCTCTGCCTGTCCCCAGCAACCAGTGCTGTAAGTACTTCCCATACCGCC
|
TCTCTCTTCCACACGATGACCCCGACAGTCCAGTGGATGATCGTGACAGTGACTACCG
|
CAGTGAAACGAGCAACAGCATCCCGCCGCCCTATTATACTACGTCACAACCCAACGCC
|
TCAGTCCACCAATATTCTGTTCGCCCACCACCCCTGCGCTCCCGGGAGTCCTACAGTG
|
ACTCCATGCACAGTTACGAGGAGTTCTCTCAGCCACAAGCCCTCAGCCCCACGGGTAG
|
CAGCCGCTATGCCTCTTCCGGGGAGCTGAGCCAGGGAAGCTCTCAGCTGAGCGAGGAC
|
TTCGACCCTGACCAGCACAGCCTGCAGGGCTCCGACATGGACGATGAGCGGCACCGGG
|
ACTCCTACCACTCCTGCCACAGCTCGGTCAGCTACCACAAAGACTCGCCTCGCTGGGA
|
CCAGGATGAGGAAGAGCTGGAGGAGGACCTGGAGGACTTCCTGGAGCAGGAGGAGCTG
|
CCTGAAGATGAGGAGCAGCTGGAGGAGGAGGAGGAGGAGGTGCCTGACCATTTGGGCA
|
GCTATGCCCAGCGTGAAGACGTAGCTGTGGCTGAGCCCAAAGACTTCAAACGCATCAG
|
CCTCCCGCCAGCTGCCCCAGGGAAGGAGGACAACGCCCCAGTCGCACCCACCGAGGCC
|
CCCGACATGGCCAAGGTGGCCCCCAAGCCAGCCACGCCCGACAAGGTGCCTGCAGCTG
|
AGCAGATCCCTGAGGCTGAGCCACCCAAGGACGAGGAGAGTTTCAGGCCGAGAGAGGA
|
TGAGGAAGGCCAGGAGGGGCAGGACTCCATGTCCAGGGCCAAGGCCAACTGGCTGCGT
|
GCCTTCAACAAGGTGCGGATGCAGCTGCAGGAGGCCCGGGGAGAAGGAGAGATGTCTA
|
AATCCCTATGGTTCAAAGGCGGGCCAGGGGGCGGTCTCATCATCATCGACAGCATGCC
|
AGACATCCGCAAGAGGAAACCTATCCCACTCGTGAGCGACTTGGTACTGTCCCTGGTC
|
CAGTCCAGGAAAGCGGGCATCACCTCGCCCTTGGCCTCCAGCACGTTGAACAACGAGG
|
AGCTGAAAAACCACGTTTACAAGAAGACCCTGCAAGCCTTAATCTACCCCATCTCGTG
|
CACGACGCCACACAACTTCGAAGTGTCGACGGCCACCACGCCCACCTACTGCTACGAG
|
TGCGAGGGGCTGCTGTGGGGCATCGCGAGGCAGGGCATGCGCTGCACCGAGTGCGGTG
|
TCAAGTGCCACGAGAAGTGCCAGGACCTGCTCAACGCCGACTGCCTGCAGCGTGCTGC
|
GGAGAAGAGCTCCAAGCACGGGGCGGAGGACCGGACACACAACATCATCATGGTGCTC
|
AAGGACCGCATGAAGATCCGGGAGCGCAACAAGCCCGAGATCTTCGAGCTCATCCAGG
|
AGATCTTCGCGGTGACCAAGACGGCGCACACGCACCACATGAAGGCGGTCAAGCAGAG
|
CGTGCTGGACGGCACGTCCAAGTGGTCCGCCAACATCAGCATCACCGTGGTCTGCGCC
|
CAGGGCTTGCAGGCAAAGGACAAGACAGGATCCAGTGACCCCTATGTCACCGTCCAGG
|
TCGGGAAGACCAAGAAACGGACAAAAACCATCTATGGGAACCTCAACCCGGTGTGGGA
|
GGAGAATTTCCACTTTGAATGTCACAATTCCTCCGACCGCATCAAGGTGCGCGTCTGG
|
GACGAGGATGACGACATCAAATCCCGCGTGAAACAGAGGTTCAAGAGGGAATCTGACG
|
ATTTCCTGGGGCAGACGATCATTGAGGTGCGGACGCTCAGCGGCGAGATGGACGTGTG
|
GTACAACCTGGACAAGCGAACTGACAAATCTGCCGTGTCGGGTGCCATCCGGCTCCAC
|
ATCAGTGTGGAGATCAAAGGCGAGGAGAAGGTGGCCCCGTACCATGTCCAGTACACCT
|
GTCTGCATGAGAACCTGTTCCACTTCGTCACCGACGTGCAGAACAATGGGGTCGTGAA
|
GATCCCAGATGCCAAGGGTGACGATGCCTGGAAGGTTTACTACGATGAGACAGCCCAG
|
GAGATTGTCGACGAGTTTGCCATGCGCTACCGCGTCGAGTCCATCTACCAAGCCATGA
|
CCCACTTTGCCTCCCTCTCCTCCAAGTATATGTCCCCAGGGGTGCCTGCCGTCATGAG
|
CACCCTGCTCGCCAACATCAATGCCTACTACGCACACACCACCGCCTCCACCAACGTG
|
TCTGCCTCCGACCGCTTCGCCGCCTCCAACTTTGGGAAAGAGCCCTTCCTGAAACTCC
|
TGGACCAGCTGCATAACTCCCTGCGGATTGACCTCTCCATGTACCGGAATAACTTCCC
|
AGCCAGCAGCCCGGAGAGACTCCAGGACCTCAAATCCACTGTGGACCTTCTCACCAGC
|
ATCACCTTCTTTCGGATGAAGGTACAAGAACTCCAGAGCCCGCCCCGAGCCAGCCAGG
|
TGCTAAAGGACTGTGTGAAAGCCTGCCTTAATTCTACCTACGACTACATCTTCAATAA
|
CTGCCATGAACTGTACAGCCGGGAGTACCAGACAGACCCGGCCAAGAAGGGGGAAGTT
|
CCCCCAGAGGAACAGGGGCCCAGCATCAAGAACCTCGACTTCTGGTCCAAGCTGATTA
|
CCCTCATAGTGTCCATCATTGACGAAGACAAGAATTCCTACACTCCCTGCCTCAACCA
|
GTTTCCCCAGGAGCTGAATGTGGGTAAAATCAGCGCTGAAGTGATGTGGAATCTGTTT
|
GCCCAAGACATGAACTACGCCATGGAGGAGCACGACAAGCATCGTCTATGCAAGAGTG
|
CCGACTACATGAACCTCCACTTCAAGGTGAAATCCCTCTACAATGAGTATGTCACGGA
|
ACTTCCCTCCTTCAAGGACCGCGTGCCTGAGTACCCTGCATGGTTTGAACCCTTCGTC
|
ATCCAGTGGCTGGATGAGAATGAGGAGGTGTCCCGGGATTTCCTGCACGGTGCCCTGG
|
AGCGAGACAAGAAGGATGGGTTCCAGCAGACCTCAGAGCATGCCCTATTCTCCTGCTC
|
CGTGGTGGATGTTTTCTCCCAACTCAACCAGAGCTTTGAAATCATCAAGAAACTCGAG
|
TGTCCCGACCCTCAGATCGTGGGGCACTACATGAGGCGCTTTGCCAAGACCATCAGTA
|
ATGTGCTCCTCCAGTATGCAGACATCATCTCCAAGGACTTTGCCTCCTACTGCTCCAA
|
GGAGAAGGACAAAGTGCCCTGCATTCTCATGAATAACACTCAACAGCTACCAGTTCAG
|
CTGGAGAAGATGTTCGAAGCCATGGGAGGAAAGGAGCTGGATGCTGAAGCCAGTGACA
|
TCCTGAAGGAGCTTCAGGTGAAACTCAATAACGTCTTGGATGAGCTCAGCCGGGTGTT
|
TGCTACCAGCTTCCAGCCGCACATTGAAGAGTGTGTCAAACAGATGGGTGACATCCTT
|
AGCCAGGTTAAGGGCACAGGCAATGTGCCAGCCAGTGCCTGCAGCAGCGTGGCCCAGG
|
ACGCGGACAATGTGTTGCAGCCCATCATGGACCTGCTGGACAGCAACCTGACCCTCTT
|
TGCCAAAATCTGTGAGAAGACTGTGCTCAAGCCAGTGCTGAAGGAGCTGTGGAAGCTG
|
GTTATGAACACCATGGAGAAAACCATCGTCCTGCCGCCCCTCACTGACCAGACGATGA
|
TCGGGAACCTCTTGAGAAAACATGGCAAGGGATTAGAAAAGGGCAGGGTGAAATTGCC
|
AAGCCACTCAGACGGAACCCAGATGATCTTCAATGCAGCCAAGGAGCTGGGTCAGCTG
|
TCCAAACTCAAGGATCACATGGTACGAGAAGAAGCCAAGAGCTTCACCCCAAACCAGT
|
GCGCGGTTGTTGAGTTGGCCCTGGACACCATCAAGCAATATTTCCACGCCGGTGGCGT
|
GGGCCTCAAGAAGACCTTCCTGGAGAAGAGCCCGGACCTGCAATCCTTGCGCTATGCC
|
CTGTCACTCTACACGCAGCCCACCGACCTGCTAATCAAGACCTTTGTACAGACGCAAT
|
CGGCCCGAGGTCCATGGTGGAAAAGGTACTAGGTTACCCTTAGTGAAGACATTTATCC
|
TGAGAAGGGTACGGGTGTAGAAGACCCTCTGGGTGAAGTCTCTCTCCATGTTGAGCTG
|
TTCACTCATCGAGGAACTGGGTAACACAAGGTCACAGTGAAAGTGGTGGCTGCCAATG
|
ACCTCAAGTGGCAGACTTCTGGCATCTTCCGGCCGTTCATCGAGGTCAACATCATTGG
|
GCCCCAGCTCAGCCACAACAAACGCAAGTTTCCCACCAAATCCAAGAACAATAGCTGG
|
GCTCCCAAGTACAATGAGAGCTTCCAGTTCACCCTGAGCGCCGACGCGGGTCCCCAGT
|
GGGGCTGGCCGTGCTGCAGCTGCGTGAGCTGGCCCAGCGCGGGAGCGCCGCCTGCTGG
|
CTGCCGCTCGGCCGCCGCATCCACATGGACGACACGGGCCTCACGGTGCTGCGAATCC
|
TCTCGCAGCGCAGCAACGACGAGGTGGCCAAGGAGTTCGTGAAGCTCAAGTCGGACAC
|
GCGCTCCGCCGAGGAGGGCGGTGCCGCGCCTGCGCCTTAGCGCGGGCGGTCGGCCGAG
|
CGGCACTGCGCCTGCGCGGACGGCGCTGCGCGGGGAGGGACGGGGCTTGCGCCTTGGT
|
GGGACCTCCCCAGGGGCGGGGCTCGGGGGGCTCCACGCCAAGGGTGGGCTGCGCCTAC
|
GCCCTTGACTCAGCTTTCCCTTTTGGGCAATTAGCAATGCAGGATGCCCCGCCCTCTC
|
GGGAGGCCACGCCCAAGGGCGCGACGAAGGAAGGAGCCACATCCCCAACTTGAGGCCA
|
CGCCCCCAGCACCTAGGGGGCATTTTGAGCTGGGATGGGGGAAACCTCGTCCCTATGG
|
AGCAGGCCACATCCCGGGGCTCTGGTACCGGGAGGCACCACCTCATGTCCCCTGGAAA
|
AGCCATAAGATGGGACCCAGACCCCTGGGACCCCAGACCAATTGCCAAGTATGGAAAT
|
CTCAGCTCCCTCGAGGGGGGGCCCTGGGCAAGGGGTAGGGCTCTCTGGAGCGCCCCTC
|
TAGGTGGCCTGGGGACTGGAGGGACCAGGATGCTGGTTGGAGGGCCCCGGAATACCGG
|
AGTCCCTTTAGATATTTGTGCAAAAAATAAATGGGCCGAGGGGGGACGATGGGATTTC
|
AAAAGCACATGCGCCCTTGGGCGCCCAAACCCTGGGGGCCGAGGGGACGGCTCTGGTT
|
CCCCACCCTGCCCCTACTTCCCTTTGGGAGTTTGCCTCTCCCTCTCCCCCAACAAACC
|
CAGTCCTCATATCATAGAGTTCAACACACCCATTTGACAGATGGCAAAACTGAGGCTT
|
AAAGAGCTGCTTCAGACTTGGCCAAGGTTCCAGGTGCCATACCCTCTGTGCCCCTCCC
|
TTAGGCCTGTGTGCCCCATGGAAGGGTGGGCTGAGATCGGGATGACCTGACACAGCTC
|
CCTATTGCTGCTAATTCCCCCTCGGCCTCCTCCAAGGGGTGGAAATTCCAGGCCAAGA
|
CCCCTACTTCGCCTTTCCTTCTCCGGCTGCCAAGCAGGACCTTTGCCCTCAGCCCTTT
|
CTCCTGGGATCTCCATGGGGGATGCCATGAGGGCCTCCCACCACAAAAGAGAATTTGG
|
GATCCCCTGGTCCCAGGTTTCTCCATCCCTTCTTCCTTTTCCAGAATTTTCCAAATAG
|
GAAAGAACAGAAGGAGACCAGAAACTCTAGGGGGGAGAAAGAGAATGAGAGAAAGAGA
|
ATGAGAGAGAGAGAAACACAAACACAGTGACACAGTGAGAGCTTAGTCTCCAAGAGCC
|
TATTCATTGATTCAAACACCCAAGCCACAGGATACCTCAGATGGCCCTCTTGCCAGCT
|
GGAAGCTCTTTCTCCAATGAGCAAAGTTACAGTGACCTGGCTGGAGTTACCTGGTGCA
|
CATAGGACCTTAGGGGAAAGTTCAGCGTGGACTACACTTGCTCTGGGATCTGCTTTTC
|
CACATGTGTCTATCGCACGCCTTTTTCTGCTGGATTGGGAAGGACAAGATTTTGCTGT
|
GCTAGGGAGAAATGAAAACGGGGTGAGCTGAGTAGCTGGGTTTCTGGAGGATAGAACA
|
TCAGATGGGGAGGCTTTCCGAGGTGAAGAATGAGAGGGAACCACTTACTAGAGAGAAA
|
AGAGCTCCAGGCCTGGGCAACAGCACGTGCGAAGGCCAGCAGAGAAGAACTGTTGAAA
|
CAACGAGAAGGGTGGCACGGCTGGAGCTGAGCCAGCAAGGGGGATCGTGAGGAGCCTT
|
GGGGTTGGGGAGATCTGCAGAAGCATCAGACCAGGCAGGGCCTCGTACGCAGTCCTGA
|
GGAGTTTTACTTTTATTCTAAGACAGTTGGGGAGCTCCAGGAGCTGTTTTAAGTTGGG
|
GAGAGACTGGATTCCAGCCTGCAAAAGCTGTTTTGTGAAGACTAAAACCAGTGAGGAG
|
AGGTGGAGGTTGCTTTGGGGACACTGAAATGGATTCTTGGAAAGATTCTGAAGGCTGT
|
GTTGAAAAGACACCTATAGCTGTGGGGACATGACTATAATCCCAGCATTTGGGGAGAC
|
CGAGGCTGGCAGATCACTTAAGGTCAGGAGTTTGAGACCAGCCTGGCCAACATGGCGA
|
AACCCCATCTCTGCTAAAAATACAAAAATTAGCTGGGTCCAGTGGTGCATGCCTGTAG
|
TCCCAGCTACTCAGGACACTGACGCGGGACAATTGTTTGAACCCTCGAGGCAGAGGTT
|
GTAGTGAGTCGTGATCACACAACTGCACTCCAGCCTGGGCAACAGAACAATACTCCAT
|
TCCCTCCCCTCTACCCCACC
|
ORF Start: at 3ORF Stop: TAG at 5142
SEQ ID NO: 1941713 aaMW at 194170.2kD
NOV74a, KKAKFDGAQEKFNTYVTLKVQNVKSTTIAVRGSQPSWEQDFMFEINRLDLGLTVEVWN
CG94302-01 Protein Sequence
KGLIWDTMVGTVWIPLRTIRQSNEEGPGEWLTLDSQVIMADSEICGTKDPTFHRILLD
|
TRFELPLDIPEEEARYWAKKLEQLNAMRDQDEYSFQDEQDKPLPVPSNQCCKYFPYRL
|
SLPDDDPDSAVDDRDSDYRSETSNSIPPPYYTTSQPNASVHQYSVRPPPLGSRESYSD
|
SMHSYEEFSEPQALSPTGSSRYASSGELSQGSSQLSEDFDPDEHSLQGSDMEDERDRD
|
SYHSCHSSVSYHKDSPRWDQDEEELEEDLEDFLEEEELPEDEEELEEEEEEVPDDLGS
|
YAQREDVAVAEPKDFKRISLPPAAPGKEDKAPVAPTEAPDMAKVAPKPATPDKVPAAE
|
QIPEAEPPKDEESFRPREDEEGQEGQDSMSRAKANWLRAFNKVRMQLQEARGEGEMSK
|
SLWFKGGPGGGLIIIDSMPDIRKRKPIPLVSDLVLSLVQSRKAGITSALASSTLNNEE
|
LKNHVYKKTLQALIYPISCTTPHNFEVWTATTPTYCYECEGLLWGIARQGMRCTECGV
|
KCHEKCQDLLNADCLQRAAEKSSKHGAEDRTQNIIMVLKDRMKIRERNKPEIFELIQE
|
IFAVTKTAHTQQMKAVKQSVLDGTSKWSAKISITVVCAQGLQAKDKTGSSDPYVTVQV
|
GKTKKRTKTIYGNLNPVWEENFHFECHNSSDRIKVRVWDEDDDIKSRVKQRFKRESDD
|
FLGQTIIEVRTLSGEMDVWYNLDKRTDKSAVSGAIRLHISVELKGEEKVAPYHVQYTC
|
LHENLFHFVTDVQNNGVVKIPDAKGDDAWKVYYDETAQEIVDEFAMRYGVESIYQAMT
|
HFACLSSKYMCPGVPAVMSTLLANINAYYAHTTASTNVSASDRFAASNFGKERFVKLL
|
DQLHNSLRIDLSMYRNNFPASSPERLQDLKSTVDLLTSITFFRMKVQELQSPPRASQV
|
VKDCVKACLNSTYEYIFNNCHELYSREYQTDPAKKGEVPPEEQGPSIKNLDFWSKLIT
|
LIVSIIEEDKNSYTPCLNQFPQELNVGKISAEVMWNLFAQDMKYAMEEHDKHRLCKSA
|
DYMNLHFKVKWLYNEYVTELPSFKDRVPEYPAWFEPFVIQWLDENEEVSRDFLHGALE
|
RDKKDGFQQTSEHALFSCSVVDVFSQLNQSFEIIKKLECPDPQIVGHYMRRFAKTISM
|
VLLQYADIISKDFASYCSKEKEKVPCILMNNTQQLRVQLEKMFEAMGGKELDAEASDI
|
LKELQVKLNNVLDELSRVFATSFQPHIEECVKQMGDILSQVKGTGNVPASACSSVAQD
|
ADNVLQPIMDLLDSNLTLFAKICEKTVLKRVLKELWKLVMNTMEKTIVLPPLTDQTIM
|
GNLLRKHGKGLEKGRVKLPSHSDGTQMIFNAAKELGQLSKLKDHMVREEAKSLTPKQC
|
AVVELALDTIKQYFHAGGVGLKKTFLEKSPDLQSLRYALSLYTQATDLLIKTFVQTQS
|
AQVHGGKGTRFTLSEDIYPEKGTGVEDPVGEVSVHVELFTHPGTGEHKVTVKVVAAND
|
LKWQTSGIFRPFIEVNIIGPQLSDKKRKFATKSKNNSWAPKYNESFQFTLSADAGPEC
|
YELQVCVKDYCFAREDRTVGLAVLQLRELAQRGSAACWLPLGRRIHMDDTGLTVLRIL
|
SQRSNDEVAKEFVKLKSDTRSAEEGGAAPAP
|
[0701] Further analysis of the NOV74a protein yielded the following properties shown in
388TABLE 74B
|
|
Protein Sequence Properties NOV74a
|
|
PSort0.7000 probability located in nucleus;
0.3000 probability located in microbody
analysis:(peroxisome); 0.1000 probability located
in mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0702] A search of the NOV74a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 74C.
389TABLE 74C
|
|
Geneseq Results for NOV74a
NOV74aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAW83431Rat Munc13-1 - Rattus sp, 1763 aa. 1 . . . 17131635/1727(94%)0.0
[JP10313866-A, 02 DEC. 1998] 37 . . . 17631665/1727(95%)
AAY27134Human munc13 (Hmunc13) 1 . . . 17071242/1717(72%)0.0
polypeptide - Homo sapiens, 1591 9 . . . 15901390/1717(80%)
aa. [WO9931134-A1, 24 JUN.
1999]
AAW83428Munc13-1-interacting domain of 801 . . . 1411600/611(98%)0.0
Doc2-alpha - Rattus sp, 611 aa. 1 . . . 611605/611(98%)
[JP10313866-A, 02 DEC. 1998]
AAB47279PN7098 - Homo sapiens, 1230 aa.119 . . . 677226/589(38%)4e−88
[WO200140794-A1, 07 JUN. 2001] 684 . . . 1230305/589(51%)
ABG02226Novel human diagnostic protein406 . . . 540131/135(97%)3e−69
#2217 - Homo sapiens, 146 aa. 12 . . . 146133/135(98%)
[WO200175067-A2, 11 OCT.
2001]
|
[0703] In a BLAST search of public sequence datbases, the NOV74a protein was found to have homology to the proteins shown in the BLASTP data in Table 74D.
390TABLE 74D
|
|
Public BLASTP Results for NOV74a
NOV74aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q62768MUNC13-1 - Rattus norvegicus1 . . . 17131635/1727 (94%)0.0
(Rat), 1735 aa.9 . . . 17351665/1727 (95%)
Q9WV40MUNC13-2 PROTEIN - Rattus1 . . . 17071236/1720 (71%)0.0
norvegicus (Rat), 1622 aa.9 . . . 16211393/1720 (80%)
O14795MUNC13 - Homo sapiens1 . . . 17071242/1717 (72%)0.0
(Human), 1591 aa.9 . . . 15901390/1717 (80%)
Q9Z1N9RENAL MUNC13 - Mus1 . . . 17071218/1719 (70%)0.0
musculus (Mouse), 1591 aa.9 . . . 15901375/1719 (79%)
Q62769MUNC13-2 - Rattus norvegicus459 . . . 1707 1016/1250 (81%)0.0
(Rat), 1985 aa.767 . . . 1984 1121/1250 (89%)
|
[0704] PFam analysis predicts that the NOV74a protein contains the domains shown in the Table 74E.
391TABLE 74E
|
|
Domain Analysis of NOV74a
Identities/
Similarities
NOV74a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
DAG_PE-bind:545 . . . 594 20/51 (39%)1.3e−19
domain 1 of 1 44/51 (86%)
C2: domain 1 of 2669 . . . 76037/106 (35%)3.3e−21
76/106 (72%)
sigma70: domain 1 of 11203 . . . 137132/242 (13%)2.6
108/242 (45%)
SPX: domain 1 of 11211 . . . 137530/351 (9%) 0.96
105/351 (30%)
NTF2: domain 1 of 11499 . . . 161429/135 (21%)2.7
78/135 (58%)
C2: domain 2 of 21557 . . . 1647 24/97 (25%)0.00069
58/97 (60%)
|
Example 75
[0705] The NOV75 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 75A.
392TABLE 75A
|
|
NOV75 Sequence Analysis
|
|
SEQ ID NO: 1951746 bp
NOV75a,ATGGTGGTGGTGCCTTTCACAATTCCATTCGATTCATCAGTCTGGCTGCTGCAGAAGT
CG94356-01 DNA Sequence
TGAACATCAAGGTCCAAGGAGGGCCTTCTGCTGAAGGGCCACAGAGGAACACCAGGCT
|
GGGATGGATTCAGGGCAAGCAAGTCACTGTGCTGGGAAGCCCTGTGCCTGTGAACGTG
|
TTCCTCGGAGTCCCCTTTGCTGCTCCCCCGCTGGGATCCCTGCGATTTACGAACCCGC
|
AGCCTGCATCGCCCTGGGATAACTTGCGAGAAGCCACCTCCTACCCTAATTTGTGCCT
|
CCAGAACTCAGAGTGGCTGCTCTTAGATCAACATATGCTCAAGGTGCATTACCCGAAA
|
TTCGGAGTGTCAGAAGACTGCCTCTACCTGAACATCTATGCGCCTGCCCACGCCGATA
|
CAGGCTCCAAGCTCCCCGTCTTGGTGTGGTTCCCAGGAGGTGCCTTCAAGACTGGCTC
|
AGCCTCCATCTTTGATGGGTCCGCCCTGGCTGCCTATGAGGACGTGCTGGTTGTGGTC
|
GTCCAGTACCGGCTAGGAATATTTGGTTTCTTCACCACATGGGATCAGCATGCTCCGG
|
GGAACTGGGCCTTCAAGGACCAGGTGGCTGCTCTGTCCTGGGTCCAGAAGAACATCGA
|
GTTCTTCGGTGGGGACCCCAGCTCTGTGACCATCTTTGGCGAGTCCGCGGGAGCCATA
|
AGTGTTTCTAGTCTTATACTGTCTCCCATGGCCAAAGGCTTATTCCACAAAGCCATCA
|
TGGAGAGTGGGGTGGCCATCATCCCTTACCTGGAGGCCCATGATTATGAGAAGAGTGA
|
GGACCTGCAGGTGGTTGCACATTTCTGTGGTAACAATGCGTCAGACTCTGAGGCCCTG
|
CTGAGGTGCCTGAGGACAAAACCCTCCAAGGAGCTGCTGACCCTCAGCCAGAAAACAA
|
AGTCTTTCACTCGAGTGGTTGATGGTGCTTTCTTTCCTAATGAGCCTCTAGATCTATT
|
GTCTCAGAAAGCATTTAAAGCAATTCCTTCCATCATCGGAGTCAATAACCACGAGTGT
|
GGCTTCCTGCTGCCTATGGTAAGAATTCTGGCTGTCCATACTGCCACTCCCTCAAACC
|
GTGATGCAGCTTTGGCTTCAACAGCTGGGCATTTCCACAGAAGACATATCCCGCCTCA
|
GTATTTGCACCTTGTGGCTAATGAATACTTCCATGACAAGCACTCCCTGACTGAAATC
|
CGAGACAGTCTTCTGGACTTGCTTGGAGATGTGTTCTTTGTGGTCCCTGCACTGATCA
|
CAGCTCGATATCACAGAGATGCTGGTGCACCTGTCTACTTCTATGAGTTTCGGCACCG
|
GCCTCAGTGCTTTGAAGACACGAAGCCAGCTTTTGTCAAAGCCGACCACGCTGATGAA
|
GTCCGCTTTGTGTTCGGTGGTGCCTTCCTGAAGGGGGACATTGTTATGTTCGAAGGAG
|
CCACGGAGGAGGAGAAGTTACTGAGCCGGAAGATGATGAAATACTGGGCTACCTTTGC
|
TCGAACCGGGAATCCTAATGGGAACGACCTGTCTCTGTGGCCAGCTTATAATCTGACT
|
GAGCAGTACCTCCAGCTGGACTTGAACATGAGCCTCGGACAGAGACTCAAAGAACCGC
|
GGAGAGATGTGTGGGTGACGGGGTATCCTCAGCCATGGAAAGCTGCCATCATCCAGAA
|
TAAAAAACCTAGAAGTCAAATTCTAGGCATCAAGGGTCGGATCAGCAATGCCAAGAAG
|
AAATGA
|
ORF Start: ATG at 1ORF Stop: TGA at 1744
SEQ ID NO: 196581 aaMWat64717.6kD
NOV75a,MVVVPFTIPFDSSVWLLQKLNIKVQGGPSAEGPQRNTRLGWIQGKQVTVLGSPVPVNV
CG94356-01 Protein Sequence
FLGVPFAAPPLGSLRFTNPQPASPWDNLREATSYPNLCLQNSEWLLLDQHMLKVHYPK
|
FGVSEDCLYLNIYAPAHADTGSKLPVLVWFPGGAFKTGSASIFDGSALAAYEDVLVVV
|
VQYRLGIFGFFTTWDQHAPGNWAFKDQVAALSWVQKNIEFFGGDPSSVTIFGESAGAI
|
SVSSLILSPMAKGLFHKAIMESGVAIIPYLEAHDYEKSEDLQVVAHFCGNNASDSEAL
|
LRCLRTKPSKELLTLSQKTKSFTRVVDGAFFPNEPLDLLSQKAFKAIPSIIGVNNHEC
|
GFLLPMVRILAVHTATPSNRDAALASTAGHFHRRHIPPQYLHLVANEYFHDKHSLTEI
|
RDSLLDLLGDVFFVVPALITARYHRDAGAPVYFYEFRHRPQCFEDTKPAFVKADHADE
|
VRFVFGGAFLKGDIVMFEGATEEEKLLSRKMMKYWATFARTGNPNGNDLSLWPAYNLT
|
EQYLQLDLNMSLGQRLKEPRRDVWVTGYPQPWKAAIIQNKKPRSQILGIKGRISNAKK
|
K
|
[0706] Further analysis of the NOV75a protein yielded the following properties shown in
393TABLE 75B
|
|
Protein Sequence Properties NOV75a
|
|
PSort0.6000 probability located in plasma membrane:
analysis:0.4109 probability located in mitochondrial inner membrane;
0.4000 probability located in Golgi body; 0.3000
probability located in endoplasmic reticulum (membrane)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0707] A search of the NOV75a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 75C.
394TABLE 75C
|
|
Geneseq Results for NOV75a
NOV75aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAB58981Breast and ovarian cancer associated15 . . . 552252/544 (46%)e−133
antigen protein sequence SEQ ID 689 - 3 . . . 534342/544 (62%)
Homo sapiens, 549 aa.
[WO200055173-A1, 21-SEP-2000]
AAB31700Protein encoded by an intestinal20 . . . 552252/538 (46%)e−132
carboxylesterase (iCE) cDNA - Homo20 . . . 544341/538 (62%)
sapiens, 559 aa. [WO200100784-A2,
04-JAN-2001]
ABG10273Novel human diagnostic protein20 . . . 552249/540 (46%)e−126
#10264 - Homo sapiens, 583 aa.42 . . . 568337/540 (62%)
[WO200175067-A2, 11-OCT-2001]
ABG10273Novel human diagnostic protein20 . . . 552249/540 (46%)e−126
#10264 - Homo sapiens, 583 aa.42 . . . 568337/540 (62%)
[WO200175067-A2, 11-OCT-2001]
AAE04101Human gene 2 encoded secreted27 . . . 554242/536 (45%)e−125
protein HWLFE89, SEQ ID NO: 87 -28 . . . 558331/536 (61%)
Homo sapiens, 571 aa.
[WO200134643-A1, 17-MAY-2001]
|
[0708] In a BLAST search of public sequence datbases, the NOV75a protein was found to have homology to the proteins shown in the BLASTP data in Table 75D.
395TABLE 75D
|
|
Public BLASTP Results for NOV75a
NOV75aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96DN9CDNA FLJ31547 FIS, CLONE27 . . . 551446/525 (84%)0.0
NT2RI2001010, WEAKLY SIMILAR25 . . . 494449/525 (84%)
TO FATTY ACYL-COA HYDROLASE
PRECURSOR, MEDIUM CHAIN (EC
3.1.2.14) - Homo sapiens (Human), 525
aa.
Q95KH3HYPOTHETICAL 41.0 KDA PROTEIN -238 . . . 551 274/314 (87%)e−155
Macaca fascicularis (Crab eating17 . . . 324282/314 (89%)
macaque) (Cynomolgus monkey), 361 aa.
O35533CARBOXYLESTERASE PRECURSOR16 . . . 552262/540 (48%)e−138
(EC 3.1.1.1) (ALI-ESTERASE) (B-18 . . . 544343/540 (63%)
ESTERASE) (MONOBUTYRASE)
(COCAINE ESTERASE) (PROCAINE
ESTERASE) (METHYLBUTYRASE) -
Mesocricetus auratus (Golden hamster),
559 aa.
Q91WG0SIMILAR TO CARBOXYLESTERASE20 . . . 552267/542 (49%)e−137
2 (INTESTINE, LIVER) - Mus musculus20 . . . 546337/542 (61%)
(Mouse), 561 aa.
O70177CARBOXYLESTERASE PRECURSOR20 . . . 552268/541 (49%)e−136
(EC 3.1.1.1) - Rattus norvegicus (Rat),20 . . . 546340/541 (62%)
561 aa.
|
[0709] PFam analysis predicts that the NOV75a protein contains the domains shown in the
396TABLE 75E
|
|
Domain Analysis of NOV75a
Identities/
NOV75aSimilarities
Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
COesterase: domain 1 of 140 . . . 546204/578 (35%)3.1e−157
388/578 (67%)
|
Example 76
[0710] The NOV76 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 76A.
397TABLE 76A
|
|
NOV76 Sequence Analysis
|
|
SEQ ID NO: 1972025 bp
NOV76aCTCAAGTTGACCTGCCACTTCACCCAAGGAGGGTAAGGAGCTGGGACCAGAGTCGCCA
CG94421-01 DNA Sequence
ATTCCTGCCTTCTAGATCCTGGGCAAGCTCTTGCCTCTGACAGTGTATTTGCCAACAT
|
GGCACCCAAAAAGAAGATTGTCAAAAAGAACAAAGGAGATATCAATGAGATGACTATA
|
ATCGTAGAAGATAGCCCCCTAAACAAACTGAATGCTTTCAATGGGCTCCTAGACGCAG
|
CCAATGGCCTTACCTGCATPTCTTCTGAACTAACAGATGCTTCTTATCCCCCCAACCT
|
CTTGGAAGGTTTAAGTAAAATGCGGCAGGAGAACTTCTTATGTGACTTAGTCATTGGT
|
ACCAAAACCAAATCCTTTGATGTTCATAAGTCAGTCATGGCTTCATCCAGTGAGTATT
|
TTTACAACATCCTAAAAAAAGACCCGTCAATTCAGAGCGTGGATCTCAATGATATCTC
|
ACCACTAGGCCTGGCCACTGTCATTGCATATGCCTACACTGGAAAGCTCACTCTCTCC
|
TTGTATACAATAGGAAGCATTATTTCTGCTGCTCTTTATCTTCAGATCCATACTCTTA
|
TAAAGATGTGCAGTGATTTTCTGATACGGGAGATGAGTGTTGAGAATTGCATGTATGT
|
TGTTAATATTGCTGAAACATACTCCCTAAAAAATGCAAAAGCAGCAGCCCAGAAATTT
|
ATTCGGGATAACTTCCTTGAATTTGCAGAATCGGATCAGTTTATGAAACTTACATTTG
|
AACAAATTAATGAACTTCTTATAGATGATGACTTACAGTTGCCTTCTGAGATAGTAGC
|
ATTCCAGATTGCAATGAAATGGTTAGAATTTGACCAAAAGAGAGTAAAATACGCTGCA
|
GATCTTTTGAGCAATATTCGCTTTGGTACCATCTCTGCACAAGACCTGGTCAATTATG
|
TTCAATCCGTACCAAGAATGATGCAAGATGCTGATTGTCACAGACTTCTCGTAGATGC
|
TATGAACTACCACTTGCTTCCATATCATCAAAACACATTCCAATCTAGGCGAACAAGA
|
ATCCGAGGTGGCTGCCGAGTCCTCGTCACTGTTGGGGGACGCCCAGGCCTTACTGAGA
|
AGTCCCTTAGCAGAGACATCCTGTATAGACACCCTGAAAATGGATGGACCAAGCTTAC
|
GGAAATGCCAGCCAAAAGTTTTAATCAGTCTGTGGCTGTGATCGATGGATTTCTTTAT
|
GTAGCCGGTGGTGAAGACCAGAATGATGCAAGAAATCAAGCCAAGCATGCAGTCAGCA
|
ATTTCTGCAGATACGATCCCCGCTTCAACACCTGGATACACCTGGCCAGCATGAACCA
|
GAAGCGCACGCACTTCAGCCTGAGCGTGTTCAACGGGCTCGTGTACGCCGCGGGCGGC
|
CGCAACGCAGAAGGAAGCCTGGCCTCGCTGGAGTGCTACGTGCCCTCCACCAATCAGT
|
GGCAGCCGAAGACGCCCCTGGAGGTGGCGCGCTGCTGCCACGCTAGCGCGGTCGCCGA
|
CGGCCCCGTGCTGGTGACCGGAGGCTACATCGCCAACGCCTACTCGCGCTCTGTGTGC
|
GCCTACGACCCGGCCAGCGACTCGTGGCAGGAGCTGCCGAACCTCAGCACACCCCGGG
|
GCTGGCACTGCGCGGTCACGCTGAGCGACAGAGTGTACGTGATGGGCGGCAGCCAGCT
|
GGGGCCGCGCGGGGAGCGCGTGGACGTGCTCACCGTCGAGTGCTACAGCCCCGCCACC
|
GGCCAGTGGAGCTACGCGGCGCCGCTGCAGGTGGGAGTGAGCACTGCGGGCGTCTCGG
|
CGCTGCATGGCCGCGCCTACCTGGTGGGGGGCTGGAACGAGGGCGAGAAGAAGTACAA
|
GAAGTGCATCCAGTGCTTCAGCCCCGAGCTCAACGAGTGGACGGAGGACCACGACCTA
|
CCCCAGGCCACTGTCGGCGTGTCCTGCTGCACCCTCTCCATGCCCAACAACGTGACTC
|
GGGAATCCCGGGCCAGTTCGGTATCTTCTGTGCCAGTCAGTATCTGAGCCCAG
|
ORF Start: ATG at 115ORF Stop: TGA at 2017
SEQ ID NO: 19863 aaMW at 70259.3kD
NOV76a,MAPKKKIVKKNKGDINEMTIIVEDSPLNKLNALNGLLEGGNGLSCISSELTDASYGPN
CG94421-01 Protein Sequence
LLEGLSKMRQENFLCDLVIGTKTKSFDVHKSVMASCSEYFYNILKKDPSIQRVDLNDI
|
SPLGLATVIAYAYTGKLTLSLYTIGSIISAAVYLQIHTLIKMCSDFLIREMSVENCMY
|
VVNIAETYSLKNAKAAAQKFIRDNFLEFAESDQFMKLTFEQINELLIDDDLQLPSEIV
|
AFQIAMKWLEFDQKRVKYAADLLSNIRFGTISAQDLVNYVQSVPRMMQDADCHRLLVD
|
AMNYHLLPYHQNTLQSRRTRIRGGCRVLVTVGGRPGLTEKSLSRDILYRDPENGWSKL
|
TEMPAKSFNQCVAVMDGFLYVAGGEDQNDARNQAKHAVSNFCRYDPRFNTWIHLASMN
|
QKRTHFSLSVFNGLVYAAGGRNAEGSLASLECYVPSTNQWQPKTPLEVARCCHASAVA
|
DGRVLVTGGYIANAYSRSVCAYDPASDSWQELPNLSTPRGWHCAVTLSDRVYVMGGSQ
|
LGPRGERVDVLTVECYSPATGQWSYAAPLQVGVSTAGVSALHGRAYLVGGWNEGEKKY
|
KKCIQCFSPELNEWTEDDELPEATVGVSCCTLSMPNNVTRESRASSVSSVPVSI
|
[0711] Further analysis of the NOV76a protein yielded the following properties shown in
398TABLE 76B
|
|
Protein Sequence Properties NOV76a
|
|
PSort0.3000 probability located in microbody (peroxisome);
analysis:0.3000 probability located in nucleus;
0.1000 probability located in mitochondrial matrix space;
0.1000 probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0712] A search of the NOV76a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 76C.
399TABLE 76C
|
|
Geneseq Results for NOV76a
NOV76aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
ABG10610Novel human diagnostic protein38 . . . 631209/607 (34%)e−104
#10601 - Homo sapiens, 604 aa. 3 . . . 604329/607 (53%)
[WO200175067-A2, 11-OCT-2001]
ABG10610Novel human diagnostic protein38 . . . 631209/607 (34%)e−104
#10601 - Homo sapiens, 604 aa. 3 . . . 604329/607 (53%)
[WO200175067-A2, 11-OCT-2001]
AAU28187Novel human secretory protein, Seq34 . . . 620209/595 (35%)e−102
ID No: 356 - Homo sapiens, 615 aa.27 . . . 614336/595 (56%)
[WO200166689-A2, 13-SEP-2001]
AAB93480Human protein sequence SEQ ID34 . . . 620209/595 (35%)e−102
NO: 12768 - Homo sapiens, 615 aa.27 . . . 614336/595 (56%)
[EP1074617-A2, 07-FEB-2001]
AAB95123Human protein sequence SEQ ID53 . . . 614184/572 (32%)2e−84
NO: 17120 - Homo sapiens, 616 aa.26 . . . 595300/572 (52%)
[EP1074617-A2, 07-FEB-2001]
|
[0713] In a BLAST search of public sequence datbases, the NOV76a protein was found to have homology to the proteins shown in the BLASTP data in Table 76D.
400TABLE 76D
|
|
Public BLASTP Results for NOV76a
NOV76aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q9H511BA345L23.2 (NOVEL PROTEIN 1 . . . 634633/634 (99%)0.0
WITH BTB/POZ (BROAD COMPLEX 1 . . . 634634/634 (99%)
TRAMTRACK BRIC-A-BRAC/POX
VIRUS AND ZINC FINGER)
DOMAIN) - Homo sapiens (Human),
634 aa.
CAD28475HYPOTHETICAL 69.4 KDA40 . . . 628216/613 (35%)e−106
PROTEIN - Homo sapiens (Human), 6 . . . 614335/613 (54%)
617 aa.
Q9P2J3Hypothetical protein KIAA1354 -40 . . . 628216/613 (35%)e−106
Homo sapiens (Human), 632 aa21 . . . 629335/613 (54%)
(fragment).
Q9DBY71200009K10RIK PROTEIN - Mus51 . . . 631207/590 (35%)e−106
musculus (Mouse), 638 aa.53 . . . 638324/590 (54%)
Q9P2N7Hypothetical protein KIAA1309 -39 . . . 631210/613 (34%)e−105
Homo sapiens (Human), 639 aa32 . . . 639333/613 (54%)
(fragment).
|
[0714] PFam analysis predicts that the NOV76a protein contains the domains shown in the Table 76E.
401TABLE 76E
|
|
Domain Analysis of NOV76a
Identities/
NOV76aSimilarities forExpect
Pfam DomainMatch Regionthe Matched RegionValue
|
BTB: domain 1 of 1 57 . . . 16733/143 (23%) 1.4e−17
85/143 (59%)
Kelch: domain 1 of 5354 . . . 40615/53 (28%)1.1e−05
37/53 (70%)
Kelch: domain 2 of 5408 . . . 45310/47 (21%)8.2e−07
36/47 (77%)
Kelch: domain 3 of 5455 . . . 50012/47 (26%) 3e−07
36/47 (77%)
Kelch: domain 4 of 5502 . . . 55212/51 (24%)1.2e−06
39/51 (76%)
Kelch: domain 5 of 5554 . . . 60113/48 (27%)0.16
32/48 (67%)
|
Example 77
[0715] The NOV77 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 77A.
402TABLE 77A
|
|
NOV77 Sequence Analysis
|
|
SEQ ID NO:1992880 bp
NOV77A,ACGAGGGCGGGGCGCGCCGGCGGGGTGACCTCACGGCCGCGCGCGGCGTCGGCGGAGC
|
CG94465-01 DNA SequenceCTCACTTTGAACCCAGTTGGCGGGAGTGGCTGCTCGCGGAGGGGCAGTGTCTGCGGGG
|
CCGCTGTATGCTGTCCAGCGATGGATCCCACCGCGGGAAGCAAGAAGGAGCCTGGAGG
|
AGGCGCGGCGACTGAGGAGGGCGTGAATAGGATCGCAGTGCCAAAACCGCCCTCCATT
|
GAGGAATTCAGCATAGTGAAGCCCATTAGCCGGGGCGCCTTCGGGAAAGTGTATCTGG
|
GGCAGAAAGGCGGCAAATTGTATGCAGTAAAGGTAGTTAAAAAAGCAGACATGATCAA
|
CAAAAATATGACTCATCAGGTCCAAGCTGACAGAGATGCACTGGCACTAAGCAAAAGC
|
CCATTCATTGTCCATTTGTATTATTCACTGCAGTCTGCAAACAATGTCTACTTGGTGA
|
TGGAATATCTTATTGGGGGAGATGTCAAGTCTCTCCTACATATATATGGTTATTTTGA
|
TGAACACATGGCTGTGAAATATATTTCTGAAGTAGCACTGGCTCTAGACTACCTTCAC
|
AGACATGGAATCATCCACAGGGACTTGAAACCGGACAATATGCTTATTTCTAATGAGG
|
GTCATATTAAACTGACGGATTTTGGCCTTTCAAAAGTTACTTTGAATAGAGATATTAA
|
TATGATCGATATCCTTACAACACCATCAATGGCAAAACCTAGACAAGATTATTCAAGA
|
ACCCCAGGACAAGTGTTATCGCTTATCAGCTCGTTGGGATTTAACACACCAATTGCAG
|
AAAAAAATCAAGACCCTGCAAACATCCTTTCAGCCTGTCTGTCTGAAACATCACAGCT
|
TTCTCAAGGACTCGTATGCCCTATGTCTGTAGATCAAAAGGACACTACGCCTTATTCT
|
AGCAAATTACTAAAATCATGTGAAACAGTTGCCTCCAACCCAGGAATGCCTGTGAAGT
|
GTCTAACTTCTAATTTACTCCAGTCTAGGAAAAGGCTGGCCACATCCAGTGCCAGTAG
|
TCAATCCCACACCTTCATATCCAGTGTGGAATCAGAATGCCACAGCAGTCCCAAATGG
|
GAAAAAGATTGCCAGGAAAGTGATGAAGCATTGGGCCCAACAATGATGAGTTGGAATG
|
CAGTTGAAAAGTTATGCGCAAAATCTGCAAATGCCATTGAGACGAAAGGTTTCAATAA
|
AAAGGATCTGGAGTTAGCTCTTTCTCCCATTCATAACAGCAGTGCCCTTCCCACCACT
|
GGACGCTCTTGTGTAAACCTTGCTAAAAAATGCTTCTCTGGGGAAGTTTCTTCGCAAG
|
CAGTAGAACTGGATGTAAATAATATAAATATGGACACTGACACAAGTCAGTTAGGTTT
|
CCATCAGTCAAATCAGTGGGCTGTGGATTCTGGTGGGATATCTGAAGAGCACCTTGGG
|
AAAAGAAGTTTAAAAAGAAATTTTGAGTTGGTTGACTCCAGTCCTTGTAAAAAAATTA
|
TACAGAATAAAAAAACTTGTGTAGAGTATAAGCATAACGAAATGACAAATTGTTATAC
|
AAATCAAAATACAGGCTTAACAGTTGAAGTGCAGGACCTTAAGCTATCAGTGCACAAA
|
AGTCAACAAAATGACTCTGCTAATAAGGAGAACATTGTCAATTCTTTTACTGATAAAC
|
AACAAACACCAGAAAAATTACCTATACCAATGATAGCAAAAAACCTTATGTGTCAACT
|
CGATGAAGACTGTGAAAAGAATAGTAAGAGGGACTACTTAAGTTCTAGTTTTCTATGT
|
TCTGATGATGATAGAGCTTCTAAAAATATTTCTATGAACTCTGATTCATCTTTTCCTG
|
GAATTTCTATAATGGAAAGTCCATTAGAAAGTCAGCCCTTAGATTCAGATACAAGCAT
|
CAAAGAATCCTCTTTTCAAGAATCAAATATTGAAGATCCACTTATTGTAACACCAGAT
|
TGCCAAGAAAAGACCTCACCAAAAGGTGTCGAGAACCCTGCTGTACAAGAGAGTAACC
|
AAAAAATGTTAGGTCCTCCTTTGGAGGTGCTGAAAACGTTAGCCTCTAAAAGAAATGC
|
TGTTGCTTTTCGAAGTTTTAACAGTCATATTAATGCATCCAATAACTCAGAACCATCC
|
AGAATGAACATGACTTCTTTAGATGCAATCGATATTTCGTGTGCCTACAGTGGTTCAT
|
ATCCCATGGCTATAACCCCTACTCAAAAAAGAAGATCCTGTATGCCACATCAGCAGAC
|
CCCAAATCAGATCAAGTCGGGAACTCCATACCGAACTCCGAAGAGTGTGAGAAGAGGG
|
GTGGCCCCCGTTGATGATGCGCCAATTCTAGGAACCCCAGACTACCTTGCACCTGACC
|
TGTTACTAGGCAGGGCCCATGGTAAGGCATGCATGCCTGCGGTAGACTGGTGGGCACT
|
TGGAGTTTGCTTGTTTGAATTTCTAACAGGAATTCCCCCTTTCAATGATGAAACACCA
|
CAACAAGTATTCCAGAATATTCTGAAAAGAGATATCCCTTGGCCAGAAGGTGAAGAAA
|
AGTTATCTGATAATGCTCAAAGTGCAGTAGAAATACTTTTAACCATTGATGATACAAA
|
GAGAGCTGGAATGAAACGTCATCCTCTCTTCAGTGATGTGGACTGGGAAAATCTGCAG
|
CATCAGACTATGCCTTTCATCCCCCAGCCAGATGATGAAACAGATACCTCCTATTTTG
|
AAGCCAGGAATACTGCTCAGCACCTGACTGTATCTGGATTTAGTCTGTAGCACAAAAA
|
TTTTCCTTTTAGTCTAGCCTTGTGTTATAGAATGAACTTGCATAATTATATACTCCTT
|
AATACTAGATTGATCTAAGGGGCAAAGATCANNNNNNN
|
ORF Staff: ATG at 137ORF Stop: TAG at 2774
SEQ ID NO: 200879 aa MW at 97267.9 kD
NOV77a,MDPTAGSKKEPGGGAATEEGVNRIAVPKPPSIEEFSIVKPISRGAFGKVYLGQKGGKL
|
CG94465-01 Protein SequenceYAVKVVKKADMINKNMTHQVQAERDALALSKSPFIVHLYYSLQSANNVYLVMEYLIGG
|
DVKSLLHIYGYFDEEMAVKYISEVALALDYLHRHGIIHRDLKPDNMLISNEGHIKLTD
|
FGLSKVTLNRDINMMDILTTPSMAKPRQDYSRTPGQVLSLISSLGFNTPIAEKNQDPA
|
NILSACLSETSQLSQGLVCPMSVDQKDTTPYSSKLLKSCETVASNPGMPVKCLTSNLL
|
QSRKRLATSSASSQSHTFISSVESECHSSPKWEKDCQESDEALGPTMMSWNAVEKLCA
|
KSANAIETKGFNKPWLELALSPIHNSSALPTTGRSCVNLAKKCFSGEVSWEAVELDVN
|
NINMDTDTSQLGFHQSNQWAVDSGGISEEHLGKRSLKRNFELVDSSPCKKIIQNKKTC
|
VEYKHNEMTNCYTNQNTGLTVEVQDLKLSVHKSQQNDCANKENIVNSFTDKQQTPEKL
|
PIPMIAKNLMCELDEDCEKNSKRDYLSSSFLCSDDDRASKNISMNSDSSFPGISIMES
|
PLESQPLDSDRSIKESSFEESNIEDPLIVTPDCQEKTSPKGVENPAVQESNQKMLGPP
|
LEVLKTLASKRNAVAFRSFNSHINASMNSEPSRNNMTSLDAMDISCAYSGSYPMAITP
|
TQKRRSCMPHQQTPNQIKSGTPYRTPKSVRRGVAPVDDGRILGTPDYLAPELLLGRAH
|
GKACMPAVDWWALGVCLFEFLTGIPPFNDETPQQVFQNILKRDIPWPEGEEKLSDNAQ
|
SAVEILLTIDDTKRAGMKRHPLFSDVDWENLQHQTMPFIPQPDDETDTSYFEARNTAQ
|
HLTVSGFSL
|
SEQ ID NO:2012702 bp
NOV77b,GAGCCTCACTTTGAACCCAGTTGGCGGGAATGGCTGCTCGCGGAGGGGCAGTGTACGC
|
CG04465-02 DNA SequenceGGGCCCGCTGTAGGCTGTCCAGCGATGGATCCCACCGCGGGAAGCAAGAAGGAGCCTG
|
GAGGAGGCGCGGCGACTGAGGAGGGCGTGAATAGGATCGCAGTGCCAAAACCGCCCCTC
|
CATTGAGGAATTCAGCATAGTGAAGCCCATTAGCCGGGGCGCCTTCGGGAAAGTGTAT
|
CTGGGGCAGAAAGGCGGCAAATTGTATGCAGTAAAGGTTGTTAAAAAAGCAGACATGA
|
TCAACAAAAATATGACTCATCAGGTCCAAGCTGAGAGAGATCCACTGGCACTAAGCAA
|
AAGCCCATTCATTGTCCATTTGTATTATTCACTGCACTCTGCAAACAATGTCTACTTG
|
GTAATGGAATATCTTATTGGGGGAGATGTCAAGTCTCTCCTACATATATATGGTTATT
|
TTGATGAAGAGATGGCTGTGAAATATATTTCTGAAGTAGCACTGGCTCTAGACTACCT
|
TCACAGACATGGAATCATCCACAGGGACTTGAAACCGGACAATATGCTTATTTCTAAT
|
GAGGGTCATATTAAACTGACGGATTTTGGCCTTTCAAAAGTTACTTTGAATAGAGATA
|
TTAATATGATGGATATCCTTACAACACCATCAATGGCAAAACCTAGACAAGATTATTC
|
AAGAACCCCAGGACAAGTGTTATCGCTTATCAGCTCGTTGGGATTTAACACACCAATT
|
GCAGAAAAAAATCAAGACCCTGCAAACATCCTTTCAGCCTGTCTCTGTGAAACATCAC
|
AGCTTTCTCAAGGACTCGTATGCCCTATGTCTGTAGATCAAAAGGACACTACGCCTTA
|
TTCTAGCAAATTACTAAAATCATGTCTTGAAACAGTTGCCTCCAACCCAGGAATGCCT
|
GTGAAGTGTCTAACTTCTAATTTACTCCAGTCTAGGAAAAGGCTGGCCACATCCAGTG
|
CCAGTAGTCAATCCCACACCTTCATATCCAGTGTGGAATCAGAATGCCACAGCAGTCC
|
CAAATGGGAAAAAGATTGCCAGGAAAGTGATGAAGCATTGGGCCCAACAATGATGAGT
|
TGGAATGCAGTTGAAAAGTTATGCGCAAAATCTGCAAATGCCATTGAGACGAAAGGTT
|
TCAATAAAAAGGATCTGGAGTTAGCTCTTTCTCCCATTCATAACAGCAGTGCCCTTCC
|
CACCACTGGACGCTCTTGTGTAAACCTTGCTAAAAAATGCTTCTCTGGGGAAGTTTCT
|
TGGGAAGCAGTAGAACTGGATGTAAATAATATAAATATGGACACTGACACAAGTCAGT
|
TAGGTTTCCATCAGTCAAATCAGTGGGCTGTGGATTCTGGTGGGATATCTGAAGAGCA
|
CCTTGGGAAAAGAAGTTTAAAAAGAAATTTTGAGTTGGTTGACTCCAGTCCTTGTAAA
|
AAAATTATACAGAATAAAAAAACTTGTGTAGAGTATAAGCATAACGAAATGGCAAATT
|
GTTATACAAATCAAAATACAGGCTTAACAGTTGAAGTGCAGGACCTTAAGCTATCAGT
|
GCACAAAAGTCAACAAAATGACTGTGCTAATAAGGAGAACATTGTCAATTCTTTTACT
|
GATAAACAACAAACACCAGAAAAATTACCTATACCAATGATAGCAAAAAACCTTATGT
|
GTGAACTCGATGAAGACTGTGAAAAGAATAGTAAGAGGGACTACTTAAGTTCTAGTTT
|
TCTATGTTCTGATGATGATAGAGCTTCTAAAAATATTTCTATGAACTCTGATTCATCT
|
TTTCCTGGAATTTCTATAATGGAAAGTCCATTAGAAAGTCAGCCCTTAGATTCAGATA
|
GAAGCATTAAAGAATCCTCTTTTGAAGAATCAAATATTGAAGATCCACTTATTGTAAC
|
ACCAGATTGCCAAGAAAACACCTCACCAAAAGGTCTCGAGAACCCTGCTGTACAAGAG
|
AGTAACCAAAAAATGTTAGGTCCTCCTTTCCAGCTGCTGAAAACGTTAGCCTCTAAAA
|
ACCAGATTGCCAAGAAAAGACCTCACCAAAAGGTGTCGAGAACCCTGCTGTACAAGAG
|
AGTAACCAAAAAATGTTAGGTCCTCCTTTGGAGGTGCTGAAAACGTTAGCCTCTAAAA
|
GAAATGCTGTTGCTTTTCGAAGTTTTAACAGTCATATTAATGCATCCAATAACTCAGA
|
ACCATCCAGAATGAACATGACTTCTTTAGATGCAATGGATATTTCGTGTGCCTACAGT
|
GGTTCATATCCCATGGCTATAACCCCTACTCAAAAAAGAAGATCCTGTATGCCACATC
|
AGACCCCAAATCAGATCAAGTCGGGAACTCCATACCGAACTCCGAAGAGTGTGAGAAG
|
AGGGGTGGCCCCCGTTGATGATGGGCGAATTCTAGGAACCCCAGACTACCTTGCACCT
|
GAGCTGTTACTAGGCAGGGCCCATGATATCCCTTGGCCAGAAGGTGAAGAAAAGTTAT
|
CTGATAATGCTCAAAGTGCAGTAGAAATACTTTTAACCATTGATGATACAAAGAGAGC
|
TGGAATGAAAGAGCTAAAACGTCATCCTCTCTTCAGTGATGTCGACTGGGAAAATCTG
|
CAGCATCAGACTATGCCTTTCATCCCCCAGCCAGATGATGAAACAGATACCTCCTATT
|
TTGAAGCCAGGAATACTGCTCAGCACCTGACCGTATCTGGATTTAGTCTGTAGCACAA
|
AAATTTTCCTTTTAGTCTAGCCTCGTGTTATAGAATGAACTTGCATAATTATATACTC
|
CTTAATACTAGATTGATCTAAGGGGGAAAGATCA
|
ORF Start: ATG at 83ORF Stop: TAG at 2603
SEQ ID NO:202840 aa MW at 92829.8 kD
NOV77b,MDPTAGSKKEPGGGAATEEGVNRIAVPKPPSIEEFSIVKPISRGAFGKVYLGQKGGKL
|
CG94465-02 DNA SequenceYAVKVVKKADMINKMTHQVQAERDALALSKSPFIVHLYYSLQSANNVYLVMEYLIGG
|
DVKSLLHIYGYFDEEMAVKYISEVALALDYLHRHGIIHRDLKPDNMLISNEGHIKLTD
|
FGLSKVTLNRDINMMDILTTPSMAKPRQDYSRTPGQVLSLISSLGFNTPIAEKNQDPA
|
NILSACLSETSQLSQGLVCPMSVDQKDTTPYSSKLLKSCLETVASNPGMPVKCLTSNL
|
LQSRKRLATSSASSQSHTFISSVESECHSSPKWEKDCQESDEALGPTMMSWNAVEKLC
|
AKSANAIETKGFNKKDLELALSPIHNSSALPTTGRSCVNLAKKCFSGEVSWEAVELDV
|
NNINMDTDTSQLGFHQSNQWAVDSGGISEEHLGKRSLKRNFELVDSSPCKKIIQNKKT
|
CVEYKHNEMANCYTNQNTGLTVEVQDLKLSVHKSQQNDCANKENIVNSFTDKQQTPEK
|
LPIPMIAKNLMCELDEDCEKNSKRDYLSSSFLCSDDDRASKNISMNSDSSFPGISIME
|
SPLESQPLDSDRSIKESSFEESNIEDPLIVTPDCQEKTSPKGVENPAVQESNQKMLGP
|
PLEVLKTLASKRNAVAFRSFNSHTNASNNSEPSRMNMTSLDAMDISCAYSGSYPMAIT
|
PTQKRRSCMPHQTPNQIKSGTPYRTPKSVRRGVAPVDDGRILGTPDYLAPELLLGRAH
|
DIPWPEGEEKLSDNAQSAVEILLTIDDTKRAGMKELKRHPLFSDVDWENLQHQTMPFI
|
PQPDDETDTSYFEARNTAQHLTVSCFSL
|
SEQ ID NO:2031415 bp
NOV77cGAGCCTCACTTTGAACCCAGTTGGCCGGAATGGCTGCTCGCGGAGGGGCAGTGTACGC
|
CG94465-03 DNA SequenceGGGCCGCTGTAGGCTGTCCAGCGATGGATCCCACCGCGGGAAGCAAGAAGGAGCCTG
|
GAGGAGGCGGGGCGACTGAGGAGGGCGTGAATAGGATCGCAGTGCCAAAACCGCCCTC
|
CATTGAGGAATTCAGCATAGTGAAGCCCATTAGCCGGGGCGCCTTCGGGAAAGTGTAT
|
CTGGGGCAGAAAGGCGGCAAATTGTATGCAGTAAAGGTTGTTAAAAAAGCAGACATGA
|
TCAACAAAAATATGACTCATCAGGTCCAAGCTGAGAGAGATGCACTGGCACTAAGCAA
|
AAGCCCATTCATTGTCCATTTGTATTATTCACTGCAGTCTGCAAACAATGTCTACTTG
|
GTAATGGAATATCTTATTGGGGGAGATGTCAAGTCTCTCCTACATATATATGGTTATT
|
TTGATGAAGAGATGGCTGTGAAATATATTTCTGAAGTAGCACTGGCTCTAGACTACCT
|
TCACAGACATGGAATCATCCACAGGGACTTGAAACCGGACAATATGCTTATTTCTAAT
|
GAGGGTCATATTAAACTGACGGATTTTGGCCTTTCAAAAGTTACTTTGAATAGAGATA
|
TTAATATGATGGATATCCTTACAACACCATCAATGGCAAAACCTAGACAAGATTATTC
|
AAGAACCCCACGACAAGTGTTATCGCTTATCAGCTCGTTGGGATTTAACACACCAATT
|
GCAGAAAAAAATCAAGACCCTGCAAACATCCTTTCAGCCTGTCTGTCTGAAACATCAC
|
AGCTTTCTCAAGGACTCGTATGCCCTATGTCTGTAGATCAAAAGGACACTACGCCTTA
|
TTCTAGCAAATTACTAAAATCATCTCTTGAAACAGTTGCCTCCAACCCAGGAATGCCT
|
GTGAAGTGTCTAACTTCTAATTTACTCCAGTCTAGGAAAAGGCTGGCCACATCCAGTG
|
CCAGTAGTCAATCCCACACCTTCATATCCAGTGTGGAATCAGAATGCCACACCAGTCC
|
CAAATGGGAAAAAGATTGCCAGCACACCCCAAATCAGATCAAGTCGGGAACTCCATAC
|
CGAACTCCGAAGAGTGTGAGAAGAGGGGTGGCCCCCGTTGATGATGGGCGAATTCTAG
|
GAACCCCAGACTACCTTGCACCTGAGCTGTTACTAGGCAGGGCCCATGAGCTAAAACG
|
TCATCCTCTCTTCAGTGATGTGGACTGGGAAAATCTGCAGCATCAGACTATGCCTTTC
|
ATCCCCCAGCCAGATGATGAACAGATACCTCCTATTTTGAAGCCAGGAATACTGCTC
|
AGCACCTGACTGTATCTCGATTTAGTCTGTAGCACAAAATTTTCCTTTTAGTCTAGCC
|
TTGTGTTATAGAATGAACTTGCA
|
ORF Start: ATG at 83ORF Stop: TAG at 1364
SEQ ID NO:204427 aa MW at 47124.2 kD
NOV77cMDPTAGSKKEPGGGAATEEGVNRIAVPKPPSIEEFSIVKPISRGAFGKVYLGQKGGKL
|
CG94465-03 Protein SequenceYAVKVVKKADMINKNMTHQVQAERDALALSKSPFIVHLYYSLQSANNVYLVMEYLIGG
|
DVKSLLHIYGYFDEEMAVKYISEVALALDYLHRHGIIHRDLKPDNMLISNEGHIKLTD
|
FGLSKVTLNRDINMMDILTTPSMAKPRQDYSRTPGQVLSLISSLGFNTPIAEKNQDPA
|
NILSACLSETSQLSQGLVCPMSVDQKDTTPYSSKLLKSCLETVASNPGMPVKCLTSNL
|
LQSRKRLATSSASSQSHTFISSVESECHSSPKWEKDCQQTPNQIKSGTPYRTPKSVRR
|
GVAPVDDGRILGTPDYLAPELLLGRAHELKRHPLFSDVDWENLQHQTMPFIPQPDDET
|
DTSYFEARNTAQHLTVSGFSL
|
[0716] Sequence comparison of the above protein sequences yields the following sequence relationships shown in Table 77B.
403TABLE 77B
|
|
Comparison of NOV77a against NOV77b and NOV77c.
NOV77a Residues/Identities/Similarities
Protein SequenceMatch Residuesfor the Matched Region
|
NOV77b1 . . . 879835/883 (94%)
1 . . . 840835/883 (94%)
NOV77c1 . . . 329327/330 (99%)
1 . . . 330329/330 (99%)
|
[0717] Further analysis of the NOV77a protein yielded the following properties shown in Table 77C.
404TABLE 77C
|
|
Protein Sequence Properties NOV77a
|
|
PSort0.4500 probability located in cytoplasm;
analysis:0.3000 probability located in microbody
(peroxisome); 0.1000 probability located
in mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0718] A search of the NOV77a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 77D.
405TABLE 77D
|
|
Geneseq Results for NOV77a
NOV77aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAG66515Human 18477 protein kinase - Homo 1 . . . 879875/883 (99%)0.0
sapiens, 879 aa. [WO200159080-A1, 1 . . . 879875/883 (99%)
16-AUG-2001]
AAB93303Human protein sequence SEQ ID 1 . . . 879872/883 (98%)0.0
NO: 12373 - Homo sapiens, 878 aa. 1 . . . 878872/883 (98%)
[EP1074617-A2, 07-FEB-2001]
AAM93339Human polypeptide, SEQ ID NO: 1 . . . 879834/883 (94%)0.0
2877 - Homo sapiens, 840 aa. 1 . . . 840835/883 (94%)
[EP1130094-A2, 05-SEP-2001]
AAB65602Novel protein kinase, SEQ ID NO: 1 . . . 327327/328 (99%)0.0
127 - Homo sapiens, 329 aa. 1 . . . 328327/328 (99%)
[WO200073469-A2, 07-DEC-2000]
AAM48169CEK1-34 protein sequence - 566 . . . 879308/317 (97%)e−177
Unidentified, 312 aa. [CN1315336- 1 . . . 312309/317 (97%)
A, 03-OCT-2001]
|
[0719] In a BLAST search of public sequence datbases, the NOV77a protein was found to have homology to the proteins shown in the BLASTP data in Table 77E.
406TABLE 77E
|
|
Public BLASTP Results for NOV77a
NOV77aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q96GX5UNKNOWN (PROTEIN FOR 1 . . . 879875/883 (99%)0.0
MGC: 17352) - Homo sapiens (Human), 1 . . . 879875/883 (99%)
879 aa.
Q96SJ5CDNA FLJ14813 FIS, CLONE 1 . . . 879872/883 (98%)0.0
NT2RP4002408, MODERATELY 1 . . . 878872/883 (98%)
SIMILAR TO PROTEIN KINASE
CEK1 (EC 2.7.1. - ) - Homo sapiens
(Human), 878 aa.
Q95K03HYPOTHETICAL 53.7 KDA162 . . . 633451/473 (95%)0.0
PROTEIN - Macaca fascicularis (Crab 1 . . . 473459/473 (96%)
eating macaque) (Cynomolgus
monkey), 487 aa.
Q9D9V02700091H24RIK PROTEIN - Mus253 . . . 879439/632 (69%)0.0
musculus (Mouse), 615 aa. 1 . . . 615508/632 (79%)
Q9CZH92700091H24RIK PROTEIN - Mus253 . . . 879439/632 (69%)0.0
musculus (Mouse), 615 aa. 1 . . . 615507/632 (79%)
|
[0720] PFam analysis predicts that the NOV77a protein contains the domains shown in the Table 77F.
407TABLE 77F
|
|
Domain Analysis of NOV77a
Identities/
Similarities
NOV77afor the
Pfam DomainMatch RegionMatched RegionExpect Value
|
pkinase: domain 35 . . . 180 53/148 (36%)1.2e−44
1 of 2122/148 (82%)
RIO1: domain 48 . . . 208 43/226 (19%)2
1 of 1 77/226 (34%)
wap: domain 1 of 1515 . . . 550 12/55 (22%)8.1
27/55 (49%)
pkinase: domain 739 . . . 835 34/138 (25%)3.2e−10
2 of 2 74/138 (54%)
pkinase_C: domain836 . . . 864 10/31 (32%)0.027
1 of 1 23/31 (74%)
|
Example 78
[0721] The NOV78 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 78A.
408TABLE 78A
|
|
NOV78 Sequence Analysis
|
|
SEQ ID NO:2051173 bp
NOV78a,CCTATCCATGAATATTATGTTCCCTTAAAAAATGTCTGTGGGCCTGCAGCCTGGTGCA
|
CG94511-01 DNA SequenceGAGTATAATAACGGGCTCACCCACAAGCCAGCTAGCAGAGTGCTGGTAGCATTCTGTA
|
ATTTTGCAAATGAGGCAAACTTGAAATTAAGAAATCTGGAAGAGGGCTCGCCTATCAC
|
AATAGTACTCACCAGGGAAGACGGACTCAAATATTACAGGATGACGCAGACTGTTCAT
|
CAAATGGAATTTAAATATATTGGCACCAAAAATGACAGAAAATATTCACATTTCTGTC
|
ATTTTTGTCATGTGAGGAACACTGCTGGGATGGGACTTGAAGCCAGGATAAATCCCAC
|
AGATCATCTCATCACAGCCTATTTAGGCTCTGGCTTTAGCTTTACTCATGAACTTTCT
|
GTCCCAGAAATTGTTCTAGAGCTTACAGGACAAAGAGGAGGTTGTGCTAAAGGAAAAG
|
TAGGATCAATGCATATGTATGCCAAGAACTTCTACAGGAGCCATGGCACTGTCAGAGC
|
TCAGGTCTCCCTGGGAGCTGGTATTGGCTTGGCCTGTGAGTATAACGGAAAAGATCAG
|
GTCGGTCTGAATGACTTTATATTTGAGGCCTACAATATAGCCGCTTTGTGGAAATTAC
|
CTCCTATTTTCATCTGTGAGAATAACTGCTATGGAATAAGAGCATCTGTTGAGACAGC
|
AGCACCTAGCACTGATTACTATAAGACAGGCAATTTTGTACCTGGACTAAGGATAGAC
|
AGAATGAATATTCTATGTGTCCATGAGGCAACAAAGTTTGCCTGTTGTATATTTGGAA
|
AGGGGACCATACTGATGAAGCTGCAGACTTACCATTATCAAGGCCACAGTGTAAGTGA
|
TCCTGGAGTCAGCTGTACACAAGAAGAAATTCAAGAAGCAAGAAGCAAGAGGACTGAC
|
TCTATTATGCTTCTCAAAGATAAAATGGTAAATAGCAATCTTGCCAATGTTAAAGAAG
|
CCTGGGCTACAGAGCGAGACTCTGTCTCAAAAAACAAAAGAAAGGAAACCGATGTTGA
|
TACAACTGATCCCAAACTACCTTTGGAAGAATTAGGCCGTTACATTTATTTACATTTA
|
TTGCATTTTGAAGTTTATGATGTAAATCAGTGGATCCAGTTTAAGTAGATTTTTATTT
|
TTTTTAAGACAGG
|
ORF Staff: ATG at 32ORF Stop: TAG at 1148
SEQ ID NO:206372 aa MW at 41908.5 kD
NOV78a,MSVGLQPGAEYNNGLTQKPASRVLVAFCNFANEANLKLRNLEEGSPITIVLTREDGLK
|
CG94511-01 Protein SequenceYYRMTQTVHQMEFKYIGTKNDRKYSHFCHFCDVRKTAGMGLEARINPTDHLITAYLGS
|
GFSFTHELSVPEILVELTGQRGGCAKGKVGSMHMYAKNFYRSHGTVRAQVSLGAGIGL
|
ACEYNGKDEVGLNDFIFEAYNIAALWKLPPIFICENNCYGIRASVETAAPSTDYYKTG
|
NFVPGLRIDRMNILCVHEATKFACCIFGKGTILMKLQTYHYQGHSVSDPGVSCTQEEI
|
QEARSKRTDSIMLLDKMVNSNLANVKEAWATERDSVSKNKRKETDVDTTDPKLPLEE
|
LGRYIYLHLLHFEVYDVNQWIQFK
|
[0722] Further analysis of the NOV78a protein yielded the following properties shown in Table 78B.
409TABLE 78B
|
|
Protein Sequence Properties NOV78a
|
|
PSort0.6500 probability located in cytoplasm; 0.1000 probability
analysis:located in mitochondrial matrix space; 0.1000 probability
located in lysosome (lumen); 0.0436 probability located
in microbody (peroxisome)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0723] A search of the NOV78a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 78C.
410TABLE 78C
|
|
Geneseq Results for NOV78a
NOV78aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [Patent #,Matchthe MatchedExpect
IdentifierDate]ResiduesRegionValue
|
AAG38280Arabidopsis thaliana protein fragment57 . . . 336114/293 (38%)7e−47
SEQ ID NO: 47202 - Arabidopsis60 . . . 346160/293 (53%)
thaliana, 389 aa.
[EP1033405-A2, 06-SEP-2000]
AAG38279Arabidopsis thaliana protein fragment57 . . . 336114/293 (38%)7e−47
SEQ ID NO: 47201 - Arabidopsis84 . . . 370160/293 (53%)
thaliana, 413 aa.
[EP1033405-A2, 06-SEP-2000]
AAG32461Arabidopsis thaliana protein fragment57 . . . 336114/293 (38%)7e−47
SEQ ID NO: 39163 - Arabidopsis60 . . . 346160/293 (53%)
thaliana, 389 aa.
[EP1033405-A2, 06-SEP-2000]
AAG38281Arabidopsis thaliana protein fragment85 . . . 336110/265 (41%)5e−46
SEQ ID NO: 47203 - Arabidopsis23 . . . 281151/265 (56%)
thaliana, 324 aa.
[EP1033405-A2, 06-SEP-2000]
AAG32463Arabidopsis thaliana protein fragment85 . . . 336110/265 (41%)5e−46
SEQ ID NO: 39165 - Arabidopsis20 . . . 278151/265 (56%)
thaliana, 321 aa.
[EP1033405-A2, 06-SEP-2000]
|
[0724] In a BLAST search of public sequence datbases, the NOV78a protein was found to have homology to the proteins shown in the BLASTP data in Table 78D.
411TABLE 78D
|
|
Public BLASTP Results for NOV78a
NOV78aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
P08559Pyruvate dehydrogenase E1 component13 . . . 372232/377 (61%)e−118
alpha subunit, somatic form,13 . . . 387269/377 (70%)
mitochondrial precursor (EC 1.2.4.1)
(PDHE1-A type I) - Homo sapiens
(Human), 390 aa.
P29804Pyruvate dehydrogenase E1 component13 . . . 372230/377 (61%)e−117
alpha subunit, somatic form,12 . . . 386268/377 (71%)
mitochondrial precursor (EC 1.2.4.1)
(PDHE1-A type I) - Sus scrofa (Pig),
389 aa (fragment).
DERTP1pyruvate dehydrogenase (lipoamide)14 . . . 372232/376 (61%)e−117
(EC 1.2.4.1) alpha chain 1 precursor - 14 . . . 387265/376 (69%)
rat, 390 aa.
P35486Pyruvate dehydrogenase E1 component14 . . . 372232/376 (61%)e−117
alpha subunit, somatic form,14 . . . 387265/376 (69%)
mitochondrial precursor (EC 1.2.4.1)
(PDHE1-A type I) - Mus musculus
(Mouse), 390 aa.
P26284Pyruvate dehydrogenase E1 component14 . . . 372229/376 (60%)e−116
alpha subunit, somatic form,14 . . . 387265/376 (69%)
mitochondrial precursor (EC 1.2.4.1)
(PDHE1-A type I) - Rattus norvegicus
(Rat), 390 aa.
|
[0725] PFam analysis predicts that the NOV78a protein contains the domains shown in the Table 78E.
412TABLE 78E
|
|
Domain Analysis of NOV78a
Identities/
Similarities
NOV78a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
E1_dehydrog: domain61 . . . 347111/329 (34%)3.2e−51
1 of 1217/329 (66%)
|
Example 79
[0726] The NOV79 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 79A.
413TABLE 79A
|
|
NOV79 Sequence Analysis
|
|
SEQ ID NO:2076757 bp
NOV79a,TCCTGATACTTGTTTACTTTTCTGGGGCAGAAAAGCTTGCACTAATTGCTCTCCATGG
|
CG94551-01 DNA SequenceTGGCTAATTTTTTCAAGAGCTTGATTTTACCTTACATTCATAAGCTTTGCAAAGGAAT
|
GTTTACAAAGAAATTGGGAAATACAAACAAAAACAAAGAGTATCGTCAGCAGAAAAAG
|
GATCAAGACTTCCCCACTGCTGGCCAGACCAAATCCCCCAAATTTTCTTACACTTTTA
|
AAAGCACTGTAAAGAAGATTGCAAAGTGTTCATCCACTCACAACTTATCCACTCAGGA
|
AGACGAGGCCAGTAAAGAGTTTTCCCTCTCACCAACATTCAGTTACCGAGTAGCTATT
|
GCCAATGGCCTACAACCCTCTCACCAACATTCAGACAATGAGGATCTGCTTCAAGAGC
|
TCTCTTCAATCGAGAGTTCCTACTCAGAATCATTAAATGAACTAAGGAGTAGCACAGA
|
AAACCAGGCACAATCAACACACACAATGCCAGTTAGACGCAACAGAAAGAGTTCAAGC
|
AGCCTTGCACCCTCTGAGGGCAGCTCTGACGGGGAGCGTACTCTACATGGCTTAAAAC
|
TGGGAGCTTTACGAAAACTGAGAAAATGGAAAAAGAGTCAAGAATGTGTCTCCTCAGA
|
CTCAGAGTTAAGCACCATGAAAAAATCCTGGGGAATAAGAAGTAAGTCTTTGGACAGA
|
ACTGTCCGAAACCCAAACACAAATGCCCTGGAGCCAGGGTTCAGTTCCTCTGGCTGCA
|
TTAGCCAAACACATGATGTCATGGAAATGATCTTTAAGGAACTTCAGGGAATAAGTCA
|
GATTGAAACAGAACTTTCTGAACTACGAGGGCACGTCAATGCTCTCAAGCACTCCATC
|
GATGAGATCTCCAGCAGTGTGGAGGTTGTACAAAGTGAAATTCAGCACTTGCGCACAG
|
GGTTTGTCCAGTCTCGGAGGGAAACTAGAGACATCCATGATTATATTAAGCACTTACG
|
TCATATGGGTAGCAAGGCAAGCCTCAGATTTTTAAATGTGACTGAAGAAAGATTTGAA
|
TATGTTGAAAGCGTGGTGTACCAAATTCTAATAGATAAAATGGGTTTTTCAGATGCAC
|
CAAATGCTATTAAAATTGAATTTGCTCAGAGGATAGGACACCAGAGACACTGCCCAAA
|
TGCAAAGCCTCGACCCATACTTGTGTACTTTGAAACCCCTCAACAAAGGGATTCTGTC
|
TTAAAAAAGTCATATAAACTCAAAGGAACAGGCATTGGAATCTCAACAGATATTCTAA
|
CTCATGACATCAGAGAAAGAAAAGAGAAAGGGATACCATCCTCCCAGACATATGAGAG
|
CATGGCTATAAAGTTGTCTACTCCAGAGCCAAAAATCAAGAAGAACAATTGGCAGTCA
|
CCTGATGACAGTGATGAAGATCTTGAATCTGACCTCAATAGAAACAGTTACGCTGTGC
|
TTTCCAAGTCAGAGCTTCTAACAAAGGGAAGTACTTCCAAGCCAAGCTCAAAATCACA
|
CAGTGCTAGATCCAAGAATAAAACTGCTAATAGCAGCAGAATTTCAAATAAATCAGAT
|
TATGATAAAATCTCCTCACAGTTGCCAGAATCAGATATCTTGGAAAAGCAAACCACAA
|
CCCATTATGCAGATGCAACACCTCTCTGGGACTCACAGAGTGATTTTTTCACTGCTAA
|
ACTTAGTCGTTCTGAATCAGATTTTTCCAAATTGTGTCAGTCTTACTCAGAAGATTTT
|
TCAGAAAATCAGTTTTTCACTAGAACTAATGGAAGCTCTCTCCTGTCATCTTCGGACC
|
GGGAGCTATGGCACAGGAAACAGGAAGGAACAGCGACCCTGTATGACAGTCCCAAGGA
|
CCAGCATTTGAATGGAGGTGTTCAGGGTATCCAAGGGCAGACTGAAACTGAAAACACA
|
GAAACTGTGGATAGTGGAATGAGTAATGGCATGGTGTGTGCATCTGGAGACCGGAGTC
|
ATTACAGTGATTCTCAGCTCTCTTTACATGAGGATCTTTCTCCATGGAAGGAATGGAA
|
TCAAGGAGCTGATTTAGGCTTGGATTCATCCACCCAGGAAGGTTTTGATTATGAAACA
|
AACAGTCTTTTTGACCAACAGCTTGATGTTTACAATAAAGACCTAGAATACTTGGGAA
|
AGTGCCACAGTGATCTTCAAGATGACTCAGAGAGCTACGACTTAACTCAAGATGACAA
|
TTCTTCTCCATGCCCTGGCTTGGATAATGAACCACAAGGCCAGTGGGTTGGCCAATAT
|
GATTCTTATCACGGAGCTAATTCTAATGAGCTATACCAAAATCAAAACCAGTTGTCCA
|
TGATGTATCGAAGTCAAAGTGAATTGCAAAGTGATGATTCAGAGGATGCCCCACCCAA
|
ATCATGGCATAGTCGATTAAGCATTGACCTTTCTGATAAGACTTTCAGCTTCCCAAAA
|
TTTGGATCTACACTGCAGAGGGCTAAATCAGCCTTGGAAGTAGTATGGAACAAAAGCA
|
CACAGAGTCTGAGTGGGTATGAGGACAGTGGCTCTTCATTAATGGGGAGATTTCGGAC
|
ATTATCTCAATCAACTGCAAATGAGTCAAGTACCACACTTGACTCTGATGTCTACACG
|
GAGCCCTATTACTATAAAGCAGAGGATGAGGAAGATTATACTGAACCACTGGCTGACA
|
ATGAAACAGATTATGTTGAAGTCATGGAACAAGTCCTTGCTAAACTAGAAAACAGGAC
|
TAGTATTACTGAAACAGATGAACAAATGCAAGCATATGATCACCTTTCATATGAAACA
|
CCTTATGAAACCCCACAAGATGAGGGTTATGATGGTCCAGCAGATGATATGGTTAGTG
|
AAGAGGGGTTAGAACCCTTAAATGAAACATCAGCTGAGATGGAAATAAGAGAAGATGA
|
AAACCAAAACATTCCTGAACAGCCAGTGGAGATCACAAAGCCAAAGAGAATTCGTCCT
|
TCTTTCAAAGAAGCAGCTTTAAGGGCCTATAAAAAGCAAATGGCAGAGTTGGAAGAGA
|
AGATCTTGGCTGGATCTTACTCTTCATTTAAGGCTCGAATAGTAAGTGGCAATGATTT
|
GGATGCTTCCAAATTTTCTGCACTCCAGGTGTGTGGTGGGGCTGGAGGTGGACTTTAT
|
GGTATTGACAGCATGCCGGATCTTCGCAGAAAAAAAACTTTGCCTATTGTCCGAGATG
|
TGGTAAGTACCCTGGCTGCCCGGAAATCTGGACTCTCCCTGGCTATGGTGATTAGGAC
|
ATCCCTAAATAATGAGGAACTGAAAATGCACGTCTTCAAGAAGACCTTGCAGGCACTG
|
ATCTACCCTATGTCTTCTACCATCCCACACAATTTTGAGGTCTGGACGGCTACCACAC
|
CCACCTACTCTTATGAGTGTGAAGGGCTCCTGTGGGGCATTGCAAGGCAAGGCATGAA
|
GTGTCTGGAGTGTGGAGTGAAATGCCACGAAAAGTGTCAGGACCTGCTAAACGCTGAC
|
TGCTTGCAGAGTGCAGCAGAAAAGAGTTCTAAACATGGTGCCGAAGACAAGACTCAGA
|
CCATTATTACAGCAATGAAAGAAAGAATGAAGATCAGGGAGAAAAACCGGCCAGAACT
|
ATTTGAAGTAATCCAGGAAATGTTTCAGATTTCTAAAGAAGATTTTGTGCAGTTTACA
|
AAGGCGGCCAAACAGAGTGTACTGGATGGGACATCTAAGTGGTCTGCAAAAATAACCA
|
TTACAGTGGTTTCTGCACAGGGTCTACAGGCAAAAGATAAAACAGGGTCTAGTGATCC
|
ATATGTTACAGTTCAAGTTGGAAAGAACAAAAGAAGAACAAAAACCATTTTTGGAAAT
|
TTGAATCCAGTATGGGATGAGAAGTTTTATTTGGAGTGTCATAACTCCACAGATCGAA
|
TCAAAGTCAGAGTATGGGATGAAGATGATGATATTAAATCCAGAGTCAAGCAACATTT
|
CAAAAAGGAGTCAGATGATTTTCTGGGACAAACAATTGTAGAAGTGAGGACCTTGAGT
|
GGAGAAATGGATGTCTGGTACAACTTAGAGAAAAGGACAGATAAGTCAGCTGTATCTG
|
GGGCCATACGATTGAAAATCAATGTGGAGATAAAAGGAGAAGAGAAGGTTGCTCCATA
|
TCATATTCAATATACATGTTTACATGAGAATCTGTTCCATTACTTGACTGAAGTGAAA
|
TCTAATGGTGGAGTGAAAATCCCAGAAGTCAAAGGGGATGAAGCCTGGAAGGTTTTCT
|
TTGATGATGCTTCCCAAGAAATAGTTGATGAATTTGCTATGCGTTATGGAATTGAATC
|
CATTTATCAAGCTATGACGCACTTTTCATGTCTGTCTTCTAAATACATGTGCCCCGGT
|
GTCCCTGCCGTCATGAGCACCTTGCTGGCTAATATAAATGCTTTTTATGCTCACACAA
|
CAGTTTCAACAAACATACAGGTTTCTGCCTCAGATCGATTTGCTGCTACCAACTTTGG
|
TAGGGAAAAATTCATAAAACTACTGGACCAGTTACATAACTCTTTGAGGATTGATCTG
|
TCAAAGTATAGGCAGGAAAACTTTCCTGCAAGCAATACTGAAAGACTGCAAGACCTGA
|
AATCAACTGTTGACCTGTTAACAAGTATCACCTTTTTTAGGATGAAGGTACTGGAGCT
|
GCAAACCCCCCCAAAAGCGAGCATGGTGGTGAAGGACTGTGTAAGGGCTTGCCTGCAT
|
TCTACATACAAGTATATTTTTGACAACTGCCATGAACTCTACTCCCAGCTAACAGACC
|
CAAGTAAGAAACAGGATATTCCTCGTGAAGATCAGGGACCAACCACCAAGAATTTGGA
|
TTTTTGGCCCCAACTTATTACACTGATGGTTACTATTATTGATGAGGATAAAACTGCC
|
TACACACCTGTCCTGAATCAGTTTCCTCAAGAGCTGAACATGGGAAAAATAAGTGCCG
|
AAATTATGTGGACTCTTTTTGCTCTGGATATGAAATATGCATTAGAAGAACATGAAAA
|
TCAGCGCTTATGCAAGAGCACCGATTATATGAATTTGCATTTCAAAGTTAAATGGTTT
|
TATAATGAATATGTGCGTGAACTTCCTGCCTTGAAGGATGCTGTTCCTCAATACTCCT
|
TGTGGTTTGAACCTTTTGTCATGCAATGCCTAGATGAAAACGAAGATGTGTCAATGGA
|
ATTCCTTCATGGAGCACTGGGAAGAGACAAAAAAGATGGATTCCAGCAGACATCTGAG
|
CATGCTCTCTTTTGTTGCTCCGTGGTTGATGTCTTTGCTCAGCTGAATCAGAGCTTTG
|
AAATTATTAAGAAACTGGAATGCCCTAATCCTGAAGCATTATCTCACTTAATGAGAAG
|
ATTTGCAAAGACTATCAATAAAGTGCTGCTCCAGTATGCTGCAATTGTATCAAGTGAT
|
TTCAGTTCACATTGTGATAAGGAAAATGTGCCCTGTATCTTGATGAACAATATTCAAC
|
AATTGCGGGTCCAGCTGGAAAAAATGTTTGAATCCATGGGAGGGAAGGAGCTAGATTC
|
TGAAGCTAGTACTATTCTAAAAGAACTTCAGGTTAAGCTCAGTGGGGTCCTGGATGAG
|
CTCAGCGTCACTTATGGTGAAAGTTTCCAGGTTATAATTGAAGAGTGTATAAAACAGA
|
TGAGTTTCGAACTAAATCAAATGAGAGCAAATGGAAACACCACATCTAATAAGAACAG
|
TGCAGCAATGGATGCAGAGATTGTGTTAAGATCTCTTATGGATTTTTTGGACAAAACG
|
AGTCTCTCAGCAAAAATCTGTGAGAAAACAGTCCTAAAGCGAGTTTTAAAAGAGTTAT
|
GGAAGCTAGTTCTCAACAAAATAGAAAAACAAATTGTTCTTCCTCCTCTGACAGATCA
|
AACAGGACCCCAGATGATTTTCATTGCAGCTAAAGATCTTGGACAATTATCCAAACTG
|
AAGCAGGAGCACATGATTCGAGAGGATGCCAGGGGTCTGACGCCAAGACAATGCGCTT
|
TAATGGAGGTAGTGCTGGCTACCATCAAGTCCTTGTACCAATATTTTCATGCAGAAGA
|
AATGGGCCTGAAGAACAATTTTCTGAGAAAAGGCCCGAATCTCCAGTTTCTTAAATGC
|
GCTCCCAGTCTTTATACCCAAACTACTGATGCCTTGATCAAGAAATTCATAGATACTC
|
AAACCTCACAGAGTCGTTCCTCCAAAGATGCCGTGGGTCAGATATCTGTTCATGTGGA
|
CATCACTCCCACCCCAGGAACGGGAGATCATAAAGTCACTGTAAAAGTGATTGCTATT
|
AATGACCTAAACTGGCAGACCACAGCAATGTTCCGCCCCTTTGTGGAAGTTTCTATAC
|
TGGGACCCAACCTTGGAGACAAGAAGAGAAAACAAGGCACAAAAACAAAAAGCAACAC
|
ATGGTCACCAAAGTACAATGAAACATTTCAGTTGATTCTCGGAAAGGAAAATCGACCA
|
GGCGCTTATGAACTTCATCTCTCAGTTAAGGATTACTGCTTTGCCACAGAAGATCGAA
|
TTATCGGAATGACAGTCATTCAGCTACAGAACATAGCAGAAAAGGGAAGCTATGGGGC
|
ATGGTATCCTCTTCTGAAAAATATCTCTATGGATGAAACTGGTTTGACTATCCTTAGA
|
ATACTCTCTCAGAGGACCAGTGATGATGTGGCTAAAGAATTTGTAAGACTTAAATCTG
|
AAACAAGATCTACTGAAGAGAGTGCTTGAAACAAAGACTGCAAGCTAAATACATAACT
|
ATAATTGTTTGACTACTGCATGCATGTGC
|
ORF Staff: ATG at 550RF Stop: TGA at 6697
SEQ ID NO:2082214 aa MW at 251058.0 kD
NOV79a,MVANFFKSLTLPYIHKLCKGMFTKKLGNTNKNKEYRQQKKDQDPPTAGQTKSPKFSYT
|
CG94551-01 Protein SequenceFKSTVKKIAKCSSTHNLSTEEDEASKEFSLSPTFSYRVAIANGLQPSHQHSDNEDLLQ
|
ELSSIESSYSESLNELRSSTEHQAQSTHTMPVRRNRKSSSSLAPSEGSSDGERTLHGL
|
KLGALRKLRKWKKSQECVSSDSELSTMKKSWGIRSKSLDRTVRNPKTNALEPGFSSSG
|
CISQTHDVMEMIFKELQGISQIETELSELRGHVNALKHSIDEISSSVEVVQSEIEQLR
|
TGFVQSRRETRDIHDYIKHLGHMGSKASLRFLNVTEERFEYVESVVYQILIDKMGFSD
|
APNAIKIEFAQRIGHQRDCPNAKPRPILVYFETPQQRDSVLKKSYKLKGTGIGISTDI
|
LTHDIRERKEKGIPSSQTYESMAIKLSTPEPKIKKNNWQSPDDSDEDLESDLNRNSYA
|
VLSKSELLTKGSTSKPSSKSHSARSKNKTANSSRISNKSDYDKISSQLPESDILEKQT
|
TTHYADATPLWHSQSDFFTAKLSRSESDFSKLCQSYSEDFSENQFFTRTNGSSLLSSS
|
DRELWQRKQEGTATLYDSPKDQHLNGGVQGIQGQTETENTETVDSGMSNGMVCASGDR
|
SHYSDSQLSLHEDLSPWKEWNQGADLGLDSSTQEGFDYETNSLFDQQLDVYNKDLEYL
|
GKCHSDLQDDSESYDLTQDDNSSPCPGLDNEPQGQWVGQYDSYQGANSNELYQNQNQL
|
SMMYRSQSELQSDDSEDAPPKSWHSRLSIDLSDKTFSFPKFGSTLQRAKSALEVVWNK
|
STQSLSGYEDSGSSLMGRFRTLSQSTANESSTTLDSDVYTEPYYYKAEDEEDYTEPVA
|
DNETDYVEVMEQVLAKLENRTSITETDEQMQAYDHLSYETPYETPQDEGYDCPADDMV
|
SEEGLEPLNETSAEMEIREDENQNIPEQPVEITKPKRIRPSFKEAALRAYKKQMAELE
|
EKILAGSYSSFKARIVSGNDLDASKFSALQVCGGAGGGLYGIDSMPDLRRKKTLPIVR
|
DVVSTLAARKSGLSLAMVIRTSLNNEELKMHVFKKTLQALIYPMSSTIPHNFEVWTAT
|
TPTYCYECEGLLWGIARQGMKCLECGVKCHEKCQDLLNADCLQSAAEKSSKHGAEDKT
|
QTIITAMKERMKIREKNRPEVFEVIQEMFQISKEDFVQFTKAAKQSVLDGTSKWSAKI
|
TITVVSAQGLQAKDKTGSSDPYVTVQVGKNKRRTKTIFGNLNPVWDEKFYLECHNSTD
|
RIKVRVWDEDDDIKSRVKQHFKKESDDFLGQTIVEVRTLSGEMDVWYNLEKRTDKSAV
|
SGAIRLKINVEIKGEEKVAPYHIQYTCLHENLFHYLTEVKSNGGVKIPEVKGDEAWKV
|
FFDDASQEIVDEFAMRYGIESIYQAMTHFSCLSSKYMCPGVPAVMSTLLANINAFYAH
|
TTVSTNIQVSASDRFAATNFGREKFIKLLDQLHNSLRIDLSKYRQENFPASNTERLQD
|
LKSTVDLLTSITFFRMKVLELQSPPKASMVVKDCVRACLDSTYKYIFDNCHELYSQLT
|
DPSKKQDIPREDQGPTTKNLDFWPQLITLMVTIIDEDKTAYTPVLNQFPQELNMGKIS
|
AEIMWTLFALDMKYALEEHENQRLCKSTDYMNLHFKVKWFYNEYVRELPAFKDAVPEY
|
SLWFEPFVMQWLDENEDVSMEFLHGALGRDKKDGFQQTSEHALFSCSVVDVFAQLNQS
|
FEIIKKLECPNPEALSHLMRRFAKTINKVLLQYAAIVSSDFSSHCDKENVPCILMNNI
|
QQLRVQLEKMFESMGGKELDSEASTILKELQVKLSGVLDELSVTYGESFQVIIEECIK
|
QMSFELNQMRANGNTTSNKESAAMDAEIVLRSLMDFLDKTSLSAKICEKTVLKRVLKE
|
LWKLVLNKIEKQIVLPPLTDQTGPQMIFIAAKDLGQLSKLKQEHMIREDARGLTPRQC
|
ALMEVVLATIKSLYQYFHAEEMGLKKNFLRKGPNLQFLKCAPSLYTQTTDALIKKFID
|
TQTSQSRSSKDAVGQISVHVDITATPGTGDHKVTVKVIAINDLNWQTTAMFRPFVEVC
|
ILGPNLGDKKRKQGTKTKSNTWSPKYNETFQLILGKENRPGAYELHLSVKDYCFARED
|
RIIGMTVIQLQNIAEKGSYGAWYPLLKNISMDETGLTILRILSQRTSDDVAKEFVRLK
|
SETRSTEESA
|
[0727] Further analysis of the NOV79a protein yielded the following properties shown in Table 79B.
414TABLE 79B
|
|
Protein Sequence Properties NOV79a
|
|
PSort0.9800 probability located in nucleus;
analysis:0.3000 probability located in microbody
(peroxisome); 0.1000 probability located
in mitochondrial matrix space; 0.1000
probability located in lysosome (lumen)
SignalPNo Known Signal Sequence Predicted
analysis:
|
[0728] A search of the NOV79a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 79C.
415TABLE 79C
|
|
Geneseq Results for NOV79a
NOV79aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAB47279PN7098 - Homo sapiens, 1230 aa. 1 . . . 12261214/1230 (98%)0.0
[WO200140794-A1, 07-JUN-2001] 1 . . . 12301216/1230 (98%)
AAY27134Human munc13 (Hmunc13)1020 . . . 2214 880/1197 (73%)0.0
polypeptide - Homo sapiens, 1591 405 . . . 15911027/1197 (85%)
aa. [WO9931134-A1, 24-JUN-1999]
AAW83431Rat Munc13-1 - Rattus sp, 1763 aa. 705 . . . 2212 948/1582 (59%)0.0
[JP10313866-A, 02-DEC-1998] 222 . . . 17561144/1582 (71%)
AAW83428Munc13-1-interacting domain of1350 . . . 1936 449/593 (75%)0.0
Doc2-alpha - Rattus sp, 611 aa. 1 . . . 590 520/593 (86%)
[JP10313866-A, 02-DEC-1998]
ABG13673Novel human diagnostic protein2037 . . . 2214 176/178 (98%)6e−98
#13664 - Homo sapiens, 183 aa. 6 . . . 183 177/178 (98%)
[WO200175067-A2, 11-OCT-2001]
|
[0729] In a BLAST search of public sequence datbases, the NOV79a protein was found to have homology to the proteins shown in the BLASTP data in Table 79D.
416TABLE 79D
|
|
Public BLASTP Results for NOV79a
NOV79a
ProteinResidues/Identities/
AccessionMatchSimilarities for theExpect
NumberProtein/Organism/LengthResiduesMatched PortionValue
|
T42759Munc13-3 protein - rat, 2207 aa. 1 . . . 22141980/2221 (89%)0.0
4 . . . 22072065/2221 (92%)
Q62770MUNC13-3 - Rattus norvegicus 1 . . . 22141980/2221 (89%)0.0
(Rat), 2207 aa. 4 . . . 22072065/2221 (92%)
O14795MUNC13 - Homo sapiens1020 . . . 2214 880/1197 (73%)0.0
(Human), 1591 aa. 405 . . . 15911027/1197 (85%)
Q62769MUNC13-2 - Rattus norvegicus1020 . . . 2214 879/1217 (72%)0.0
(Rat), 1985 aa. 779 . . . 19851026/1217 (84%)
Q9WV40MUNC13-2 PROTEIN - Rattus1020 . . . 2214 879/1217 (72%)0.0
norvegicus (Rat), 1622 aa. 416 . . . 16221026/1217 (84%)
|
[0730] PFam analysis predicts that the NOV79a protein contains the domains shown in the Table 79E.
417TABLE 79E
|
|
Domain Analysis of NOV79a
Identities/
Similarities
NOV79a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Transposase_22:146 . . . 492 73/394 (19%)0.46
domain 1 of 1146/394 (37%)
DAG_PE-bind:1094 . . . 1143 21/51 (41%)1.8e−18
domain 1 of 1 43/51 (84%)
C2: domain 1 of 21218 . . . 1309 33/106 (31%)2.3e−19
73/106 (69%)
C2: domain 2 of 22063 . . . 2153 26/97 (27%)0.0054
57/97 (59%)
|
Example 80
[0731] The NOV80 clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 80A.
418TABLE 80A
|
|
NOV80 Sequence Analysis
|
|
SEQ ID NO:2091683 bp
NOV 80aGTCAGGCCTATGGCCATGGCCTTCACAGACCTGCTGGATGCTCTGGGCAGCATGGCCC
|
CG94682-02 DNA SequenceGCTTCCAGCTCAACCACACAGCCCTGCTGCTGCTGCCCTGCGGCCTCCTGGCCTGCCA
|
CAACTTCCTGCAGAACTTCACCGCCGCTGTCCCCCCCCACCACTGCCGGGGCCCTGCC
|
AACCACACTGAGGCCTCCACCAACGACTCGGGGGCCTGGCTCAGGCCCACCATACCCC
|
TGGACCAGCTTGGGGCCCCTCAGCCCTGCCGGCGCTTCACCAACCCTCAGTGGGCCCT
|
GCTGAGCCCCAACTCCTCCATCCCGGGCGCGGCCACGGAGGGCTGCAAGGACGGCTGG
|
GTCTATAACCGCAGTGTTTTCCCGTCCACCATCGTGATGGAGTGGGATCTGGTGTGTG
|
AGGCCCGCACTCTCCGAGACCTGGCGCAGTCCGTCTACATTGCCGGGGTGCTGGTGGG
|
GGCTGCCGTCTTTGGCAGCTTGGCAGACAGGCTGGGCTGCAAGGGCCCCCTGGTCTGG
|
TCCTACCTGCAGCTGGCAGCTTCGGGGGCCGCCACAGCGTATTTCAGCTCCTTCAGTG
|
CCTATTGCGTCTTCCGGTTCCTGATGGGCATGACCTTCTCTGGCATCATCCTCAACTC
|
CGTCTCCCTGATTGTGGAGTGGATGCCCACACGGGGCCGGACTGTGGCGGGTATTTTG
|
CTGGGGTATTCCTTCACCCTGGCCCAGCTCATCCTGGCTGGGGTAGCCTACCTGATTC
|
GCCCCTGGCGGTGCCTGCAGTTTGCCATCTCTGCTCCTTTCCTGATCTTTTTCCTCTA
|
TTCTTGGTGGCTTCCAGAGTCATCCCGCTGGCTCCTCCTGCATGGCAAGTCCCACTTA
|
GCTGTACAGAATCTGCAGAAGGTGGCTGCAATGAACGGGAGGAAGCAGGAAGGGGAAA
|
GGCTGACCAAGGAGGTGATGAGCTCCTACATCCAAAGCGAGTTTGCAAGTGTCTGCAC
|
CTCCAACTCAATCTTGGACCTCTTCCGAACCCCGGCCATCCGCAAGGTCACATGCTGT
|
CTCATGGTGATTTGGTGGGGCCATTCTGTGGCTTACTATGGCCTGGCCATGGACCTGC
|
AGAAGTTTGGGCTCAGCCTATACCTGGTGCAGGCCCTGTTTGGAATCATCAACATCCC
|
GGCCATGCTGGTGGCCACCGCCACCATGATTTACGTGGGCCGCCGTGCCACGGTGGCC
|
TCCTTCCTCATCCTGGCCGGGCTCATGGTGATCGCCAACATGTTTGTGCCAGAAGGCA
|
CGCAGATCCTGTGCACAGCCCAGGCAGCGCTGGGCAAAGOCTGCCTGGCCAGCTCCTT
|
CATCTGTGTGTACCTGTTTACCGGCGAGCTGTACCCCACCGAGATCAGGCAGATGGGG
|
ATGGGCTTTGCCTCTGTCCACGCCCGCCTCGGGGGCCTGACGGCGCCCCTGGTTACCA
|
CACTTGGGGAATACAGCACCATCCTGCCACCCGTGAGCTTTGGGGCCACCGCAATCCT
|
GGCTGGGCTGGCCGTCTGCTTCCTGACTGAGACCCGCAACATGCCCCTGGTGGAGACC
|
ATCGCAGCCATGGAGAGGAGGGTCAAAGAAGGCTCTTCCAAGAAACATGTAGAAGAGA
|
AGAGTGAAGAAATTTCTCTTCAGCAGCTGAGAGCATCTCCCCTCAAAGAGACCATCTA
|
A
|
ORF Start: ATG at 10ORF Stop: TAA at 1681
SEQ ID NO:210557 aa MW at 60747.4 kD
NOV80a,MAMAFTDLLDALGSMGRFQLNHTALLLLPCGLLACHNFLQNPTAAVPPHHCRGPANHT
|
CG94682-02 Protein SequenceEASTNDSGAWLRATIPLDQLGAPEPCRRFTKPQWALLSPNSSIPGAATEGCKDGWVYN
|
RSVFPSTIVMEWDLVCEARTLRDLAQSVYIAGVLVCAAVFGSLADRLGCKGPLVWSYL
|
QLAASGAATAYFSSFSAYCVFRFLMGMTFSGIILNSVSLIVEWMPTRCRTVAGILLGY
|
SFTLGQLILAGVAYLIRPWRCLQFAISAPFLIFFLYSWWLPESSRWLLLHGKSQLAVQ
|
NLQKVAAMNGRKQEGERLTKEVMSSYIQSEFASVCTSNSILDLFRTPAIRKVTCCLMV
|
IWWGHSVAYYGLAMDLQKFGLSLYLVQALFGIINIPAMLVATATMIYVGRRATVASFL
|
ILAGLMVIANMFVPEGTQILCTAQAALGKGCLASSFICVYLFTGELYPTEIRQMGMGF
|
ASVHARLGGLTAPLVTTLGEYSTILPPVSFGATAILAGLAVCFLTETRNMPLVETIAA
|
MERRVKEGSSKSSEEKSEEISLQQLRASPLKETI
|
[0732] Further analysis of the NOV80a protein yielded the following properties shown in Table 80B.
419TABLE 80B
|
|
Protein Sequence Properties NOV80a
|
|
PSort0.6000 probability located in plasma membrane;
analysis:0.4000 probability located in Golgi body;
0.3142 probability located in mitochondrial inner membrane;
0.3000 probability located in endoplasmic
reticulum (membrane)
SignalPCleavage site between residues 35 and 36
analysis:
|
[0733] A search of the NOV80a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 80C.
420TABLE 80C
|
|
Geneseq Results for NOV80a
NOV80aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/Length [PatentMatchthe MatchedExpect
Identifier#, Date]ResiduesRegionValue
|
AAE13280Human transporters and ion channels 1 . . . 557548/558 (98%)0.0
(TRICH)-7 - Homo sapiens, 589 aa. 34 . . . 589551/558 (98%)
[WO200177174-A2, 18-OCT-2001]
AAB47271hOAT1 - Homo sapiens, 550 aa. 3 . . . 546253/546 (46%)e−141
[WO200104283-A2, 18-JAN-2001] 1 . . . 541343/546 (62%)
AAY44278Human organic anion transporter - 3 . . . 546253/546 (46%)e−141
Homo sapiens, 550 aa. [WO9964459- 1 . . . 541343/546 (62%)
A2, 16-DEC-1999]
AAW88489Human organic anion transporter 3 . . . 526249/526 (47%)e−141
OAT-1 - Homo sapiens, 563 aa. 1 . . . 522334/526 (63%)
[WO9853064-A1, 26-NOV-1998]
AAW88488Rat organic anion transporter OAT-1 - 3 . . . 546247/546 (45%)e−141
Rattus sp. 551 aa. [WO9853064-A1, 1 . . . 542344/546 (62%)
26-NOV-1998]
|
[0734] In a BLAST search of public sequence datbases, the NOV80a protein was found to have homology to the proteins shown in the BLASTP data in Table 80D.
421TABLE 80D
|
|
Public BLASTP Results for NOV80a
NOV80aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
O57379RENAL ORGANIC ANION3 . . . 556261/565 (46%)e−144
TRANSPORTER -1 . . . 561361/565 (63%)
Pseudopleuronecta americanus
(Winter flounder), 562 aa.
O35956RENAL ORGANIC ANION3 . . . 546248/546 (45%)e−141
TRANSPORT PROTEIN 1 - Rattus1 . . . 542346/546 (62%)
norvegicus (Rat), 551 aa.
Q9TSY7RENAL ORGANIC ANION3 . . . 546252/546 (46%)e−141
TRANSPORTER 1 (RBOAT1) -1 . . . 542342/546 (62%)
Oryctolagus cuniculus (Rabbit), 551
aa.
O95742RENAL ORGANIC ANION3 . . . 526249/526 (47%)e−141
TRANSPORT PROTEIN 1 - Homo1 . . . 522334/526 (63%)
sapiens (Human), 563 aa.
Q9R1U7ORGANIC ANION3 . . . 546255/550 (46%)e−139
TRANSPORTER 3 - Rattus1 . . . 533350/550 (63%)
norvegicus (Rat), 536 aa.
|
[0735] PFam analysis predicts that the NOV80a protein contains the domains shown in the Table 80E.
422TABLE 80E
|
|
Domain Analysis of NOV80a
Identities/
Similarities
NOV80a Matchfor the MatchedExpect
Pfam DomainRegionRegionValue
|
Chal_stil_syntC:200 . . . 212 5/13 (38%)9.8
domain 1 of 1 11/13 (85%)
sugar_tr:100 . . . 521 95/504 (19%)1.3e−07
domain 1 of 1284/504 (56%)
|
Example 81
[0736] The clone was analyzed, and the nucleotide and encoded polypeptide sequences are shown in Table 81A.
423TABLE 81A
|
|
NOV81 Sequence Analysis
|
|
SEQ ID NO:2111537 bp
NOV81a,AGCTCGAGATTTCTGTGGCTCCTCAAGATATTGGTCATAATCCTGGTACTTGGCATTG
|
CG90214-01 DNA SequenceTTGGATTTATGTTCGGAAGCATGTTCCTTCAAGCAGTGTTCAGCAGCCCCAAGCCAGA
|
ACTCCCAAGTCCTGCCCCGGGTGTCCAGAAGCTGAAGCTTCTGCCTGAGGAACGTCTC
|
AGGAACCTCTTTTCCTACGATGGAATCTGGCTGTTCCCGAAAAATCAGTGCAAATGTG
|
AGGAACCTCTTTTCCTACGATGGAATCTGGCTGTTCCCGAAAAATCAGTGCAAATGTG
|
AAGCCAACAAAGAGCAGGGAGGTTACAACTTTCAGGATGCCTATGGCCAGAGCGACCT
|
CCCAGCGGTGAAAGCGAGGAGACAGGCTGAATTTGAACACTTTCAGAGGAGGTGCAGA
|
GAAGGCCTGCCCCGCCCACTGCCCCTGCTGGTCCAGCCCAACCTCCCCTTTGGGTACC
|
CAGTCCACGGAGTGGAGGTGATGCCCCTGCACACGGTTCCCATCCCAGGTAAGTACAT
|
CCACATACCAAGAGACCCCGTCACCCTGACAGCTTCTCTGGGGACACTGAACACCCTT
|
GCTGATGTCCCAGACAGTGTGGTGCAGGGCAGAGGCCAGAAGCAGCTGATCATTTCTA
|
CCAGTGACCGGAAGCTGTTGAAGTTCATTCTTCAGCACGTGACATACACCAGCACGGG
|
GTACCAGCACCAGAAGGTAGACATAGGTGTGAGTCTGGAGTCCAGGTCCTCAGTGGCC
|
AAGTTTCCAGTGACCATCCGCCATCCTGTCATACCCAAGCTATACGACCCTGGACCAG
|
AGAGGAAGCTCAGAAACCTGGTTACCATTGCTACCAAGACTTTCCTCCGCCCCCACAA
|
GCTCATGATCATGCTCCGGAGTATTCGAGAGTATTACCCAGACTTGACCGTAATAGTG
|
GCTGATGACAGCCAGAAGCCCCTGGAAATTAAAGACAACCACGTGGAGTATTACACTA
|
TGCCCTTTGGGAAGGGTTGGTTTGCTGGTAGGAACCTGGCCATATCTCAGGTCACCAC
|
CAAATACGTTCTCTGGGTGGACGATGATTTTCTCTTCAACGAGGAGACCAACATTGAG
|
GTGCTGGTGGATGTCCTGGAGAAAACAGAACTGGACGTGGTAGGGGCCTGCCTTCACA
|
AGAGGATGGGATTTTTCCAACCCCTGGATGGCTTCCCCAGCTGCGTGGTGACCAGTGG
|
CGTGGTCAACTTCTTCCTGGCCCACACGGAGCGACTCCAAAGAGTTGGCTTTGATCCC
|
CGCCTGCAACGAGTGGCTCACGAATTCTTCATTGATGGGCTAGGGACCCTACTCGTGG
|
GGTCATGCCCAGAAGTGATTATAGGTCACCAGTCTCGGTCTCCAGTGGTGGACTCAGA
|
ACTGGCTGCCCTAGAGAAGACCTACAATACATACCGGTCCAACACCCTCACCCGGGTC
|
CAGTTCAAGCTGGCCCTCCACTACTTCAAGAACCATCTCCAATGTGCCGCATAAACGT
|
GTGAGGGCATAGCAGAPACACTAGCCTGGCTGGTTATGGTATCTATAGCAGGCCACCA
|
AAAACTGGACTCCTGATAGGTGAACGTTG
|
ORF Start: AGC at 1ORF Stop: TAA at 1444
SEQ ID NO:212481 aa MW at 54526.8 kD
NOV81a,SSRFLWLLKILVIILVLGIVGFMFGSMFLQAVFSSPKPELPSPAPGVQKLKLLPEERL
|
CG90214-01 Protein SequenceRNLFSYDGIWLFPKNQCKCEANKEQGGYNFQDAYGQSDLPAVKARRQAEFEHFQRRCR
|
EGLPRPLPLLVQPNLPFGYPVHGVEVMPLHTVPIPGKYTHIPRDPVTLTASLGTLNTL
|
ADVPDSVVQGRGQKQLIISTSDRKLLKFILQHVTYTSTGYQHQKVDIGVSLESRSSVA
|
KFPVTIRHPVIPKLYDPGPERKLRNLVTIATKTFLRPHKLMIMLRSIREYYPDLTVIV
|
ADDSQKPLEIKDNHVEYYTMPFGKGWFAGRNLAISQVTTKYVLWVDDDFLFNEETKIE
|
VLVDVLEKTELDVVGACLHKRMGFFQPLDGFPSCVVTSGVVNFFLAHTERLQRVCFDP
|
RLQRVAHEFFIDGLGTLLVCSCPEVIIGHQSRSPVVDSELAALEKTYNTYRSNTLTRV
|
QFKLALHYFKNHLQCAA
|
[0737] Further analysis of the NOV81 a protein yielded the following properties shown in Table 81B.
424TABLE 81B
|
|
Protein Sequence Properties NOV81a
|
|
PSort analysis:0.4600 probability located in plasma membrane
SignalP analysis:Cleavage site between residues 34 and 35
|
[0738] A search of the NOV81 a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 81C.
425TABLE 81C
|
|
Geneseq Results for NOV81a
NOV81aIdentities/
Residues/Similarities for
GeneseqProtein/Organism/LengthMatchthe MatchedExpect
Identifier[Patent #, Date]ResiduesRegionValue
|
AAE03702Novel human transferase-related 2 . . . 363352/362 (97%)1.2 e−247
protein #1, 506 aa. 6 . . . 367352/362 (97%)
AAE03719Novel human transferase-related 2 . . . 363352/362 (97%)1.2 e−247
protein #18, 566 aa. 66 . . . 427352/362 (97%)
AAE03717Novel human transferase-related 2 . . . 363351/368 (95%)3.0 e−244
protein #16, 572 aa. 66 . . . 433351/368 (95%)
AAE03718Novel human transferase-related 2 . . . 363351/368 (95%)3.0 e−244
protein #17, 512 aa. 6 . . . 373351/368 (95%)
AAE03711Novel human transferase-related 2 . . . 362351/361 (97%)1.1 e−183
protein #10, 448 aa. 66 . . . 426351/368 (97%)
|
[0739] In a BLAST search of public sequence datbases, the NOV81a protein was found to have homology to the proteins shown in the BLASTP data in Table 81D.
426TABLE 81D
|
|
Public BLASTP Results for NOV81a
NOV81aIdentities/
ProteinResidues/Similarities for
AccessionMatchthe MatchedExpect
NumberProtein/Organism/LengthResiduesPortionValue
|
Q09199Beta-1,4 N-22 . . . 480341/483 (70%)0.0
acetylgalactosaminyltransferase - Mus30 . . . 509378/483 (77%)
musculus (Mouse), 510 aa.
Q00973Beta-1,4 N-34 . . . 479205/496 (41%)1e−90
acetylgalactosaminyltransferase (EC45 . . . 529274/496 (54%)
2.4.1.92) ((N-acetylneuraminyl)-
galactosylglucosylceramide) (GM2/GD2
synthase) (GalNAc-T) - Homo sapiens
(Human), 533 aa.
Q09200Beta-1,4 N-29 . . . 479209/514 (40%)3e−90
acetylgalactosaminyltransferase (EC20 . . . 529281/514 (54%)
2.4.1.92) ((N-acetylneuraminyl)-
galactosylglucosylceramide) (GM2/GD2
synthase) (GalNAc-T) - Mus musculus
(Mouse), 533 aa.
Q10468Beta-1,4 N-41 . . . 479200/491 (40%)5e−89
acetylgalactosaminyltransferase (EC43 . . . 529272/491 (54%)
2.4.1.92) ((N-acetylneuraminyl)-
galactosylglucosylceramide) (GM2/GD2
synthase) (GalNAc-T) - Rattus
norvegicus (Rat), 533 aa.
BC022180UDP-N-acetyl-alpha-D-29 . . . 22265/222 (29%)2e−13
galactosamine: (N-acetylneuraminyl)-20 . . . 23898/222 (43%)
galactosylglucosylceramide-beta-1,
4-N-acetylgalactosaminyltransferase -
Mus musculus (Mouse), 244 aa.
|
[0740] PFam analysis predicts that the NOV81 a protein contains the domains shown in the Table 81E.
427TABLE 81E
|
|
Domain Analysis of NOV81a
Expect
Pfam DomainNOV81a Match RegionScoreValue
|
gnl|Pfam|pfam00535;260 . . . 42049.64.2e−12
Glycos_transf_2, Glycosyl
transferase
|
Sequencing Methodology and Identofication of NOVX Clones
[0741] 1. GeneCalling™ Technology: This is a proprietary method of performing differential gene expression profiling between two or more samples developed at CuraGen and described by Shimkets, et al., “Gene expression analysis by transcript profiling coupled to a gene database query” Nature Biotechnology 17:198-803 (1999). cDNA was derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then digested with up to as many as 120 pairs of restriction enzymes and pairs of linker-adaptors specific for each pair of restriction enzymes were ligated to the appropriate end. The restriction digestion generates a mixture of unique cDNA gene fragments. Limited PCR amplification is performed with primers homologous to the linker adapter sequence where one primer is biotinylated and the other is fluorescently labeled. The doubly labeled material is isolated and the fluorescently labeled single strand is resolved by capillary gel electrophoresis. A computer algorithm compares the electropherograms from an experimental and control group for each of the restriction digestions. This and additional sequence-derived information is used to predict the identity of each differentially expressed gene fragment using a variety of genetic databases. The identity of the gene fragment is confirmed by additional, gene-specific competitive PCR or by isolation and sequencing of the gene fragment.
[0742] 2. SeqCalling Technology: cDNA was derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then sequenced using CuraGen's proprietary SeqCalling technology. Sequence traces were evaluated manually and edited for corrections if appropriate. cDNA sequences from all samples were assembled together, sometimes including public human sequences, using bioinformatic programs to produce a consensus sequence for each assembly. Each assembly is included in CuraGen Corporation's database. Sequences were included as components for assembly when the extent of identity with another component was at least 95% over 50 bp. Each assembly represents a gene or portion thereof and includes information on variants, such as splice forms single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations.
[0743] 3. PathCalling™ Technology:
[0744] The NOVX nucleic acid sequences are derived by laboratory screening of cDNA library by the two-hybrid approach. cDNA fragments covering either the full length of the DNA sequence, or part of the sequence, or both, are sequenced. In silico prediction was based on sequences available in CuraGen Corporation's proprietary sequence databases or in the public human sequence databases, and provided either the full length DNA sequence, or some portion thereof.
[0745] The laboratory screening was performed using the methods summarized below:
[0746] cDNA libraries were derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then directionally cloned into the appropriate two-hybrid vector (Gal4-activation domain (Gal4-AD) fusion). Such cDNA libraries as well as commercially available cDNA libraries from Clontech (Palo Alto, Calif.) were then transferred from E.coli into a CuraGen Corporation proprietary yeast strain (disclosed in U.S. Pat. Nos. 6,057,101 and 6,083,693, incorporated herein by reference in their entireties).
[0747] Gal4-binding domain (Gal4-BD) fusions of a CuraGen Corportion proprietary library of human sequences was used to screen multiple Gal4-AD fusion cDNA libraries resulting in the selection of yeast hybrid diploids in each of which the Gal4-AD fusion contains an individual cDNA. Each sample was amplified using the polymerase chain reaction (PCR) using non-specific primers at the cDNA insert boundaries. Such PCR product was sequenced; sequence traces were evaluated manually and edited for corrections if appropriate. cDNA sequences from all samples were assembled together, sometimes including public human sequences, using bioinformatic programs to produce a consensus sequence for each assembly. Each assembly is included in CuraGen Corporation's database. Sequences were included as components for assembly when the extent of identity with another component was at least 95% over 50 bp. Each assembly represents a gene or portion thereof and includes information on variants, such as splice forms single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations.
[0748] Physical clone: the cDNA fragment derived by the screening procedure, covering the entire open reading frame is, as a recombinant DNA, cloned into pACT2 plasmid (Clontech) used to make the cDNA library. The recombinant plasmid is inserted into the host and selected by the yeast hybrid diploid generated during the screening procedure by the mating of both CuraGen Corporation proprietary yeast strains N106′ and YULH (U.S. Pat. Nos. 6,057,101 and 6,083,693).
[0749] 4. RACE: Techniques based on the polymerase chain reaction such as rapid amplification of cDNA ends (RACE), were used to isolate or complete the predicted sequence of the cDNA of the invention. Usually multiple clones were sequenced from one or more human samples to derive the sequences for fragments. Various human tissue samples from different donors were used for the RACE reaction. The sequences derived from these procedures were included in the SeqCalling Assembly process described in preceding paragraphs.
[0750] 5. Exon Linking: The NOVX target sequences identified in the present invention were subjected to the exon linking process to confirm the sequence. PCR primers were designed by starting at the most upstream sequence available, for the forward primer, and at the most downstream sequence available for the reverse primer. In each case, the sequence was examined, walking inward from the respective termini toward the coding sequence, until a suitable sequence that is either unique or highly selective was encountered, or, in the case of the reverse primer, until the stop codon was reached. Such primers were designed based on in silico predictions for the full length cDNA, part (one or more exons) of the DNA or protein sequence of the target sequence, or by translated homology of the predicted exons to closely related human sequences from other species. These primers were then employed in PCR amplification based on the following pool of human cDNAs: adrenal gland, bone marrow, brain—amygdala, brain—cerebellum, brain—hippocampus, brain—substantia nigra, brain—thalamus, brain—whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma—Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea, uterus. Usually the resulting amplicons were gel purified, cloned and sequenced to high redundancy. The PCR product derived from exon linking was cloned into the pCR2.1 vector from Invitrogen. The resulting bacterial clone has an insert covering the entire open reading frame cloned into the pCR2.1 vector. The resulting sequences from all clones were assembled with themselves, with other fragments in CuraGen Corporation's database and with public ESTs. Fragments and ESTs were included as components for an assembly when the extent of their identity with another component of the assembly was at least 95% over 50 bp. In addition, sequence traces were evaluated manually and edited for corrections if appropriate. These procedures provide the sequence reported herein.
[0751] 6. Physical Clone:
[0752] Exons were predicted by homology and the intron/exon boundaries were determined using standard genetic rules. Exons were further selected and refined by means of similarity determination using multiple BLAST (for example, tBlastN, BlastX, and BlastN) searches, and, in some instances, GeneScan and Grail. Expressed sequences from both public and proprietary databases were also added when available to further define and complete the gene sequence. The DNA sequence was then manually corrected for apparent inconsistencies thereby obtaining the sequences encoding the full-length protein.
[0753] The PCR product derived by exon linking, covering the entire open reading frame, was cloned into the pCR2.1 vector from Invitrogen to provide clones used for expression and screening purposes.
Quantitative Expression Analysis of Clones in Various Cells and Tissues
[0754] The quantitative expression of various clones was assessed using microtiter plates containing RNA samples from a variety of normal and pathology-derived cells, cell lines and tissues using real time quantitative PCR (RTQ PCR). RTQ PCR was performed on an Applied Biosystems ABI PRISM® 7700 or an ABI PRISM® 7900 HT Sequence Detection System. Various collections of samples are assembled on the plates, and referred to as Panel 1 (containing normal tissues and cancer cell lines), Panel 2 (containing samples derived from tissues from normal and cancer sources), Panel 3 (containing cancer cell lines), Panel 4 (containing cells and cell lines from normal tissues and cells related to inflammatory conditions), Panel 5D/51 (containing human tissues and cell lines with an emphasis on metabolic diseases), AI_comprehensive_panel (containing normal tissue and samples from autoimmune diseases), Panel CNSD.01 (containing central nervous system samples from normal and diseased brains) and CNS_neurodegeneration_panel (containing samples from normal and Alzheimer's diseased brains).
[0755] RNA integrity from all samples is controlled for quality by visual assessment of agarose gel electropherograms using 28S and 18S ribosomal RNA staining intensity ratio as a guide (2:1 to 2.5:128s: 18s) and the absence of low molecular weight RNAs that would be indicative of degradation products. Samples are controlled against genomic DNA contamination by RTQ PCR reactions run in the absence of reverse transcriptase using probe and primer sets designed to amplify across the span of a single exon.
[0756] First, the RNA samples were normalized to reference nucleic acids such as constitutively expressed genes (for example, O-actin and GAPDH). Normalized RNA (5 ul) was converted to cDNA and analyzed by RTQ-PCR using One Step RT-PCR Master Mix Reagents (Applied Biosystems; Catalog No. 4309169) and gene-specific primers according to the manufacturer's instructions.
[0757] In other cases, non-normalized RNA samples were converted to single strand cDNA (sscDNA) using Superscript II (Invitrogen Corporation; Catalog No. 18064-147) and random hexamers according to the manufacturer's instructions. Reactions containing up to 10 gg of total RNA were performed in a volume of 20 ul and incubated for 60 minutes at 42° C. This reaction can be scaled up to 50 μg of total RNA in a final volume of 100 μl. sscDNA samples are then normalized to reference nucleic acids as described previously, using 1×TaqMang Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instructions.
[0758] Probes and primers were designed for each assay according to Applied Biosystems Primer Express Software package (version I for Apple Computer's Macintosh Power PC) or a similar algorithm using the target sequence as input. Default settings were used for reaction conditions and the following parameters were set before selecting primers: primer concentration=250 nM, primer melting temperature (Tm) range=58°-60° C., primer optimal Tm=59° C., maximum primer difference=2° C., probe does not have 5′G, probe Tm must be 10° C. greater than primer Tm, amplicon size 75 bp to 100 bp. The probes and primers selected (see below) were synthesized by Synthegen (Houston, Tex., USA). Probes were double purified by HPLC to remove uncoupled dye and evaluated by mass spectroscopy to verify coupling of reporter and quencher dyes to the 5′ and 3′ ends of the probe, respectively. Their final concentrations were: forward and reverse primers, 900 nM each, and probe, 200 nM.
[0759] PCR conditions: When working with RNA samples, normalized RNA from each tissue and each cell line was spotted in each well of either a 96 well or a 384-well PCR plate (Applied Biosystems). PCR cocktails included either a single gene specific probe and primers set, or two multiplexed probe and primers sets (a set specific for the target clone and another gene-specific set multiplexed with the target probe). PCR reactions were set up using TaqMan (V One-Step RT-PCR Master Mix (Applied Biosystems, Catalog No. 4313803) following manufacturer's instructions. Reverse transcription was performed at 48° C. for 30 minutes followed by amplification/PCR cycles as follows: 95° C. 10 min, then 40 cycles of 95° C. for 15 seconds, 60° C. for 1 minute. Results were recorded as CT values (cycle at which a given sample crosses a threshold level of fluorescence) using a log scale, with the difference in RNA concentration between a given sample and the sample with the lowest CT value being represented as 2 to the power of delta CT. The percent relative expression is then obtained by taking the reciprocal of this RNA difference and multiplying by 100.
[0760] When working with sscDNA samples, normalized sscDNA was used as described previously for RNA samples. PCR reactions containing one or two sets of probe and primers were set up as described previously, using 1×TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instructions. PCR amplification was performed as follows: 95° C. 10 min, then 40 cycles of 95° C. for 15 seconds, 60° C. for 1 minute. Results were analyzed and processed as described previously.
[0761] Panels 1, 1.1, 1.2, and 1.3D
[0762] The plates for Panels 1, 1.1, 1.2 and 1.3D include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in these panels are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers of the following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in these panels are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on these panels are comprised of samples derived from all major organ systems from single adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions of the brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and adipose.
[0763] In the results for Panels 1, 1.1, 1.2 and 1.3D, the following abbreviations are used:
[0764] ca.=carcinoma,
[0765] *=established from metastasis,
[0766] met=metastasis,
[0767] s cell var=small cell variant,
[0768] non-s=non-sm=non-small,
[0769] squam=squamous,
[0770] pl. eff=pl effusion=pleural effusion,
[0771] glio=glioma,
[0772] astro=astrocytoma, and
[0773] neuro=neuroblastoma.
[0774] General_Screening_Panel_v1.4 and General_Screening_Panel_v1.5
[0775] The plates for Panels 1.4 and 1.5 include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in Panels 1.4 and 1.5 are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers of the following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in Panel 1.4 are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on Panels 1.4 and 1.5 are comprised of pools of samples derived from all major organ systems from 2 to 5 different adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions of the brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and adipose. Abbreviations are as described for Panels 1, 1.1, 1.2, and 1.3D.
[0776] Panels 2D and 2.2
[0777] The plates for Panels 2D and 2.2 generally include 2 control wells and 94 test samples composed of RNA or cDNA isolated from human tissue procured by surgeons working in close cooperation with the National Cancer Institute's Cooperative Human Tissue Network (CHTN) or the National Disease Research Initiative (NDR1). The tissues are derived from human malignancies and in cases where indicated many malignant tissues have “matched margins” obtained from noncancerous tissue just adjacent to the tumor. These are termed normal adjacent tissues and are denoted “NAT” in the results below. The tumor tissue and the “matched margins” are evaluated by two independent pathologists (the surgical pathologists and again by a pathologist at NDRI or CHTN). This analysis provides a gross histopathological assessment of tumor differentiation grade. Moreover, most samples include the original surgical pathology report that provides information regarding the clinical stage of the patient. These matched margins are taken from the tissue surrounding (i.e. immediately proximal) to the zone of surgery (designated “NAT”, for normal adjacent tissue, in Table RR). In addition, RNA and cDNA samples were obtained from various human tissues derived from autopsies performed on elderly people or sudden death victims (accidents, etc.). These tissues were ascertained to be free of disease and were purchased from various commercial sources such as Clontech (Palo Alto, Calif.), Research Genetics, and Invitrogen.
[0778] Panel 3D
[0779] The plates of Panel 3D are comprised of 94 cDNA samples and two control samples. Specifically, 92 of these samples are derived from cultured human cancer cell lines, 2 samples of human primary cerebellar tissue and 2 controls. The human cell lines are generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: Squamous cell carcinoma of the tongue, breast cancer, prostate cancer, melanoma, epidermoid carcinoma, sarcomas, bladder carcinomas, pancreatic cancers, kidney cancers, leukemias/lymphomas, ovarian/uterine/cervical, gastric, colon, lung and CNS cancer cell lines. In addition, there are two independent samples of cerebellum. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. The cell lines in panel 3D and 1.3D are of the most common cell lines used in the scientific literature.
[0780] Panels 4D, 4R, and 4.1D
[0781] Panel 4 includes samples on a 96 well plate (2 control wells, 94 test samples) composed of RNA (Panel 4R) or cDNA (Panels 4D/4.1 D) isolated from various human cell lines or tissues related to inflammatory conditions. Total RNA from control normal tissues such as colon and lung (Stratagene, La Jolla, Calif.) and thymus and kidney (Clontech) was employed. Total RNA from liver tissue from cirrhosis patients and kidney from lupus patients was obtained from BioChain (Biochain Institute, Inc., Hayward, Calif.). Intestinal tissue for RNA preparation from patients diagnosed as having Crohn's disease and ulcerative colitis was obtained from the National Disease Research Interchange (NDR1) (Philadelphia, Pa.).
[0782] Astrocytes, lung fibroblasts, dermal fibroblasts, coronary artery smooth muscle cells, small airway epithelium, bronchial epithelium, microvascular dermal endothelial cells, microvascular lung endothelial cells, human pulmonary aortic endothelial cells, human umbilical vein endothelial cells were all purchased from Clonetics (Walkersville, MD) and grown in the media supplied for these cell types by Clonetics. These primary cell types were activated with various cytokines or combinations of cytokines for 6 and/or 12-14 hours, as indicated. The following cytokines were used; IL-1 beta at approximately 1-5 ng/ml, TNF alpha at approximately 5-10 ng/ml, IFN gamma at approximately 20-50 ng/ml, IL-4 at approximately 5-10 ng/ml, IL-9 at approximately 5-10 ng/ml, IL-13 at approximately 5-10 ng/ml. Endothelial cells were sometimes starved for various times by culture in the basal media from Clonetics with 0.1% serum.
[0783] Mononuclear cells were prepared from blood of employees at CuraGen Corporation, using Ficoll. LAK cells were prepared from these cells by culture in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco/Life Technologies, Rockville, Md.), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) and Interleukin 2 for 4-6 days. Cells were then either activated with 10-20 ng/ml PMA and 1-2ftg/ml ionomycin, IL-12 at 5-10 ng/ml, IFN gamma at 20-SOng/ml and IL-18 at 5-10 ng/ml for 6 hours. In some cases, mononuclear cells were cultured for 4-5 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) with PHA (phytohemagglutinin) or PWM (pokeweed mitogen) at approximately 5 μg/ml. Samples were taken at 24, 48 and 72 hours for RNA preparation. MLR (mixed lymphocyte reaction) samples were obtained by taking blood from two donors, isolating the mononuclear cells using Ficoll and mixing the isolated mononuclear cells 1:1 at a final concentration of approximately 2×106cells/ml in DMEM 5% FCS (Hyclone), 100gM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol (5.5×105M) (Gibco), and 10 mM Hepes (Gibco). The MLR was cultured and samples taken at various time points ranging from 1-7 days for RNA preparation.
[0784] Monocytes were isolated from mononuclear cells using CD14 Miltenyi Beads, +ve VS selection columns and a Vario Magnet according to the manufacturer's instructions. Monocytes were differentiated into dendritic cells by culture in DMEM 5% fetal calf serum (FCS) (Hyclone, Logan, Utah), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×105M (Gibco), and 10 mM Hepes (Gibco), 50 ng/ml GMCSF and 5 ng/ml IL-4 for 5-7 days. Macrophages were prepared by culture of monocytes for 5-7 days in DMEM 5% FCS (Hyclone), 100 uM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), 10 mM Hepes (Gibco) and 10% AB Human Serum or MCSF at approximately 50 ng/ml. Monocytes, macrophages and dendritic cells were stimulated for 6 and 12-14 hours with lipopolysaccharide (LPS) at 100 ng/ml. Dendritic cells were also stimulated with anti-CD40 monoclonal antibody (Pharmingen) at 10 g/ml for 6 and 12-14 hours.
[0785] CD4 lymphocytes, CD8 lymphocytes and NK cells were also isolated from mononuclear cells using CD4, CD8 and CD56 Miltenyi beads, positive VS selection columns and a Vario Magnet according to the manufacturer's instructions. CD45RA and CD45RO CD4 lymphocytes were isolated by depleting mononuclear cells of CD8, CD56, CD14 and CD19 cells using CD8, CD56, CD14 and CDl9 Miltenyi beads and positive selection. CD45RO beads were then used to isolate the CD45RO CD4 lymphocytes with the remaining cells being CD45RA CD4 lymphocytes. CD45RA CD4, CD45RO CD4 and CD8 lymphocytes were placed in DMEM 5% FCS (Hyclone), 100M non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) and plated at 106cells/ml onto Falcon 6 well tissue culture plates that had been coated overnight with 0.5gg/ml anti-CD28 (Pharmingen) and 3 ug/ml anti-CD3 (OKT3, ATCC) in PBS. After 6 and 24 hours, the cells were harvested for RNA preparation. To prepare chronically activated CD8 lymphocytes, we activated the isolated CD8 lymphocytes for 4 days on anti-CD28 and anti-CD3 coated plates and then harvested the cells and expanded them in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×105M (Gibco), and 10 mM Hepes (Gibco) and IL-2. The expanded CD8 cells were then activated again with plate bound anti-CD3 and anti-CD28 for 4 days and expanded as before. RNA was isolated 6 and 24 hours after the second activation and after 4 days of the second expansion culture. The isolated NK cells were cultured in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×105M (Gibco), and 10 mM Hepes (Gibco) and IL-2 for 4-6 days before RNA was prepared.
[0786] To obtain B cells, tonsils were procured from NDR1. The tonsil was cut up with sterile dissecting scissors and then passed through a sieve. Tonsil cells were then spun down and resupended at 106cells/ml in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco). To activate the cells, we used PWM at 5gg/ml or anti-CD40 (Pharmingen) at approximately 10 μg/ml and IL-4 at 5-10 ng/ml. Cells were harvested for RNA preparation at 24,48 and 72 hours.
[0787] To prepare the primary and secondary Th1/Th2 and Trl cells, six-well Falcon plates were coated overnight with 10 μg/ml anti-CD28 (Pharmingen) and 2 μg/ml OKT3 (ATCC), and then washed twice with PBS. Umbilical cord blood CD4 lymphocytes (Poietic Systems, German Town, Md.) were cultured at 105-106cells/ml in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), 10 mM Hepes (Gibco) and IL-2 (4 ng/ml). IL-12 (5 ng/ml) and anti-IL4 (1 μg/ml) were used to direct to Th1, while IL-4 (5 ng/ml) and anti-IFN gamma (1 μg/ml) were used to direct to Th2 and IL-10 at 5 ng/ml was used to direct to Tr1. After 4-5 days, the activated Th1, Th2 and Tr1 lymphocytes were washed once in DMEM and expanded for 4-7 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), 10 mM Hepes (Gibco) and IL-2 (1 ng/ml). Following this, the activated Th1, Th2 and Tr1 lymphocytes were re-stimulated for 5 days with anti-CD28/OKT3 and cytokines as described above, but with the addition of anti-CD95L (1 μg/ml) to prevent apoptosis. After 4-5 days, the Th1, Th2 and Tr1 lymphocytes were washed and then expanded again with IL-2 for 4-7 days. Activated Th1 and Th2 lymphocytes were maintained in this way for a maximum of three cycles. RNA was prepared from primary and secondary Th1, Th2 and Tr1 after 6 and 24 hours following the second and third activations with plate bound anti-CD3 and anti-CD28 mAbs and 4 days into the second and third expansion cultures in Interleukin 2.
[0788] The following leukocyte cells lines were obtained from the ATCC: Ramos, EOL-1, KU-812. EOL cells were further differentiated by culture in 0.1 mM dbcAMP at 5×105cells/ml for 8 days, changing the media every 3 days and adjusting the cell concentration to 5×105cells/ml. For the culture of these cells, we used DMEM or RPMI (as recommended by the ATCC), with the addition of 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×105M (Gibco), 10 mM Hepes (Gibco). RNA was either prepared from resting cells or cells activated with PMA at 10 ng/ml and ionomycin at 1 μg/ml for 6 and 14 hours. Keratinocyte line CCD106 and an airway epithelial tumor line NCI-H292 were also obtained from the ATCC. Both were cultured in DMEM 5% FCS (Hyclone), 100 M non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco). CCD1106 cells were activated for 6 and 14 hours with approximately 5 ng/ml TNF alpha and 1 ng/ml IL-1 beta, while NCI-H292 cells were activated for 6 and 14 hours with the following cytokines: 5 ng/ml IL-4, 5 ng/ml IL-9, 5 ng/ml IL-13 and 25 ng/ml IFN gamma.
[0789] For these cell lines and blood cells, RNA was prepared by lysing approximately 107cells/ml using Trizol (Gibco BRL). Briefly, {fraction (1/10)} volume of bromochloropropane (Molecular Research Corporation) was added to the RNA sample, vortexed and after 10 minutes at room temperature, the tubes were spun at 14,000 rpm in a Sorvall SS34 rotor. The aqueous phase was removed and placed in a 15 ml Falcon Tube. An equal volume of isopropanol was added and left at −20° C. overnight. The precipitated RNA was spun down at 9,000 rpm for 15 min in a Sorvall SS34 rotor and washed in 70% ethanol. The pellet was redissolved in 3001 μl of RNAse-free water and 351l buffer (Promega) 5 μl DTT, 7 μl RNAsin and 8 g1 DNAse were added. The tube was incubated at 37° C. for 30 minutes to remove contaminating genomic DNA, extracted once with phenol chloroform and re-precipitated with {fraction (1/10)} volume of 3M sodium acetate and 2 volumes of 100% ethanol. The RNA was spun down and placed in RNAse free water. RNA was stored at −80° C.
[0790] AI_Comprehensive Panel_v1.0
[0791] The plates for AI_comprehensive panel_v1.0 include two control wells and 89 test samples comprised of cDNA isolated from surgical and postmortem human tissues obtained from the Backus Hospital and Clinomics (Frederick, Md.). Total RNA was extracted from tissue samples from the Backus Hospital in the Facility at CuraGen. Total RNA from other tissues was obtained from Clinomics.
[0792] Joint tissues including synovial fluid, synovium, bone and cartilage were obtained from patients undergoing total knee or hip replacement surgery at the Backus Hospital. Tissue samples were immediately snap frozen in liquid nitrogen to ensure that isolated RNA was of optimal quality and not degraded. Additional samples of osteoarthritis and rheumatoid arthritis joint tissues were obtained from Clinomics. Normal control tissues were supplied by Clinomics and were obtained during autopsy of trauma victims.
[0793] Surgical specimens of psoriatic tissues and adjacent matched tissues were provided as total RNA by Clinomics. Two male and two female patients were selected between the ages of 25 and 47. None of the patients were taking prescription drugs at the time samples were isolated.
[0794] Surgical specimens of diseased colon from patients with ulcerative colitis and Crohns disease and adjacent matched tissues were obtained from Clinomics. Bowel tissue from three female and three male Crohn's patients between the ages of 41-69 were used. Two patients were not on prescription medication while the others were taking dexamethasone, phenobarbital, or tylenol. Ulcerative colitis tissue was from three male and four female patients. Four of the patients were taking lebvid and two were on phenobarbital.
[0795] Total RNA from post mortem lung tissue from trauma victims with no disease or with emphysema, asthma or COPD was purchased from Clinomics. Emphysema patients ranged in age from 40-70 and all were smokers, this age range was chosen to focus on patients with cigarette-linked emphysema and to avoid those patients with alpha-lanti-trypsin deficiencies. Asthma patients ranged in age from 36-75, and excluded smokers to prevent those patients that could also have COPD. COPD patients ranged in age from 35-80 and included both smokers and non-smokers. Most patients were taking corticosteroids, and bronchodilators.
[0796] In the labels employed to identify tissues in the AI13 comprehensive panel_v1.0 panel, the following abbreviations are used:
[0797] Al=Autoimmunity
[0798] Syn=Synovial
[0799] Normal=No apparent disease
[0800] Rep22/Rep20=individual patients
[0801] RA=Rheumatoid arthritis
[0802] Backus=From Backus Hospital
[0803] OA=Osteoarthritis
[0804] (SS)(BA)(MF)=Individual patients
[0805] Adj=Adjacent tissue
[0806] Match control=adjacent tissues
[0807] −M=Male
[0808] −F=Female
[0809] COPD=Chronic obstructive pulmonary disease
[0810] Panels 5D and 5I
[0811] The plates for Panel 5D and 5I include two control wells and a variety of cDNAs isolated from human tissues and cell lines with an emphasis on metabolic diseases. Metabolic tissues were obtained from patients enrolled in the Gestational Diabetes study. Cells were obtained during different stages in the differentiation of adipocytes from human mesenchymal stem cells. Human pancreatic islets were also obtained.
[0812] In the Gestational Diabetes study subjects are young (18-40 years), otherwise healthy women with and without gestational diabetes undergoing routine (elective) Caesarean section. After delivery of the infant, when the surgical incisions were being repaired/closed, the obstetrician removed a small sample (<1 cc) of the exposed metabolic tissues during the closure of each surgical level. The biopsy material was rinsed in sterile saline, blotted and fast frozen within 5 minutes from the time of removal. The tissue was then flash frozen in liquid nitrogen and stored, individually, in sterile screw-top tubes and kept on dry ice for shipment to or to be picked up by CuraGen. The metabolic tissues of interest include uterine wall (smooth muscle), visceral adipose, skeletal muscle (rectus) and subcutaneous adipose. Patient descriptions are as follows:
[0813] Patient 2: Diabetic Hispanic, overweight, not on insulin
[0814] Patient 7-9: Nondiabetic Caucasian and obese (BMI>30)
[0815] Patient 10: Diabetic Hispanic, overweight, on insulin
[0816] Patient 11: Nondiabetic African American and overweight
[0817] Patient 12: Diabetic Hispanic on insulin
[0818] Adipocyte differentiation was induced in donor progenitor cells obtained from Osirus (a division of Clonetics/BioWhittaker) in triplicate, except for Donor 3U which had only two replicates. Scientists at Clonetics isolated, grew and differentiated human mesenchymal stem cells (HuMSCs) for CuraGen based on the published protocol found in Mark F. Pittenger, et al., Multilineage Potential of Adult Human Mesenchymal Stem Cells Science Apr. 2, 1999: 143-147. Clonetics provided Trizol lysates or frozen pellets suitable for mRNA isolation and ds cDNA production. A general description of each donor is as follows:
[0819] Donor 2 and 3 U: Mesenchymal Stem cells, Undifferentiated Adipose
[0820] Donor 2 and 3 AM: Adipose, AdiposeMidway Differentiated
[0821] Donor 2 and 3 AD: Adipose, Adipose Differentiated
[0822] Human cell lines were generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: kidney proximal convoluted tubule, uterine smooth muscle cells, small intestine, liver HepG2 cancer cells, heart primary stromal cells, and adrenal cortical adenoma cells. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. All samples were processed at CuraGen to produce single stranded cDNA.
[0823] Panel 5I contains all samples previously described with the addition of pancreatic islets from a 58 year old female patient obtained from the Diabetes Research Institute at the University of Miami School of Medicine. Islet tissue was processed to total RNA at an outside source and delivered to CuraGen for addition to panel 5I.
[0824] In the labels employed to identify tissues in the 5D and 5I panels, the following abbreviations are used:
[0825] GO Adipose=Greater Omentum Adipose
[0826] SK=Skeletal Muscle
[0827] UT=Uterus
[0828] PL=Placenta
[0829] AD=Adipose Differentiated
[0830] AM=Adipose Midway Differentiated
[0831] U=Undifferentiated Stem Cells
[0832] Panel CNSD.01
[0833] The plates for Panel CNSD.01 include two control wells and 94 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center. Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at −80° C. in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.
[0834] Disease diagnoses are taken from patient records. The panel contains two brains from each of the following diagnoses: Alzheimer's disease, Parkinson's disease, Huntington's disease, Progressive Supemuclear Palsy, Depression, and “Normal controls”. Within each of these brains, the following regions are represented: cingulate gyrus, temporal pole, globus palladus, substantia nigra, Brodman Area 4 (primary motor strip), Brodman Area 7 (parietal cortex), Brodman Area 9 (prefrontal cortex), and Brodman area 17 (occipital cortex). Not all brain regions are represented in all cases; e.g., Huntington's disease is characterized in part by neurodegeneration in the globus palladus, thus this region is impossible to obtain from confirmed Huntington's cases. Likewise Parkinson's disease is characterized by degeneration of the substantia nigra making this region more difficult to obtain. Normal control brains were examined for neuropathology and found to be free of any pathology consistent with neurodegeneration.
[0835] In the labels employed to identify tissues in the CNS panel, the following abbreviations are used:
[0836] PSP=Progressive supranuclear palsy
[0837] Sub Nigra=Substantia nigra
[0838] Glob Palladus=Globus palladus
[0839] Temp Pole=Temporal pole
[0840] Cing Gyr=Cingulate gyrus
[0841] BA 4=Brodman Area 4
[0842] Panel CNS_Neurodegeneration_V1.0
[0843] The plates for Panel CNS_Neurodegeneration_V1.0 include two control wells and 47 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center (McLean Hospital) and the Human Brain and Spinal Fluid Resource Center (VA Greater Los Angeles Healthcare System). Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at −80° C. in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.
[0844] Disease diagnoses are taken from patient records. The panel contains six brains from Alzheimer's disease (AD) patients, and eight brains from “Normal controls” who showed no evidence of dementia prior to death. The eight normal control brains are divided into two categories: Controls with no dementia and no Alzheimer's like pathology (Controls) and controls with no dementia but evidence of severe Alzheimer's like pathology, (specifically senile plaque load rated as level 3 on a scale of 0-3; 0=no evidence of plaques, 3=severe AD senile plaque load). Within each of these brains, the following regions are represented: hippocampus, temporal cortex (Brodman Area 21), parietal cortex (Brodman area 7), and occipital cortex (Brodman area 17). These regions were chosen to encompass all levels of neurodegeneration in AD. The hippocampus is a region of early and severe neuronal loss in AD; the temporal cortex is known to show neurodegeneration in AD after the hippocampus;
[0845] the parietal cortex shows moderate neuronal death in the late stages of the disease; the occipital cortex is spared in AD and therefore acts as a “control” region within AD patients. Not all brain regions are represented in all cases.
[0846] In the labels employed to identify tissues in the CNS_Neurodegeneration_V1.0 panel, the following abbreviations are used:
[0847] AD=Alzheimer's disease brain; patient was demented and showed AD-like pathology upon autopsy
[0848] Control=Control brains; patient not demented, showing no neuropathology
[0849] Control (Path)=Control brains; pateint not demented but showing sever AD-like pathology
[0850] SupTemporal Ctx=Superior Temporal Cortex
[0851] Inf Temporal Ctx=Inferior Temporal Cortex
[0852] A. CG55912-01: CACNG4
[0853] Expression of gene CG55912-01 was assessed using the primer-probe set Ag2841, described in Table AA. Results of the RTQ-PCR runs are shown in Tables AB, AC, and AD.
428TABLE AA
|
|
Probe Name Ag2841
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-gtctgcgtgaagatcaatcatt-3′22283213
ProbeTET-5′-aggacacggactacgaccacgacag-25311214
3′-TAMRA
Reverse5′-cggaccgtacggagtagatact-3′22341215
|
[0854]
429
TABLE AB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag2841, Run
Rel. Exp. (%) Ag2841, Run
|
Tissue Name
209779166
Tissue Name
209779166
|
|
AD 1 Hippo
15.8
Control (Path) 3
0.0
|
Temporal Ctx
|
AD 2 Hippo
34.6
Control (Path) 4
25.9
|
Temporal Ctx
|
AD 3 Hippo
15.9
AD 1 Occipital Ctx
6.8
|
AD 4 Hippo
17.0
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
85.3
AD 3 Occipital Ctx
24.0
|
AD 6 Hippo
51.1
AD 4 Occipital Ctx
21.8
|
Control 2 Hippo
21.0
AD 5 Occipital Ctx
0.0
|
Control 4 Hippo
0.0
AD 6 Occipital Ctx
51.8
|
Control (Path) 3 Hippo
0.0
Control 1 Occipital Ctx
6.0
|
AD 1 Temporal Ctx
6.3
Control 2 Occipital Ctx
100.0
|
AD 2 Temporal Ctx
16.3
Control 3 Occipital Ctx
39.2
|
AD 3 Temporal Ctx
5.0
Control 4 Occipital Ctx
5.1
|
AD 4 Temporal Ctx
7.9
Control (Path) 1
46.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
64.6
Control (Path) 2
23.0
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
38.4
Control (Path) 3
6.3
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
46.3
Control (Path) 4
25.3
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
88.3
Control 1 Parietal Ctx
0.0
|
Control 1 Temporal Ctx
10.4
Control 2 Parietal Ctx
51.4
|
Control 2 Temporal Ctx
42.9
Control 3 Parietal Ctx
18.9
|
Control 3 Temporal Ctx
20.3
Control (Path) 1
86.5
|
Parietal Ctx
|
Control 4 Temporal Ctx
8.4
Control (Path) 2
31.0
|
Parietal Ctx
|
Control (Path) 1
58.2
Control (Path) 3
7.5
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
49.7
Control (Path) 4
63.7
|
Temporal Ctx
Parietal Ctx
|
|
[0855]
430
TABLE AC
|
|
|
Panel 1.3D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2841, Run
Ag2841, Run
Ag2841, Run
Ag2841, Run
|
Tissue Name
161922470
165721032
Tissue Name
161922470
165721032
|
|
Liver
3.1
7.2
Kidney (fetal)
0.0
0.0
|
adenocarcinoma
|
Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0
|
Pancreatic ca.
5.7
5.9
Renal ca. A498
0.0
0.0
|
CAPAN 2
|
Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0
|
393
|
Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0
|
Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0
|
Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0
|
Brain (fetal)
18.3
27.0
Liver
0.0
0.0
|
Brain (whole)
27.0
19.5
Liver (fetal)
0.0
0.0
|
Brain (amygdala)
77.4
54.3
Liver ca.
0.0
0.0
|
(hepatoblast)
|
HepG2
|
Brain (cerebellum)
0.0
0.0
Lung
3.0
9.2
|
Brain (hippocampus)
100.0
100.0
Lung (fetal)
0.0
0.0
|
Brain (substantia
0.0
3.5
Lung ca. (small
0.0
0.0
|
nigra)
cell) LX-1
|
Brain (thalamus)
7.9
0.0
Lung ca. (small
0.0
0.0
|
cell) NCI-H69
|
Cerebral Cortex
90.8
8.4
Lung ca. (s.cell
0.0
3.9
|
var.) SHP-77
|
Spinal cord
3.9
0.0
Lung ca. (large
0.0
0.0
|
cell)NCI-H460
|
glio/astro U87-MG
2.7
6.5
Lung ca. (non-
15.0
0.0
|
sm. cell) A549
|
glio/astro U-118-
0.0
0.0
Lung ca. (non-
7.2
12.5
|
MG
s.cell) NCI-H23
|
astrocytoma
0.0
6.6
Lung ca. (non-
0.0
0.0
|
SW1783
s.cell) HOP-62
|
neuro*; met SK-N-
8.4
0.0
Lung ca. (non-
12.9
3.2
|
AS
s.cl) NCI-H522
|
astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.0
|
(squam.) SW 900
|
astrocytoma SNB-75
0.0
0.0
Lung ca.
0.0
0.0
|
(squam.) NCI-
|
H596
|
glioma SNB-19
27.2
6.2
Mammary gland
0.0
0.0
|
glioma U251
0.0
0.0
Breast ca.*
0.0
0.0
|
(pl.ef) MCF-7
|
glioma SF-295
0.0
3.4
Breast ca.*
0.0
0.0
|
(pl.ef) MDA-
|
MB-231
|
Heart (fetal)
0.0
0.0
Breast ca.*
0.0
0.0
|
(pl.ef) T47D
|
Heart
0.0
0.0
Breast ca. BT-
0.0
0.0
|
549
|
Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0
|
(fetal)
|
Skeletal muscle
0.0
0.0
Ovary
0.0
0.0
|
Bone marrow
3.8
0.0
Ovarian ca.
0.0
0.0
|
OVCAR-3
|
Thymus
0.0
0.0
Ovarian ca.
0.0
22.1
|
OVCAR-4
|
Spleen
0.0
0.0
Ovarian ca.
7.0
0.0
|
OVCAR-5
|
Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0
|
OVCAR-8
|
Colorectal
0.0
0.0
Ovarian ca.
0.0
0.0
|
IGROV-1
|
Stomach
0.0
0.0
Ovarian ca.*
0.0
0.0
|
(ascites) SK-OV-3
|
Small intestine
0.0
0.0
Uterus
0.0
2.6
|
Colon ca. SW480
0.0
2.6
Placenta
0.0
0.0
|
Colon ca.*
0.0
0.0
Prostate
0.0
0.0
|
SW620(SW480 met)
|
Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
2.0
|
(bone met)PC-3
|
Colon ca. HCT-116
0.0
0.0
Testis
21.0
0.0
|
Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0
|
Hs688(A).T
|
Colon ca.
0.0
0.0
Melanoma*
0.0
0.0
|
tissue(ODO3866)
(met)
|
Hs688(B).T
|
Colon ca. HCC-2998
8.4
0.0
Melanoma
0.0
0.0
|
UACC-62
|
Gastric ca.* (liver
18.0
4.2
Melanoma M14
0.0
0.0
|
met) NCI-N87
|
Bladder
0.0
0.0
Melanoma LOX
0.0
11.0
|
IMVI
|
Trachea
0.0
0.0
Melanoma*
0.0
0.0
|
(met) SK-MEL-5
|
Kidney
0.0
0.0
Adipose
0.0
0.0
|
|
[0856]
431
TABLE AD
|
|
|
Panel 4D
|
Rel. Exp. (%) Ag2841,
Rel. Exp. (%) Ag2841,
|
Tissue Name
Run 159616564
Tissue Name
Run 159616564
|
|
Secondary Th1 act
0.0
HUVEC IL-1 beta
2.7
|
Secondary Th2 act
0.0
HUVEC IFN gamma
8.8
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
15.9
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
2.2
|
Secondary Th2 rest
0.0
HUVEC IL-11
13.6
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
21.5
|
Primary Th1 act
0.0
Lung Microvascular EC
33.7
|
TNF alpha + IL-1 beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
12.1
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0
|
IL1 beta
|
Primary Th2 rest
0.0
Small airway epithelium none
2.4
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
10.8
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
8.3
|
act
IL-1 beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
9.8
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
0.0
Liver cirrhosis
15.1
|
LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0
|
LAK cells
0.0
NCI-H292 IL-9
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0
|
Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 5 day
0.0
HPAEC none
19.6
|
Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
7.0
|
PBMC rest
5.9
Lung fibroblast none
0.0
|
PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1 beta
|
PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0
|
B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
3.6
|
and IL-4
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0
|
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
PMA/ionomycin
1 beta
|
Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0
|
Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0
|
Monocytes rest
0.0
IBD Crohn's
0.0
|
Monocytes LPS
0.0
Colon
100.0
|
Macrophages rest
0.0
Lung
40.1
|
Macrophages LPS
0.0
Thymus
0.0
|
HUVEC none
5.8
Kidney
0.0
|
HUVEC starved
6.9
|
|
[0857] CNS_neurodegeneration_v1.0 Summary: Ag2841 Expression of the CG55912-01 gene is low (CTs>35) across all of the samples on this panel. Although levels are low for this gene, there is a significant difference in expression levels between non-demented controls and patients suffering from Alzheimer's disease, such that the levels of mRNA appear to be downregulated 2-fold in the postmortem AD brain (p=0.0018 when analyzed by ANCOVA; estimate of RNA loaded per well used as a covariate). This gene may therefore represent a drug target for the treatment of Alzheimer's disease or other dementias.
[0858] Panel 1.3D Summary: Ag2841 Two experiments with same primer and probe set are in excellent agreement, with highest expression of the CG55912-01 gene in the hippocampus region of the brain (CT=34). In addition, expression of this gene is exclusive to brain region. Therefore, expression of this gene can be used to distinguish this sample from other samples in the panel. In addition, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[0859] The CG55912-01 gene encodes a homolog of a neuronal voltage-gated calcium channel. In Caenorhabditis elegans voltage-gated calcium channels have been shown to direct neuronal migration. In C. elegans mutants carrying loss-of-function alleles of the calcium channel gene unc-2, the touch receptor neuron AVM and the interneuron SDQR often migrated inappropriately, leading to misplacement of their cell bodies (Tam T, Mathews E, Snutch T P, Schafer W R. (2000) Voltage-gated calcium channels direct neuronal migration in Caenorhabditis elegans. Dev Biol 226(1):104-17). Therefore, in analogy with C. elegan unc-2, neuronal voltage-gated calcium channel encoded by the CG55912-01 gene may also play a role in directing neuronal migration. In addition, calcium channels have been implicated in number of neurological diseases such as familial hemiplegic migraine, episodic ataxia type 2, spinocerebellar ataxia 6, and Lambert-Eaton myasthenic syndrome and other diseases (Greenberg DA. (1997) Calcium channels in neurological disease. Ann Neurol 42(3):275-82). Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of the different neurological diseases.
[0860] Panel 2D Summary: Ag2841 Expression of the CG55912-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[0861] Panel 3D Summary: Ag2841 Expression of the CG55912-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[0862] Panel 4D Summary: Ag2841 Low but significant expression of the CG55912-01 gene is detected exclusively in colon. Therefore, expression of this gene may be used to distinguish colon from the other tissues on this panel. Furthermore, expression of this gene is decreased in colon samples from patients with IBD colitis and Crohn's disease relative to normal colon. Therefore, therapeutic modulation of the activity of the calcium channel encoded by this gene may be useful in the treatment of inflammatory bowel disease.
[0863] Panel CNS—1 Summary: Ag2841 Expression of the CG55912-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[0864] B. CG55918-01 and CG55918-O2: ZINC TRANSPORTER 2 (ZNT-2)
[0865] Expression of gene CG55918-01 and full length clone CG55918-02 was assessed using the primer-probe set Ag2845, described in Table BA. Results of the RTQ-PCR runs are shown in Tables BB, BC, BD and BE. Please note that CG55918-02 represents a full-length physical clone of the CG55918-01 gene, validating the prediction of the gene sequence.
432TABLE BA
|
|
Probe Name Ag2845
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-aagggtcctgacagtcactgt-3′21123216
ProbeTET-5′-cagcgccagctgtatgtagcctctg-3′-25165217
TAMRA
Reverse5′-atcatgaacaacaggcagatg-3′21191218
|
[0866]
433
TABLE BB
|
|
|
Panel 1.3D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2845, Run
Ag2845, Run
Ag2845, Run
Ag2845, Run
|
Tissue Name
161590530
165701936
Tissue Name
161590530
165701936
|
|
Liver
0.8
0.4
Kidney (fetal)
3.4
4.5
|
adenocarcinoma
|
Pancreas
4.9
16.3
Renal ca. 786-0
0.0
0.0
|
Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0
|
CAPAN 2
|
Adrenal gland
0.5
0.7
Renal ca. RXF
0.0
0.0
|
393
|
Thyroid
4.7
7.8
Renal ca. ACHN
0.0
0.0
|
Salivary gland
0.4
0.7
Renal ca. UO-31
0.0
0.0
|
Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0
|
Brain (fetal)
0.0
0.0
Liver
0.0
0.0
|
Brain (whole)
0.0
0.0
Liver (fetal)
0.0
0.0
|
Brain (amygdala)
0.0
1.1
Liver ca.
0.2
0.0
|
(hepatoblast)
|
HepG2
|
Brain (cerebellum)
0.0
0.0
Lung
0.0
0.0
|
Brain (hippocampus)
0.1
0.0
Lung (fetal)
0.0
0.0
|
Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0
|
nigra)
cell) LX-1
|
Brain (thalamus)
0.0
0.0
Lung ca. (small
0.0
0.0
|
cell) NCI-H69
|
Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
0.0
0.0
|
var.) SHP-77
|
Spinal cord
0.0
0.0
Lung ca. (large
0.0
0.0
|
cell)NCI-H460
|
glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0
|
sm. cell) A549
|
glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.9
|
MG
s.cell) NCI-H23
|
astrocytoma
0.1
0.5
Lung ca. (non-
0.0
0.0
|
SW1783
s.cell) HOP-62
|
neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.3
0.0
|
AS
s.cl) NCI-H522
|
astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.2
|
(squam.) SW 900
|
astrocytoma SNB-75
0.0
0.5
Lung ca.
0.0
0.0
|
(squam.) NCI-
|
H596
|
glioma SNB-19
0.1
0.0
Mammary gland
0.3
0.9
|
glioma U251
0.0
0.0
Breast ca.*
0.0
0.0
|
(pl.ef) MCF-7
|
glioma SF-295
0.0
0.2
Breast ca.*
0.0
0.0
|
(pl.ef) MDA-
|
MB-231
|
Heart (fetal)
2.1
1.3
Breast ca.*
0.0
0.0
|
(pl.ef) T47D
|
Heart
2.8
2.1
Breast ca. BT-
0.0
1.0
|
549
|
Skeletal muscle
2.9
0.0
Breast ca. MDA-N
0.0
0.0
|
(fetal)
|
Skeletal muscle
0.4
2.5
Ovary
0.3
0.0
|
Bone marrow
0.0
0.0
Ovarian ca.
0.0
1.1
|
OVCAR-3
|
Thymus
0.0
0.0
Ovarian ca.
0.0
0.0
|
OVCAR-4
|
Spleen
0.0
0.0
Ovarian ca.
0.0
0.0
|
OVCAR-5
|
Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0
|
OVCAR-8
|
Colorectal
0.3
0.0
Ovarian ca.
0.0
0.0
|
IGROV-1
|
Stomach
0.0
0.2
Ovarian ca.*
0.0
0.0
|
(ascites) SK-OV-3
|
Small intestine
0.0
0.6
Uterus
0.1
0.6
|
Colon ca. SW480
1.6
3.7
Placenta
30.6
80.7
|
Colon ca.*
0.0
0.4
Prostate
0.0
0.0
|
SW620(SW480 met)
|
Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
0.0
|
(bone met)PC-3
|
Colon ca. HCT-116
0.2
0.0
Testis
0.5
2.3
|
Colon ca. CaCo-2
1.6
2.5
Melanoma
0.0
0.0
|
Hs688(A).T
|
Colon ca.
0.3
1.8
Melanoma*
0.0
0.0
|
tissue(ODO3866)
(met)
|
Hs688(B).T
|
Colon ca. HCC-2998
6.7
16.8
Melanoma
0.0
0.0
|
UACC-62
|
Gastric ca.* (liver
0.0
0.0
Melanoma M14
0.1
0.0
|
met) NCI-N87
|
Bladder
100.0
100.0
Melanoma LOX
0.0
0.0
|
IMVI
|
Trachea
0.5
0.7
Melanoma*
0.0
0.0
|
(met) SK-MEL-5
|
Kidney
10.9
11.8
Adipose
0.1
0.3
|
|
[0867]
434
TABLE BC
|
|
|
Panel 2D
|
Rel. Exp. (%) Ag2845,
Rel. Exp. (%) Ag2845,
|
Tissue Name
Run 161590724
Tissue Name
Run 161590724
|
|
Normal Colon
0.1
Kidney Margin 8120608
50.7
|
CC Well to Mod Diff
0.5
Kidney Cancer 8120613
0.0
|
(ODO3866)
|
CC Margin (ODO3866)
0.0
Kidney Margin 8120614
30.1
|
CC Gr.2 rectosigmoid
0.0
Kidney Cancer 9010320
0.0
|
(ODO3868)
|
CC Margin (ODO3868)
0.0
Kidney Margin 9010321
72.7
|
CC Mod Diff (ODO3920)
1.6
Normal Uterus
0.0
|
CC Margin (ODO3920)
0.1
Uterus Cancer 064011
0.0
|
CC Gr.2 ascend colon
0.3
Normal Thyroid
7.6
|
(ODO3921)
|
CC Margin (ODO3921)
0.3
Thyroid Cancer 064010
42.9
|
CC from Partial Hepatectomy
1.2
Thyroid Cancer A302152
14.4
|
(ODO4309) Mets
|
Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
1.1
|
Colon mets to lung (OD04451-
0.0
Normal Breast
0.3
|
01)
|
Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
0.7
|
Normal Prostate 6546-1
0.1
Breast Cancer (OD04590-
0.0
|
01)
|
Prostate Cancer (OD04410)
0.1
Breast Cancer Mets
0.0
|
(OD04590-03)
|
Prostate Margin (OD04410)
0.0
Breast Cancer Metastasis
0.0
|
(OD04655-05)
|
Prostate Cancer (OD04720-01)
0.2
Breast Cancer 064006
0.3
|
Prostate Margin (OD04720-02)
0.2
Breast Cancer 1024
2.0
|
Normal Lung 061010
0.4
Breast Cancer 9100266
0.1
|
Lung Met to Muscle
0.2
Breast Margin 9100265
0.1
|
(ODO4286)
|
Muscle Margin (ODO4286)
0.7
Breast Cancer A209073
0.1
|
Lung Malignant Cancer
0.0
Breast Margin A209073
0.2
|
(OD03126)
|
Lung Margin (OD03126)
0.0
Normal Liver
0.1
|
Lung Cancer (OD04404)
0.0
Liver Cancer 064003
0.1
|
Lung Margin (OD04404)
0.2
Liver Cancer 1025
0.0
|
Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.8
|
Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
0.0
|
Lung Cancer (OD04237-01)
0.2
Liver Tissue 6004-N
0.1
|
Lung Margin (OD04237-02)
0.3
Liver Cancer 6005-T
0.5
|
Ocular Mel Met to Liver
2.6
Liver Tissue 6005-N
0.1
|
(ODO4310)
|
Liver Margin (ODO4310)
0.0
Normal Bladder
100.0
|
Melanoma Mets to Lung
0.2
Bladder Cancer 1023
0.0
|
(OD04321)
|
Lung Margin (OD04321)
0.0
Bladder Cancer A302173
0.1
|
Normal Kidney
12.2
Bladder Cancer
0.3
|
(OD04718-01)
|
Kidney Ca, Nuclear grade 2
3.6
Bladder Normal Adjacent
0.2
|
(OD04338)
(OD04718-03)
|
Kidney Margin (OD04338)
15.9
Normal Ovary
0.0
|
Kidney Ca, Nuclear grade 1/2
0.1
Ovarian Cancer 064008
0.1
|
(OD04339)
|
Kidney Margin (OD04339)
20.9
Ovarian Cancer
0.2
|
(OD04768-07)
|
Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0
|
(OD04340)
08)
|
Kidney Margin (OD04340)
16.0
Normal Stomach
0.0
|
Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 9060358
0.2
|
(OD04348)
|
Kidney Margin (OD04348)
8.8
Stomach Margin 9060359
0.0
|
Kidney Cancer (OD04622-01)
0.2
Gastric Cancer 9060395
1.0
|
Kidney Margin (OD04622-03)
5.8
Stomach Margin 9060394
0.8
|
Kidney Cancer (OD04450-01)
1.2
Gastric Cancer 9060397
0.2
|
Kidney Margin (OD04450-03)
2.6
Stomach Margin 9060396
0.0
|
Kidney Cancer 8120607
0.3
Gastric Cancer 064005
0.0
|
|
[0868]
435
TABLE BD
|
|
|
Panel 3D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2845, Run
Ag2845, Run
|
Tissue Name
164843786
Tissue Name
164843786
|
|
Daoy-Medulloblastoma
0.0
Ca Ski-Cervical epidermoid
0.0
|
carcinoma (metastasis)
|
TE671-Medulloblastoma
0.4
ES-2-Ovarian clear cell carcinoma
0.0
|
D283 Med-Medulloblastoma
0.0
Ramos-Stimulated with
0.0
|
PMA/ionomycin 6 h
|
PFSK-1-Primitive
0.0
Ramos-Stimulated with
0.0
|
Neuroectodermal
PMA/ionomycin 14 h
|
XF-498-CNS
0.0
MEG-01-Chronic myelogenous
1.0
|
leukemia (megokaryoblast)
|
SNB-78-Glioma
0.0
Raji-Burkitt's lymphoma
0.0
|
SF-268-Glioblastoma
0.0
Daudi-Burkitt's lymphoma
0.0
|
T98G-Glioblastoma
1.0
U266-B-cell plasmacytoma
0.0
|
SK-N-SH-Neuroblastoma
0.0
CA46-Burkitt's lymphoma
0.0
|
(metastasis)
|
SF-295-Glioblastoma
0.0
RL-non-Hodgkin's B-cell
0.0
|
lymphoma
|
Cerebellum
0.0
JM1-pre-B-cell lymphoma
0.0
|
Cerebellum
0.0
Jurkat-T cell leukemia
0.0
|
NCI-H292-Mucoepidermoid
0.0
TF-1-Erythroleukemia
0.0
|
lung carcinoma
|
DMS-114-Small cell lung
2.4
HUT 78-T-cell lymphoma
0.0
|
cancer
|
DMS-79-Small cell lung
84.1
U937-Histiocytic lymphoma
0.0
|
cancer
|
NCI-H146-Small cell lung
0.0
KU-812-Myelogenous leukemia
0.0
|
cancer
|
NCI-H526-Small cell lung
1.0
769-P-Clear cell renal carcinoma
0.0
|
cancer
|
NCI-N417-Small cell lung
0.1
Caki-2-Clear cell renal carcinoma
0.2
|
cancer
|
NCI-H82-Small cell lung
0.0
SW 839-Clear cell renal carcinoma
0.0
|
cancer
|
NCI-H157-Squamous cell
0.0
G401-Wilms' tumor
0.9
|
lung cancer (metastasis)
|
NCI-H1155-Large cell lung
3.4
Hs766T-Pancreatic carcinoma (LN
0.0
|
cancer
metastasis)
|
NCI-H1299-Large cell lung
0.0
CAPAN-1-Pancreatic
0.6
|
cancer
adenocarcinoma (liver metastasis)
|
NCI-H727-Lung carcinoid
0.0
SU86.86-Pancreatic carcinoma
1.7
|
(liver metastasis)
|
NCI-UMC-11-Lung
0.0
BxPC-3-Pancreatic
0.0
|
carcinoid
adenocarcinoma
|
LX-1-Small cell lung cancer
0.0
HPAC-Pancreatic adenocarcinoma
0.5
|
Colo-205-Colon cancer
0.0
MIA PaCa-2-Pancreatic carcinoma
0.0
|
KM12-Colon cancer
0.0
CFPAC-1-Pancreatic ductal
100.0
|
adenocarcinoma
|
KM20L2-Colon cancer
1.9
PANC-1-Pancreatic epithelioid
0.0
|
ductal carcinoma
|
NCI-H716-Colon cancer
0.0
T24-Bladder carcinma (transitional
0.0
|
cell)
|
SW-48-Colon
0.0
5637-Bladder carcinoma
0.0
|
adenocarcinoma
|
SW1116-Colon
0.0
HT-1197-Bladder carcinoma
54.7
|
adenocarcinoma
|
LS 174T-Colon
2.7
UM-UC-3-Bladder carcinma
0.0
|
adenocarcinoma
(transitional cell)
|
SW-948-Colon
0.0
A204-Rhabdomyosarcoma
0.0
|
adenocarcinoma
|
SW-480-Colon
0.0
HT-1080-Fibrosarcoma
0.0
|
adenocarcinoma
|
NCI-SNU-5-Gastric
15.8
MG-63-Osteosarcoma
0.0
|
carcinoma
|
KATO III-Gastric carcinoma
0.6
SK-LMS-1-Leiomyosarcoma
0.0
|
(vulva)
|
NCI-SNU-16-Gastric
0.0
SJRH30-Rhabdomyosarcoma (met
0.0
|
carcinoma
to bone marrow)
|
NCI-SNU-1-Gastric
0.0
A431-Epidermoid carcinoma
0.0
|
carcinoma
|
RF-1-Gastric
0.0
WM266-4-Melanoma
1.0
|
adenocarcinoma
|
RF-48-Gastric
0.0
DU 145-Prostate carcinoma (brain
0.0
|
adenocarcinoma
metastasis)
|
MKN-45-Gastric carcinoma
0.0
MDA-MB-468-Breast
0.5
|
adenocarcinoma
|
NCI-N87-Gastric carcinoma
0.0
SCC-4-Squamous cell carcinoma
0.0
|
of tongue
|
OVCAR-5-Ovarian
0.0
SCC-9-Squamous cell carcinoma
0.0
|
carcinoma
of tongue
|
RL95-2-Uterine carcinoma
0.0
SCC-15-Squamous cell carcinoma
0.0
|
of tongue
|
HelaS3-Cervical
0.0
CAL 27-Squamous cell carcinoma
0.0
|
adenocarcinoma
of tongue
|
|
[0869]
436
TABLE BE
|
|
|
Panel 4D
|
Rel. Exp. (%) Ag2845,
Rel. Exp. (%) Ag2845,
|
Tissue Name
Run 159841918
Tissue Name
Run 159841918
|
|
Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1 beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.7
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0
|
IL1 beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1 beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.7
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
1.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
0.0
Liver cirrhosis
6.3
|
LAK cells IL-2 + IL-12
0.0
Lupus kidney
6.2
|
LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0
|
LAK cells
0.0
NCI-H292 IL-9
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0
|
Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.7
|
Two Way MLR 5 day
0.0
HPAEC none
0.0
|
Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
1.1
|
PBMC rest
0.0
Lung fibroblast none
0.0
|
PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1 beta
|
PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0
|
B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0
|
and IL-4
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0
|
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
PMA/ionomycin
1 beta
|
Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0
|
Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.5
|
Monocytes rest
0.0
IBD Crohn's
0.5
|
Monocytes LPS
0.0
Colon
12.8
|
Macrophages rest
0.0
Lung
5.1
|
Macrophages LPS
0.0
Thymus
100.0
|
HUVEC none
0.0
Kidney
0.7
|
HUVEC starved
0.0
|
|
[0870] Panel 1.3D Summary: Ag2845 Two experiments with same primer and probe sets are in excellent agreement, with highest expression of the CG55918-01 gene in bladder (CTs=27-30). In addition, high expression of this gene is also seen in placenta. Thus, expression of this gene can be used to distinguish these tissue samples from other samples in the panel. Furthermore, therapeutic modulation of the activity of the zinc transporter encoded by this gene may be useful in the treatment diseases associated with bladder and placenta including fertility and reproductive disorders.
[0871] Among tissues with metabolic or endocrine function, this gene is expressed at low levels in pancreas, adrenal gland, thyroid, skeletal muscle, and heart. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[0872] Panel 2D Summary: Ag2845 Highest expression of the CG55918-01 gene is detected in bladder (CT=26.7). Interestingly, expression of this gene is down regulated in bladder and kidney cancer samples (CTs>35) as compared to the corresponding control margin samples (CTs=27-30). Therefore, therapeutic modulation of the activity of the zinc transporter encoded by this gene may be useful in the treatment bladder and kidney cancers.
[0873] In addition, expression of this gene is up-regulated in thyroid cancer A302152 (CT=29.6) as compared to control margin sample (CT=33.4). Therefore, expression of this gene can be used as a diagnostic marker for thyroid cancer and therapeutic modulation of the activity of the zinc transporter encoded by this gene may be useful in the treatment this cancer.
[0874] Panel 3D Summary: Ag2845 Highest expression of the CG55918-01 gene is detected in the CFPAC-1-pancreatic ductal adenocarcinoma cell line (CT=29.6). In addition, high expression of this gene is also detected in HT-1197-bladder carcinoma, and DMS-79-small cell lung cancer cell line. Therefore, expression of this gene can be used as a diagnostic marker for these cancers and also, therapeutic modulation of the activity of the zinc transporter encoded by this gene may be useful in the treatment of these cancers.
[0875] Panel 4D Summary: Ag2845 Highest expression of the CG55918-01 gene is detected in thymus (CT=29.6). The zinc transporter encoded for by this gene could therefore play an important role in T cell development. Small molecule therapeutics, or antibody therapeutics designed against the protein encoded for by this gene could be utilized to modulate immune function (T cell development) and be important for organ transplant, AIDS treatment or post chemotherapy immune reconstitution.
[0876] In addition, low but significant expression of this gene is also seen in colon (CT=33). Furthermore, expression of this gene is decreased in colon samples from patients with IBD colitis and Crohn's disease relative to normal colon. Therefore, therapeutic modulation of the activity of the protein encoded by this gene may be useful in the treatment of inflammatory bowel disease.
[0877] Low expression of this gene is also observed in lung, liver cirrhosis and lupus kidney samples (CTs=34). Therefore, therapeutic modulation of the activity of the zinc transporter encoded by this gene may be useful in the treatment of disease associated with lung, liver cirrhosis and lupus disease.
[0878] C. CG56832-01 and CG56832-02 and CG56832-03: Guanylate Kinase
[0879] Expression of gene CG56832-01, variant CG56832-03 and clone CG56832-02 was assessed using the primer-probe sets Ag3O33 and Ag3718, described in Tables CA and CB. Results of the RTQ-PCR runs are shown in Tables CC, CD and CE.
437TABLE CA
|
|
Probe Name Ag3033
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-tgtggacatgaagttcaaggat-3′221154219
ProbeTET-5′-cctacaagagatggaaaatttagccca-271181220
3′-TAMRA
Reverse5′-gccaaactgagtttccattctt-3′221208221
|
[0880]
438
TABLE CB
|
|
|
Probe Name Ag3718
|
SEQ
|
Start
ID
|
Primers
Sequences
Length
Position
NO:
|
|
Forward
5′-tgtggacatgaagttcaaggat-3′
22
1154
222
|
Probe
TET-5′-cctacaagagatggaaaatttagccca-
27
1181
223
|
3′-TAMRA
|
Reverse
5′-gccaaactgagtttccattctt-3′
22
1208
224
|
|
[0881]
439
TABLE CC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp.(%) Ag3718, Run
Rel. Exp.(%) Ag3718, Run
|
Tissue Name
218267394
Tissue Name
218267394
|
|
Adipose
0.4
Renal ca. TK-10
0.6
|
Melanoma* Hs688(A).T
1.9
Bladder
0.2
|
Melanoma* Hs688(B).T
2.3
Gastric ca. (liver met.)
0.6
|
NCI-N87
|
Melanoma* M14
0.8
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
100.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
0.2
Colon ca. SW480
1.4
|
Squamous cell
0.3
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
0.3
SW620
|
Testis Pool
0.0
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
1.2
Colon ca. HCT-116
0.4
|
PC-3
|
Prostate Pool
0.0
Colon ca. CaCo-2
0.1
|
Placenta
0.0
Colon cancer tissue
0.2
|
Uterus Pool
0.0
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
1.0
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
0.0
|
Ovarian ca. OVCAR-5
0.6
Small Intestine Pool
0.0
|
Ovarian ca. IGROV-1
2.3
Stomach Pool
0.0
|
Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
0.1
|
Ovary
0.2
Fetal Heart
0.2
|
Breast ca. MCF-7
0.0
Heart Pool
0.0
|
Breast ca. MDA-MB-
2.3
Lymph Node Pool
0.2
|
231
|
Breast ca. BT 549
1.1
Fetal Skeletal Muscle
0.2
|
Breast ca. T47D
0.5
Skeletal Muscle Pool
0.4
|
Breast ca. MDA-N
0.0
Spleen Pool
4.0
|
Breast Pool
0.2
Thymus Pool
0.2
|
Trachea
0.2
CNS cancer (glio/astro)
42.9
|
U87-MG
|
Lung
0.3
CNS cancer (glio/astro) U-
33.9
|
118-MG
|
Fetal Lung
0.0
CNS cancer (neuro; met)
4.2
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.2
|
Lung Ca. LX-1
0.0
CNS cancer (astro) SNB-75
14.9
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
2.4
|
Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
0.8
|
Lung ca. A549
0.0
Brain (Amygdala) Pool
0.2
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
1.3
|
Lung ca. NCI-H23
0.2
Brain (fetal)
0.3
|
Lung ca. NCI-H460
4.5
Brain (Hippocampus) Pool
0.8
|
Lung ca. HOP-62
0.7
Cerebral Cortex Pool
1.0
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
0.1
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
1.1
|
Fetal Liver
0.0
Brain (whole)
0.5
|
Liver ca. HepG2
0.0
Spinal Cord Pool
0.3
|
Kidney Pool
0.0
Adrenal Gland
0.0
|
Fetal Kidney
0.4
Pituitary gland Pool
0.2
|
Renal ca. 786-0
0.3
Salivary Gland
0.0
|
Renal ca. A498
0.4
Thyroid (female)
0.0
|
Renal Ca. ACHN
0.4
Pancreatic ca. CAPAN2
0.2
|
Renal ca. UO-31
5.8
Pancreas Pool
0.0
|
|
[0882]
440
TABLE CD
|
|
|
Panel 1.3D
|
Rel. Exp. (%) Ag3033, Run
Rel. Exp. (%) Ag3033, Run
|
Tissue Name
167962289
Tissue Name
167962289
|
|
Liver adenocarcinoma
2.0
Kidney (fetal)
0.0
|
Pancreas
0.0
Renal ca. 786-0
1.0
|
Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
6.5
|
Adrenal gland
0.0
Renal ca. RXF 393
0.0
|
Thyroid
0.0
Renal ca. ACHN
0.0
|
Salivary gland
0.0
Renal ca. UO-31
19.9
|
Pituitary gland
1.5
Renal ca. TK-10
0.0
|
Brain (fetal)
4.0
Liver
0.0
|
Brain (whole)
5.1
Liver (fetal)
0.0
|
Brain (amygdala)
3.6
Liver ca. (hepatoblast)
0.0
|
HepG2
|
Brain (cerebellum)
13.1
Lung
0.0
|
Brain (hippocampus)
4.3
Lung (fetal)
2.3
|
Brain (substantia nigra)
1.0
Lung ca. (small cell) LX-1
0.0
|
Brain (thalamus)
1.4
Lung ca. (small cell)
8.5
|
NCI-H69
|
Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0
|
SHP-77
|
Spinal cord
2.8
Lung ca. (large cell)NCI-
2.5
|
H460
|
glio/astro U87-MG
40.9
Lung ca. (non-sm. cell)
0.0
|
A549
|
glio/astro U-118-MG
31.4
Lung ca. (non-s.cell)
0.9
|
NCI-H23
|
astrocytoma SW1783
27.7
Lung ca. (non-s.cell)
0.9
|
HOP-62
|
neuro*; met SK-N-AS
1.0
Lung ca. (non-s.cl) NCI-
0.0
|
H522
|
astrocytoma SF-539
1.1
Lung ca. (squam.) SW
0.0
|
900
|
astrocytoma SNB-75
9.2
Lung ca. (squam.) NCI-
0.0
|
H596
|
glioma SNB-19
3.7
Mammary gland
0.0
|
glioma U251
0.3
Breast ca.* (pl.ef) MCF-7
0.0
|
glioma SF-295
1.0
Breast ca.* (pl.ef) MDA
3.8
|
MB-231
|
Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0
|
Heart
0.0
Breast ca. BT-549
3.6
|
Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
3.3
|
Skeletal muscle
2.0
Ovary
0.0
|
Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0
|
Thymus
0.0
Ovarian ca. OVCAR-4
0.0
|
Spleen
5.4
Ovarian ca. OVCAR-5
3.7
|
Lymph node
0.0
Ovarian ca. OVCAR-8
1.0
|
Colorectal
0.0
Ovarian ca. IGROV-1
0.0
|
Stomach
0.0
Ovarian ca.* (ascites)
7.7
|
SK-OV-3
|
Small intestine
0.0
Uterus
0.0
|
Colon ca. SW480
0.0
Placenta
0.0
|
Colon ca.* SW620(SW480
1.1
Prostate
0.0
|
met)
|
Colon ca. HT29
0.0
Prostate ca.* (bone
2.5
|
met)PC-3
|
Colon ca. HCT-116
0.0
Testis
2.7
|
Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
3.0
|
Colon ca.
0.0
Melanoma* (met)
2.0
|
tissue(ODO3866)
Hs688(B).T
|
Colon ca. HCC-2998
2.1
Melanoma UACC-62
0.0
|
Gastric ca.* (liver met)
1.0
Melanoma M14
0.0
|
NCI-N87
|
Bladder
0.0
Melanoma LOX IMVI
100.0
|
Trachea
1.3
Melanoma* (met) SK-
0.0
|
MEL-5
|
Kidney
0.0
Adipose
1.1
|
|
[0883]
441
TABLE CE
|
|
|
Panel 4D
|
Rel. Exp. (%) Ag3033,
Rel. Exp. (%) Ag3033,
|
Tissue Name
Run 162427946
Tissue Name
Run 162427946
|
|
Secondary Th1 act
0.0
HUVEC IL-1 beta
19.3
|
Secondary Th2 act
0.0
HUVEC IFN gamma
4.1
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
18.4
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
18.3
|
Secondary Th2 rest
0.0
HUVEC IL-11
7.2
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
10.8
|
Primary Th1 act
0.0
Lung Microvascular EC
12.3
|
TNF alpha + IL-1 beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
30.1
|
Primary Tr1 act
0.4
Microsvasular Dermal EC
18.7
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.7
|
IL1 beta
|
Primary Th2 rest
0.0
Small airway epithelium none
1.9
|
Primary Tr1 rest
0.0
Small airway epithelium
1.3
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
2.7
Coronery artery SMC rest
36.6
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
16.8
|
act
IL-1 beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
8.8
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
7.1
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.5
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0
|
LAK cells IL-2 + IFN
0.5
NCI-H292 none
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.4
|
LAK cells
0.0
NCI-H292 IL-9
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
1.2
NCI-H292 IL-13
0.0
|
Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 5 day
0.0
HPAEC none
10.4
|
Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
36.6
|
PBMC rest
0.0
Lung fibroblast none
0.9
|
PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.4
|
1 beta
|
PBMC PHA-L
0.0
Lung fibroblast IL-4
1.2
|
Ramos (B cell) none
0.0
Lung fibroblast IL-9
5.6
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.3
|
B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.4
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
13.5
|
and IL-4
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
38.2
|
TNF alpha
|
EOL-1 dbcAMP
0.4
Dermal fibroblast CCD1070 IL-
19.9
|
PMA/ionomycin
1 beta
|
Dendritic cells none
0.0
Dermal fibroblast IFN gamma
1.2
|
Dendritic cells LPS
0.0
Dermal fibroblast IL-4
2.2
|
Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0
|
Monocytes rest
0.0
IBD Crohn's
0.0
|
Monocytes LPS
0.0
Colon
0.5
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
0.0
Thymus
0.0
|
HUVEC none
62.9
Kidney
0.4
|
HUVEC starved
100.0
|
|
[0884] CNS neurodegeneration v1.0 Summary: Ag3718 Expression of the CG56832-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[0885] General_screeningpanel_v1.4 Summary: Ag3718 Highest expression of the CG56832-01 gene is detected in melanoma LOXIMVI cell line (CT=28.6). Thus, expression of this gene can be used to distinguish this sample from other samples in this panel. In addition, significant expression of this gene is associated with number of CNS cancer, renal cancer UO-31, lung cancer NCI-H460, breast cancer MDA-MB-231, and ovarian cancer IGROV-1 cell lines. The CG56832-01 gene codes for a homologue of Drosophila the tumour suppressor protein Dlg, a membrane-associated guanylate kinase homolog (MAGUK). In Drosophila, genetic loss of the tumour suppressor protein Dlg (in dIg mutants) or p 127 (in 1 gl mutants) leads to loss of epithelial structure and excess proliferation in the imaginal discs and brain of the developing larva. These phenotypes show most of the characteristic features of human neoplasia (De Lorenzo C, Mechler B M, Bryant P J. (1999) What is Drosophila telling us about cancer? Cancer Metastasis Rev 18(2):295-311). Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, or antibodies, might be beneficial in the treatment of these cancers.
[0886] Panel 1.3D Summary: Ag3033 Highest expression of the CG56832-01 gene is detected in melanoma LOXIMVI cell line (CT=31.3). Thus, expression of this gene can be used to distinguish this sample from other samples in this panel. In addition, significant expression of this gene is associated with number of CNS cancer, renal cancer UO-31, lung cancer NCI-H69, and ovarian cancer SK-OV-3 cell lines. Please see Panel 1.4 for a discussion of the potential utility of this gene.
[0887] In addition, low but significant expression of this gene is also seen in brain cerebellum sample (CT=34.2). Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, might be beneficial in the treatment of ataxia and autism.
[0888] Panel 4D Summary: Ag3033 Highest expression of the CG56832-01 gene is detected in starved HUVEC cells (CT=30). In addition, significant expression of this gene is seen in dermal fibroblasts, IL-9 treated lung fibroblast, HPAEC, astrocytes, coronery artery SMC, microvascular dermal EC, lung microvascular EC, and HUVEC. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, might be beneficial in the treatment of autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[0889] CG59580-01: GPCR
[0890] Expression of gene CG59580-01 was assessed using the primer-probe set Ag3481, described in Table DA. Results of the RTQ-PCR runs are shown in Table DB.
442TABLE DA
|
|
Probe Name Ag3481
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-ccatgtacttcttcctctccaa-3′22273225
ProbeTET-5′-tgtctaccactgtcccaaagatgctg-3′-26321226
TAMRA
Reverse5′-gctctgtgtctggatgttcac-3′21347227
|
[0891]
443
TABLE DB
|
|
|
Panel 4D
|
Rel. Exp. (%) Ag3481,
Rel. Exp. (%) Ag3481,
|
Tissue Name
Run 166441541
Tissue Name
Run 166441541
|
|
Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0
|
Secondary Th2 act
0.3
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1 beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microvasular Dermal EC
0.0
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0
|
IL1 beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1 beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.4
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
0.0
Liver cirrhosis
100.0
|
LAK cells IL-2 + IL-12
0.0
Lupus kidney
8.4
|
LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0
|
LAK cells
0.0
NCI-H292 IL-9
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0
|
Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 5 day
0.0
HPAEC none
0.0
|
Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0
|
PBMC rest
0.0
Lung fibroblast none
0.0
|
PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1 beta
|
PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0
|
B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0
|
and IL-4
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0
|
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
PMA/ionomycin
1 beta
|
Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0
|
Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells anti-CD40
0.0
IBD Colitis 2
6.3
|
Monocytes rest
0.0
IBD Crohn's
0.0
|
Monocytes LPS
0.0
Colon
9.2
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
0.0
Thymus
0.4
|
HUVEC none
0.0
Kidney
0.0
|
HUVEC starved
0.0
|
|
[0892] CNS_neurodegeneration_v1.0 Summary: Ag3481 Expression of the CG59580-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[0893] General_screening_panel_v1.4 Summary: Ag3481 Expression of the CG59580-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[0894] Panel 4D Summary: Ag3481 Expression of the CG59580-01 gene is detected in a liver cirrhosis sample (CT 31.3). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.
[0895] E. CG59611-01: ODORANT RECEPTOR
[0896] Expression of gene CG59611-01 was assessed using the primer-probe set Ag3495, described in Table EA.
444TABLE EA
|
|
Probe Name Ag3495
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-ctatccacctcacaccttcact-3′22152228
ProbeTET-5′-cacgtacttcctgctcagcaacctgt-3′-26178229
TAMRA
Reverse5′-gaggacaggcacatgtcaatat-3′22207230
|
[0897] CNS_neurodegeneration_v1.0 Summary: Ag3495 Expression of the CG59611-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.) The amp plot indicates that there is a high probability of a probe failure.
[0898] General_screening_panel_v1.4 Summary: Ag3495 Results from one experiment with the CG59611-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[0899] Panel 4.1D Summary: Ag3495 Expression of the CG59611-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.) The amp plot indicates that there is a high probability of a probe failure.
[0900] F. CG59617-01: GPCR
[0901] Expression of gene CG59617-01 was assessed using the primer-probe set Ag3497, described in Table FA.
445TABLE FA
|
|
Probe Name Ag3497
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-gccagttgcttcatgttaattc-3′22649231
ProbeTET-5′-tgccaacatagtaaatgccatcctga-3′-26681232
TAMRA
Reverse5′-cccatcagtggtgcgtatc-3′19708233
|
[0902] CNS_neurodegeneration_v1.0 Summary: Ag3497 Expression of the CG59617-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[0903] General_screening_panel_v1.4 Summary: Ag3497 Expression of the CG59617-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[0904] Panel 4.1D Summary: Ag3497 Expression of the CG59617-01 gene is limited to a few samples, with highest expression in a 7 day post MLR (CT=28.4).
[0905] G. CG59826-01: Transformation-Sensitive Protein IEF SSP like Protein
[0906] Expression of gene CG59826-01 was assessed using the primer-probe set Ag3599, described in Table GA. Results of the RTQ-PCR runs are shown in Tables GB, GC and GD.
446TABLE GA
|
|
Probe Name Ag3599
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-agctgcctgttacaccaaact-3′211213234
ProbeTET-5′-atttccagctggcactcaaggactgt-3′-261239235
TAMRA
Reverse5′-aaggtcggttctagctggatac-3′221272236
|
[0907]
447
TABLE GB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3599, Run
Ag3599, Run
Ag3599, Run
Ag3599, Run
|
Tissue Name
211010105
224079046
Tissue Name
211010105
224079046
|
|
AD 1 Hippo
28.1
26.8
Control (Path)
5.3
5.8
|
3 Temporal
|
Ctx
|
AD 2 Hippo
29.5
33.0
Control (Path)
21.6
26.8
|
4 Temporal
|
Ctx
|
AD 3 Hippo
24.8
25.3
AD 1 Occipital
35.4
37.9
|
Ctx
|
AD 4 Hippo
6.2
7.3
AD 2 Occipital
0.0
0.0
|
Ctx (Missing)
|
AD 5 hippo
87.7
100.0
AD 3 Occipital
24.3
29.3
|
Ctx
|
AD 6 Hippo
71.2
85.9
AD 4 Occipital
24.3
18.7
|
Ctx
|
Control 2 Hippo
36.9
44.8
AD 5 Occipital
37.6
76.8
|
Ctx
|
Control 4 Hippo
9.6
10.7
AD 6 Occipital
61.1
41.5
|
Ctx
|
Control (Path) 3
7.4
8.4
Control 1
6.0
7.2
|
Hippo
Occipital Ctx
|
AD 1 Temporal
28.7
28.1
Control 2
69.3
65.5
|
Ctx
Occipital Ctx
|
AD 2 Temporal
35.4
40.6
Control 3
13.0
14.7
|
Ctx
Occipital Ctx
|
AD 3 Temporal
16.8
18.4
Control 4
7.3
8.2
|
Ctx
Occipital Ctx
|
AD 4 Temporal
20.0
20.3
Control (Path)
57.4
67.4
|
Ctx
1 Occipital Ctx
|
AD 5 Inf
100.0
95.9
Control (Path)
8.4
11.3
|
Temporal Ctx
2 Occipital Ctx
|
AD 5
47.0
49.0
Control (Path)
3.2
3.3
|
SupTemporal Ctx
3 Occipital Ctx
|
AD 6 Inf
60.7
77.9
Control (Path)
9.9
11.0
|
Temporal Ctx
4 Occipital Ctx
|
AD 6 Sup
61.6
73.2
Control 1
9.0
10.7
|
Temporal Ctx
Parietal Ctx
|
Control 1
7.9
9.4
Control 2
33.2
40.3
|
Temporal Ctx
Parietal Ctx
|
Control 2
40.1
47.0
Control 3
18.0
21.6
|
Temporal Ctx
Parietal Ctx
|
Control 3
15.8
17.8
Control (Path)
59.9
72.2
|
Temporal Ctx
1 Parietal Ctx
|
Control 4
6.2
7.4
Control (Path)
21.6
24.3
|
Temporal Ctx
2 Parietal Ctx
|
Control (Path) 1
41.5
50.3
Control (Path)
4.5
4.4
|
Temporal Ctx
3 Parietal Ctx
|
Control (Path) 2
28.9
34.2
Control (Path)
33.7
40.3
|
Temporal Ctx
4 Parietal Ctx
|
|
[0908]
448
TABLE GC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3599, Run
Ag3599, Run
|
Tissue Name
217676501
Tissue Name
217676501
|
|
Adipose
3.9
Renal ca. TK-10
33.0
|
Melanoma* Hs688(A).T
14.9
Bladder
7.6
|
Melanoma* Hs688(B).T
14.3
Gastric ca. (liver met.)
29.7
|
NCI-N87
|
Melanoma* M14
50.0
Gastric ca. KATO III
97.3
|
Melanoma* LOXIMVI
54.3
Colon ca. SW-948
9.7
|
Melanoma* SK-MEL-5
56.6
Colon ca. SW480
88.3
|
Squamous cell
54.7
Colon ca.* (SW480 met)
39.8
|
carcinoma SCC-4
SW620
|
Testis Pool
10.3
Colon ca. HT29
29.7
|
Prostate ca.* (bone met)
34.2
Colon ca. HCT-116
100.0
|
PC-3
|
Prostate Pool
3.0
Colon ca. CaCo-2
33.9
|
Placenta
6.4
Colon cancer tissue
10.4
|
Uterus Pool
1.7
Colon ca. SW1116
7.9
|
Ovarian ca. OVCAR-3
39.5
Colon ca. Colo-205
11.3
|
Ovarian ca. SK-OV-3
37.9
Colon ca. SW-48
15.1
|
Ovarian ca. OVCAR-4
29.3
Colon Pool
2.2
|
Ovarian ca. OVCAR-5
33.0
Small Intestine Pool
4.6
|
Ovarian ca. IGROV-1
17.0
Stomach Pool
3.5
|
Ovarian ca. OVCAR-8
13.0
Bone Marrow Pool
1.7
|
Ovary
4.8
Fetal Heart
5.9
|
Breast ca. MCF-7
19.8
Heart Pool
2.7
|
Breast ca. MDA-MB-
60.7
Lymph Node Pool
6.0
|
231
|
Breast ca. BT 549
65.1
Fetal Skeletal Muscle
4.3
|
Breast ca. T47D
57.4
Skeletal Muscle Pool
15.7
|
Breast ca. MDA-N
28.3
Spleen Pool
4.2
|
Breast Pool
5.3
Thymus Pool
5.1
|
Trachea
5.0
CNS cancer (glio/astro)
41.2
|
U87-MG
|
Lung
1.0
CNS cancer (glio/astro) U-
74.2
|
118-MG
|
Fetal Lung
11.3
CNS cancer (neuro; met)
34.4
|
SK-N-AS
|
Lung ca. NCI-N417
8.9
CNS cancer (astro) SF-539
24.1
|
Lung ca. LX-1
33.2
CNS cancer (astro) SNB-75
36.3
|
Lung ca. NCI-H146
18.0
CNS cancer (glio) SNB-19
14.7
|
Lung ca. SHP-77
42.0
CNS cancer (glio) SF-295
39.0
|
Lung ca. A549
43.5
Brain (Amygdala) Pool
6.0
|
Lung ca. NCI-H526
19.6
Brain (cerebellum)
26.4
|
Lung ca. NCI-H23
27.5
Brain (fetal)
11.8
|
Lung ca. NCI-H460
17.1
Brain (Hippocampus) Pool
6.3
|
Lung ca. HOP-62
12.5
Cerebral Cortex Pool
9.1
|
Lung ca. NCI-H522
38.4
Brain (Substantia nigra)
7.9
|
Pool
|
Liver
1.3
Brain (Thalamus) Pool
9.5
|
Fetal Liver
13.6
Brain (whole)
9.2
|
Liver ca. HepG2
10.0
Spinal Cord Pool
7.2
|
Kidney Pool
5.8
Adrenal Gland
15.0
|
Fetal Kidney
5.3
Pituitary gland Pool
1.5
|
Renal ca. 786-0
17.7
Salivary Gland
3.0
|
Renal ca. A498
8.7
Thyroid (female)
4.3
|
Renal ca. ACHN
9.9
Pancreatic ca. CAPAN2
28.1
|
Renal ca. UO-31
26.2
Pancreas Pool
7.3
|
|
[0909]
449
TABLE GD
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3599,
Ag3599,
|
Tissue Name
Run 169910500
Tissue Name
Run 169910500
|
|
Secondary Th1 act
95.9
HUVEC IL-1beta
46.0
|
Secondary Th2 act
100.0
HUVEC IFN gamma
32.5
|
Secondary Tr1 act
91.4
HUVEC TNF alpha + IFN
26.8
|
gamma
|
Secondary Th1 rest
6.1
HUVEC TNF alpha + IL4
30.4
|
Secondary Th2 rest
14.5
HUVEC IL-11
17.1
|
Secondary Tr1 rest
9.5
Lung Microvascular EC none
28.3
|
Primary Th1 act
90.1
Lung Microvascular EC
25.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
54.7
Microvascular Dermal EC none
25.5
|
Primary Tr1 act
84.1
Microsvasular Dermal EC
26.1
|
TNF alpha + IL-1beta
|
Primary Th1 rest
14.2
Bronchial epithelium
18.7
|
TNF alpha + IL1beta
|
Primary Th2 rest
10.4
Small airway epithelium none
12.3
|
Primary Tr1 rest
22.5
Small airway epithelium
19.5
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
51.4
Coronery artery SMC rest
25.5
|
act
|
CD45RO CD4 lymphocyte
55.9
Coronery artery SMC
20.0
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
68.3
Astrocytes rest
19.2
|
Secondary CD8
68.3
Astrocytes TNF alpha + IL-1beta
13.8
|
lymphocyte rest
|
Secondary CD8
41.5
KU-812 (Basophil) rest
47.3
|
lymphocyte act
|
CD4 lymphocyte none
3.1
KU-812 (Basophil)
75.8
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
10.1
CCD1106 (Keratinocytes) none
29.1
|
CD95 CH11
|
LAK cells rest
32.8
CCD1106 (Keratinocytes)
27.7
|
TNF alpha + IL-1beta
|
LAK cells IL-2
44.4
Liver cirrhosis
2.9
|
LAK cells IL-2 + IL-12
61.6
NCI-H292 none
22.7
|
LAK cells IL-2 + IFN
53.2
NCI-H292 IL-4
42.0
|
gamma
|
LAK cells IL-2 + IL-18
50.3
NCI-H292 IL-9
53.6
|
LAK cells
44.8
NCI-H292 IL-13
45.7
|
PMA/ionomycin
|
NK Cells IL-2 rest
28.7
NCI-H292 IFN gamma
42.6
|
Two Way MLR 3 day
27.7
HPAEC none
22.1
|
Two Way MLR 5 day
51.1
HPAEC TNF alpha + IL-1beta
39.5
|
Two Way MLR 7 day
28.7
Lung fibroblast none
21.6
|
PBMC rest
4.9
Lung fibroblast TNF alpha + IL-
15.5
|
1beta
|
PBMC PWM
59.9
Lung fibroblast IL-4
31.2
|
PBMC PHA-L
50.0
Lung fibroblast IL-9
34.4
|
Ramos (B cell) none
44.1
Lung fibroblast IL-13
25.3
|
Ramos (B cell) ionomycin
39.0
Lung fibroblast IFN gamma
25.3
|
B lymphocytes PWM
48.6
Dermal fibroblast CCD1070 rest
37.6
|
B lymphocytes CD40L
27.7
Dermal fibroblast CCD1070
52.9
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
25.7
Dermal fibroblast CCD1070 IL-
24.7
|
1beta
|
EOL-1 dbcAMP
28.3
Dermal fibroblast IFN gamma
15.8
|
PMA/ionomycin
|
Dendritic cells none
43.8
Dermal fibroblast IL-4
35.8
|
Dendritic cells LPS
17.8
Dermal Fibroblast rest
20.6
|
Dendritic cells anti-CD40
47.0
Neutrophils TNFa + LPS
2.8
|
Monocytes rest
8.5
Neutrophils rest
1.9
|
Monocytes LPS
18.4
Colon
5.0
|
Macrophages rest
50.7
Lung
10.7
|
Macrophages LPS
13.8
Thymus
11.5
|
HUVEC none
25.2
Kidney
9.4
|
HUVEC starved
29.1
|
|
[0910] CNS_neurodegeneration_v1.0 Summary: Ag3599 Two experiments with the same probe and primer produce results that are in excellent agreement. The CG59826-01 gene appears to be slightly upregulated in the temporal cortex of Alzheimer's disease patients. Therefore, blockade of this receptor may decrease neuronal death and be of use in the treatment of Alzheimer's disease and other neurodegenerative disorders.
[0911] General_screening_panel_v1.4 Summary: Ag3599 Highest expression of the CG59826-01 gene is seen in a colon cancer cell line (CT=23.8). Significant levels of expression are seen in a cluster of samples derived from ovarian, breast, brain, colon, gastric, melanoma, pancreatic and lung cancer cell lines. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of ovarian, breast, brain, colon, gastric, melanoma, pancreatic and lung cancers. Overall, this gene is more highly expressed in cancer cell lines and some fetal cell lines than in the samples derived from normal tissues. This expression profile suggests a role for this gene in cellular growth and differentiation.
[0912] In addition, this gene is expressed at much higher levels in fetal lung and liver and skeletal muscle tissue (CTs=26-27) when compared to expression in the adult counterpart (CTs=30). Thus, expression of this gene may be used to differentiate between the fetal and adult source of these tissues.
[0913] Among tissues with metabolic function, this gene is expressed at moderate to high levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[0914] This molecule is also expressed at high levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy. Please see Panel CNS_neurodegeneration_v1.0 for further discussion of utility of this gene in the central nervous system.
[0915] Panel 4.1D Summary: Ag3599 The CG59826-01 gene is ubiquitously expressed in this panel, with highest expression in activated secondary Th2 cells (CT=26). This gene appears to be more highly expressed in activated T cells than in resting T cells. Thus, expression of this gene could be used to identify activated T cells. Furthermore, therapeutic regulation of the transcript or the protein encoded by the transcript could be important in the treatment of T cell-mediated diseases such as asthma, arthritis, psoriasis, IBD, and lupus.
[0916] H. CG59839-01: CATION-TRANSPORTING ATPASE
[0917] Expression of gene CG59839-01 was assessed using the primer-probe sets Ag1417 and Ag3604, described in Tables HA and HB. Results of the RTQ-PCR runs are shown in Tables HC, HD and HE.
450TABLE HA
|
|
Probe Name Ag1417
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ataggaaaatggacgcctacat-3′22730237
ProbeTET-5′-ccattgccggtctctgtaaacctgaa-26769238
3′-TAMRA
Reverse5′-ttttgaaaatcgacaggaactg-3′22796239
|
[0918]
451
TABLE HB
|
|
|
Probe Name Ag3604
|
Start
SEQ ID
|
Primers
Sequences
Length
Position
NO:
|
|
Forward
5′-gcaattgagaacaacatggatt-3′
22
924
240
|
Probe
TET-5′-
26
971
241
|
caaattaaagcaagaaacccctgcag-
|
3′-TAMRA
|
Reverse
5′-tgttggctttatgcaaatcttc-3′
22
1002
242
|
|
[0919]
452
TABLE HC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3604, Run
Ag3604,
|
Tissue Name
210997046
Tissue Name
Run 210997046
|
|
AD 1 Hippo
8.8
Control (Path) 3
9.1
|
Temporal Ctx
|
AD 2 Hippo
26.8
Control (Path) 4
39.8
|
Temporal Ctx
|
AD 3 Hippo
7.3
AD 1 Occipital Ctx
14.3
|
AD 4 Hippo
10.7
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
97.9
AD 3 Occipital Ctx
5.0
|
AD 6 Hippo
87.7
AD 4 Occipital Ctx
23.3
|
Control 2 Hippo
28.9
AD 5 Occipital Ctx
47.3
|
Control 4 Hippo
18.9
AD 6 Occipital Ctx
48.6
|
Control (Path) 3 Hippo
11.3
Control 1 Occipital Ctx
5.8
|
AD 1 Temporal Ctx
15.5
Control 2 Occipital Ctx
74.7
|
AD 2 Temporal Ctx
35.4
Control 3 Occipital Ctx
26.6
|
AD 3 Temporal Ctx
6.0
Control 4 Occipital Ctx
6.8
|
AD 4 Temporal Ctx
23.8
Control (Path) 1
100.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
94.0
Control (Path) 2
13.9
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
55.1
Control (Path) 3
5.0
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
65.5
Control (Path) 4
30.8
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
66.0
Control 1 Parietal Ctx
10.7
|
Control 1 Temporal Ctx
9.3
Control 2 Parietal Ctx
46.7
|
Control 2 Temporal Ctx
42.3
Control 3 Parietal Ctx
16.5
|
Control 3 Temporal Ctx
15.6
Control (Path) 1
88.9
|
Parietal Ctx
|
Control 4 Temporal Ctx
12.8
Control (Path) 2
25.7
|
Parietal Ctx
|
Control (Path) 1
52.9
Control (Path) 3
6.3
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
48.3
Control (Path) 4
52.5
|
Temporal Ctx
Parietal Ctx
|
|
[0920]
453
TABLE HD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3604, Run
Ag3604, Run
|
Tissue Name
217674539
Tissue Name
217674539
|
|
Adipose
5.6
Renal ca. TK-10
17.9
|
Melanoma* Hs688(A).T
17.9
Bladder
10.9
|
Melanoma* Hs688(B).T
24.0
Gastric ca. (liver met.)
17.0
|
NCI-N87
|
Melanoma* M14
12.3
Gastric ca. KATO III
38.7
|
Melanoma* LOXIMVI
13.4
Colon ca. SW-948
4.4
|
Melanoma* SK-MEL-5
17.8
Colon ca. SW480
31.9
|
Squamous cell
11.9
Colon ca.* (SW480 met)
17.0
|
carcinoma SCC-4
SW620
|
Testis Pool
1.3
Colon ca. HT29
9.1
|
Prostate ca.* (bone met)
15.5
Colon ca. HCT-116
27.9
|
PC-3
|
Prostate Pool
1.4
Colon ca. CaCo-2
14.8
|
Placenta
0.9
Colon cancer tissue
10.2
|
Uterus Pool
1.4
Colon ca. SW1116
1.5
|
Ovarian ca. OVCAR-3
12.4
Colon ca. Colo-205
4.1
|
Ovarian ca. SK-OV-3
24.3
Colon ca. SW-48
5.8
|
Ovarian ca. OVCAR-4
10.8
Colon Pool
4.0
|
Ovarian ca. OVCAR-5
50.3
Small Intestine Pool
2.5
|
Ovarian ca. IGROV-1
9.0
Stomach Pool
3.0
|
Ovarian ca. OVCAR-8
5.4
Bone Marrow Pool
1.2
|
Ovary
2.1
Fetal Heart
5.6
|
Breast ca. MCF-7
12.0
Heart Pool
2.1
|
Breast ca. MDA-MB-
15.3
Lymph Node Pool
4.7
|
231
|
Breast ca. BT 549
9.2
Fetal Skeletal Muscle
0.6
|
Breast ca. T47D
100.0
Skeletal Muscle Pool
1.7
|
Breast ca. MDA-N
15.2
Spleen Pool
4.8
|
Breast Pool
3.9
Thymus Pool
2.9
|
Trachea
3.0
CNS cancer (glio/astro)
84.7
|
U87-MG
|
Lung
0.5
CNS cancer (glio/astro) U-
30.8
|
118-MG
|
Fetal Lung
8.0
CNS cancer (neuro; met)
14.5
|
SK-N-AS
|
Lung ca. NCI-N417
1.5
CNS cancer (astro) SF-539
13.1
|
Lung ca. LX-1
10.9
CNS cancer (astro) SNB-75
39.8
|
Lung ca. NCI-H146
11.7
CNS cancer (glio) SNB-19
9.8
|
Lung ca. SHP-77
5.3
CNS cancer (glio) SF-295
30.6
|
Lung ca. A549
9.6
Brain (Amygdala) Pool
1.9
|
Lung ca. NCI-H526
4.5
Brain (cerebellum)
1.4
|
Lung ca. NCI-H23
25.7
Brain (fetal)
4.4
|
Lung ca. NCI-H460
5.9
Brain (Hippocampus) Pool
2.1
|
Lung ca. HOP-62
5.8
Cerebral Cortex Pool
2.7
|
Lung ca. NCI-H522
8.8
Brain (Substantia nigra)
1.9
|
Pool
|
Liver
0.6
Brain (Thalamus) Pool
2.8
|
Fetal Liver
11.1
Brain (whole)
2.4
|
Liver ca. HepG2
6.2
Spinal Cord Pool
1.9
|
Kidney Pool
5.2
Adrenal Gland
2.5
|
Fetal Kidney
4.2
Pituitary gland Pool
0.7
|
Renal ca. 786-0
44.1
Salivary Gland
0.8
|
Renal ca. A498
10.2
Thyroid (female)
5.0
|
Renal ca. ACHN
6.4
Pancreatic ca. CAPAN2
12.0
|
Renal ca. UO-31
37.9
Pancreas Pool
5.6
|
|
[0921]
454
TABLE HE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3604,
Ag3604,
|
Tissue Name
Run 169910577
Tissue Name
Run 169910577
|
|
Secondary Th1 act
14.2
HUVEC IL-1beta
8.5
|
Secondary Th2 act
18.0
HUVEC IFN gamma
5.2
|
Secondary Tr1 act
17.9
HUVEC TNF alpha + IFN
7.4
|
gamma
|
Secondary Th1 rest
1.6
HUVEC TNF alpha + IL4
11.3
|
Secondary Th2 rest
3.8
HUVEC IL-11
1.8
|
Secondary Tr1 rest
2.5
Lung Microvascular EC none
8.0
|
Primary Th1 act
11.8
Lung Microvascular EC
24.1
|
TNF alpha + IL-1beta
|
Primary Th2 act
13.6
Microvascular Dermal EC none
4.1
|
Primary Tr1 act
12.1
Microsvascular Dermal EC
12.2
|
TNF alpha + IL-1beta
|
Primary Th1 rest
3.6
Bronchial epithelium
11.7
|
TNF alpha + 1beta
|
Primary Th2 rest
3.4
Small airway epithelium none
4.2
|
Primary Tr1 rest
3.4
Small airway epithelium
13.6
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
13.5
Coronery artery SMC rest
37.1
|
act
|
CD45RO CD4 lymphocyte
14.8
Coronery artery SMC
48.6
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
14.1
Astrocytes rest
6.7
|
Secondary CD8
11.9
Astrocytes TNF alpha + IL-1beta
15.1
|
lymphocyte rest
|
Secondary CD8
7.2
KU-812 (Basophil) rest
9.3
|
lymphocyte act
|
CD4 lymphocyte none
1.6
KU-812 (Basophil)
23.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
2.8
CCD1106 (Keratinocytes) none
10.6
|
CD95 CH11
|
LAK cells rest
15.7
CCD1106 (Keratinocytes)
16.2
|
TNF alpha + IL-1beta
|
LAK cells IL-2
6.7
Liver cirrhosis
3.5
|
LAK cells IL-2 + IL-12
7.2
NCI-H292 none
6.0
|
LAK cells IL-2 + IFN
10.4
NCI-H292 IL-4
13.3
|
gamma
|
LAK cells IL-2 + IL-18
9.4
NCI-H292 IL-9
13.6
|
LAK cells
60.7
NCI-H292 IL-13
12.5
|
PMA/ionomycin
|
NK Cells IL-2 rest
7.2
NCI-H292 IFN gamma
13.7
|
Two Way MLR 3 day
15.1
HPAEC none
5.3
|
Two Way MLR 5 day
13.1
HPAEC TNF alpha + IL-1beta
54.7
|
Two Way MLR 7 day
8.7
Lung fibroblast none
11.1
|
PBMC rest
1.6
Lung fibroblast TNF alpha + IL-
7.4
|
1beta
|
PBMC PWM
12.8
Lung fibroblast IL-4
18.6
|
PBMC PHA-L
10.1
Lung fibroblast IL-9
24.7
|
Ramos (B cell) none
10.0
Lung fibroblast IL-13
13.8
|
Ramos (B cell) ionomycin
8.4
Lung fibroblast IFN gamma
20.4
|
B lymphocytes PWM
9.7
Dermal fibroblast CCD1070 rest
11.8
|
B lymphocytes CD40L
6.7
Dermal fibroblast CCD1070
23.2
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
7.9
Dermal fibroblast CCD1070 IL-
25.7
|
1beta
|
EOL-1 dbcAMP
24.0
Dermal fibroblast IFN gamma
12.2
|
PMA/ionomycin
|
Dendritic cells none
23.3
Dermal fibroblast IL-4
12.6
|
Dendritic cells LPS
28.7
Dermal fibroblasts rest
8.7
|
Dendritic cells anti-CD40
18.6
Neutrophils TNFa + LPS
7.5
|
Monocytes rest
2.8
Neutrophils rest
0.6
|
Monocytes LPS
100.0
Colon
1.6
|
Macrophages rest
27.7
Lung
3.7
|
Macrophages LPS
24.8
Thymus
5.7
|
HUVEC none
3.5
Kidney
6.6
|
HUVEC starved
4.2
|
|
[0922] CNS_neurodegeneration_v1.0 Summary: Ag3604 This panel does not show differential expression of the CG59839-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the occipital cortex of a control patient (CT=28.5). Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[0923] General_screening_panel_v1.4 Summary: Ag3604 Highest expression of the CG59839-01 gene is seen in a breast cancer cell line (CT=24). High levels of expression are also seen in all the cell lines on this panel. In addition, higher levels of expression are seen in the fetal tissue samples. Expression in fetal liver and lung (CTs=27) is significantly higher than in the adult liver and lung (CTs=31.5). Therefore, expression of this gene could be used to differentiate between the adult and fetal sources of these tissues. Furthermore, this expression profile suggests a role for this gene product in cell growth and proliferation.
[0924] Among tissues with metabolic function, this gene is expressed at moderate to low levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[0925] This gene is also expressed at moderate levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[0926] Panel 4.1D Summary: Ag3604 Highest expression of the CG59839-01 gene is seen in LPS stimulated monocytes (CT=25.3). The protein encoded by this gene may therefore be involved in the activation of monocytes in their function as antigen-presenting cells. This suggests that therapeutics that block the function of this membrane protein may be useful as anti-inflammatory therapeutics for the treatment of autoimmune and inflammatory diseases. Furthermore, antibodies or small molecule therapeutics that stimulate the function of this protein may be useful therapeutics for the treatment of immunosupressed individuals.
[0927] This gene is also expressed at moderate to low levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[0928] I. CG59847-01: Novel Intracellular Protein
[0929] Expression of gene CG59847-01 was assessed using the primer-probe set Ag3608, described in Table IA. Results of the RTQ-PCR runs are shown in Tables IB, IC and ID.
455TABLE IA
|
|
Probe Name Ag3608
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-cataacgaaatgaaggcagaaa-3′22786243
ProbeTET-5′-28814244
tgttctttgaaaccaacaagaacaaaga-
3′-TAMRA
Reverse5′-cccagagattctggtatgttgt-3′22843245
|
[0930]
456
TABLE IB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3608, Run
Ag3608,
|
Tissue Name
210998067
Tissue Name
Run 210998067
|
|
AD 1 Hippo
16.7
Control (Path) 3
16.0
|
Temporal Ctx
|
AD 2 Hippo
32.5
Control (Path) 4
83.5
|
Temporal Ctx
|
AD 3 Hippo
22.1
AD 1 Occipital Ctx
37.4
|
AD 4 Hippo
0.0
AD 2 Occipital Ctx
0.9
|
(Missing)
|
AD 5 hippo
95.9
AD 3 Occipital Ctx
14.8
|
AD 6 Hippo
80.7
AD 4 Occipital Ctx
43.5
|
Control 2 Hippo
16.0
AD 5 Occipital Ctx
38.2
|
Control 4 Hippo
24.3
AD 6 Occipital Ctx
40.3
|
Control (Path) 3 Hippo
12.9
Control 1 Occipital Ctx
7.4
|
AD 1 Temporal Ctx
38.7
Control 2 Occipital Ctx
39.0
|
AD 2 Temporal Ctx
43.2
Control 3 Occipital Ctx
58.6
|
AD 3 Temporal Ctx
23.8
Control 4 Occipital Ctx
14.8
|
AD 4 Temporal Ctx
46.0
Control (Path) 1
97.9
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
98.6
Control (Path) 2
51.1
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
91.4
Control (Path) 3
4.6
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
87.7
Control (Path) 4
44.4
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
100.0
Control 1 Parietal Ctx
20.7
|
Control 1 Temporal Ctx
18.2
Control 2 Parietal Ctx
87.7
|
Control 2 Temporal Ctx
21.5
Control 3 Parietal Ctx
20.7
|
Control 3 Temporal Ctx
42.6
Control (Path) 1
87.1
|
Parietal Ctx
|
Control 4 Temporal Ctx
27.0
Control (Path) 2
0.0
|
Parietal Ctx
|
Control (Path) 1
90.1
Control (Path) 3
12.9
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
93.3
Control (Path) 4
71.2
|
Temporal Ctx
Parietal Ctx
|
|
[0931]
457
TABLE IC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3608, Run
Ag3608, Run
|
Tissue Name
217675976
Tissue Name
217675976
|
|
Adipose
19.5
Renal ca. TK-10
39.8
|
Melanoma* Hs688(A).T
16.5
Bladder
45.1
|
Melanoma* Hs688(B).T
11.8
Gastric ca. (liver met.)
52.5
|
NCI-N87
|
Melanoma* M14
11.3
Gastric ca. KATO III
18.3
|
Melanoma* LOXIMVI
11.6
Colon ca. SW-948
3.0
|
Melanoma* SK-MEL-5
44.1
Colon ca. SW480
26.6
|
Squamous cell
17.0
Colon ca.* (SW480 met)
16.6
|
carcinoma SCC-4
SW620
|
Testis Pool
17.7
Colon ca. HT29
28.5
|
Prostate ca.* (bone met)
30.6
Colon ca. HCT-116
24.3
|
PC-3
|
Prostate Pool
15.1
Colon ca. CaCo-2
30.1
|
Placenta
4.0
Colon cancer tissue
25.0
|
Uterus Pool
9.7
Colon ca. SW1116
5.3
|
Ovarian ca. OVCAR-3
50.0
Colon ca. Colo-205
4.4
|
Ovarian ca. SK-OV-3
24.3
Colon ca. SW-48
3.2
|
Ovarian ca. OVCAR-4
3.6
Colon Pool
39.0
|
Ovarian ca. OVCAR-5
65.1
Small Intestine Pool
53.2
|
Ovarian ca. IGROV-1
9.5
Stomach Pool
18.6
|
Ovarian ca. OVCAR-8
5.6
Bone Marrow Pool
26.2
|
Ovary
20.9
Fetal Heart
58.6
|
Breast ca. MCF-7
18.2
Heart Pool
17.6
|
Breast ca. MDA-MB-
17.2
Lymph Node Pool
55.1
|
231
|
Breast ca. BT 549
38.2
Fetal Skeletal Muscle
16.2
|
Breast ca. T47D
87.7
Skeletal Muscle Pool
22.1
|
Breast ca. MDA-N
16.6
Spleen Pool
27.2
|
Breast Pool
50.0
Thymus Pool
41.8
|
Trachea
13.4
CNS cancer (glio/astro)
31.2
|
U87-MG
|
Lung
52.5
CNS cancer (glio/astro) U-
27.0
|
118-MG
|
Fetal Lung
62.9
CNS cancer (neuro; met)
21.9
|
SK-N-AS
|
Lung ca. NCI-N417
54.3
CNS cancer (astro) SF-539
13.0
|
Lung ca. LX-1
27.2
CNS cancer (astro) SNB-75
36.6
|
Lung ca. NCI-H146
13.5
CNS cancer (glio) SNB-19
11.6
|
Lung ca. SHP-77
43.8
CNS cancer (glio) SF-295
85.9
|
Lung ca. A549
17.9
Brain (Amygdala) Pool
8.3
|
Lung ca. NCI-H526
7.2
Brain (cerebellum)
15.8
|
Lung ca. NCI-H23
26.8
Brain (fetal)
24.3
|
Lung ca. NCI-H460
15.2
Brain (Hippocampus) Pool
14.1
|
Lung ca. HOP-62
18.7
Cerebral Cortex Pool
17.8
|
Lung ca. NCI-H522
26.8
Brain (Substantia nigra)
9.8
|
Pool
|
Liver
0.9
Brain (Thalamus) Pool
26.1
|
Fetal Liver
15.5
Brain (whole)
10.4
|
Liver ca. HepG2
11.6
Spinal Cord Pool
15.1
|
Kidney Pool
55.5
Adrenal Gland
8.5
|
Fetal Kidney
100.0
Pituitary gland Pool
9.2
|
Renal ca. 786-0
23.7
Salivary Gland
6.0
|
Renal ca. A498
17.8
Thyroid (female)
5.4
|
Renal ca. ACHN
28.9
Pancreatic ca. CAPAN2
27.0
|
Renal ca. UO-31
15.3
Pancreas Pool
44.4
|
|
[0932]
458
TABLE ID
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3608,
Ag3608,
|
Tissue Name
Run 169943567
Tissue Name
Run 169943567
|
|
Secondary Th1 act
16.8
HUVEC IL-1beta
18.8
|
Secondary Th2 act
33.2
HUVEC IFN gamma
27.4
|
Secondary Tr1 act
24.8
HUVEC TNF alpha + IFN
7.9
|
gamma
|
Secondary Th1 rest
23.2
HUVEC TNF alpha + IL4
9.8
|
Secondary Th2 rest
24.7
HUVEC IL-11
15.6
|
Secondary Tr1 rest
32.1
Lung Microvascular EC none
19.1
|
Primary Th1 act
32.8
Lung Microvascular EC
19.9
|
TNF alpha + IL-1beta
|
Primary Th2 act
40.9
Microvascular Dermal EC none
17.6
|
Primary Tr1 act
34.4
Microsvasular Dermal EC
9.7
|
TNF alpha + IL-1beta
|
Primary Th1 rest
24.5
Bronchial epithelium
18.4
|
TNF alpha + IL1beta
|
Primary Th2 rest
28.1
Small airway epithelium none
6.3
|
Primary Tr1 rest
29.1
Small airway epithelium
15.4
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
20.7
Coronery artery SMC rest
9.6
|
act
|
CD45RO CD4 lymphocyte
37.1
Coronery artery SMC
12.2
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
27.9
Astrocytes rest
12.3
|
Secondary CD8
28.9
Astrocytes TNF alpha + IL-1beta
11.8
|
lymphocyte rest
|
Secondary CD8
14.2
KU-812 (Basophil) rest
18.7
|
lymphocyte act
|
CD4 lymphocyte none
24.3
KU-812 (Basophil)
12.1
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
16.3
CCD1106 (Keratinocytes) none
15.6
|
CD95 CH11
|
LAK cells rest
3.9
CCD1106 (Keratinocytes)
14.1
|
TNF alpha + IL-1beta
|
LAK cells IL-2
32.3
Liver cirrhosis
61.6
|
LAK cells IL-2 + IL-12
32.5
NCI-H292 none
20.7
|
LAK cells IL-2 + IFN
41.5
NCI-H292 IL-4
30.4
|
gamma
|
LAK cells IL-2 + IL-18
36.3
NCI-H292 IL-9
42.6
|
LAK cells
15.0
NCI-H292 IL-13
22.8
|
PMA/ionomycin
|
NK Cells IL-2 rest
24.1
NCI-H292 IFN gamma
24.3
|
Two Way MLR 3 day
38.2
HPAEC none
23.8
|
Two Way MLR 5 day
19.2
HPAEC TNF alpha + IL-1beta
29.9
|
Two Way MLR 7 day
20.7
Lung fibroblast none
14.8
|
PBMC rest
11.7
Lung fibroblast TNF alpha + IL-
9.2
|
1beta
|
PBMC PWM
22.4
Lung fibroblast IL-4
15.7
|
PBMC PHA-L
16.4
Lung fibroblast IL-9
19.1
|
Ramos (B cell) none
21.2
Lung fibroblast IL-13
16.6
|
Ramos (B cell) ionomycin
24.8
Lung fibroblast IFN gamma
12.2
|
B lymphocytes PWM
18.3
Dermal fibroblast CCD1070 rest
26.8
|
B lymphocytes CD40L
24.8
Dermal fibroblast CCD1070
37.6
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
49.3
Dermal fibroblast CCD1070 IL-
11.4
|
1beta
|
EOL-1 dbcAMP
31.4
Dermal fibroblast IFN gamma
10.9
|
PMA/ionomycin
|
Dendritic cells none
16.4
Dermal fibroblast IL-4
50.3
|
Dendritic cells LPS
10.3
Dermal Fibroblasts rest
14.3
|
Dendritic cells anti-CD40
17.7
Neutrophils TNFa + LPS
13.0
|
Monocytes rest
10.7
Neutrophils rest
21.0
|
Monocytes LPS
29.5
Colon
27.9
|
Macrophages rest
8.2
Lung
15.6
|
Macrophages LPS
11.1
Thymus
100.0
|
HUVEC none
11.7
Kidney
40.9
|
HUVEC starved
27.7
|
|
[0933] CNS_neurodegeneration_v1.0 Summary: Ag3608 This panel does not show differential expression of the CG59847-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[0934] General_screeningpanel_v1.4 Summary: Ag3608 The CG59847-01 gene is expressed ubiquitously throughout this panel, with highest expression in fetal kidney (CT=25.4). Overall, this expression profile suggests that this gene product may be involved in cellular growth and proliferation.
[0935] Among tissues with metabolic function, this gene is expressed at high to moderate levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[0936] In addition, this gene is expressed at much higher levels in fetal liver tissue (CT=27) when compared to expression in the adult counterpart (CT=31). Thus, expression of this gene may be used to differentiate between the fetal and adult source of this tissue.
[0937] This gene is also expressed at high levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[0938] Panel 4.1D Summary: Ag3608 The CG59847-01 gene is expressed ubiquitously throughout this panel, with highest expression in the thymus (CT=28). In addition, this gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[0939] J. CG59905-01: Sushi Containing Membrane Protein
[0940] Expression of gene CG59905-01 was assessed using the primer-probe set Ag2443, described in Table JA. Results of the RTQ-PCR runs are shown in Tables JB, JC, JD, JE and JF.
459TABLE JA
|
|
Probe Name Ag2443
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-cagtttgggaccgacttca-3′19652246
ProbeTET-5′-28678247
caagactgtgagctatcagtgtaaccca-
3′-TAMRA
Reverse5′-tgactgcttccatgacatagc-3′21707248
|
[0941]
460
TABLE JB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2443, Run
Ag2443,
|
Tissue Name
228397047
Tissue Name
Run 228397047
|
|
AD 1 Hippo
14.6
Control (Path) 3
6.9
|
Temporal Ctx
|
AD 2 Hippo
31.6
Control (Path) 4
41.2
|
Temporal Ctx
|
AD 3 Hippo
9.1
AD 1 Occipital Ctx
23.5
|
AD 4 Hippo
9.5
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
8.1
|
AD 6 Hippo
34.6
AD 4 Occipital Ctx
31.0
|
Control 2 Hippo
22.7
AD 5 Occipital Ctx
39.2
|
Control 4 Hippo
7.4
AD 6 Occipital Ctx
19.8
|
Control (Path) 3 Hippo
5.8
Control 1 Occipital Ctx
3.0
|
AD 1 Temporal Ctx
13.8
Control 2 Occipital Ctx
52.5
|
AD 2 Temporal Ctx
36.1
Control 3 Occipital Ctx
36.9
|
AD 3 Temporal Ctx
11.3
Control 4 Occipital Ctx
8.7
|
AD 4 Temporal Ctx
33.7
Control (Path) 1
94.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
76.3
Control (Path) 2
27.9
|
Occipital Ctx
|
AD 5 Sup Temporal
40.9
Control (Path) 3
2.2
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
43.5
Control (Path) 4
25.9
|
Occipital Ctx
|
AD 6 Sup Temporal
52.5
Control 1 Parietal Ctx
8.7
|
Ctx
|
Control 1 Temporal Ctx
10.1
Control 2 Parietal Ctx
37.4
|
Control 2 Temporal Ctx
36.1
Control 3 Parietal Ctx
25.5
|
Control 3 Temporal Ctx
27.7
Control (Path) 1
81.8
|
Parietal Ctx
|
Control 3 Temporal Ctx
15.7
Control (Path) 2
32.8
|
Parietal Ctx
|
Control (Path) 1
65.1
Control (Path) 3
5.5
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
50.7
Control (Path) 4
62.0
|
Temporal Ctx
Parietal Ctx
|
|
[0942]
461
TABLE JC
|
|
|
Panel 1.3D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2443, Run
Ag2443, Run
|
Tissue Name
165517738
Tissue Name
165517738
|
|
Liver adenocarcinoma
16.6
Kidney (fetal)
2.5
|
Pancreas
0.0
Renal ca. 786-0
0.0
|
Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0
|
Adrenal gland
1.5
Renal ca. RXF 393
0.0
|
Thyroid
0.0
Renal ca. ACHN
0.0
|
Salivary gland
0.0
Renal ca. UO-31
0.0
|
Pituitary gland
2.2
Renal ca. TK-10
0.0
|
Brain (fetal)
51.4
Liver
0.0
|
Brain (whole)
92.0
Liver (fetal)
0.0
|
Brain (amygdala)
52.1
Liver ca. (hepatoblast)
0.0
|
HepG2
|
Brain (cerebellum)
47.6
Lung
0.0
|
Brain (hippocampus)
78.5
Lung (fetal)
1.4
|
Brain (substantia nigra)
8.0
Lung ca. (small cell) LX-1
0.0
|
Brain (thalamus)
12.9
Lung ca. (small cell)
0.0
|
NCI-H69
|
Cerebral Cortex
100.0
Lung ca. (s. cell var.)
16.8
|
SHP-77
|
Spinal cord
6.5
Lung ca. (large cell)NCI-
1.6
|
H460
|
glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0
|
A549
|
glio/astro U-118-MG
0.0
Lung ca. (non-s. cell)
11.2
|
NCI-H23
|
astrocytoma SW1783
0.0
Lung ca. (non-s. cell)
0.0
|
HOP-62
|
neuro*; met SK-N-AS
0.0
Lung ca. (non-s. cl) NCI-
0.0
|
H522
|
astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0
|
900
|
astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
4.8
|
H596
|
glioma SNB-19
0.6
Mammary gland
2.1
|
glioma U251
29.9
Breast ca.* (pl. ef) MCF-7
0.0
|
glioma SF-295
1.9
Breast ca.* (pl. ef) MDA-
0.0
|
MB-231
|
Heart (fetal)
0.0
Breast ca.* (pl. ef) T47D
10.9
|
Heart
0.0
Breast ca. BT-549
0.0
|
Skeletal muscle (fetal)
6.2
Breast ca. MDA-N
9.3
|
Skeletal muscle
0.8
Ovary
1.1
|
Bone marrow
0.0
Ovarian ca. OVCAR-3
11.5
|
Thymus
0.0
Ovarian ca. OVCAR-4
0.0
|
Spleen
5.4
Ovarian ca. OVCAR-5
0.0
|
Lymph node
0.5
Ovarian ca. OVCAR-8
0.0
|
Colorectal
0.7
Ovarian ca. IGROV-1
0.0
|
Stomach
0.0
Ovarian ca.* (ascites)
0.8
|
SK-OV-3
|
Small intestine
1.5
Uterus
2.8
|
Colon ca. SW480
0.0
Placenta
2.9
|
Colon ca.* SW620(SW480
0.0
Prostate
0.9
|
met)
|
Colon ca. HT29
0.0
Prostate ca.* (bone
0.0
|
met)PC-3
|
Colon ca. HCT-116
0.0
Testis
20.9
|
Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0
|
Colon ca.
0.0
Melanoma* (met)
0.6
|
tissue(ODO3866)
Hs688(B).T
|
Colon ca. HCC-2998
0.0
Melanoma UACC-62
19.5
|
Gastric ca.* (liver met)
0.0
Melanoma M14
15.1
|
NCI-N87
|
Bladder
1.2
Melanoma LOX IMVI
0.7
|
Trachea
2.0
Melanoma* (met) SK-
7.3
|
MEL-5
|
Kidney
4.1
Adipose
0.4
|
|
[0943]
462
TABLE JD
|
|
|
Panel 2D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2443,
Ag2443,
|
Tissue Name
Run 164988396
Tissue Name
Run 164988396
|
|
Normal Colon
12.6
Kidney Margin 8120608
37.1
|
CC Well to Mod Diff
1.2
Kidney Cancer 8120613
0.0
|
(ODO3866)
|
CC Margin (ODO3866)
2.9
Kidney Margin 8120614
31.6
|
CC Gr.2 rectosigmoid
1.9
Kidney Cancer 9010320
4.2
|
(ODO3868)
|
CC Margin (ODO3868)
3.2
Kidney Margin 9010321
62.9
|
CC Mod Diff (ODO3920)
1.5
Normal Uterus
1.4
|
CC Margin (ODO3920)
2.6
Uterus Cancer 064011
19.6
|
CC Gr.2 ascend colon
9.3
Normal Thyroid
0.6
|
(ODO3921)
|
CC Margin (ODO3921)
2.9
Thyroid Cancer 064010
6.6
|
CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0
|
(ODO4309) Mets
|
Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
1.9
|
Colon mets to lung (OD04451-
2.5
Normal Breast
22.2
|
01)
|
Lung Margin (OD04451-02)
18.0
Breast Cancer (OD04566)
0.8
|
Normal Prostate 6546-1
4.2
Breast Cancer (OD04590-
2.6
|
01)
|
Prostate Cancer (OD04410)
0.0
Breast Cancer Mets
2.0
|
(OD04590-03)
|
Prostate Margin (OD04410)
5.3
Breast Cancer Metastasis
2.9
|
(OD04655-05)
|
Prostate Cancer (OD04720-01)
6.0
Breast Cancer 064006
4.3
|
Prostate Margin (OD04720-02)
4.5
Breast Cancer 1024
6.5
|
Normal Lung 061010
29.9
Breast Cancer 9100266
6.5
|
Lung Met to Muscle
2.4
Breast Margin 9100265
15.7
|
(ODO4286)
|
Muscle Margin (ODO4286)
3.1
Breast Cancer A209073
10.7
|
Lung Malignant Cancer
12.2
Breast Margin A209073
31.4
|
(OD03126)
|
Lung Margin (OD03126)
20.4
Normal Liver
0.0
|
Lung Cancer (OD04404)
3.0
Liver Cancer 064003
26.6
|
Lung Margin (OD04404)
30.1
Liver Cancer 1025
0.0
|
Lung Cancer (OD04565)
0.8
Liver Cancer 1026
0.0
|
Lung Margin (OD04565)
9.0
Liver Cancer 6004-T
0.0
|
Lung Cancer (OD04237-01)
7.8
Liver Tissue 6004-N
29.1
|
Lung Margin (OD04237-02)
3.1
Liver Cancer 6005-T
0.0
|
Ocular Mel Met to Liver
3.2
Liver Tissue 6005-N
0.0
|
(ODO4310)
|
Liver Margin (ODO4310)
1.1
Normal Bladder
0.4
|
Melanoma Mets to Lung
0.0
Bladder Cancer 1023
0.0
|
(OD04321)
|
Lung Margin (OD04321)
17.2
Bladder Cancer A302173
39.0
|
Normal Kidney
100.0
Bladder Cancer
0.0
|
(OD04718-01)
|
Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
4.5
|
(OD04338)
(OD04718-03)
|
Kidney Margin (OD04338)
16.8
Normal Ovary
2.6
|
Kidney Ca Nuclear grade 1/2
6.3
Ovarian Cancer 064008
0.0
|
(OD04339)
|
Kidney Margin (OD04339)
59.0
Ovarian Cancer
0.0
|
(OD04768-07)
|
Kidney Ca, Clear cell type
10.7
Ovary Margin (OD04768-
0.0
|
(OD04340)
08)
|
Kidney Margin (OD04340)
38.4
Normal Stomach
2.2
|
Kidney Ca, Nuclear grade 3
0.8
Gastric Cancer 9060358
0.0
|
(OD04348)
|
Kidney Margin (OD04348)
18.4
Stomach Margin 9060359
1.3
|
Kidney Cancer (OD04622-01)
1.8
Gastric Cancer 9060395
4.6
|
Kidney Margin (OD04622-03)
8.9
Stomach Margin 9060394
5.3
|
Kidney Cancer (OD04450-01)
1.6
Gastric Cancer 9060397
2.5
|
Kidney Margin (OD04450-03)
21.3
Stomach Margin 9060396
0.8
|
Kidney Cancer 8120607
0.0
Gastric Cancer 064005
0.0
|
|
[0944]
463
TABLE JE
|
|
|
Panel 4D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag2443,
Ag2443,
|
Tissue Name
Run 164337611
Tissue Name
Run 164337611
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium
0.0
|
TNF alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC
0.0
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.7
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.2
|
LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.1
|
LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0
|
LAK cells
0.0
NCI-H292 IL-9
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0
|
Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 5 day
0.0
HPAEC none
0.2
|
Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
0.1
|
PBMC rest
0.1
Lung fibroblast none
0.0
|
PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0
|
Ramos (B cell) none
35.6
Lung fibroblast IL-9
0.0
|
Ramos (B cell) ionomycin
100.0
Lung fibroblast IL-13
0.0
|
B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0
|
and IL-4
|
EOL-1 dbcAMP
0.5
Dermal fibroblast CCD1070
0.0
|
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
PMA/ionomycin
1beta
|
Dendritic cells none
0.2
Dermal fibroblast IFN gamma
0.0
|
Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0
|
Monocytes rest
0.1
IBD Crohn's
0.0
|
Monocytes LPS
0.0
Colon
0.1
|
Macrophages rest
0.0
Lung
0.1
|
Macrophages LPS
0.0
Thymus
2.5
|
HUVEC none
0.0
Kidney
0.1
|
HUVEC starved
0.0
|
|
[0945]
464
TABLE JF
|
|
|
Panel CNS_1
|
Rel. Exp.
Rel.
|
(%) Ag2443, Run
Exp. (%) Ag2443, Run
|
Tissue Name
171656290
Tissue Name
171656290
|
|
BA4 Control
21.0
BA17 PSP
14.1
|
BA4 Control2
17.2
BA17 PSP2
12.0
|
BA4 Alzheimer's2
7.6
Sub Nigra Control
12.6
|
BA4 Parkinson's
42.9
Sub Nigra Control2
6.5
|
BA4 Parkinson's2
46.3
Sub Nigra Alzheimer's2
2.3
|
BA4 Huntington's
18.8
Sub Nigra Parkinson's2
9.7
|
BA4
17.0
Sub Nigra Huntington's
11.7
|
Huntington's2
|
BA4 PSP
6.1
Sub Nigra
9.2
|
Huntington's2
|
BA4 PSP2
19.1
Sub Nigra PSP2
0.0
|
BA4 Depression
9.5
Sub Nigra Depression
0.0
|
BA4 Depression2
2.9
Sub Nigra Depression2
0.5
|
BA7 Control
27.9
Glob Palladus Control
5.9
|
BA7 Control2
22.8
Glob Palladus Control2
3.7
|
BA7 Alzheimer's2
15.7
Glob Palladus
4.5
|
Alzheimer's
|
BA7 Parkinson's
19.6
Glob Palladus
3.2
|
Alzheimer's2
|
BA7 Parkinson's2
28.1
Glob Palladus
49.7
|
Parkinson's
|
BA7 Huntington's
27.9
Glob Palladus
5.3
|
Parkinson's2
|
BA7
0.0
Glob Palladus PSP
1.0
|
Huntington's2
|
BA7 PSP
24.7
Glob Palladus PSP2
0.6
|
BA7 PSP2
19.3
Glob Palladus
1.6
|
Depression
|
BA7 Depression
13.0
Temp Pole Control
14.1
|
BA9 Control
14.2
Temp Pole Control2
23.8
|
BA9 Control2
31.6
Temp Pole Alzheimer's
4.4
|
BA9 Alzheimer's
3.3
Temp Pole Alzheimer's2
7.8
|
BA9 Alzheimer's2
22.5
Temp Pole Parkinson's
29.9
|
BA9 Parkinson's
31.4
Temp Pole Parkinson's2
35.4
|
BA9 Parkinson's2
38.7
Temp Pole Huntington's
24.7
|
BA9 Huntington's
29.1
Temp Pole PSP
1.8
|
BA9
21.8
Temp Pole PSP2
5.6
|
Huntington's2
|
BA9 PSP
7.4
Temp Pole Depression2
8.5
|
BA9 PSP2
4.6
Cing Gyr Control
46.7
|
BA9 Depression
6.0
Cing Gyr Control2
17.9
|
BA9 Depression2
8.6
Cing Gyr Alzheimer's
8.7
|
BA17 Control
54.3
Cing Gyr Alzheimer's2
15.7
|
BA17 Control2
28.5
Cing Gyr Parkinson's
13.1
|
BA17
21.9
Cing Gyr Parkinson's2
21.9
|
Alzheimer's2
|
BA17 Parkinson's
47.3
Cing Gyr Huntington's
24.3
|
BA17
100.0
Cing Gyr Huntington's2
13.4
|
Parkinson's2
|
BA17
34.9
Cing Gyr PSP
10.4
|
Huntington's
|
BA17
33.4
Cing Gyr PSP2
4.4
|
Huntington's2
|
BA17 Depression
13.8
Cing Gyr Depression
5.3
|
BA17 Depression2
32.5
Cing Gyr Depression2
4.9
|
|
[0946] CNS_neurodegeneration_v1.0 Summary: Ag2443 This panel confirms the expression of the CG59905-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders. Results from a second experiment with the same probe and primer set are not included. The data suggests that there were experimental difficulties with this run.
[0947] Panel 1.3D Summary: Ag2443 Highest expression of the CG59905-01 gene is detected in cerebral cortex of brain (CT=30.9). In addition, high expression of this gene is observed exclusively in all the brain regions (CTs=31). Thus, expression of this gene can be used to distinguish these brain samples from other samples used in this panel. Low but significant expression is also seen in spinal cord region. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[0948] The CG59905-01 gene encodes a variant of cub and sushi multiple domains 1 protein (CSMD 1). A related protein, SEZ-6, has recently been shown to be associated with neuronal bursting activity of seizures (Shimizu-Nishikawa K, Kajiwara K, Kimura M, Katsuki M, Sugaya E. (1995) Cloning and expression of SEZ-6, a brain-specific and seizure-related cDNA. Brain Res Mol Brain Res 28(2):201-10). Thus, the protein encoded by the CG59905-01 may also play a role in neuronal siezures.
[0949] In addition, low to moderate expression of this gene is seen in melanoma, ovarian cancer OVCAR-3, breast cancer, lung cancer, liver adenocarcinoma and glioma U251 cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, or antibodies, might be beneficial in the treatment of these cancers.
[0950] Significant expression is also detected in fetal skeletal muscle. Interestingly, this gene is expressed at much higher levels in fetal (CT=34.9) when compared to adult skeletal muscle (CT=37.9). This observation suggests that expression of this gene can be used to distinguish fetal from adult skeletal muscle. In addition, the relative overexpression of this gene in fetal skeletal muscle suggests that the protein product may enhance muscular growth or development in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the GPCR encoded by this gene could be useful in treatment of muscle related diseases. More specifically, treatment of weak or dystrophic muscle with the protein encoded by this gene could restore muscle mass or function.
[0951] Panel 2D Summary: Ag2443 Highest expression of the CG59905-01 gene is detected in kidney (CT=31). Expression of this gene is down-regulated in kidney and lung cancer. In addition, there appears to be substantial expression in other samples derived from bladder cancer, liver cancer, uterine cancer, breast cancer, kidney cancer and lung cancer. The CG59905-01 gene encodes a variant of cub and sushi multiple domains 1 protein (CSMD1). Recently, CSMD1 has been shown to be a candidate for tumor suppressor of multiple types of cancer (Sun P C, Uppaluri R, Schmidt A P, Pashia M E, Quant E C, Sunwoo J B, Gollin S M, Scholnick S B. (2001) Transcript map of the 8p23 putative tumor suppressor region. Genomics 75(1-3): 17-25). Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, or antibodies, might be beneficial in the treatment of bladder, liver, uterine, breast, kidney and lung cancers.
[0952] Panel 4D Summary: Ag2443 Highest expression of the CG59905-01 gene is seen y in Ramos B cells (CTs=26-27). Thus, expression of this gene can be used to distinguish these samples from other samples in this panel. In addition, expression of this gene in B cells suggests that this gene may be involved in rheumatic disease including rheumatoid arthritis, lupus, osteoarthritis, and hyperproliferative B cell disorders.
[0953] Low but significant expression of this gene is also detected in thymus. Therefore, drugs that inhibit the function of this protein may regulate T cell development in the thymus and reduce or eliminate the symptoms of T cell mediated autoimmune or inflammatory diseases, including asthma, allergies, inflammatory bowel disease, lupus erythematosus, or rheumatoid arthritis. Additionally, small molecule or antibody therapeutics designed against this putative protein may disrupt T cell development in the thymus and function as an immunosuppresant for tissue transplant.
[0954] Panel CNS—1 Summary: Ag2443 This panel confirms the expression of the CG59905-01 gene at low levels in the brains of an independent group of individuals. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[0955] K. CG59930-01 and CG59930-O2: LEUCINE-RICH REPEAT-CONTAINING PROTEIN
[0956] Expression of gene CG59930-01 and full length clone CG59930-02 was assessed using the primer-probe sets Agl 106 and Agl614, described in Tables KA and KB. Results of the RTQ-PCR runs are shown in Tables KC, KD, KE, KF and KG. Please note that CG59930-02 represents a full-length physical clone of the CG59930-01 gene, validating the prediction of the gene sequence.
465TABLE KA
|
|
Probe Name Ag1106
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ccagggattggactgcttac-3′20614249
ProbeTET-5′-27661250
caacaaaatcaagactgtccccaagga-
3′-TAMRA
Reverse5′-tctccaagctggcacaatta-3′20694251
|
[0957]
466
TABLE KB
|
|
|
Probe Name Ag1614
|
SEQ
|
Start
ID
|
Primers
Sequences
Length
Position
NO:
|
|
Forward
5′-tgaaaattcctgaattcattgg-3′
22
534
252
|
Probe
TET-5′-ccagaacctcattgtgttagatttatctcg-
30
562
253
|
3′-TAMRA
|
Reverse
5′-ccctggtggtatctctgaaatt-3′
22
598
254
|
|
[0958]
467
TABLE KC
|
|
|
Panel 1.3D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag1614, Run
Ag1614, Run
|
Tissue Name
165920458
Tissue Name
165920458
|
|
Liver adenocarcinoma
0.0
Kidney (fetal)
0.1
|
Pancreas
0.2
Renal ca. 786-0
0.0
|
Pancreatic ca. CAPAN 2
0.3
Renal ca. A498
0.1
|
Adrenal gland
0.3
Renal ca. RXF 393
0.0
|
Thyroid
0.4
Renal ca. ACHN
0.1
|
Salivary gland
0.5
Renal ca. UO-31
0.0
|
Pituitary gland
0.5
Renal ca. TK-10
0.1
|
Brain (fetal)
0.6
Liver
0.2
|
Brain (whole)
1.8
Liver (fetal)
0.1
|
Brain (amygdala)
1.3
Liver ca. (hepatoblast)
0.0
|
HepG2
|
Brain (cerebellum)
1.2
Lung
0.1
|
Brain (hippocampus)
1.1
Lung (fetal)
3.0
|
Brain (substantia nigra)
0.9
Lung ca. (small cell) LX-1
0.1
|
Brain (thalamus)
2.4
Lung ca. (small cell)
0.1
|
NCI-H69
|
Cerebral Cortex
0.1
Lung ca. (s.cell var.)
0.0
|
SHP-77
|
Spinal cord
0.4
Lung ca. (large cell) NCI-
0.0
|
H460
|
glio/astro U87-MG
0.1
Lung ca. (non-sm. cell)
0.0
|
A549
|
glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.2
|
NCI-H23
|
astrocytoma SW1783
0.2
Lung ca. (non-s.cell)
0.1
|
HOP-62
|
neuro; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0
|
H522
|
astrocytoma SF-539
0.4
Lung ca. (squam.) SW
0.2
|
900
|
astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.1
|
H596
|
glioma SNB-19
0.4
Mammary gland
0.0
|
glioma U251
0.3
Breast ca.* (pl.ef) MCF-7
0.2
|
glioma SF-295
0.1
Breast ca.* (pl.ef) MDA-
0.0
|
MB-231
|
Heart (fetal)
3.1
Breast ca.* (pl.ef) T47D
0.3
|
Heart
18.0
Breast ca. BT-549
0.0
|
Skeletal muscle (fetal)
1.5
Breast ca. MDA-N
0.0
|
Skeletal muscle
100.0
Ovary
0.0
|
Bone marrow
0.1
Ovarian ca. OVCAR-3
0.1
|
Thymus
0.5
Ovarian ca. OVCAR-4
0.1
|
Spleen
1.0
Ovarian ca. OVCAR-5
0.0
|
Lymph node
0.2
Ovarian ca. OVCAR-8
0.0
|
Colorectal
0.0
Ovarian ca. IGROV-1
0.1
|
Stomach
0.3
Ovarian ca.* (ascites)
0.2
|
SK-OV-3
|
Small intestine
0.2
Uterus
0.0
|
Colon ca. SW480
0.1
Placenta
0.2
|
Colon ca.* SW620(SW480
0.2
Prostate
0.2
|
met)
|
Colon ca. HT29
0.0
Prostate ca.* (bone
0.2
|
met)PC-3
|
Colon ca. HCT-116
0.0
Testis
0.8
|
Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0
|
Colon ca.
0.1
Melanoma* (met)
0.1
|
tissue(ODO3866)
Hs688(B).T
|
Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0
|
Gastric ca.* (liver met)
0.1
Melanoma M14
0.1
|
NCI-N87
|
Bladder
0.3
Melanoma LOX IMVI
0.0
|
Trachea
0.1
Melanoma* (met) SK-
0.9
|
MEL-5
|
Kidney
0.2
Adipose
0.8
|
|
[0959]
468
TABLE KD
|
|
|
Panel 2.2
|
Rel. Exp. (%) Ag1614,
Rel. Exp. (%) Ag1614,
|
Tissue Name
Run 173968961
Tissue Name
Run 173968961
|
|
Normal Colon
3.5
Kidney Margin (OD04348)
20.9
|
Colon cancer (OD06064)
1.9
Kidney malignant cancer
0.0
|
(OD06204B)
|
Colon Margin (OD06064)
0.5
Kidney normal adjacent
1.9
|
tissue (OD06204E)
|
Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
6.7
|
01)
|
Colon Margin (OD06159)
2.0
Kidney Margin (OD04450-
8.8
|
03)
|
Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0
|
Colon Margin (OD06297-05)
5.7
Kidney Margin 8120614
1.2
|
CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0
|
(ODO3921)
|
CC Margin (ODO3921)
2.8
Kidney Margin 9010321
1.4
|
Colon cancer metastasis
1.2
Kidney Cancer 8120607
0.0
|
(OD06104)
|
Lung Margin (OD06104)
23.7
Kidney Margin 8120608
0.0
|
Colon mets to lung
26.6
Normal Uterus
10.7
|
(OD04451-01)
|
Lung Margin (OD04451-02)
7.2
Uterine Cancer 064011
1.1
|
Normal Prostate
5.6
Normal Thyroid
5.2
|
Prostate Cancer (OD04410)
2.0
Thyroid Cancer 064010
0.5
|
Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
3.8
|
Normal Ovary
0.0
Thyroid Margin A302153
2.2
|
Ovarian cancer (OD06283-
0.0
Normal Breast
7.0
|
03)
|
Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
3.2
|
07)
|
Ovarian Cancer 064008
4.0
Breast Cancer 1024
3.8
|
Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
2.5
|
01)
|
Ovarian Margin (OD06145)
2.1
Breast Cancer Mets
1.5
|
(OD04590-03)
|
Ovarian cancer (OD06455-
3.6
Breast Cancer Metastasis
8.1
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
10.2
|
07)
|
Normal Lung
6.1
Breast Cancer 9100266
0.0
|
Invasive poor diff. lung
5.6
Breast Margin 9100265
1.5
|
adeno (ODO4945-01
|
Lung Margin (ODO4945-03)
25.7
Breast Cancer A209073
0.5
|
Lung Malignant Cancer
2.4
Breast Margin A2090734
1.7
|
(OD03126)
|
Lung Margin (OD03126)
4.3
Breast cancer (OD06083)
15.5
|
Lung Cancer (OD05014A)
0.5
Breast cancer node
5.2
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
7.3
Normal Liver
6.8
|
Lung cancer (OD06081)
4.6
Liver Cancer 1026
0.0
|
Lung Margin (OD06081)
16.6
Liver Cancer 1025
4.2
|
Lung Cancer (OD04237-01)
3.1
Liver Cancer 6004-T
2.2
|
Lung Margin (OD04237-02)
6.0
Liver Tissue 6004-N
1.7
|
Ocular Melanoma Metastasis
100.0
Liver Cancer 6005-T
0.0
|
Ocular Melanoma Margin
8.8
Liver Tissue 6005-N
0.0
|
(Liver)
|
Melanoma Metastasis
3.2
Liver Cancer 064003
5.3
|
Melanoma Margin (Lung)
4.7
Normal Bladder
4.7
|
Normal Kidney
6.3
Bladder Cancer 1023
0.0
|
Kidney Ca, Nuclear grade 2
26.1
Bladder Cancer A302173
1.3
|
(OD04338)
|
Kidney Margin (OD04338)
5.6
Normal Stomach
5.8
|
Kidney Ca Nuclear grade 1/2
20.4
Gastric Cancer 9060397
0.0
|
(OD04339)
|
Kidney Margin (OD04339)
3.8
Stomach Margin 9060396
0.0
|
Kidney Ca, Clear cell type
1.2
Gastric Cancer 9060395
1.6
|
(OD04340)
|
Kidney Margin (OD04340)
11.0
Stomach Margin 9060394
1.4
|
Kidney Ca, Nuclear grade 3
1.2
Gastric Cancer 064005
1.2
|
(OD04348)
|
|
[0960]
469
TABLE KE
|
|
|
Panel 3D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag1614, Run
Ag1614, Run
|
Tissue Name
170188914
Tissue Name
170188914
|
|
Daoy-Medulloblastoma
0.0
Ca Ski-Cervical epidermoid
0.1
|
carcinoma (metastasis)
|
TE671-Medulloblastoma
0.0
ES-2-Ovarian clear cell carcinoma
0.0
|
D283 Med-Medulloblastoma
0.1
Ramos-Stimulated with
0.0
|
PMA/ionomycin 6 h
|
PFSK-1-Primitive
0.0
Ramos-Stimulated with
0.0
|
Neuroectodermal
PMA/ionomycin 14 h
|
XF-498-CNS
0.0
MEG-01-Chronic myelogenous
0.1
|
leukemia (megokaryoblast)
|
SNB-78-Glioma
0.0
Raji-Burkitt's lymphoma
0.0
|
SF-268-Glioblastoma
0.0
Daudi-Burkitt's lymphoma
0.0
|
T98G-Glioblastoma
0.0
U266-B-cell plasmacytoma
0.1
|
SK-N-SH-Neuroblastoma
0.0
CA46-Burkitt's lymphoma
0.0
|
(metastasis)
|
SF-295-Glioblastoma
0.0
RL-non-Hodgkin's B-cell
0.0
|
lymphoma
|
Cerebellum
0.3
JM1-pre-B-cell lymphoma
0.0
|
Cerebellum
0.0
Jurkat-T cell leukemia
0.0
|
NCI-H292-Mucoepidermoid
100.0
TF-1-Erythroleukemia
0.1
|
lung carcinoma
|
DMS-114-Small cell lung
0.0
HUT 78-T-cell lymphoma
0.0
|
cancer
|
DMS-79-Small cell lung
0.6
U937-Histiocytic lymphoma
0.0
|
cancer
|
NCI-H146-Small cell lung
0.2
KU-812-Myelogenous leukemia
0.1
|
cancer
|
NCI-H526-Small cell lung
0.1
769-P-Clear cell renal carcinoma
0.0
|
cancer
|
NCI-N417-Small cell lung
0.0
Caki-2-Clear cell renal carcinoma
0.0
|
cancer
|
NCI-H82-Small cell lung
0.0
SW 839-Clear cell renal carcinoma
0.0
|
cancer
|
NCI-H157-Squamous cell
0.0
G401-Wilms' tumor
0.0
|
lung cancer (metastasis)
|
NCI-H1155-Large cell lung
0.0
Hs766T-Pancreatic carcinoma (LN
0.0
|
cancer
metastasis)
|
NCI-H1299-Large cell lung
0.0
CAPAN-1-Pancreatic
0.0
|
cancer
adenocarcinoma (liver metastasis)
|
NCI-H727-Lung carcinoid
0.0
SU86.86-Pancreatic carcinoma
0.1
|
(liver metastasis)
|
NCI-UMC-11-Lung
0.0
BxPC-3-Pancreatic
0.0
|
carcinoid
adenocarcinoma
|
LX-1-Small cell lung cancer
0.1
HPAC-Pancreatic adenocarcinoma
0.0
|
Colo-205-Colon cancer
0.0
MIA PaCa-2-Pancreatic carcinoma
0.0
|
KM12-Colon cancer
0.0
CFPAC-1-Pancreatic ductal
0.0
|
adenocarcinoma
|
KM20L2-Colon cancer
0.0
PANC-1-Pancreatic epithelioid
0.0
|
ductal carcinoma
|
NCI-H716-Colon cancer
0.0
T24-Bladder carcinma (transitional
0.1
|
cell)
|
SW-48-Colon
0.0
5637-Bladder carcinoma
0.0
|
adenocarcinoma
|
SW1116-Colon
0.0
HT-1197-Bladder carcinoma
0.0
|
adenocarcinoma
|
LS 174T-Colon
0.0
UM-UC-3-Bladder carcinma
0.0
|
adenocarcinoma
(transitional cell)
|
SW-948-Colon
0.0
A204-Rhabdomyosarcoma
0.0
|
adenocarcinoma
|
SW-480-Colon
0.0
HT-1080-Fibrosarcoma
0.0
|
adenocarcinoma
|
NCI-SNU-5-Gastric
0.0
MG-63-Osteosarcoma
0.0
|
carcinoma
|
KATO III-Gastric carcinoma
0.0
SK-LMS-1-Leiomyosarcoma
0.1
|
(vulva)
|
NCI-SNU-16-Gastric
0.0
SJRH30-Rhabdomyosarcoma (met
0.0
|
carcinoma
to bone marrow)
|
NCI-SNU-1-Gastric
0.0
A431-Epidermoid carcinoma
0.0
|
carcinoma
|
RF-1-Gastric
0.0
WM266-4-Melanoma
0.0
|
adenocarcinoma
|
RF-48-Gastric
0.0
DU 145-Prostate carcinoma (brain
0.0
|
adenocarcinoma
metastasis)
|
MKN-45-Gastric carcinoma
0.0
MDA-MB-468-Breast
0.1
|
adenocarcinoma
|
NCI-N87-Gastric carcinoma
0.0
SCC-4-Squamous cell carcinoma
0.0
|
of tongue
|
OVCAR-5-Ovarian
0.0
SCC-9-Squamous cell carcinoma
0.0
|
carcinoma
of tongue
|
RL95-2-Uterine carcinoma
0.0
SCC-15-Squamous cell carcinoma
0.0
|
of tongue
|
HelaS3-Cervical
0.0
CAL 27-Squamous cell carcinoma
0.0
|
adenocarcinoma
of tongue
|
|
[0961]
470
TABLE KF
|
|
|
Panel 4D
|
Rel. Exp. (%) Ag1614,
Rel. Exp. (%) Ag1614,
|
Tissue Name
Run 165762852
Tissue Name
Run 165762852
|
|
Secondary Th1 act
13.9
HUVEC IL-1beta
11.5
|
Secondary Th2 act
19.5
HUVEC IFN gamma
9.0
|
Secondary Tr1 act
25.5
HUVEC TNF alpha + IFN
3.6
|
gamma
|
Secondary Th1 rest
39.2
HUVEC TNF alpha + IL4
1.4
|
Secondary Th2 rest
17.0
HUVEC IL-11
3.1
|
Secondary Tr1 rest
19.5
Lung Microvascular EC none
5.2
|
Primary Th1 act
10.8
Lung Microvascular EC
3.1
|
TNF alpha + IL-1beta
|
Primary Th2 act
12.8
Microvascular Dermal EC none
12.7
|
Primary Tr1 act
25.7
Microsvasular Dermal EC
5.7
|
TNF alpha + IL-1beta
|
Primary Th1 rest
77.4
Bronchial epithelium TNF
10.9
|
alpha + IL1beta
|
Primary Th2 rest
40.3
Small airway epithelium none
6.7
|
Primary Tr1 rest
17.2
Small airway epithelium
25.2
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
21.0
Coronery artery SMC rest
6.0
|
act
|
CD45RO CD4 lymphocyte
28.3
Coronery artery SMC TNF
6.7
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
34.2
Astrocytes rest
19.9
|
Secondary CD8
19.8
Astrocytes TNF alpha + IL-1beta
18.3
|
lymphocyte rest
|
Secondary CD8
11.7
KU-812 (Basophil) rest
15.2
|
lymphocyte act
|
CD4 lymphocyte none
33.9
KU-812 (Basophil)
67.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
40.6
CCD1106 (Keratinocytes) none
2.9
|
CD95 CH11
|
LAK cells rest
8.1
CCD1106 (Keratinocytes)
10.9
|
TNF alpha + IL-1beta
|
LAK cells IL-2
33.2
Liver cirrhosis
28.5
|
LAK cells IL-2 + IL-12
13.8
Lupus kidney
8.5
|
LAK cells IL-2 + IFN
39.8
NCI-H292 none
27.7
|
gamma
|
LAK cells IL-2 + IL-18
25.3
NCI-H292 IL-4
28.1
|
LAK cells
3.0
NCI-H292 IL-9
19.5
|
PMA/ionomycin
|
NK Cells IL-2 rest
19.2
NCI-H292 IL-13
20.3
|
Two Way MLR 3 day
22.1
NCI-H292 IFN gamma
12.1
|
Two Way MLR 5 day
16.3
HPAEC none
9.1
|
Two Way MLR 7 day
15.5
HPAEC TNF alpha + IL-1beta
3.3
|
PBMC rest
16.3
Lung fibroblast none
12.3
|
PBMC PWM
14.1
Lung fibroblast TNF alpha + IL-
11.5
|
1beta
|
PBMC PHA-L
18.9
Lung fibroblast IL-4
7.3
|
Ramos (B cell) none
12.9
Lung fibroblast IL-9
10.2
|
Ramos (B cell) ionomycin
22.1
Lung fibroblast IL-13
8.7
|
B lymphocytes PWM
26.4
Lung fibroblast IFN gamma
5.6
|
B lymphocytes CD40L
100.0
Dermal fibroblast CCD1070 rest
15.4
|
and IL-4
|
EOL-1 dbcAMP
10.4
Dermal fibroblast CCD1070
20.9
|
TNF alpha
|
EOL-1 dbcAMP
8.0
Dermal fibroblast CCD1070 IL-
2.3
|
PMA/ionomycin
1beta
|
Dendritic cells none
16.0
Dermal fibroblast IFN gamma
4.6
|
Dendritic cells LPS
9.3
Dermal fibroblast IL-4
7.1
|
Dendritic cells anti-CD40
18.0
IBD Colitis 2
3.9
|
Monocytes rest
33.0
IBD Crohn's
19.1
|
Monocytes LPS
2.9
Colon
27.4
|
Macrophages rest
26.4
Lung
13.0
|
Macrophages LPS
13.2
Thymus
33.4
|
HUVEC none
8.6
Kidney
51.8
|
HUVEC starved
12.0
|
|
[0962]
471
TABLE KG
|
|
|
Panel 5D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag1614, Run
Ag1614, Run
|
Tissue Name
169269227
Tissue Name
169269227
|
|
97457_Patient-
0.3
94709_Donor 2 AM - A_adipose
0.2
|
02go_adipose
|
97476_Patient-07sk_skeletal
10.0
94710_Donor 2 AM - B_adipose
0.1
|
muscle
|
97477_Patient-07ut_uterus
0.3
94711_Donor 2 AM - C_adipose
0.3
|
97478_Patient-
1.3
94712_Donor 2 AD - A_adipose
0.5
|
07pl_placenta
|
97481_Patient-08sk_skeletal
17.4
94713_Donor 2 AD - B_adipose
0.3
|
muscle
|
97482_Patient-08ut_uterus
0.0
94714_Donor 2 AD - C_adipose
0.4
|
97483_Patient-
1.1
94742_Donor 3 U - A_Mesenchymal
0.0
|
08pl_placenta
Stem Cells
|
97486_Patient-09sk_skeletal
23.3
94743_Donor 3 U - B_Mesenchymal
0.1
|
muscle
Stem Cells
|
97487_Patient-09ut_uterus
0.4
94730_Donor 3 AM - A_adipose
0.0
|
97488_Patient-
0.7
94731_Donor 3 AM - B_adipose
0.1
|
09pl_placenta
|
97492_Patient-10ut_uterus
0.0
94732_Donor 3 AM - C_adipose
0.4
|
97493_Patient-
1.6
94733_Donor 3 AD - A_adipose
0.3
|
10pl_placenta
|
97495_Patient-
0.3
94734_Donor 3 AD - B_adipose
0.1
|
11go_adipose
|
97496_Patient-11sk_skeletal
88.3
94735_Donor 3 AD - C_adipose
0.1
|
muscle
|
97497_Patient-11ut_uterus
0.1
77138_Liver_HepG2untreated
0.2
|
97498_Patient-
0.5
73556_Heart_Cardiac stromal cells
0.0
|
11pl_placenta
(primary)
|
97500_Patient-
0.3
81735_Small Intestine
0.5
|
12go_adipose
|
97501_Patient-12sk_skeletal
100.0
72409_Kidney_Proximal Convoluted
0.1
|
muscle
Tubule
|
97502_Patient-12ut_uterus
0.3
82685_Small intestine_Duodenum
0.1
|
97503_Patient-
1.1
90650_Adrenal_Adrenocortical
0.6
|
12pl_placenta
adenoma
|
94721_Donor 2 U -
0.0
72410_Kidney_HRCE
0.4
|
A_Mesenchymal Stem Cells
|
94722_Donor 2 U -
0.2
72411_Kidney_HRE
0.1
|
B_Mesenchymal Stem Cells
|
94723_Donor 2 U -
0.1
73139_Uterus_Uterine smooth
0.2
|
C_Mesenchymal Stem Cells
muscle cells
|
|
[0963] Panel 1.3D Summary: Ag1614 Expression of the CG59930-01 gene is limited to a few samples with highest expression in skeletal muscle (CT=27). Moderate levels of expression are seen in heart and low but significant levels are seen in fetal heart, fetal skeletal muscle, amygdala, thalamus, cerebellum, substantia nigra and hippocampus. Thus, expression of this gene could be used to differentiate skeletal muscle from other samples on this panel, particularly from fetal skeletal muscle (CT=33.8), and as a marker of skeletal tissue.
[0964] This gene encodes a putative leucine-rich repeat protein. Leucine rich repeats (LRR) mediate reversible protein-protein interactions and have diverse cellular functions, including cellular adhesion and signaling. Several of these proteins, such as connectin, slit, chaoptin, and Toll have pivotal roles in neuronal development in Drosophila and may play significant but distinct roles in neural development and in the adult nervous system of humans (Ref. 2). In Drosophilia, the LRR region of axon guidance proteins has been shown to be critical for their function (especially in axon repulsion). Therefore, therapeutic modulation of the levels of this protein, or possible signaling via this protein, may be of utility in enhancing/directing compensatory synaptogenesis and fiber growth in the CNS in response to neuronal death (stroke, head trauma), axon lesion (spinal cord injury), or neurodegeneration (Alzheimer's, Parkinson's, Huntington's, vascular dementia or any neurodegenerative disease).
[0965] Panel 2.2 Summary: Agl614 Highest expression of the CG59930-01 gene is seen in an ocular melanoma metastasis to the liver (CT=31.9). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel.
[0966] Panel 3D Summary: Ag1614 Expression of the CG59930-01 gene is highest in a lung cancer cell line (CT=25.6). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel and as a marker to detect the presence of lung cancer. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of lung cancer.
[0967] Panel 4D Summary: Ag1614 The CG59930-01 gene is expressed ubiquitously in this panel, with highest expression in CD40/IL-4 treated B lymphocytes (CT=31.3). In addition, this gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[0968] Panel 5D Summary: Ag1614 Expression of the CG59930-01 gene is in agreement with the profile in Panel 1.3D. Highest expression is in skeletal muscle (CT=27.8) and several samples of skeletal muscle show prominent expression of this gene. Thus, this gene may be involved in the pathogenesis and/or treatment of metabolic disorders that involve skeletal muscle, including obesity and diabetes.
[0969] L. CG59934-01 and CG59934-O2: Leucine-Rich Repeat Protein
[0970] Expression of gene CG59934-01 and full length clone CG59934-02 was assessed using the primer-probe sets Ag3633 and Ag4659, described in Tables LA and LB. Results of the RTQ-PCR runs are shown in Tables LC, LD and LE. Please note that the Ag3633 probe and primer set is specific to CG59934-01 only.
[0971] Table LA. Probe Name Ag3633
472|
|
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-gctgaacgtccagctctct-3′191701255
ProbeTET-5′-ctccctgggactgcagtacaatgct-251731256
3′-TAMRA
Reverse5′-ctcaggtcccactcctgact-3′201756257
|
[0972]
473
TABLE LB
|
|
|
Probe Name Ag4659
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-cacaggcagtagcttcttgaac-3′
22
510
258
|
Probe
TET-5′-cagcaccaacatcttctccaacctga-
26
534
259
|
3′-TAMRA
|
Reverse
5′-cctccagcatgttgaagttg-3′
20
582
260
|
|
[0973]
474
TABLE LC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag4659,
Rel. Exp. (%) Ag4659,
|
Tissue Name
Run 224723494
Tissue Name
Run 224723494
|
|
AD 1 Hippo
0.0
Control (Path) 3
0.0
|
Temporal Ctx
|
AD 2 Hippo
54.0
Control (Path) 4
0.0
|
Temporal Ctx
|
AD 3 Hippo
100.0
AD 1 Occipital Ctx
0.0
|
AD 4 Hippo
0.0
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
57.4
AD 3 Occipital Ctx
0.0
|
AD 6 Hippo
0.0
AD 4 Occipital Ctx
0.0
|
Control 2 Hippo
0.0
AD 5 Occipital Ctx
0.0
|
Control 4 Hippo
0.0
AD 6 Occipital Ctx
0.0
|
Control (Path) 3 Hippo
0.0
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
0.0
Control 2 Occipital Ctx
0.0
|
AD 2 Temporal Ctx
80.7
Control 3 Occipital Ctx
0.0
|
AD 3 Temporal Ctx
0.0
Control 4 Occipital Ctx
0.0
|
AD 4 Temporal Ctx
0.0
Control (Path) 1
0.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
0.0
Control (Path) 2
0.0
|
Occipital Ctx
|
AD 5 SupTemporal
0.0
Control (Path) 3
0.0
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
0.0
Control (Path) 4
0.0
|
Occipital Ctx
|
AD 6 Sup Temporal
0.0
Control 1 Parietal Ctx
0.0
|
Ctx
|
Control 1 Temporal Ctx
0.0
Control 2 Parietal Ctx
0.0
|
Control 2 Temporal Ctx
0.0
Control 3 Parietal Ctx
0.0
|
Control 3 Temporal Ctx
0.0
Control (Path) 1
0.0
|
Parietal Ctx
|
Control 3 Temporal Ctx
0.0
Control (Path) 2
0.0
|
Parietal Ctx
|
Control (Path) 1
0.0
Control (Path) 3
0.0
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
0.0
Control (Path) 4
0.0
|
Temporal Ctx
Parietal Ctx
|
|
[0974]
475
TABLE LD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3633, Run
Ag3633, Run
Ag3633, Run
Ag3633, Run
|
Tissue Name
217218047
218233757
Tissue Name
217218047
218233757
|
|
Adipose
0.0
0.0
Renal ca. TK-10
8.7
5.8
|
Melanoma*
0.8
1.1
Bladder
0.0
0.9
|
Hs688(A).T
|
Melanoma*
0.4
0.0
Gastric ca. (liver
0.2
0.0
|
Hs688(B).T
met.) NCI-N87
|
Melanoma*
0.0
0.0
Gastric ca. KATO
0.0
0.0
|
M14
III
|
Melanoma*
0.0
0.0
Colon ca. SW-948
0.0
0.0
|
LOXIMVI
|
Melanoma* SK-
0.0
0.0
Colon ca. SW480
0.0
0.0
|
MEL-5
|
Squamous cell
0.0
0.0
Colon ca.* (SW480
0.0
0.0
|
carcinoma SCC-4
met) SW620
|
Testis Pool
0.7
0.0
Colon ca. HT29
0.0
0.0
|
Prostate ca.*
0.0
0.0
Colon ca. HCT-116
0.2
0.0
|
(bone met) PC-3
|
Prostate Pool
0.0
0.0
Colon ca. CaCo-2
0.4
0.3
|
Placenta
0.0
0.0
Colon cancer tissue
0.0
0.6
|
Uterus Pool
0.0
0.0
Colon ca. SW1116
0.0
0.3
|
Ovarian ca.
0.0
0.0
Colon ca. Colo-205
0.0
0.0
|
OVCAR-3
|
Ovarian ca. SK-
0.0
1.6
Colon ca. SW-48
0.0
0.0
|
OV-3
|
Ovarian ca.
0.0
0.0
Colon Pool
0.6
0.0
|
OVCAR-4
|
Ovarian ca.
0.0
0.6
Small Intestine Pool
0.0
0.5
|
OVCAR-5
|
Ovarian ca.
0.0
0.0
Stomach Pool
0.0
0.4
|
IGROV-1
|
Ovarian ca.
0.0
0.0
Bone Marrow Pool
0.4
0.0
|
OVCAR-8
|
Ovary
0.0
0.0
Fetal Heart
0.1
0.0
|
Breast ca. MCF-7
0.0
0.0
Heart Pool
0.5
0.1
|
Breast ca. MDA-
0.7
0.0
Lymph Node Pool
0.0
1.1
|
MB-231
|
Breast ca. BT
0.4
0.0
Fetal Skeletal
0.0
0.0
|
549
Muscle
|
Breast ca. T47D
0.0
0.4
Skeletal Muscle
0.0
0.0
|
Pool
|
Breast ca. MDA-N
0.0
0.0
Spleen Pool
0.1
0.0
|
Breast Pool
0.3
0.4
Thymus Pool
0.3
0.3
|
Trachea
0.0
0.0
CNS cancer
3.3
1.4
|
(glio/astro) U87-
|
MG
|
Lung
0.4
0.0
CNS cancer
1.7
1.9
|
(glio/astro) U-118-
|
MG
|
Fetal Lung
0.5
0.0
CNS cancer
0.0
0.0
|
(neuro; met) SK-N-
|
AS
|
Lung ca. NCI-
0.0
0.0
CNS cancer (astro)
0.0
0.0
|
N417
SF-539
|
Lung ca. LX-1
0.0
0.0
CNS cancer (astro)
0.0
0.4
|
SNB-75
|
Lung ca. NCI-
0.0
0.0
CNS cancer (glio)
0.0
0.0
|
H146
SNB-19
|
Lung ca. SHP-
0.0
0.0
CNS cancer (glio)
0.0
0.4
|
77
SF-295
|
Lung ca. A549
0.0
0.0
Brain (Amygdala)
0.0
0.0
|
Pool
|
Lung ca. NCI-
0.0
0.0
Brain (cerebellum)
0.0
0.0
|
H526
|
Lung ca. NCI-
0.6
0.0
Brain (fetal)
0.0
0.0
|
H23
|
Lung ca. NCI-
0.0
0.0
Brain
0.0
0.0
|
H460
(Hippocampus)
|
Pool
|
Lung ca. HOP-
0.0
0.0
Cerebral Cortex
0.0
0.0
|
62
Pool
|
Lung ca. NCI-
0.0
0.0
Brain (Substantia
0.4
0.0
|
H522
nigra) Pool
|
Liver
100.0
100.0
Brain (Thalamus)
0.0
0.6
|
Pool
|
Fetal Liver
48.6
54.0
Brain (whole)
2.7
2.0
|
Liver ca. HepG2
23.2
21.8
Spinal Cord Pool
0.4
0.0
|
Kidney Pool
0.5
0.5
Adrenal Gland
3.3
3.1
|
Fetal Kidney
0.5
0.0
Pituitary gland Pool
0.0
0.0
|
Renal ca. 786-0
0.0
0.0
Salivary Gland
0.0
0.0
|
Renal ca. A498
0.0
0.5
Thyroid (female)
0.8
0.0
|
Renal ca. ACHN
0.0
0.0
Pancreatic ca.
0.0
0.0
|
CAPAN2
|
Renal ca. UO-31
0.0
0.4
Pancreas Pool
0.5
2.2
|
|
[0975]
476
TABLE LE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3633, Run
Ag4659, Run
Ag3633, Run
Ag4659, Run
|
Tissue Name
169960871
200690190
Tissue Name
169960871
200690190
|
|
Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0
|
Secondary Th2 act
0.0
1.1
HUVEC IFN gamma
0.0
0.0
|
Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0
|
IFN gamma
|
Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0
|
IL4
|
Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0
|
Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0
|
EC none
|
Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0
|
EC TNF alpha + IL-
|
1beta
|
Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0
|
Dermal EC none
|
Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0
|
EC TNF alpha + IL-
|
1beta
|
Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0
|
TNF alpha + IL1beta
|
Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0
|
epithelium none
|
Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0
|
epithelium TNF
|
alpha + IL-1beta
|
CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0
|
lymphocyte act
rest
|
CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0
|
lymphocyte act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0
|
Secondary CD8
0.0
0.0
Astrocytes TNF
0.0
0.0
|
lymphocyte rest
alpha + IL-1beta
|
Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0
|
lymphocyte act
rest
|
CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0
|
none
PMA/ionomycin
|
2ry
0.0
0.0
CCD1106
0.0
0.0
|
Th1/Th2/Tr1_anti-
(Keratinocytes) none
|
CD95 CH11
|
LAK cells rest
0.0
0.0
CCD1106
0.0
0.0
|
(Keratinocytes)
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
0.0
Liver cirrhosis
87.1
47.3
|
LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 none
0.0
0.0
|
12
|
LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 IL-4
0.0
0.0
|
gamma
|
LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-9
0.0
0.0
|
18
|
LAK cells
0.0
0.0
NCI-H292 IL-13
0.0
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
0.0
NCI-H292 IFN
0.0
0.0
|
gamma
|
Two Way MLR 3
0.0
0.0
HPAEC none
0.0
0.0
|
day
|
Two Way MLR 5
0.0
0.0
HPAEC TNF alpha +
0.0
0.0
|
day
IL-1beta
|
Two Way MLR 7
0.0
0.0
Lung fibroblast none
0.0
0.0
|
day
|
PBMC rest
0.0
0.0
Lung fibroblast TNF
0.0
0.5
|
alpha + IL-1beta
|
PBMC PWM
0.0
0.0
Lung fibroblast IL-4
0.0
0.0
|
PBMC PHA-L
0.0
0.0
Lung fibroblast IL-9
0.0
0.0
|
Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-13
0.0
0.0
|
Ramos (B cell)
0.0
0.0
Lung fibroblast IFN
0.0
0.0
|
ionomycin
gamma
|
B lymphocytes PWM
0.0
0.0
Dermal fibroblast
0.0
0.0
|
CCD1070 rest
|
B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0
|
CD40L and IL-4
CCD1070 TNF alpha
|
EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
5.7
0.0
|
CCD1070 IL-1beta
|
EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast IFN
0.0
0.0
|
PMA/ionomycin
gamma
|
Dendritic cells none
5.9
0.0
Dermal fibroblast IL-4
0.0
0.8
|
Dendritic cells LPS
0.0
0.0
Dermal Fibroblasts
0.0
0.5
|
rest
|
Dendritic cells anti-
0.0
0.0
Neutrophils
0.0
0.0
|
CD40
TNFa + LPS
|
Monocytes rest
0.0
0.0
Neutrophils rest
0.0
0.0
|
Monocytes LPS
1.6
0.0
Colon
0.0
0.0
|
Macrophages rest
0.0
0.0
Lung
0.0
0.0
|
Macrophages LPS
0.0
0.0
Thymus
0.0
6.5
|
HUVEC none
0.0
0.0
Kidney
100.0
100.0
|
HUVEC starved
0.0
0.0
|
|
[0976] CNS_neurodegeneration_v1.0 Summary: Ag3633/Ag4659 Expression of the CG59934-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[0977] General_screening_panel_v1.4 Summary: Ag3633/Ag4659 Two experiments with two different probe and primer sets produce results that are in excellent agreement. Significant expression of the CG59934-01 gene is primarily limited to liver derived samples, with moderate expression in fetal and adult liver and liver cancer (CTs=28-30). Thus, expression of this gene could be used to differentiate between these liver derived samples and other samples on this panel and as a marker of liver tissue.
[0978] Panel 4.1D Summary: Ag3633/Ag4659 Two experiments with two different probe and primer sets produce results that are in excellent agreement. Significant expression of the CG59934-01 gene is limited to kidney and liver cirrhosis (CTs=30-33). The expression in liver cirrhosis is consistent with the liver specific expression in Panel 1.4. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker of kidney and liver tissue.
[0979] M. CG88565-01: UDP N-acetylglucosamine Transporter
[0980] Expression of gene CG88565-01 was assessed using the primer-probe set Ag3646, described in Table MA. Results of the RTQ-PCR runs are shown in Tables MB, MC and MD.
477TABLE MA
|
|
Probe Name Ag3646
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ttcgtcttcttggcttctgata-3′22646261
ProbeTET-5′-30672262
tgaccatgcagttctattcaataagtctga-
3′-TAMRA
Reverse5′-cgatgacgggaatttcatataa-3′22703263
|
[0981]
478
TABLE MB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3646,
Rel. Exp. (%) Ag3646,
|
Tissue Name
Run 211019282
Tissue Name
Run 211019282
|
|
AD 1 Hippo
8.4
Control (Path) 3
2.3
|
Temporal Ctx
|
AD 2 Hippo
28.9
Control (Path) 4
25.7
|
Temporal Ctx
|
AD 3 Hippo
2.6
AD 1 Occipital Ctx
6.2
|
AD 4 Hippo
3.8
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
1.9
|
AD 6 Hippo
31.6
AD 4 Occipital Ctx
15.7
|
Control 2 Hippo
27.7
AD 5 Occipital Ctx
57.0
|
Control 4 Hippo
7.6
AD 6 Occipital Ctx
11.7
|
Control (Path) 3 Hippo
2.4
Control 1 Occipital Ctx
1.5
|
AD 1 Temporal Ctx
7.5
Control 2 Occipital Ctx
74.7
|
AD 2 Temporal Ctx
21.9
Control 3 Occipital Ctx
9.5
|
AD 3 Temporal Ctx
2.0
Control 4 Occipital Ctx
5.4
|
AD 4 Temporal Ctx
14.7
Control (Path) 1
94.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
94.0
Control (Path) 2
7.0
|
Occipital Ctx
|
AD 5 Sup Temporal
36.9
Control (Path) 3
1.1
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
27.0
Control (Path) 4
14.1
|
Occipital Ctx
|
AD 6 Sup Temporal
26.2
Control 1 Parietal Ctx
2.9
|
Ctx
|
Control 1 Temporal Ctx
3.2
Control 2 Parietal Ctx
26.8
|
Control 2 Temporal Ctx
71.2
Control 3 Parietal Ctx
16.6
|
Control 3 Temporal Ctx
11.0
Control (Path) 1
74.2
|
Parietal Ctx
|
Control 3 Temporal Ctx
4.5
Control (Path) 2
16.8
|
Parietal Ctx
|
Control (Path) 1
69.3
Control (Path) 3
2.2
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
35.8
Control (Path) 4
47.0
|
Temporal Ctx
Parietal Ctx
|
|
[0982]
479
TABLE MC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3646,
Rel. Exp. (%) Ag3646,
|
Tissue Name
Run 218341951
Tissue Name
Run 218341951
|
|
Adipose
1.8
Renal ca. TK-10
13.7
|
Melanoma* Hs688(A).T
11.0
Bladder
3.3
|
Melanoma* Hs688(B).T
12.3
Gastric ca. (liver met.)
2.9
|
NCI-N87
|
Melanoma* M14
43.2
Gastric ca. KATO III
23.7
|
Melanoma* LOXIMVI
18.9
Colon ca. SW-948
2.2
|
Melanoma* SK-MEL-5
100.0
Colon ca. SW480
28.7
|
Squamous cell
4.5
Colon ca.* (SW480 met)
22.2
|
carcinoma SCC-4
SW620
|
Testis Pool
3.3
Colon ca. HT29
3.8
|
Prostate ca.* (bone met)
31.2
Colon ca. HCT-116
20.3
|
PC-3
|
Prostate Pool
1.9
Colon ca. CaCo-2
17.2
|
Placenta
6.4
Colon cancer tissue
11.9
|
Uterus Pool
0.8
Colon ca. SW1116
3.2
|
Ovarian ca. OVCAR-3
7.8
Colon ca. Colo-205
1.4
|
Ovarian ca. SK-OV-3
13.9
Colon ca. SW-48
1.7
|
Ovarian ca. OVCAR-4
5.5
Colon Pool
5.5
|
Ovarian ca. OVCAR-5
9.7
Small Intestine Pool
3.4
|
Ovarian ca. IGROV-1
9.7
Stomach Pool
2.5
|
Ovarian ca. OVCAR-8
8.4
Bone Marrow Pool
1.9
|
Ovary
5.9
Fetal Heart
1.5
|
Breast ca. MCF-7
6.7
Heart Pool
2.1
|
Breast ca. MDA-MB-
39.8
Lymph Node Pool
7.8
|
231
|
Breast ca. BT 549
24.0
Fetal Skeletal Muscle
1.6
|
Breast ca. T47D
23.2
Skeletal Muscle Pool
2.0
|
Breast ca. MDA-N
41.8
Spleen Pool
2.7
|
Breast Pool
7.0
Thymus Pool
3.8
|
Trachea
3.0
CNS cancer (glio/astro)
69.3
|
U87-MG
|
Lung
1.4
CNS cancer (glio/astro) U-
39.2
|
118-MG
|
Fetal Lung
7.0
CNS cancer (neuro; met)
28.3
|
SK-N-AS
|
Lung ca. NCI-N417
11.9
CNS cancer (astro) SF-539
13.3
|
Lung ca. LX-1
7.7
CNS cancer (astro) SNB-75
59.0
|
Lung ca. NCI-H146
5.0
CNS cancer (glio) SNB-19
8.8
|
Lung ca. SHP-77
25.0
CNS cancer (glio) SF-295
29.7
|
Lung ca. A549
41.5
Brain (Amygdala) Pool
7.3
|
Lung ca. NCI-H526
1.4
Brain (cerebellum)
20.0
|
Lung ca. NCI-H23
20.0
Brain (fetal)
9.2
|
Lung ca. NCI-H460
13.0
Brain (Hippocampus) Pool
9.1
|
Lung ca. HOP-62
9.7
Cerebral Cortex Pool
14.3
|
Lung ca. NCI-H522
23.0
Brain (Substantia nigra)
11.1
|
Pool
|
Liver
1.1
Brain (Thalamus) Pool
15.3
|
Fetal Liver
6.0
Brain (whole)
13.8
|
Liver ca. HepG2
6.0
Spinal Cord Pool
5.3
|
Kidney Pool
8.2
Adrenal Gland
9.3
|
Fetal Kidney
3.6
Pituitary gland Pool
1.7
|
Renal ca. 786-0
19.5
Salivary Gland
0.8
|
Renal ca. A498
8.2
Thyroid (female)
1.4
|
Renal ca. ACHN
7.4
Pancreatic ca. CAPAN2
8.4
|
Renal ca. UO-31
13.0
Pancreas Pool
7.1
|
|
[0983]
480
TABLE MD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3646,
Rel. Exp. (%) Ag3646,
|
Tissue Name
Run 169975208
Tissue Name
Run 169975208
|
|
Secondary Th1 act
16.0
HUVEC IL-1beta
15.7
|
Secondary Th2 act
22.5
HUVEC IFN gamma
11.0
|
Secondary Tr1 act
19.1
HUVEC TNF alpha + IFN
14.1
|
gamma
|
Secondary Th1 rest
4.1
HUVEC TNF alpha + IL4
11.1
|
Secondary Th2 rest
5.6
HUVEC IL-11
5.0
|
Secondary Tr1 rest
3.7
Lung Microvascular EC none
20.4
|
Primary Th1 act
12.2
Lung Microvascular EC
18.2
|
TNF alpha + IL-1beta
|
Primary Th2 act
13.5
Microvascular Dermal EC none
10.8
|
Primary Tr1 act
12.1
Microsvasular Dermal EC
8.6
|
TNF alpha + IL-1beta
|
Primary Th1 rest
4.7
Bronchial epithelium TNF
6.5
|
alpha + IL1beta
|
Primary Th2 rest
5.1
Small airway epithelium none
4.5
|
Primary Tr1 rest
9.7
Small airway epithelium
7.6
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
14.1
Coronery artery SMC rest
7.4
|
act
|
CD45RO CD4 lymphocyte
12.7
Coronery artery SMC TNF
9.2
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
12.5
Astrocytes rest
6.3
|
Secondary CD8
10.7
Astrocytes TNF alpha + IL-1beta
4.1
|
lymphocyte rest
|
Secondary CD8
8.7
KU-812 (Basophil) rest
4.6
|
lymphocyte act
|
CD4 lymphocyte none
1.7
KU-812 (Basophil)
5.3
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
4.8
CCD1106 (Keratinocytes) none
14.6
|
CD95 CH11
|
LAK cells rest
9.6
CCD1106 (Keratinocytes)
8.4
|
TNF alpha + IL-1beta
|
LAK cells IL-2
9.7
Liver cirrhosis
1.5
|
LAK cells IL-2 + IL-12
10.3
NCI-H292 none
7.3
|
LAK cells IL-2 + IFN
12.5
NCI-H292 IL-4
8.2
|
gamma
|
LAK cells IL-2 + IL-18
13.2
NCI-H292 IL-9
13.8
|
LAK cells
3.3
NCI-H292 IL-13
8.8
|
PMA/ionomycin
|
NK Cells IL-2 rest
11.6
NCI-H292 IFN gamma
6.6
|
Two Way MLR 3 day
8.3
HPAEC none
7.4
|
Two Way MLR 5 day
9.4
HPAEC TNF alpha + IL-1beta
9.6
|
Two Way MLR 7 day
6.7
Lung fibroblast none
10.2
|
PBMC rest
2.2
Lung fibroblast TNF alpha + IL-
8.3
|
1beta
|
PBMC PWM
14.2
Lung fibroblast IL-4
14.9
|
PBMC PHA-L
9.5
Lung fibroblast IL-9
21.2
|
Ramos (B cell) none
77.9
Lung fibroblast IL-13
13.2
|
Ramos (B cell) ionomycin
100.0
Lung fibroblast IFN gamma
11.6
|
B lymphocytes PWM
9.0
Dermal fibroblast CCD1070 rest
16.6
|
B lymphocytes CD40L
10.1
Dermal fibroblast CCD1070
19.3
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
24.3
Dermal fibroblast CCD1070 IL-
11.0
|
1beta
|
EOL-1 dbcAMP
11.1
Dermal fibroblast IFN gamma
8.8
|
PMA/ionomycin
|
Dendritic cells none
17.7
Dermal fibroblast IL-4
14.0
|
Dendritic cells LPS
10.2
Dermal Fibroblasts rest
11.3
|
Dendritic cells anti-CD40
18.2
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
4.5
Neutrophils rest
0.4
|
Monocytes LPS
4.2
Colon
2.6
|
Macrophages rest
14.0
Lung
5.5
|
Macrophages LPS
13.7
Thymus
8.1
|
HUVEC none
10.3
Kidney
15.9
|
HUVEC starved
15.2
|
|
[0984] CNS_neurodegeneration_v1.0 Summary: Ag3646 This panel confirms the expression of the CG88565-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[0985] General_screening_panel_v1.4 Summary: Ag3646 Highest expression of the CG88565-01 gene is detected in melanoma SK-MEL-5 cell line (CT=28.3). In addition, high to moderate expression of this gene is also seen in number of cancer cell lines (CNS, colon, renal, lung, breast, ovarian, prostate, and melanomas). Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.
[0986] Among tissues with metabolic or endocrine function, this gene is expressed at low to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[0987] In addition, this gene is expressed at moderate levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[0988] Panel 4.1D Summary: Ag3646 Highest expression of the CG88565-01 gene is detected in Ramos B cells (CTs=29). This gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[0989] N. CG88623-01: POTASSIUM CHANNEL, ALPHA SUBUNIT like Gene
[0990] Expression of gene CG88623-01 was assessed using the primer-probe set Ag3648, described in Table NA. Results of the RTQ-PCR runs are shown in Tables NB, NC and ND.
481TABLE NA
|
|
Probe Name Ag3648
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-cggagacatgtacccagaga-3′201186264
ProbeTET-5′-ttttgccttcctctgcattgcttttg-261222265
3′-TAMRA
Reverse5′-gcatcccgttgagaatgat-3′191250266
|
[0991]
482
TABLE NB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3648,
Rel. Exp. (%) Ag3648,
|
Tissue Name
Run 211019463
Tissue Name
Run 211019463
|
|
AD 1 Hippo
31.2
Control (Path) 3
12.1
|
Temporal Ctx
|
AD 2 Hippo
36.6
Control (Path) 4
49.0
|
Temporal Ctx
|
AD 3 Hippo
9.0
AD 1 Occipital Ctx
6.9
|
AD 4 Hippo
24.8
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
77.9
AD 3 Occipital Ctx
6.9
|
AD 6 Hippo
39.2
AD 4 Occipital Ctx
23.7
|
Control 2 Hippo
26.4
AD 5 Occipital Ctx
24.7
|
Control 4 Hippo
25.5
AD 6 Occipital Ctx
10.5
|
Control (Path) 3 Hippo
9.1
Control 1 Occipital Ctx
3.6
|
AD 1 Temporal Ctx
35.8
Control 2 Occipital Ctx
17.0
|
AD 2 Temporal Ctx
31.2
Control 3 Occipital Ctx
16.0
|
AD 3 Temporal Ctx
8.9
Control 4 Occipital Ctx
8.2
|
AD 4 Temporal Ctx
24.1
Control (Path) 1
68.3
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
78.5
Control (Path) 2
10.7
|
Occipital Ctx
|
AD 5 Sup Temporal
51.1
Control (Path) 3
1.2
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
43.5
Control (Path) 4
21.9
|
Occipital Ctx
|
AD 6 Sup Temporal
41.8
Control 1 Parietal Ctx
6.5
|
Ctx
|
Control 1 Temporal Ctx
8.4
Control 2 Parietal Ctx
40.1
|
Control 2 Temporal Ctx
32.1
Control 3 Parietal Ctx
21.5
|
Control 3 Temporal Ctx
9.2
Control (Path) 1
100.0
|
Parietal Ctx
|
Control 3 Temporal Ctx
22.4
Control (Path) 2
33.2
|
Parietal Ctx
|
Control (Path) 1
76.8
Control (Path) 3
10.9
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
61.1
Control (Path) 4
57.8
|
Temporal Ctx
Parietal Ctx
|
|
[0992]
483
TABLE NC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3648,
Rel. Exp. (%) Ag3648,
|
Tissue Name
Run 21978045
Tissue Name
Run 219798045
|
|
Adipose
2.3
Renal ca. TK-10
33.2
|
Melanoma* Hs688(A).T
6.8
Bladder
17.1
|
Melanoma* Hs688(B).T
7.2
Gastric ca. (liver met.)
50.0
|
NCI-N87
|
Melanoma* M14
5.8
Gastric ca. KATO III
32.8
|
Melanoma* LOXIMVI
4.2
Colon ca. SW-948
4.4
|
Melanoma* SK-MEL-5
32.5
Colon ca. SW480
18.6
|
Squamous cell
6.7
Colon ca.* (SW480 met)
30.1
|
carcinoma SCC-4
SW620
|
Testis Pool
26.2
Colon ca. HT29
4.4
|
Prostate ca.* (bone met)
11.3
Colon ca. HCT-116
79.0
|
PC-3
|
Prostate Pool
4.2
Colon ca. CaCo-2
13.6
|
Placenta
0.5
Colon cancer tissue
4.3
|
Uterus Pool
7.1
Colon ca. SW1116
0.5
|
Ovarian ca. OVCAR-3
29.5
Colon ca. Colo-205
1.4
|
Ovarian ca. SK-OV-3
21.6
Colon ca. SW-48
0.3
|
Ovarian ca. OVCAR-4
8.8
Colon Pool
7.9
|
Ovarian ca. OVCAR-5
13.8
Small Intestine Pool
15.0
|
Ovarian ca. IGROV-1
8.8
Stomach Pool
13.6
|
Ovarian ca. OVCAR-8
2.1
Bone Marrow Pool
6.2
|
Ovary
9.5
Fetal Heart
2.3
|
Breast ca. MCF-7
14.6
Heart Pool
3.1
|
Breast ca. MDA-MB-
30.6
Lymph Node Pool
16.4
|
231
|
Breast ca. BT 549
100.0
Fetal Skeletal Muscle
7.6
|
Breast ca. T47D
13.6
Skeletal Muscle Pool
15.0
|
Breast ca. MDA-N
7.9
Spleen Pool
5.1
|
Breast Pool
13.5
Thymus Pool
10.2
|
Trachea
6.2
CNS cancer (glio/astro)
35.6
|
U87-MG
|
Lung
6.6
CNS cancer (glio/astro) U-
26.4
|
118-MG
|
Fetal Lung
9.0
CNS cancer (neuro; met)
7.3
|
SK-N-AS
|
Lung ca. NCI-N417
1.6
CNS cancer (astro) SF-539
6.6
|
Lung ca. LX-1
15.6
CNS cancer (astro) SNB-75
56.3
|
Lung ca. NCI-H146
9.0
CNS cancer (glio) SNB-19
6.1
|
Lung ca. SHP-77
3.2
CNS cancer (glio) SF-295
15.7
|
Lung ca. A549
23.5
Brain (Amygdala) Pool
8.3
|
Lung ca. NCI-H526
2.0
Brain (cerebellum)
2.9
|
Lung ca. NCI-H23
40.1
Brain (fetal)
14.3
|
Lung ca. NCI-H460
29.7
Brain (Hippocampus) Pool
11.4
|
Lung ca. HOP-62
12.2
Cerebral Cortex Pool
9.2
|
Lung ca. NCI-H522
4.5
Brain (Substantia nigra)
5.8
|
Pool
|
Liver
0.2
Brain (Thalamus) Pool
13.8
|
Fetal Liver
5.5
Brain (whole)
7.2
|
Liver ca. HepG2
8.7
Spinal Cord Pool
7.4
|
Kidney Pool
25.7
Adrenal Gland
6.2
|
Fetal Kidney
14.7
Pituitary gland Pool
1.7
|
Renal ca. 786-0
11.0
Salivary Gland
1.7
|
Renal ca. A498
5.1
Thyroid (female)
1.4
|
Renal ca. ACHN
15.1
Pancreatic ca. CAPAN2
1.3
|
Renal ca. UO-31
3.6
Pancreas Pool
19.2
|
|
[0993]
484
TABLE ND
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3648,
Rel. Exp. (%) Ag3648,
|
Tissue Name
Run 169975756
Tissue Name
Run 169975756
|
|
Secondary Th1 act
12.9
HUVEC IL-1beta
11.3
|
Secondary Th2 act
15.2
HUVEC IFN gamma
23.0
|
Secondary Tr1 act
16.3
HUVEC TNF alpha + IFN
4.9
|
gamma
|
Secondary Th1 rest
1.2
HUVEC TNF alpha + IL4
6.0
|
Secondary Th2 rest
5.8
HUVEC IL-11
2.7
|
Secondary Tr1 rest
1.6
Lung Microvascular EC none
48.6
|
Primary Th1 act
15.2
Lung Microvascular EC
21.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
29.7
Microvascular Dermal EC none
6.1
|
Primary Tr1 act
19.8
Microsvasular Dermal EC
10.7
|
TNF alpha + IL-1beta
|
Primary Th1 rest
12.2
Bronchial epithelium TNF
20.9
|
alpha + IL1beta
|
Primary Th2 rest
15.2
Small airway epithelium none
2.7
|
Primary Tr1 rest
19.8
Small airway epithelium
14.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
39.0
Coronery artery SMC rest
4.8
|
act
|
CD45RO CD4 lymphocyte
55.5
Coronery artery SMC TNF
2.7
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
44.1
Astrocytes rest
19.5
|
Secondary CD8
47.0
Astrocytes TNF alpha + IL-1beta
12.1
|
lymphocyte rest
|
Secondary CD8
10.2
KU-812 (Basophil) rest
29.7
|
lymphocyte act
|
CD4 lymphocyte none
4.2
KU-812 (Basophil)
25.3
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
3.2
CCD1106 (Keratinocytes) none
48.0
|
CD95 CH11
|
LAK cells rest
15.2
CCD1106 (Keratinocytes)
55.1
|
TNF alpha + IL-1beta
|
LAK cells IL-2
22.1
Liver cirrhosis
5.4
|
LAK cells IL-2 + IL-12
18.7
NCI-H292 none
11.0
|
LAK cells IL-2 + IFN
20.4
NCI-H292 IL-4
29.1
|
gamma
|
LAK cells IL-2 + IL-18
37.9
NCI-H292 IL-9
25.5
|
LAK cells
21.6
NCI-H292 IL-13
34.6
|
PMA/ionomycin
|
NK Cells IL-2 rest
14.3
NCI-H292 IFN gamma
36.1
|
Two Way MLR 3 day
35.1
HPAEC none
15.1
|
Two Way MLR 5 day
14.8
HPAEC TNF alpha + IL-1beta
23.7
|
Two Way MLR 7 day
4.4
Lung fibroblast none
19.5
|
PBMC rest
4.2
Lung fibroblast TNF alpha + IL-
4.4
|
1beta
|
PBMC PWM
13.5
Lung fibroblast IL-4
45.4
|
PBMC PHA-L
14.6
Lung fibroblast IL-9
52.5
|
Ramos (B cell) none
54.0
Lung fibroblast IL-13
29.5
|
Ramos (B cell) ionomycin
48.3
Lung fibroblast IFN gamma
24.0
|
B lymphocytes PWM
25.0
Dermal fibroblast CCD1070 rest
14.6
|
B lymphocytes CD40L
13.5
Dermal fibroblast CCD1070
15.4
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
100.0
Dermal fibroblast CCD1070 IL-
9.0
|
1beta
|
EOL-1 dbcAMP
89.5
Dermal fibroblast IFN gamma
20.9
|
PMA/ionomycin
|
Dendritic cells none
16.6
Dermal fibroblast IL-4
42.3
|
Dendritic cells LPS
11.4
Dermal fibroblasts rest
7.4
|
Dendritic cells anti-CD40
5.9
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
1.4
Neutrophils rest
0.0
|
Monocytes LPS
3.1
Colon
5.3
|
Macrophages rest
11.0
Lung
4.5
|
Macrophages LPS
1.4
Thymus
9.2
|
HUVEC none
7.0
Kidney
13.8
|
HUVEC starved
12.2
|
|
[0994] CNS_neurodegeneration_v1.0 Summary: Ag3648 This panel does not show differential expression of the CG99623-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[0995] General_screening_panel_v1.4 Summary: Ag3648 Highest expression of the CG88623-01 gene is seen in a breast cancer cell line (CT=29.3). Moderate levels of expression are also seen in samples derived from colon, gastric, brain, lung, ovarian and melanoma cancer cell lines. Thus, expression of this gene could be used to differentiate the breast cancer sample from the other samples on this panel and as a marker for breast cancer. Furthermore, the consistent levels of expression in the cancer cell lines on this panel suggest that this gene product may be involved in cellular growth and proliferation. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of these cancers.
[0996] Among tissues with metabolic function, this gene is expressed at low but significant levels in adipose, adrenal gland, pancreas, fetal liver and adult and fetal skeletal muscle and heart. This expression pattern suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and type 2 diabetes.
[0997] This gene, a potassium channel homolog, also shows low but significant expression in the brain, including the hippocampus, cortex, amygdala, substantia nigra and thalamus. These regions are succeptable to the neurodegeneration associated with Alzheimer's disease, Parkinson's disease, Huntington's disease and other pathological neurodegenerative conditions. In fact, potassium channels have been implicated in neurodegenerative diseases, including Alzheimer's disease. It has been suggested that modulating these channels to reduce outward K+ current may provide an approach to reducing neuronal degeneration in patients with Alzheimer's disease. Therefore, agents that modulate the function of this gene product could potentially reduce neuronal degeneration in patients with Alzheimer's disease and other neurodegenerative diseases.
[0998] In addition, defective potassium channels are known to cause several CNS disorders, including epilepsy and episodic ataxia with myokymia. Therefore, modulation of the expression or function of this gene product may potentially be useful as a treatment for the symptoms produced by ataxia and epilepsy (Jhamandas J H, Cho C, Jassar B, Harris K, MacTavish D, Easaw J.Cellular Mechanisms for Amyloid beta-Protein Activation of Rat Cholinergic Basal Forebrain Neurons. J Neurophysiol 2001 Sep;86(3):1312-20; Chi X, Sutton E T, Hellermann G, Price J M. Potassium channel openers prevent beta-amyloid toxicity in bovine vascular endothelial cells. Neurosci Lett Aug. 18, 2000;290(1):9-12; Piccini A, Ciotti M T, Vitolo OV, Calissano P, Tabaton M, Galli C. Endogenous APP derivatives oppositely modulate apoptosis through an autocrine loop. Neuroreport May 15, 2000;11(7):1375-9; Yu S P, Farhangrazi Z S, Ying H S, Yeh C H, Choi D W. Enhancement of outward potassium current may participate in beta-amyloid peptide-induced cortical neuronal death. Neurobiol Dis 1998 August;5(2):81-8; Colom L V, Diaz M E, Beers D R, Neely A, Xie W J, Appel S H. Role of potassium channels in amyloid-induced cell death. J Neurochem 1998 May;70(5):1925-34).
[0999] Panel 4.1D Summary: Ag3648 Highest expression of the CG88623-01 gene is seen in untreated eosinophils and eosinophils stimulated with PMA/ionomycin (CTs=31.5). Thus, expression of this gene could be used to differentiate between the eosinophil derived samples and other samples on this panel and as a marker for eosinophils. Furthermore, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of hematopoietic disorders involving eosinphils, parasitic infections and asthma.
[1000] Low but significant levels of expression are also seen in other cells important to the immune respones, including lung and dermal fibroblasts, basophils, the B cell line Ramos, activated and resting primary Th1, Th2 and Tr1 cells, activated CD4 and CD8 lymphocytes, and resting secondary CD8 lymphocytes. Since many of these cells play an important role in lung pathology, inflammatory bowel disease and autoimmune disorders, therapies designed with the protein encoded by this gene may block or inhibit inflammation or tissue damage due to lung conditions including asthma, allergies, hypersensitivity reactions, inflammatory bowel disease, rheumatoid arthritis, viral infections and autoimmune disease.
[1001] O. CG88645-01: CARDIAC POTASSIUM CHANNEL SUBUNIT (KV6.2)-like Protein
[1002] Expression of gene CG88645-01 was assessed using the primer-probe set Ag3650, described in Table OA. Results of the RTQ-PCR runs are shown in Tables OB, OC and OD.
485TABLE OA
|
|
Probe Name Ag3650
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forwards5′-ctggagtcaccttgagggtact-3′22 980267
ProbeTET-5′-261027268
attaagcttgcccgtcacttcattgg-
3′-TAMRA
Reverse5′-agagtcaaaccgagtgtctgaa-3′221055269
|
[1003]
486
TABLE OB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3650, Run
Rel. Exp. (%) Ag3650, Run
|
Tissue Name
211019100
Tissue Name
211019100
|
|
AD 1 Hippo
17.0
Control (Path) 3
0.7
|
Temporal Ctx
|
AD 2 Hippo
15.7
Control (Path) 4
21.8
|
Temporal Ctx
|
AD 3 Hippo
6.7
AD 1 Occipital Ctx
5.1
|
AD 4 Hippo
4.4
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
40.1
AD 3 Occipital Ctx
1.4
|
AD 6 Hippo
100.0
AD 4 Occipital Ctx
9.2
|
Control 2 Hippo
26.8
AD 5 Occipital Ctx
19.3
|
Control 4 Hippo
3.5
AD 6 Occipital Ctx
9.8
|
Control (Path) 3 Hippo
1.7
Control 1 Occipital Ctx
0.3
|
AD 1 Temporal Ctx
6.2
Control 2 Occipital Ctx
11.4
|
AD 2 Temporal Ctx
24.3
Control 3 Occipital Ctx
6.7
|
AD 3 Temporal Ctx
3.1
Control 4 Occipital Ctx
0.0
|
AD 4 Temporal Ctx
11.3
Control (Path) 1
46.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
64.2
Control (Path) 2
3.6
|
Occipital Ctx
|
AD 5 Sup Temporal
53.6
Control (Path) 3
0.1
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
35.1
Control (Path) 4
2.7
|
Occipital Ctx
|
AD 6 Sup Temporal
47.6
Control 1 Parietal Ctx
3.0
|
Ctx
|
Control 1 Temporal Ctx
1.7
Control 2 Parietal Ctx
26.8
|
Control 2 Temporal Ctx
24.7
Control 3 Parietal Ctx
10.4
|
Control 3 Temporal Ctx
12.1
Control (Path) 1
58.6
|
Parietal Ctx
|
Control 3 Temporal Ctx
4.1
Control (Path) 2
13.7
|
Parietal Ctx
|
Control (Path) 1
61.1
Control (Path) 3
0.0
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
23.7
Control (Path) 4
19.3
|
Temporal Ctx
Parietal Ctx
|
|
[1004]
487
TABLE OC
|
|
|
General_screening_panel_V1.4
|
Rel. Exp. (%) Ag3650, Run
Rel. Exp. (%) Ag3650, Run
|
Tissue Name
218952325
Tissue Name
218952325
|
|
Adipose
0.8
Renal ca. TK-10
38.7
|
Melanoma* Hs688(A).T
0.0
Bladder
12.3
|
Melanoma* Hs688(B).T
0.3
Gastric ca. (liver met.)
10.3
|
NCI-N87
|
Melanoma* M14
11.7
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
40.6
Colon ca. SW480
2.3
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
SW620
|
Testis Pool
15.5
Colon ca. HT29
1.1
|
Prostate ca.* (bone met)
0.6
Colon ca. HCT-116
36.6
|
PC-3
|
Prostate Pool
3.9
Colon ca. CaCo-2
17.7
|
Placenta
0.0
Colon cancer tissue
0.5
|
Uterus Pool
0.7
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
16.4
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
100.0
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
16.3
Colon Pool
6.0
|
Ovarian ca. OVCAR-5
33.4
Small Intestine Pool
4.2
|
Ovarian ca. IGROV-1
9.0
Stomach Pool
5.4
|
Ovarian ca. OVCAR-8
5.6
Bone Marrow Pool
0.6
|
Ovary
30.4
Fetal Heart
0.0
|
Breast ca. MCF-7
1.3
Heart Pool
1.2
|
Breast ca. MDA-MB-
0.4
Lymph Node Pool
10.6
|
231
|
Breast ca. BT 549
41.8
Fetal Skeletal Muscle
0.7
|
Breast ca. T47D
46.7
Skeletal Muscle Pool
0.4
|
Breast ca. MDA-N
0.0
Spleen Pool
2.5
|
Breast Pool
5.1
Thymus Pool
23.0
|
Trachea
4.9
CNS cancer (glio/astro)
1.8
|
U87-MG
|
Lung
4.0
CNS cancer (glio/astro) U-
0.0
|
118-MG
|
Fetal Lung
2.8
CNS cancer (neuro; met)
7.9
|
SK-N-AS
|
Lung ca. NCI-N417
30.4
CNS cancer (astro) SF-539
1.7
|
Lung ca. LX-1
0.8
CNS cancer (astro) SNB-75
2.0
|
Lung ca. NCI-H146
56.6
CNS cancer (glio) SNB-19
6.8
|
Lung ca. SHP-77
85.9
CNS cancer (glio) SF-295
11.1
|
Lung ca. A549
16.0
Brain (Amygdala) Pool
24.5
|
Lung ca. NCI-H526
23.5
Brain (cerebellum)
0.4
|
Lung ca. NCI-H23
38.4
Brain (fetal)
10.7
|
Lung ca. NCI-H460
5.6
Brain (Hippocampus) Pool
42.3
|
Lung ca. HOP-62
0.9
Cerebral Cortex Pool
38.7
|
Lung ca. NCI-H522
67.8
Brain (Substantia nigra)
33.2
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
51.8
|
Fetal Liver
0.7
Brain (whole)
41.2
|
Liver ca. HepG2
12.9
Spinal Cord Pool
5.4
|
Kidney Pool
1.9
Adrenal Gland
48.3
|
Fetal Kidney
21.5
Pituitary gland Pool
10.3
|
Renal ca. 786-0
0.0
Salivary Gland
0.0
|
Renal ca. A498
4.0
Thyroid (female)
0.1
|
Renal ca. ACHN
41.8
Pancreatic ca. CAPAN2
5.2
|
Renal ca. UO-31
2.7
Pancreas Pool
21.2
|
|
[1005]
488
TABLE OD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3650,
Rel. Exp. (%) Ag3650,
|
Tissue Name
Run 169975768
Tissue Name
Run 169975768
|
|
Secondary Th1 act
7.1
HUVEC IL-1beta
0.0
|
Secondary Th2 act
13.8
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
10.2
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
1.7
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
1.7
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
16.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
18.9
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
8.8
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
3.1
Bronchial epithelium TNF alpha +
0.0
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
4.5
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
5.2
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
13.1
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
3.2
Astrocytes rest
0.0
|
Secondary CD8
1.3
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.8
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
1.4
CCD1106 (Keratinocytes)
0.6
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
1.6
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
9.5
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
7.1
NCI-H292 IL-9
3.3
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
3.4
|
Two Way MLR 3 day
3.0
HPAEC none
0.0
|
Two Way MLR 5 day
2.8
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
1.6
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
3.3
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
3.4
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
4.2
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
13.2
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.7
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
1.7
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
6.6
|
Macrophages rest
4.6
Lung
3.2
|
Macrophages LPS
0.0
Thymus
100.0
|
HUVEC none
0.0
Kidney
11.8
|
HUVEC starved
0.0
|
|
[1006] CNS_neurodegeneration_v1.0 Summary: Ag3650 This panel confirms the expression of the CG88645-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1007] General_screening13 panel_v1.4 Summary: Ag3650 Highest expression of the CG88645-01 gene is detected in ovarian cancer SK-OV-3 cell line (CT=29.9). In addition high expression of this gene is also seen in cluster of lung, ovarian, colon, renal, and breast cancer cell line. The CG88645-01 gene codes for a potassium channel subunit. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.
[1008] Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, pituitary gland and the gastrointestinal tract. Potassium channel in pancrease have been implicated in regulation of insulin secretion (Ref. 1). Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1009] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression. Furthermore, mutations in K+ channel genes have been implicated in diseases as diverse as persistent hyperinsulinemia of infancy, cardiac long QT syndrome, cerebellar degeneration, epilepsy, and certain ataxias (Dukes I D, Philipson L H. (1996) K+ channels: generating excitement in pancreatic beta-cells. Diabetes 45(7):845-53; Moulard B, Picard F, le Hellard S, Agulhon C, Weiland S, Favre I, Bertrand S, Malafosse A, Bertrand D. (2001) Ion channel variation causes epilepsies. Brain Res Brain Res Rev 36(2-3):275-84). Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of these diseases.
[1010] Panel 4.1D Summary: Ag3650 Highest expression of the CG88645-01 gene is detected in thymus (CT=32). Thus, expression of this gene may be used to identify thymic tissue. In addition, expression of this gene is stimulated in activated primary Th1 and Th2 cells (CTs—34.5-34.8). Therefore, drugs that inhibit the function of this protein may regulate T cell development in the thymus and reduce or eliminate the symptoms of T cell mediated autoimmune or inflammatory diseases, including asthma, allergies, inflammatory bowel disease, lupus erythematosus, or rheumatoid arthritis. Additionally, small molecule or antibody therapeutics designed against this putative protein may disrupt T cell development in the thymus and function as an immunosuppresant for tissue transplant.
[1011] P. CG88738-01: SYNAPTOTAGMIN INTERACTING PROTEIN STIP1
[1012] Expression of gene CG88738-01 was assessed using the primer-probe set Ag3676, described in Table PA.
489TABLE PA
|
|
Probe Name Ag3676
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-tcgatcttcctcgacgtaagta-3′22137270
ProbeTET-5′-30179271
tcctttgaagaattcaaagcatattttgca-3′-
TAMRA
Reverse5′-tcttctccactgagaacaccat-3′22210272
|
[1013] CNS_neurodegeneration_v1.0 Summary: Ag3676 Expression of the CG88738-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1014] General_screening_panel_v1.4 Summary: Ag3676 Expression of the CG88738-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1015] Panel 4.1D Summary: Ag3676 Expression of the CG88738-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1016] Q. CG88902-01 and CG88902-O2: UDP GLUCURONOSYLTRANSFERASE
[1017] Expression of gene CG88902-01 and full length clone CG88902-02 was assessed using the primer-probe set Ag3680, described in Table QA. Results of the RTQ-PCR runs are shown in Tables QB and QC. Please note that CG88902-02 represents a full-length physical clone of the CG88902-01 gene, validating the prediction of the gene sequence.
490TABLE QA
|
|
Probe Name Ag3680
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-tttactgagggctttgagaaca-3′221366273
ProbeTET-5′-301395274
ccgattcctcttataaagagaatgctatga-3′-
TAMRA
Reverse5′-caggttgatcatggtgaattct-3′221433275
|
[1018]
491
TABLE QB
|
|
|
Panel 2.2
|
Rel. Exp. (%) Ag3680,
Rel. Exp. (%) Ag3680,
|
Tissue Name
Run 174441543
Tissue Name
Run 174441543
|
|
Normal Colon
19.2
Kidney Margin (OD04348)
19.5
|
Colon cancer (OD06064)
1.2
Kidney malignant cancer
0.0
|
(OD06204B)
|
Colon Margin (OD06064)
30.4
Kidney normal adjacent
10.9
|
tissue (OD06204E)
|
Colon cancer (OD06159)
0.4
Kidney Cancer (OD04450-
100.0
|
01)
|
Colon Margin (OD06159)
19.3
Kidney Margin (OD04450-
6.3
|
03)
|
Colon cancer (OD06297-04)
1.1
Kidney Cancer 8120613
0.0
|
Colon Margin (OD06297-05)
16.3
Kidney Margin 8120614
7.7
|
CC Gr.2 ascend colon
1.7
Kidney Cancer 9010320
0.9
|
(ODO3921)
|
CC Margin (ODO3921)
7.8
Kidney Margin 9010321
3.2
|
Colon cancer metastasis
2.8
Kidney Cancer 8120607
0.6
|
(OD06104)
|
Lung Margin (OD06104)
37.6
Kidney Margin 8120608
5.1
|
Colon mets to lung
0.5
Normal Uterus
0.0
|
(OD04451-01)
|
Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0
|
Normal Prostate
0.0
Normal Thyroid
0.0
|
Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
0.0
|
Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0
|
Normal Ovary
0.0
Thyroid Margin A302153
0.0
|
Ovarian cancer (OD06283-
0.0
Normal Breast
0.0
|
03)
|
Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
0.0
|
07)
|
Ovarian Cancer 064008
0.3
Breast Cancer 1024
0.0
|
Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0
|
01)
|
Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
0.0
|
(OD04590-03)
|
Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
0.0
|
07)
|
Normal Lung
0.0
Breast Cancer 9100266
0.0
|
Invasive poor diff. lung
0.0
Breast Margin 9100265
0.0
|
adeno (ODO4945-01)
|
Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0
|
Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0
|
(OD03126)
|
Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
0.0
|
Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
0.0
Normal Liver
6.0
|
Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.8
|
Lung Margin (OD06081)
0.0
Liver Cancer 1025
4.8
|
Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
2.6
|
Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.5
|
Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
2.3
|
Ocular Melanoma Margin
3.9
Liver Tissue 6005-N
4.0
|
(Liver)
|
Melanoma Metastasis
0.0
Liver Cancer 064003
1.7
|
Melanoma Margin (Lung)
0.0
Normal Bladder
2.9
|
Normal Kidney
12.0
Bladder Cancer 1023
0.0
|
Kidney Ca, Nuclear grade 2
8.2
Bladder Cancer A302173
0.6
|
(OD04338)
|
Kidney Margin (OD04338)
8.0
Normal Stomach
0.3
|
Kidney Ca Nuclear grade 1/2
54.0
Gastric Cancer 9060397
0.0
|
(OD04339)
|
Kidney Margin (OD04339)
27.9
Stomach Margin 9060396
0.0
|
Kidney Ca, Clear cell type
23.8
Gastric Cancer 9060395
0.0
|
(OD04340)
|
Kidney Margin (OD04340)
9.9
Stomach Margin 9060394
2.5
|
Kidney Ca, Nuclear grade 3
0.5
Gastric Cancer 064005
2.6
|
(OD04348)
|
|
[1019]
492
TABLE QC
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3680,
Rel. Exp. (%) Ag3680,
|
Tissue Name
Run 169988038
Tissue Name
Run 169988038
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.1
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
7.8
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
37.1
|
Macrophages rest
0.0
Lung
0.1
|
Macrophages LPS
0.0
Thymus
0.0
|
HUVEC none
0.0
Kidney
100.0
|
HUVEC starved
0.0
|
|
[1020] CNS_neurodegeneration_v1.0 Summary: Ag3680 Expression of the CG88902-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1021] General_screening_panel_v1.4 Summary: Ag3680 Results from one experiment with the CG88902-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1022] Panel 2.2 Summary: Ag3680 Highest expression of the CG88902-01 gene is detected in kidney cancer (0D04450-01) sample (CT=31.2). Interestingly, expression of this gene is higher in 0D04450-01 cancer sample as compared to the control margin (0D04450-03) sample (CT=35.2). Thus, expression of this gene can be used as diagnostic marker to detection of kidney cancer and also, therapeutic modulation of the activity of the protein encoded by this gene may be beneficial in the treatment of kidney cancer.
[1023] In addition, expression of this gene is downregulated in colon cancer and colon cancer metastasis to lung. Therefore, therapeutic modulation to increase the activity of the protein encoded by this gene may be beneficial in the treatment of colon cancer.
[1024] Panel 4.1D Summary: Ag3680 Highest expression of the CG88902-01 gene is detected in kidney (CT=39.4). Significant expression of this gene is also seen in colon sample (Ct30.8). Thus, expression of this gene can be used to distinguish colon and kidney sample from other samples in this panel. In addition, therapeutic modulation of the activity of the protein encoded by this gene may be useful in the treatment of inflammatory bowel disease and inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis.
[1025] Moderate expression of this gene is also detected in liver cirrhosis sample (Ct33). The presence of this gene in liver cirrhosis (a component of which involves liver inflammation and fibrosis) suggests that antibodies to the protein encoded by this gene could also be used for the diagnosis of liver cirrhosis. Furthermore, therapeutic agents involving this gene may be useful in reducing or inhibiting the inflammation associated with fibrotic and inflammatory diseases.
[1026] R. CG89098-01: F-Box Protein
[1027] Expression of gene CG89098-01 was assessed using the primer-probe set Ag3688, described in Table RA. Results of the RTQ-PCR runs are shown in Tables RB, RC and RD.
493TABLE RA
|
|
Probe Name Ag3688
Start
PrimersSequencesLengthPositionSEQ ID NO:
|
Forward5′-acagcggctgtatgtacagact-3′22607276
ProbeTET-5′-26635277
ccaacttctagacgccaaccagactg-
3′-TAMRA
Reverse5′-tcaggcacagcagagaatttat-3′22667278
|
[1028]
494
TABLE RB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3688, Run
Rel. Exp. (%) Ag3688, Run
|
Tissue Name
211144702
Tissue Name
211144702
|
|
AD 1 Hippo
14.9
Control (Path) 3
8.0
|
Temporal Ctx
|
AD 2 Hippo
38.4
Control (Path) 4
34.2
|
Temporal Ctx
|
AD 3 Hippo
7.5
AD 1 Occipital Ctx
13.3
|
AD 4 Hippo
5.5
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
94.6
AD 3 Occipital Ctx
9.7
|
AD 6 Hippo
33.7
AD 4 Occipital Ctx
12.2
|
Control 2 Hippo
28.3
AD 5 Occipital Ctx
52.1
|
Control 4 Hippo
12.9
AD 6 Occipital Ctx
16.8
|
Control (Path) 3 Hippo
10.7
Control 1 Occipital Ctx
4.4
|
AD 1 Temporal Ctx
16.4
Control 2 Occipital Ctx
65.1
|
AD 2 Temporal Ctx
40.1
Control 3 Occipital Ctx
15.7
|
AD 3 Temporal Ctx
9.8
Control 4 Occipital Ctx
8.1
|
AD 4 Temporal Ctx
19.8
Control (Path) 1
84.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
14.3
|
Occipital Ctx
|
AD 5 Sup Temporal
43.2
Control (Path) 3
4.7
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
47.3
Control (Path) 4
15.8
|
Occipital Ctx
|
AD 6 Sup Temporal
49.7
Control 1 Parietal Ctx
7.8
|
Ctx
|
Control 1 Temporal Ctx
7.6
Control 2 Parietal Ctx
50.7
|
Control 2 Temporal Ctx
49.3
Control 3 Parietal Ctx
17.3
|
Control 3 Temporal Ctx
15.5
Control (Path) 1
90.8
|
Parietal Ctx
|
Control 3 Temporal Ctx
13.5
Control (Path) 2
26.2
|
Parietal Ctx
|
Control (Path) 1
83.5
Control (Path) 3
4.5
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
36.6
Control (Path) 4
43.5
|
Temporal Ctx
Parietal Ctx
|
|
[1029]
495
TABLE RC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3688, Run
Rel. Exp. (%) Ag3688, Run
|
Tissue Name
217130888
Tissue Name
217130888
|
|
Adipose
10.8
Renal ca. TK-10
20.7
|
Melanoma* Hs688(A).T
10.2
Bladder
2.7
|
Melanoma* Hs688(B).T
8.4
Gastric ca. (liver met.)
1.3
|
NCI-N87
|
Melanoma* M14
13.0
Gastric ca. KATO III
0.2
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
6.8
Colon ca. SW480
100.0
|
Squamous cell
49.7
Colon ca.* (SW480 met)
25.2
|
carcinoma SCC-4
SW620
|
Testis Pool
6.1
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
45.4
Colon ca. HCT-116
43.8
|
PC-3
|
Prostate Pool
6.2
Colon ca. CaCo-2
10.6
|
Placenta
1.3
Colon cancer tissue
1.4
|
Uterus Pool
1.9
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
19.1
Colon ca. Colo-205
0.1
|
Ovarian ca. SK-OV-3
6.3
Colon ca. SW-48
0.1
|
Ovarian ca. OVCAR-4
45.1
Colon Pool
9.4
|
Ovarian ca. OVCAR-5
23.7
Small Intestine Pool
5.8
|
Ovarian ca. IGROV-1
14.0
Stomach Pool
3.8
|
Ovarian ca. OVCAR-8
12.7
Bone Marrow Pool
5.6
|
Ovary
10.9
Fetal Heart
7.7
|
Breast ca. MCF-7
19.6
Heart Pool
4.2
|
Breast ca. MDA-MB-
57.0
Lymph Node Pool
10.5
|
231
|
Breast ca. BT 549
3.6
Fetal Skeletal Muscle
2.4
|
Breast ca. T47D
50.3
Skeletal Muscle Pool
0.5
|
Breast ca. MDA-N
0.0
Spleen Pool
0.8
|
Breast Pool
9.4
Thymus Pool
4.1
|
Trachea
8.7
CNS cancer (glio/astro)
50.7
|
U87-MG
|
Lung
4.1
CNS cancer (glio/astro) U-
16.3
|
118-MG
|
Fetal Lung
7.3
CNS cancer (neuro; met)
0.3
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.9
|
Lung ca. LX-1
35.4
CNS cancer (astro) SNB-75
7.5
|
Lung ca. NCI-H146
20.7
CNS cancer (glio) SNB-19
12.3
|
Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
5.8
|
Lung ca. A549
12.1
Brain (Amygdala) Pool
14.5
|
Lung ca. NCI-H526
5.3
Brain (cerebellum)
90.8
|
Lung ca. NCI-H23
65.1
Brain (fetal)
24.3
|
Lung ca. NCI-H460
0.1
Brain (Hippocampus) Pool
16.5
|
Lung ca. HOP-62
10.0
Cerebral Cortex Pool
27.5
|
Lung ca. NCI-H522
18.2
Brain (Substantia nigra)
22.4
|
Pool
|
Liver
1.8
Brain (Thalamus) Pool
21.6
|
Fetal Liver
11.8
Brain (whole)
27.9
|
Liver ca. HepG2
1.3
Spinal Cord Pool
10.3
|
Kidney Pool
9.2
Adrenal Gland
3.6
|
Fetal Kidney
7.1
Pituitary gland Pool
2.8
|
Renal ca. 786-0
17.7
Salivary Gland
6.4
|
Renal ca. A498
11.7
Thyroid (female)
2.1
|
Renal ca. ACHN
35.1
Pancreatic ca. CAPAN2
2.9
|
Renal ca. UO-31
33.7
Pancreas Pool
7.3
|
|
[1030]
496
TABLE RD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3688,
Rel. Exp. (%) Ag3688,
|
Tissue Name
Run 169988046
Tissue Name
Run 169988046
|
|
Secondary Th1 act
1.6
HUVEC IL-1beta
12.3
|
Secondary Th2 act
4.6
HUVEC IFN gamma
16.4
|
Secondary Tr1 act
3.1
HUVEC TNF alpha + IFN
5.4
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
3.9
|
Secondary Th2 rest
1.6
HUVEC IL-11
11.9
|
Secondary Tr1 rest
0.5
Lung Microvascular EC none
16.7
|
Primary Th1 act
1.4
Lung Microvascular EC
4.7
|
TNF alpha + IL-1beta
|
Primary Th2 act
5.6
Microvascular Dermal EC none
9.6
|
Primary Tr1 act
1.3
Microsvasular Dermal EC
2.8
|
TNF alpha + IL-1beta
|
Primary Th1 rest
2.6
Bronchial epithelium TNF alpha +
34.9
|
IL1beta
|
Primary Th2 rest
1.5
Small airway epithelium none
11.0
|
Primary Tr1 rest
0.4
Small airway epithelium
19.1
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
6.4
Coronery artery SMC rest
7.9
|
act
|
CD45RO CD4 lymphocyte
10.8
Coronery artery SMC TNF alpha +
6.3
|
act
IL-1beta
|
CD8 lymphocyte act
11.7
Astrocytes rest
6.7
|
Secondary CD8
1.6
Astrocytes TNF alpha + IL-1beta
9.9
|
lymphocyte rest
|
Secondary CD8
2.8
KU-812 (Basophil) rest
2.5
|
lymphocyte act
|
CD4 lymphocyte none
1.2
KU-812 (Basophil)
4.6
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.4
CCD1106 (Keratinocytes) none
58.6
|
CD95 CH11
|
LAK cells rest
0.3
CCD1106 (Keratinocytes)
45.1
|
TNF alpha + IL-1beta
|
LAK cells IL-2
2.7
Liver cirrhosis
6.7
|
LAK cells IL-2 + IL-12
1.2
NCI-H292 none
72.2
|
LAK cells IL-2 + IFN
2.0
NCI-H292 IL-4
85.9
|
gamma
|
LAK cells IL-2 + IL-18
0.7
NCI-H292 IL-9
100.0
|
LAK cells
0.4
NCI-H292 IL-13
87.1
|
PMA/ionomycin
|
NK Cells IL-2 rest
1.4
NCI-H292 IFN gamma
98.6
|
Two Way MLR 3 day
0.0
HPAEC none
10.4
|
Two Way MLR 5 day
6.2
HPAEC TNF alpha + IL-1beta
8.9
|
Two Way MLR 7 day
19.2
Lung fibroblast none
14.8
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
6.9
|
1beta
|
PBMC PWM
1.5
Lung fibroblast IL-4
7.7
|
PBMC PHA-L
6.2
Lung fibroblast IL-9
18.2
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
10.5
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
12.3
|
B lymphocytes PWM
2.7
Dermal fibroblast CCD1070 rest
15.8
|
B lymphocytes CD40L
4.0
Dermal fibroblast CCD1070
6.3
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
10.2
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
5.4
|
PMA/ionomycin
|
Dendritic cells none
2.3
Dermal fibroblast IL-4
13.3
|
Dendritic cells LPS
1.4
Dermal Fibroblasts rest
21.2
|
Dendritic cells anti-CD40
0.4
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
14.1
|
Macrophages rest
0.6
Lung
7.8
|
Macrophages LPS
0.4
Thymus
7.2
|
HUVEC none
11.7
Kidney
32.3
|
HUVEC starved
9.1
|
|
[1031] CNS_neurodegeneration_v1.0 Summary: Ag3688 This panel does not show differential expression of the CG89098-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1032] General_screening_panel_v1.4 Summary: Ag3688 Highest expression of the CG89098-01 gene, which encodes a putative F box protein, is seen in a colon cancer cell line (CT=27). This gene also shows high to moderate levels of expression in many of the cancer cell lines on this panel, including samples derived from brain, renal, lung, breast, ovarian and prostate cancer cell lines. Thus, expression of this gene could be used as a marker for colon cancer. It has been suggested that F-box proteins are involved in the regulation of many processes, including cell division. Therefore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of these cancers.
[1033] Among tissues with metabolic function, this gene is expressed at moderate to low levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1034] This gene is also expressed at high to moderate levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1035] Overall, this gene shows widespread expression in the samples on this panel, perhaps reflecting a broader role for this gene product in cellular processes, as has been suggested for F box proteins.
[1036] Panel 4.1D Summary: Ag3688 Expression of the CG89098-01 gene is highest in a cluster of treated and untreated samples from the NCI-H292 mucoepidermoid cell line (CTs=30). Significant levels of expression are also seen in treated and untreated keratinocytes. Low but significant levels of expression are also seen in a variety of nonhematopoietic cell samples including lung and dermal fibroblasts, small airway and bronchial epithelium, lung and dermal microvasculature, human pulmonary aortic endothelial cells, HUVECs, and smooth muscle cells. Given this distribution of expression in cells involved in inflammatory processes of the lung and skin, therapeutic modulation of the expression or function of this gene product may reduce or eliminate symptoms in patients suffering from asthma, allergy, emphysema and psoriasis.
[1037] S. CG89126-01: cytochrome P450
[1038] Expression of gene CG89126-01 was assessed using the primer-probe set Ag3689, described in Table SA. Results of the RTQ-PCR runs are shown in Table SB.
497TABLE SA
|
|
Probe Name Ag3689
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-cgccttccatttcaacatc-3′19477279
ProbeTET-5′-30502280
tcctatataacgatcttcaacaagagtgca-3′-
TAMRA
Reverse5′-ccacttgtcaagcatgatgtt-3′21532281
|
[1039]
498
TABLE SB
|
|
|
Panel 2.2
|
Rel. Exp. (%) Ag3689,
Rel. Exp. (%) Ag3689,
|
Tissue Name
Run 174441628
Tissue Name
Run 174441628
|
|
Normal Colon
0.1
Kidney Margin (OD04348)
0.0
|
Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0
|
(OD06204B)
|
Colon Margin (OD06064)
30.4
Kidney normal adjacent
0.0
|
tissue (OD06204E)
|
Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0
|
01)
|
Colon Margin (OD06159)
0.9
Kidney Margin (OD04450-
0.0
|
03)
|
Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0
|
Colon Margin (OD06297-05)
0.0
Kidney Margin 8120614
0.0
|
CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0
|
(ODO3921)
|
CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0
|
Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0
|
(OD06104)
|
Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0
|
Colon mets to lung
0.0
Normal Uterus
0.0
|
(OD04451-01)
|
Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0
|
Normal Prostate
0.0
Normal Thyroid
0.0
|
Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
0.0
|
Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0
|
Normal Ovary
0.0
Thyroid Margin A302153
0.0
|
Ovarian cancer (OD06283-
0.0
Normal Breast
0.0
|
03)
|
Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
0.0
|
07)
|
Ovarian Cancer 064008
0.0
Breast Cancer 1024
0.0
|
Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0
|
01)
|
Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
0.0
|
(OD04590-03)
|
Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
1.4
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
0.0
|
07)
|
Normal Lung
0.0
Breast Cancer 9100266
0.0
|
Invasive poor diff. lung
0.0
Breast Margin 9100265
0.0
|
adeno (ODO4945-01
|
Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0
|
Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0
|
(OD03126)
|
Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
0.0
|
Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
0.0
Normal Liver
22.2
|
Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0
|
Lung Margin (OD06081)
0.0
Liver Cancer 1025
80.7
|
Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
88.3
|
Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0
|
Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0
|
Ocular Melanoma Margin
0.7
Liver Tissue 6005-N
100.0
|
(Liver)
|
Melanoma Metastasis
0.0
Liver Cancer 064003
0.0
|
Melanoma Margin (Lung)
0.0
Normal Bladder
0.0
|
Normal Kidney
0.0
Bladder Cancer 1023
0.0
|
Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer A302173
0.0
|
(OD04338)
|
Kidney Margin (OD04338)
0.0
Normal Stomach
0.1
|
Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0
|
(OD04339)
|
Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
2.0
|
Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0
|
(OD04340)
|
Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
3.2
|
Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0
|
(OD04348)
|
|
[1040] CNS_neurodegeneration v1.0 Summary: Ag3689 Expression of the CG89126-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1041] General_screening_panel_v1.4 Summary: Ag3689 Expression of the CG89126-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1042] Panel 2.2 Summary: Ag3689 Expression of the CG89126-01 gene is essentially restricted to liver derived tissue, with highest expression in normal liver tissue (CT=29.8). Moderate levels of expression of this gene are also seen in samples derived from liver cancer. Thus, expression of this gene could be used as a marker of liver tissue and to differentiate between liver derived samples and other samples on this panel.
[1043] Panel 4.1D Summary: Ag3689 Expression of the CG89126-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1044] T. CG89677-01: arylsulfatase
[1045] Expression of gene CG89677-01 was assessed using the primer-probe set Ag3695, described in Table TA. Results of the RTQ-PCR runs are shown in Tables TB, and TC.
499TABLE TA
|
|
Probe Name Ag3695
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-gtacgctacccagctgagaac-3′211474282
ProbeTET-5′-ccccgggctcatcctgactttaat-3′-241495283
TAMRA
Reverse5′-cttcctcctcttcctcatcact-3′221543284
|
[1046]
500
TABLE TB
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3695, Run
Rel. Exp. (%) Ag3695, Run
|
Tissue Name
213406518
Tissue Name
213406518
|
|
Adipose
0.6
Renal ca. TK-10
17.9
|
Melanoma* Hs688(A).T
9.3
Bladder
3.5
|
Melanoma* Hs688(B).T
7.7
Gastric ca. (liver met.)
0.0
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.0
|
Squamous cell
4.0
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
SW620
|
Testis Pool
0.0
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
2.1
Colon ca. HCT-116
0.4
|
PC-3
|
Prostate Pool
1.8
Colon ca. CaCo-2
0.6
|
Placenta
21.9
Colon cancer tissue
13.2
|
Uterus Pool
1.8
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
3.4
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
4.5
Colon ca. SW-48
0.3
|
Ovarian ca. OVCAR-4
2.5
Colon Pool
8.0
|
Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
0.4
|
Ovarian ca. IGROV-1
0.0
Stomach Pool
2.2
|
Ovarian ca. OVCAR-8
16.5
Bone Marrow Pool
1.3
|
Ovary
0.5
Fetal Heart
0.4
|
Breast ca. MCF-7
0.0
Heart Pool
2.6
|
Breast ca. MDA-MB-
30.8
Lymph Node Pool
6.0
|
231
|
Breast ca. BT 549
9.0
Fetal Skeletal Muscle
2.3
|
Breast ca. T47D
0.9
Skeletal Muscle Pool
0.6
|
Breast ca. MDA-N
0.0
Spleen Pool
2.4
|
Breast Pool
5.2
Thymus Pool
4.3
|
Trachea
1.9
CNS cancer (glio/astro)
100.0
|
U87-MG
|
Lung
0.0
CNS cancer (glio/astro) U-
4.9
|
118-MG
|
Fetal Lung
26.6
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
24.8
|
Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
74.7
|
Lung ca. NCI-H146
1.3
CNS cancer (glio) SNB-19
0.0
|
Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
0.8
|
Lung ca. A549
9.5
Brain (Amygdala) Pool
2.6
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0
|
Lung ca. NCI-H23
9.9
Brain (fetal)
0.0
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
1.7
|
Lung ca. HOP-62
87.1
Cerebral Cortex Pool
2.0
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
4.5
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
2.2
|
Fetal Liver
0.0
Brain (whole)
0.5
|
Liver ca. HepG2
22.2
Spinal Cord Pool
4.2
|
Kidney Pool
9.0
Adrenal Gland
0.0
|
Fetal Kidney
2.8
Pituitary gland Pool
0.0
|
Renal ca. 786-0
0.0
Salivary Gland
0.0
|
Renal ca. A498
1.6
Thyroid (female)
0.0
|
Renal ca. ACHN
42.9
Pancreatic ca. CAPAN2
3.9
|
Renal ca. UO-31
9.3
Pancreas Pool
4.6
|
|
[1047]
501
TABLE TC
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3695,
Rel. Exp. (%) Ag3695,
|
Tissue Name
Run 169987358
Tissue Name
Run 169987358
|
|
Secondary Th1 act
8.5
HUVEC IL-1beta
1.5
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvascular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
32.5
|
1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
2.8
|
Primary Tr1 rest
0.0
Small airway epithelium
1.9
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
19.6
Coronery artery SMC rest
2.6
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
12.8
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
15.3
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
12.6
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
100.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
69.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
19.9
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
24.8
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
15.2
|
LAK cells
0.0
NCI-H292 IL-13
26.8
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
24.7
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
5.4
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
4.4
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
6.8
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
12.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
1.6
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
35.8
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
10.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
40.1
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
2.6
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
1.7
|
Dendritic cells LPS
0.0
Dermal Fibroblast rest
2.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
3.0
|
Monocytes LPS
0.0
Colon
1.3
|
Macrophages rest
0.0
Lung
23.5
|
Macrophages LPS
0.0
Thymus
1.4
|
HUVEC none
0.0
Kidney
6.3
|
HUVEC starved
0.0
|
|
[1048] CNS_neurodegeneration_v1.0 Summary: Ag3695 Expression of the CG89677-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1049] General_screening13 panel_v1.4 Summary: Ag3695 Expression of teh CG89677-01 gene is limited to a few samples in this panel, with highest expression in a brain cancer cell line (CT=31.6). Moderate levels of expression are also seen in a lung cancer cell line. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel, and as a marker for these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of lung or brain cancer.
[1050] Panel 2.2 Summary: Ag3695 Expression of the CG89677-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1051] Panel 4.1D Summary: Ag3695 Expression of the CG89677-01 gene is restricted to untreated keratinocytes and keratinocytes treated with the inflammatory cytokines TNF-a and IL-1b (CTs=33-34). Therefore, modulation of the expression or activity of the protein encoded by this transcript through the application of small molecule therapeutics may be useful in the treatment of psoriasis and wound healing.
[1052] U. CG89697-01 and CG89697-O2: RIS
[1053] Expression of gene CG89697-01 and full length clone CG89697-02 was assessed using the primer-probe sets Ag4699 and Ag4523, described in Tables UA and UB. Results of the RTQ-PCR runs are shown in Tables UC, UD and UE. Please note that CG89697-02 represents a full-length physical clone of the CG89697-01 gene, validating the prediction of the gene sequence.
502TABLE UA
|
|
Probe Name Ag4699
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ctggtcaccgtgaagtcatc-3′20766285
ProbeTET-5′-cctaccctgactctcctgaagggctt-26811286
3′-TAMRA
Reverse5′-ggggcctcagaagatcttg-3′19837287
|
[1054]
503
TABLE UB
|
|
|
Probe Name Ag4523
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-ggctcagtacaggcaagtca-3′
20
462
288
|
Probe
TET-5′-aggcaggtttgggtgcctgtttt-3′-
23
507
289
|
TAMRA
|
Reverse
5′-caaagtccagacaggcagag-3′
20
537
290
|
|
[1055]
504
TABLE UC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel.Exp. (%)
Rel.Exp. (%)
Rel.Exp. (%)
Rel.Exp. (%)
|
Avg4699, Run
Ag4699, Run
Ag4699, Run
Ag4699, Run
|
Tissue Name
224710805
230510316
Tissue Name
224710805
230510316
|
|
AD 1 Hippo
14.0
4.6
Control (Path)
1.0
3.2
|
3 Temporal
1.0
|
Ctx
|
AD 2 Hippo
21.9
19.1
Control (Path)
3.4
3.7
|
4 Temporal
|
Ctx
|
AD 3 Hippo
4.9
6.1
AD 1 Occipital
3.4
2.1
|
Ctx
|
AD 4 Hippo
9.6
9.8
AD 2 Occipital
0.0
0.0
|
Ctx (Missing)
|
AD 5 hippo
11.5
12.1
AD 3 Occipital
2.0
2.9
|
Ctx
|
AD 6 Hippo
100.0
100.0
AD 4 Occipital
8.2
6.4
|
Ctx
|
Control 2 Hippo
10.0
6.6
AD 5 Occipital
3.5
0.0
|
Ctx
|
Control 4 Hippo
23.7
17.8
AD 6 Occipital
3.4
3.8
|
Ctx
|
Control (Path) 3
3.9
1.4
Control 1
3.1
2.0
|
Hippo
Occipital Ctx
|
AD 1 Temporal
7.4
5.6
Control 2
6.3
3.9
|
Ctx
Occipital CTx
|
AD 2 Temporal
7.3
9.9
Control 3
2.5
2.8
|
Ctx
Occipital Ctx
|
AD 3 Temporal
4.9
3.4
Control 4
6.6
5.3
|
Ctx
Occipital Ctx
|
AD 4 Temporal
7.9
3.0
Control (Path)
12.2
10.7
|
Ctx
1 Occipital Ctx
|
AD 5 Inf
23.3
27.52
Control (Path)
2.8
0.0
|
Temporal Ctx
2 Occipital Ctx
|
AD 5
40.6
44.4
Control (Path)
0.7
1.4
|
SupTemporal Ctx
3 Occipital Ctx
|
AD 6 Inf
36.9
53.2
Control (Path)
4.1
5.1
|
Temporal Ctx
4 Occipital Ctx
|
AD 6 Sup
23.2
31.0
Control 1
4.0
4.9
|
Temporal Ctx
Parietal Ctx
|
Control 1
2.3
3.1
Control 2
16.4
19.8
|
Temporal Ctx
Parietal Ctx
|
Control 2
4.3
2.0
Control 3
3.4
0.5
|
Temporal Ctx
Parietal Ctx
|
Control 3
1.6
2.5
Control (Path)
4.3
4.8
|
Temporal Ctx
1 Parietal Ctx
|
Control 4
5.3
6.2
Control (Path)
2.3
4.4
|
Temporal Ctx
2 Parietal Ctx
|
Control (Path) 1
8.0
4.5
Control (Path)
1.3
1.4
|
Temporal Ctx
3 Parietal Ctx
|
Control (Path) 2
1.7
3.1
Control (Path)
6.4
3.3
|
Temporal Ctx
4 Parietal Ctx
|
|
[1056]
505
TABLE UD
|
|
|
General_screening_panel_v1.4
|
Rel.Exp.(%)
Rel.Exp.(%)
Rel.Exp.(%)
Rel.Exp.(%)
|
Ag4523, Run
Ag4699, Run
Ag4523, Run
Ag4699, Run
|
Tissue Name
222714393
222812001
Tissue Name
222714393
222812001
|
|
Adipose
6.9
3.8
Renal ca. TK-10
0.0
0.1
|
Melanoma*
0.0
0.0
Bladder
7.1
6.5
|
Hs688(A).T
|
Melanoma*
0.0
0.1
Gastric Ca. (liver
0.4
0.0
|
Hs688(B).T
met.) NCI-N87
|
Melanoma*
0.0
0.0
Gastric ca. KATO
0.0
0.2
|
M14
III
|
Melanoma*
0.4
0.0
Colon ca. SW-948
0.0
0.0
|
LOXIMVI
|
Melanoma* SK-
0.2
0.0
Colon ca. SW480
0.0
0.0
|
MEL-5
|
Squamous cell
0.3
0.0
Colon ca.* (SW480
0.0
0.0
|
carcinoma SCC-
met) SW620
|
4
|
Testis Pool
17.7
17.7
Colon ca. HT29
0.0
0.1
|
Prostate ca.*
0.0
0.0
Colon ca. HCT-116
0.0
0.0
|
(bone met) PC-3
|
Prostate Pool
16.3
14.7
Colon ca. CaCo-2
10.2
8.6
|
Placenta
11.9
9.9
Colon cancer tissue
4.9
5.1
|
Uterus Pool
7.5
16.7
Colon ca. SW1116
0.0
0.2
|
Ovarian ca.
4.5
3.6
Colon ca. Colo-205
0.0
0.0
|
OVCAR-3
|
Ovarian ca. SK-
12.7
13.3
Colon ca. SW-48
1.7
2.1
|
OV-3
|
Ovarian ca.
1.6
1.1
Colon Pool
44.1
45.1
|
OVCAR-4
|
Ovarian ca.
0.4
0.4
Small Intestine Pool
18.9
18.3
|
OVCAR-5
|
Ovarian ca.
0.0
0.1
Stomach Pool
12.8
15.9
|
IGROV-1
|
Ovarian ca.
0.0
0.0
Bone Marrow Pool
12.6
12.7
|
OVCAR-8
|
Ovary
6.4
7.4
Fetal Heart
23.2
18.4
|
Breast ca. MCF-
0.0
0.0
Heart Pool
11.8
10.7
|
7
|
Breast ca. MDA-
0.0
0.0
Lymph Node Pool
52.5
58.2
|
MB-231
|
Breast ca. BT
0.4
0.5
Fetal Skeletal
5.3
7.3
|
549
Muscle
|
Breast ca. T47D
0.9
0.4
Skeletal Muscle
9.3
9.1
|
Pool
|
Breast ca. MDA-
0.0
0.0
Spleen Pool
3.5
5.0
|
N
|
Breast Pool
42.0
40.6
Thymus Pool
12.2
11.1
|
Trachea
18.8
18.6
CNS cancer
0.0
0.0
|
(glio/astro) U87-
|
MG
|
Lung
3.2
4.3
CNS cancer
0.3
0.2
|
(glio/astro) U-118-
|
MG
|
Fetal Lung
100.0
100.0
CNS cancer
0.2
0.1
|
(neuro;met) SK-N-
|
AS
|
Lung ca. NCI-
0.0
0.0
CNS cancer (astro)
|
N417
SF-539
|
Lung ca. LX-1
0.4
0.5
CNS cancer (astro)
0.5
0.3
|
SNB-75
|
Lung ca. NCI-
0.7
0.5
CNS cancer (glio)
0.0
0.3
|
H146
SNB-19
|
Lung ca. SHP-
0.0
0.1
CNS cancer (glio)
|
77
SF-295
|
Lung ca. A549
0.4
1.0
Brain (Amygdala)
6.0
9.7
|
Pool
|
Lung ca. NCI-
0.0
0.0
Brian (cerebellum)
9.8
11.4
|
H526
|
Lung ca. NCI-
5.4
3.3
Brain (fetal)
3.7
2.4
|
H23
|
Lung ca. NCI-
0.0
0.0
Brain)
13.3
13.7
|
H460
(Hippocampus)
|
Pool
|
Lung ca. HOP-
0.7
0.2
Cerebral Cortex
7.0
4.7
|
62
Pool
|
Lung ca. NCI-
0.7
0.0
Brain (Substantia
9.1
7.9
|
H522
nigra) Pool
|
Liver
0.1
0.1
Brain (Thalamus)
11.4
12.7
|
Pool
|
Fetal Liver
1.7
1.3
Brain (whole)
6.7
6.1
|
Liver ca. HepG2
0.0
0.1
Spinal Cord Pool
18.8
16.5
|
Kidney Pool
50.3
64.2
Adrenal Gland
4.4
2.2
|
Fetal Kidney
3.3
3.5
Pituitary gland Pool
0.5
0.7
|
Renal ca. 786-0
0.0
0.2
Salivary Gland
3.4
3.0
|
Renal ca. A498
0.0
0.0
Thyroid (female)
2.5
2.5
|
Renal ca. ACHN
1.2
0.0
Pancreatic ca.
0.5
0.0
|
CAPAN2
|
Renal ca. UO-31
0.2
0.0
Pancreas Pool
26.6
31.6
|
|
[1057]
506
TABLE UE
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag4699,
Rel. Exp. (%) Ag4699,
|
Tissue Name
Run 200924185
Tissue Name
Run 200924185
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
2.6
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
1.7
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
1.4
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
15.5
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
70.7
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
39.2
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
26.4
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
9.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
27.7
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
34.4
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
3.7
|
B lymphocytes CD40L
2.9
Dermal fibroblast CCD1070
5.1
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
1.2
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
11.7
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
8.4
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
5.1
|
Dendritic cells anti-CD40
1.1
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
5.2
|
Macrophages rest
3.1
Lung
100.0
|
Macrophages LPS
1.9
Thymus
7.4
|
HUVEC none
0.0
Kidney
39.8
|
HUVEC starved
0.0
|
|
[1058] CNS_neurodegeneration_v1.0 Summary: Ag4699 Two experiments with the same probe and primer set produce results that are in excellent agreement, with highest expression of the CG89697-01 gene in the hippocampus of an Alzheimer's patient (CTs=29-30). In both cases, the expression of this gene is up-regulated in the temporal cortex of Alzheimer's disease patients when compared to non-demented controls. This difference is apparent when data are analyzed via ANCOVA, using overall RNA quality and/or quantity as a covariate. The temporal cortex is a region that shows degeneration at the mid-stages of this disease. Thus, it is likely that the phenomenon of neurodegeneration is captured in this region, as opposed to the hippocampus and entorhinal cortex where a large number of neurons are already lost by the time of death in AD. Furthermore, in the occipital cortex (where neurodegeneration does not occur in Alzheimer's) this gene is not found to be up-regulated in the same patients. Taken together, these data suggest that this gene is at least a marker of Alzheimer's-like neurodegeneration, and is probably involved in the process of neurodegeneration.
[1059] General_screening13 panel_v1.4 Summary: Ag4253/Ag4699 Two experiments with the same probe and primer set produce results that are in excellent agreement, with highest expression of the CG89697-01 gene in the fetal lung (CTs=28). In addition, this gene is expressed at much higher levels in fetal when compared to expression in the adult counterpart (CTs=32-33). Thus, expression of this gene may be used to differentiate between the fetal and adult source of this tissue.
[1060] Among tissues with metabolic function, this gene is expressed at moderate to low levels in adipose, adrenal gland, pancreas, thyroid, fetal liver and adult and fetal skeletal muscle and heart. This expression suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1061] This gene is also expressed at low levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1062] Panel 4.1D Summary: Ag4699 Highest expression of the CG89697-01 gene is seen in the lung (CT=31.2), in agreement with expression in Panel 1.4. In addition, this gene shows a very restricted pattern of expression in lung fibroblasts, with higher expression in untreated lung fibroblasts. This lung prominent expression suggests that expression of this gene could be used to identify lung derived tissue. In addition, this gene product may be involved in the normal homeostasis of this organ. Therefore, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of pathological and inflammatory lung conditions such as asthma, emphysema, and allergy.
[1063] V. CG90001-01: PEPTIDYLPROLYL ISOMERASE A
[1064] Expression of gene CG9000l-01 was assessed using the primer-probe set Ag3699, described in Table VA. Results of the RTQ-PCR runs are shown in Table VB.
507TABLE VA
|
|
Probe Name Ag3699
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-caacacaaatgattcccagttt-3′22317291
ProbeTET-5′-ttcatctgcactgccaagactgagtg-26339292
3′-TAMRA
Reverse5′-atctttcaccttgccaaagac-3′21384293
|
[1065]
508
TABLE VB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3699, Run
Rel. Exp. (%) Ag3699, Run
|
Tissue Name
211291161
Tissue Name
211291161
|
|
AD 1 Hippo
8.0
Control (Path) 3
3.5
|
Temporal Ctx
|
AD 2 Hippo
34.4
Control (Path) 4
40.1
|
Temporal Ctx
|
AD 3 Hippo
3.7
AD 1 Occipital Ctx
21.6
|
AD 4 Hippo
8.9
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
60.3
AD 3 Occipital Ctx
4.5
|
AD 6 Hippo
58.2
AD 4 Occipital Ctx
25.7
|
Control 2 Hippo
19.2
AD 5 Occipital Ctx
20.2
|
Control 4 Hippo
5.2
AD 6 Occipital Ctx
17.9
|
Control (Path) 3 Hippo
3.8
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
10.0
Control 2 Occipital Ctx
18.7
|
AD 2 Temporal Ctx
22.2
Control 3 Occipital Ctx
17.7
|
AD 3 Temporal Ctx
5.4
Control 4 Occipital Ctx
4.8
|
AD 4 Temporal Ctx
30.1
Control (Path) 1
100.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
79.6
Control (Path) 2
17.9
|
Occipital Ctx
|
AD 5 Sup Temporal
27.4
Control (Path) 3
2.6
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
31.0
Control (Path) 4
16.5
|
Occipital Ctx
|
AD 6 Sup Temporal
71.7
Control 1 Parietal Ctx
2.4
|
Ctx
|
Control 1 Temporal Ctx
15.5
Control 2 Parietal Ctx
39.5
|
Control 2 Temporal Ctx
17.8
Control 3 Parietal Ctx
18.4
|
Control 3 Temporal Ctx
19.9
Control (Path) 1
69.3
|
Parietal Ctx
|
Control 3 Temporal Ctx
10.7
Control (Path) 2
29.3
|
Parietal Ctx
|
Control (Path) 1
47.6
Control (Path) 3
3.3
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
42.3
Control (Path) 4
42.9
|
Temporal Ctx
Parietal Ctx
|
|
[1066] CNS_neurodegeneration_vl.0 Summary: Ag3699 Expression of the CG90001-01 gene is low seen at low levels in the brain in this panel, with highest expression in occipital cortex of a control patient (CT=33.7). Low but significant expression of this gene is also seen in the hippocampus. Thus, this gene product may be involved in the function of the CNS.
[1067] General_screening_panel_v1.4 Summary: Ag3699 Expression of the CG90001-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.) Panel 4.1D Summary: Ag3699 Expression of the CG90001-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1068] W. CG90011-01: ATP SPECIFIC SUCCINYL COA SYNTHETASE BETA SUBUNIT PRECURSOR
[1069] Expression of gene CG90011-01 was assessed using the primer-probe set Ag3700, described in Table WA.
509TABLE WA
|
|
Probe Name Ag3700
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-tggtgctacagtccatcaagt-3′211034294
ProbeTET-5′-281057295
cagaagcatttaagcctatcacttcaga-
3′-TAMRA
Reverse5′-ctccacaaatgttgaccagaat-3′221101296
|
[1070] CNS_neurodegeneration_v1.0 Summary: Ag3700 Expression of the CG90011-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1071] General_screening_panel_v1.4 Summary: Ag3700 Expression of the CG90011-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1072] Panel 4.1D Summary: Ag3700 Expression of the CG90011-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1073] Panel 5 Islet Summary: Ag3700 Expression of the CG90Ol 1-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1074] X. CG90204-01: Semaphorin Cytoplasmic Domain-Associated Protein
[1075] Expression of gene CG90204-01 was assessed using the primer-probe set Ag3706, described in Table XA. Results of the RTQ-PCR runs are shown in Tables XB, XC and XD.
510TABLE XA
|
|
Probe Name Ag3706
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-caagaagaagaagaaggcacaa-3′221599297
ProbeTET-5′-261627298
ctgcaacatcctcatccaacaaccat-
3′-TAMRA
Reverse5′-atttcgcaagctttcatctgt-3′211677299
|
[1076]
511
TABLE XB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3706, Run
Ag3706, Run
|
Tissue Name
211291164
Tissue Name
211291164
|
|
AD 1 Hippo
11.0
Control (Path) 3
7.2
|
Temporal Ctx
|
AD 2 Hippo
17.4
Control (Path) 4
59.9
|
Temporal Ctx
|
AD 3 Hippo
5.3
AD 1 Occipital Ctx
37.9
|
AD 4 Hippo
7.8
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
64.6
AD 3 Occipital Ctx
9.2
|
AD 6 Hippo
21.5
AD 4 Occipital Ctx
22.8
|
Control 2 Hippo
25.3
AD 5 Occipital Ctx
25.3
|
Control 4 Hippo
9.5
AD 6 Occipital Ctx
48.0
|
Control (Path) 3 Hippo
11.0
Control 1 Occipital Ctx
2.9
|
AD 1 Temporal Ctx
19.9
Control 2 Occipital Ctx
70.2
|
AD 2 Temporal Ctx
37.4
Control 3 Occipital Ctx
2.7
|
AD 3 Temporal Ctx
9.2
Control 4 Occipital Ctx
5.4
|
AD 4 Temporal Ctx
27.0
Control (Path) 1
100.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
66.0
Control (Path) 2
16.2
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
24.8
Control (Path) 3
3.1
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
74.7
Control (Path) 4
41.2
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
56.6
Control 1 Parietal Ctx
6.6
|
Control 1 Temporal Ctx
8.6
Control 2 Parietal Ctx
44.8
|
Control 2 Temporal Ctx
32.3
Control 3 Parietal Ctx
12.8
|
Control 3 Temporal Ctx
33.0
Control (Path) 1
69.3
|
Parietal Ctx
|
Control 4 Temporal Ctx
12.2
Control (Path) 2
23.0
|
Parietal Ctx
|
Control (Path) 1
55.1
Control (Path) 3
4.2
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
42.3
Control (Path) 4
57.0
|
Temporal Ctx
Parietal Ctx
|
|
[1077]
512
TABLE XC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3706, Run
Ag3706, Run
|
Tissue Name
218253760
Tissue Name
218253760
|
|
Adipose
14.0
Renal ca. TK-10
0.0
|
Melanoma* Hs688(A).T
0.0
Bladder
4.7
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.0
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
8.2
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.0
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
SW620
|
Testis Pool
80.1
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
0.2
Colon ca. HCT-116
0.0
|
PC-3
|
Prostate Pool
94.0
Colon ca. CaCo-2
0.4
|
Placenta
0.9
Colon cancer tissue
6.1
|
Uterus Pool
9.7
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
6.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
0.2
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
7.6
|
Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
84.1
|
Ovarian ca. IGROV-1
0.0
Stomach Pool
24.3
|
Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
5.6
|
Ovary
44.8
Fetal Heart
2.8
|
Breast ca. MCF-7
0.4
Heart Pool
8.5
|
Breast ca. MDA-MB-
0.0
Lymph Node Pool
12.6
|
231
|
Breast ca. BT 549
0.0
Fetal Skeletal Muscle
5.4
|
Breast ca. T47D
0.2
Skeletal Muscle Pool
1.5
|
Breast ca. MDA-N
0.0
Spleen Pool
6.2
|
Breast Pool
6.2
Thymus Pool
6.6
|
Trachea
13.4
CNS cancer (glio/astro)
0.0
|
U87-MG
|
Lung
29.5
CNS cancer (glio/astro) U-
0.0
|
118-MG
|
Fetal Lung
4.2
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
1.4
|
Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
0.0
|
Lung ca. NCI-H146
2.5
CNS cancer (glio) SNB-19
0.0
|
Lung ca. SHP-77
0.2
CNS cancer (glio) SF-295
0.3
|
Lung ca. A549
0.3
Brain (Amygdala) Pool
19.5
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
8.1
|
Lung ca. NCI-H23
1.9
Brain (fetal)
100.0
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
25.7
|
Lung ca. HOP-62
0.0
Cerebral Cortex Pool
36.1
|
Lung ca. NCI-H522
0.2
Brain (Substantia nigra)
19.2
|
Pool
|
Liver
0.2
Brain (Thalamus) Pool
35.1
|
Fetal Liver
1.7
Brain (whole)
22.2
|
Liver ca. HepG2
0.0
Spinal Cord Pool
29.1
|
Kidney Pool
40.3
Adrenal Gland
2.1
|
Fetal Kidney
24.7
Pituitary gland Pool
2.9
|
Renal ca. 786-0
0.7
Salivary Gland
0.9
|
Renal ca. A498
0.0
Thyroid (female)
3.2
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.0
|
Renal ca. UO-31
0.3
Pancreas Pool
4.0
|
|
[1078]
513
TABLE XD
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3706, Run
Ag3706, Run
|
Tissue Name
169987426
Tissue Name
169987426
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
1.7
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
0.0
|
alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
0.0
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-
0.0
|
lymphocyte rest
1beta
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
1.2
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
6.6
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
87.7
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
1.7
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
12.9
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
4.1
Neutrophils TNF a + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
100.0
|
Macrophages rest
0.0
Lung
12.9
|
Macrophages LPS
0.0
Thymus
1.7
|
HUVEC none
0.0
Kidney
17.1
|
HUVEC starved
0.0
|
|
[1079] CNS_neurodegeneration_v1.0 Summary: Ag3706 This panel does not show differential expression of the CG90204-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1080] General_screening_panel_v1.4 Summary: Ag3706 The CG90204-01 gene shows moderate levels of expression in most of the CNS regions examined, with highest expression in the fetal brain (CT=30). Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1081] Moderate levels of expression are also expressed in other normal tissue samples, including prostate, ovary and testis.
[1082] Panel 4.1D Summary: Ag3706 Expression of the CG90204-01 gene is restricted to a few samples, with highest expression in the colon and liver cirrhosis (CTs=31.7). Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel. In addition, expression in liver cirrhosis, but not liver on Panel 1.4 suggests that this gene product may be useful as in the diagnosis and/or treatment of liver cirrhosis.
[1083] Y. CG90214-01: BETA-1,4 N ACETYLGALACTOSAMINYLTRANSFERASE
[1084] Expression of gene CG90214-01 was assessed using the primer-probe set Ag3708, described in Table YA. Results of the RTQ-PCR runs are shown in Tables YB and YC.
514TABLE YA
|
|
Probe Name Ag3708
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-cccagacttgaccgtaatagtg-3′22849300
ProbeTET-5′-26878301
acagccagaagcccctggaaattaaa-
3′-TAMRA
Reverse5′-tcccaaagggcatagtgtaata-3′22919302
|
[1085]
515
TABLE YB
|
|
|
Panel 2.2
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3708, Run
Ag3708, Run
|
Tissue Name
173764403
Tissue Name
173764403
|
|
Normal Colon
29.5
Kidney Margin (OD04348)
21.0
|
Colon cancer (OD06064)
11.8
Kidney malignant cancer
0.0
|
(OD06204B)
|
Colon Margin (OD06064)
66.9
Kidney normal adjacent
2.2
|
tissue (OD06204E)
|
Colon cancer (OD06159)
3.6
Kidney Cancer (OD04450-
0.0
|
01)
|
Colon Margin (OD06159)
100.0
Kidney Margin (OD04450-
9.0
|
03)
|
Colon cancer (OD06297-04)
2.6
Kidney Cancer 8120613
5.3
|
Colon Margin (OD06297-05)
24.1
Kidney Margin 8120614
2.0
|
CC Gr.2 ascend colon
12.0
Kidney Cancer 9010320
0.6
|
(ODO3921)
|
CC Margin (ODO3921)
29.1
Kidney Margin 9010321
3.3
|
Colon cancer metastasis
4.5
Kidney Cancer 8120607
0.0
|
(OD06104)
|
Lung Margin (OD06104)
6.9
Kidney Margin 8120608
1.4
|
Colon mets to lung
0.0
Normal Uterus
0.0
|
(OD04451-01)
|
Lung Margin (OD04451-02)
2.0
Uterine Cancer 064011
0.0
|
Normal Prostate
0.0
Normal Thyroid
0.7
|
Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
4.6
|
Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
1.5
|
Normal Ovary
0.0
Thyroid Margin A302153
5.0
|
Ovarian cancer (OD06283-
2.5
Normal Breast
0.0
|
03)
|
Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
1.7
|
07)
|
Ovarian Cancer 064008
8.7
Breast Cancer 1024
17.8
|
Ovarian cancer (OD06145)
1.3
Breast Cancer (OD04590-
0.7
|
01)
|
Ovarian Margin (OD06145)
0.6
Breast Cancer Mets
0.0
|
(OD04590-03)
|
Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
2.2
|
07)
|
Normal Lung
0.5
Breast Cancer 9100266
1.2
|
Invasive poor diff. lung
2.0
Breast Margin 9100265
0.4
|
adeno (ODO4945-01)
|
Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
3.7
|
Lung Malignant Cancer
8.5
Breast Margin A2090734
1.0
|
(OD03126)
|
Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
0.8
|
Lung Cancer (OD05014A)
9.0
Breast cancer node
1.2
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
0.0
Normal Liver
0.0
|
Lung cancer (OD06081)
0.6
Liver Cancer 1026
22.7
|
Lung Margin (OD06081)
0.5
Liver Cancer 1025
0.0
|
Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
1.1
|
Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
1.3
|
Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
35.4
|
Ocular Melanoma Margin
0.0
Liver Tissue 6005-N
0.0
|
(Liver)
|
Melanoma Metastasis
0.0
Liver Cancer 064003
0.0
|
Melanoma Margin (Lung)
0.0
Normal Bladder
0.0
|
Normal Kidney
4.0
Bladder Cancer 1023
1.1
|
Kidney Ca, Nuclear grade 2
36.6
Bladder Cancer A302173
0.0
|
(OD04338)
|
Kidney Margin (OD04338)
1.6
Normal Stomach
16.2
|
Kidney Ca Nuclear grade 1/2
6.1
Gastric Cancer 9060397
0.0
|
(OD04339)
|
Kidney Margin (OD04339)
1.4
Stomach Margin 9060396
0.6
|
Kidney Ca, Clear cell type
2.5
Gastric Cancer 9060395
5.3
|
(OD04340)
|
Kidney Margin (OD04340)
31.0
Stomach Margin 9060394
4.3
|
Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0
|
(OD04348)
|
|
[1086]
516
TABLE YC
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3708, Run
Ag3708, Run
|
Tissue Name
169960432
Tissue Name
169960432
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
1.5
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
17.7
|
alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
34.6
|
Primary Tr1 rest
0.0
Small airway epithelium
30.4
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
1.1
Coronery artery SMC TNF
0.0
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-
0.0
|
lymphocyte rest
1beta
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
18.7
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
33.4
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
3.7
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
2.3
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
1.3
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
1.6
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
1.8
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNF a + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
71.7
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
0.0
Thymus
0.0
|
HUVEC none
0.0
Kidney
100.0
|
HUVEC starved
0.0
|
|
[1087] CNS_neurodegeneration_v1.0 Summary: Ag3708 Expression of the CG90214-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1088] General_screening_panel_v1.4 Summary: Ag3708 Results from one experiment with the CG90214-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1089] Panel 2.2 Summary: Ag3708 Highest expression of the CG90214-01 gene is detected in colon margin (OD06159) sample (CT=31.6). Interestingly, expression of this gene is down-regulated in colon and kidney cancer. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drug, might be beneficial in the treatment of these cancers.
[1090] In addition, expression of this gene is higher in two of the liver cancer (1026, 6005-T) and kidney cancer nuclear grade 2 (OD04338) samples. Therefore, expression of this gene can be used as diagnostic marker for kidney and liver cancer. Furthermore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, might be beneficial in the treatment of these cancers.
[1091] Panel 4.1D Summary: Ag3708 Highest expression of the CG90214-01 gene is detected in kidney sample (CT=33). Significant expression of this gene is also seen in colon (CT=33.6). Thus, expression of this gene can be used to distinguish this sample from other samples used in this panel. In addition, low but significant expression of this gene is also seen in small airway epithelium and TNFalpha+IL-1beta treated keratinocytes. Therefore, therapeutic modulation of the activity of the protein encoded by this gene may be useful in the treatment of inflammatory bowel disease, psoriasis, wound healing, asthma, COPD, and emphysema, and inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis.
[1092] Panel CNS—1 Summary: Ag3708 Expression of the CG90214-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1093] Z. CG90385-01: Mitogen Activated Protein Kinase
[1094] Expression of gene CG90385-01 was assessed using the primer-probe sets Ag658 and Ag3979, described in Tables ZA and ZB. Results of the RTQ-PCR runs are shown in Tables ZC, ZD and ZE.
517TABLE ZA
|
|
Probe Name Ag658
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-gctccttcaagacggtgtatc-3′21512303
ProbeTET-5′-26538304
ctagacaccgacaccacagtggaggt-
3′-TAMRA
Reverse5′-ccgctcagctctagacagttt-3′21589305
|
[1095]
518
TABLE ZB
|
|
|
Probe Name Ag3979
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-gctccttcaagacggtgtatc-3′
21
512
306
|
Probe
TET-5′-
26
538
307
|
ctagacaccgacaccacagtggaggt-
|
3′-TAMRA
|
Reverse
5′-ccgctcagctctagacagttt-3′
21
589
308
|
|
[1096]
519
TABLE ZC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3979, Run
Ag3979, Run
|
Tissue Name
217534174
Tissue Name
217534174
|
|
Adipose
1.3
Renal ca. TK-10
14.5
|
Melanoma* Hs688(A).T
20.7
Bladder
0.6
|
Melanoma* Hs688(B).T
91.4
Gastric ca. (liver met.)
6.7
|
NCI-N87
|
Melanoma* M14
8.6
Gastric ca. KATO III
0.3
|
Melanoma* LOXIMVI
4.2
Colon ca. SW-948
3.4
|
Melanoma* SK-MEL-5
0.8
Colon ca. SW480
4.5
|
Squamous cell
0.5
Colon ca.* (SW480 met)
5.9
|
carcinoma SCC-4
SW620
|
Testis Pool
0.8
Colon ca. HT29
24.3
|
Prostate ca.* (bone met)
100.0
Colon ca. HCT-116
5.1
|
PC-3
|
Prostate Pool
15.4
Colon ca. CaCo-2
39.8
|
Placenta
0.0
Colon cancer tissue
24.1
|
Uterus Pool
0.3
Colon ca. SW1116
0.6
|
Ovarian ca. OVCAR-3
0.5
Colon ca. Colo-205
0.2
|
Ovarian ca. SK-OV-3
0.7
Colon ca. SW-48
15.8
|
Ovarian ca. OVCAR-4
0.5
Colon Pool
0.0
|
Ovarian ca. OVCAR-5
11.5
Small Intestine Pool
2.9
|
Ovarian ca. IGROV-1
1.3
Stomach Pool
0.5
|
Ovarian ca. OVCAR-8
2.4
Bone Marrow Pool
0.6
|
Ovary
3.2
Fetal Heart
0.5
|
Breast ca. MCF-7
0.8
Heart Pool
0.1
|
Breast ca. MDA-MB-
6.1
Lymph Node Pool
0.3
|
231
|
Breast ca. BT 549
21.8
Fetal Skeletal Muscle
0.0
|
Breast ca. T47D
16.0
Skeletal Muscle Pool
0.0
|
Breast ca. MDA-N
0.9
Spleen Pool
0.3
|
Breast Pool
0.2
Thymus Pool
0.1
|
Trachea
6.3
CNS cancer (glio/astro)
0.3
|
U87-MG
|
Lung
1.2
CNS cancer (glio/astro) U-
0.3
|
118-MG
|
Fetal Lung
2.9
CNS cancer (neuro; met)
0.1
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0
|
Lung ca. LX-1
6.9
CNS cancer (astro) SNB-75
1.7
|
Lung ca. NCI-H146
0.1
CNS cancer (glio) SNB-19
0.7
|
Lung ca. SHP-77
1.0
CNS cancer (glio) SF-295
11.4
|
Lung ca. A549
5.4
Brain (Amygdala) Pool
0.3
|
Lung ca. NCI-H526
0.1
Brain (cerebellum)
1.7
|
Lung ca. NCI-H23
3.1
Brain (fetal)
2.1
|
Lung ca. NCI-H460
0.8
Brain (Hippocampus) Pool
1.3
|
Lung ca. HOP-62
12.8
Cerebral Cortex Pool
0.5
|
Lung ca. NCI-H522
5.6
Brain (Substantia nigra)
0.2
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
0.3
|
Fetal Liver
52.5
Brain (whole)
1.0
|
Liver ca. HepG2
28.7
Spinal Cord Pool
0.4
|
Kidney Pool
0.0
Adrenal Gland
0.2
|
Fetal Kidney
24.5
Pituitary gland Pool
0.6
|
Renal ca. 786-0
0.9
Salivary Gland
0.6
|
Renal ca. A498
1.6
Thyroid (female)
0.0
|
Renal ca. ACHN
32.1
Pancreatic ca. CAPAN2
1.5
|
Renal ca. UO-31
20.7
Pancreas Pool
1.6
|
|
[1097]
520
TABLE ZD
|
|
|
Panel 2.1
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3979, Run
Ag3979, Run
|
Tissue Name
170721574
Tissue Name
170721574
|
|
Normal Colon
9.9
Kidney Cancer 9010320
0.0
|
Colon cancer (OD06064)
0.2
Kidney margin 9010321
44.4
|
Colon cancer margin
2.6
Kidney Cancer 8120607
1.5
|
(OD06064)
|
Colon cancer (OD06159)
3.5
Kidney margin 8120608
8.7
|
Colon cancer margin
2.4
Normal Uterus
0.0
|
(OD06159)
|
Colon cancer (OD06298-08)
30.8
Uterus Cancer
0.6
|
Colon cancer margin
12.7
Normal Thyroid
0.0
|
(OD06298-018)
|
Colon Cancer Gr.2 ascend colon
4.7
Thyroid Cancer
0.0
|
(ODO3921)
|
Colon Cancer margin
4.1
Thyroid Cancer
0.0
|
(ODO3921)
A302152
|
Colon cancer metastasis
0.8
Thyroid margin
0.0
|
(OD06104)
A302153
|
Lung margin (OD06104)
1.2
Normal Breast
3.2
|
Colon mets to lung (OD04451-
9.5
Breast Cancer
3.4
|
01)
|
Lung margin (OD04451-02)
0.0
Breast Cancer
2.9
|
Normal Prostate
9.3
Breast Cancer
1.3
|
(OD04590-01)
|
Prostate Cancer (OD04410)
6.4
Breast Cancer Mets
2.3
|
(OD04590-03)
|
Prostate margin (OD04410)
9.0
Breast Cancer
100.0
|
Metastasis
|
Normal Lung
0.2
Breast Cancer
0.0
|
Invasive poor diff. lung adeno 1
0.3
Breast Cancer 9100266
2.8
|
(ODO4945-01)
|
Lung margin (ODO4945-03)
0.0
Breast margin 9100265
2.9
|
Lung Malignant Cancer
2.8
Breast Cancer A209073
0.5
|
(OD03126)
|
Lung margin (OD03126)
0.6
Breast margin
3.5
|
A2090734
|
Lung Cancer (OD05014A)
0.0
Normal Liver
0.0
|
Lung margin (OD05014B)
0.0
Liver Cancer 1026
2.6
|
Lung Cancer (OD04237-01)
0.0
Liver Cancer 1025
0.3
|
Lung margin (OD04237-02)
0.0
Liver Cancer 6004-T
0.6
|
Ocular Mel Met to Liver
3.9
Liver Tissue 6004-N
1.4
|
(ODO4310)
|
Liver margin (ODO4310)
0.0
Liver Cancer 6005-T
4.4
|
Melanoma Mets to Lung
13.9
Liver Tissue 6005-N
0.0
|
(OD04321)
|
Lung margin (OD04321)
0.0
Liver Cancer
1.7
|
Normal Kidney
19.9
Normal Bladder
0.0
|
Kidney Ca, Nuclear grade 2
76.8
Bladder Cancer
6.7
|
(OD04338)
|
Kidney margin (OD04338)
1.5
Bladder Cancer
0.0
|
Kidney Ca Nuclear grade 1/2
0.7
Normal Ovary
1.8
|
(OD04339)
|
Kidney margin (OD04339)
19.1
Ovarian Cancer
0.0
|
Kidney Ca, Clear cell type
0.0
Ovarian cancer
0.0
|
(OD04340)
(OD06145)
|
Kidney margin (OD04340)
15.4
Ovarian cancer margin
0.0
|
(OD06145)
|
Kidney Ca, Nuclear grade 3
0.0
Normal Stomach
1.8
|
(OD04348)
|
Kidney margin (OD04348)
20.7
Gastric Cancer 9060397
2.5
|
Kidney Cancer (OD04450-01)
1.4
Stomach margin
1.2
|
9060396
|
Kidney margin (OD04450-03)
42.9
Gastric Cancer 9060395
1.0
|
Kidney Cancer 8120613
0.0
Stomach margin
2.2
|
9060394
|
Kidney margin 8120614
9.9
Gastric Cancer 064005
1.0
|
|
[1098]
521
TABLE ZE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3979, Run
Ag3979, Run
|
Tissue Name
170721251
Tissue Name
170721251
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
1.2
|
Secondary Th2 act
0.0
HUVEC IFN gamma
1.8
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.2
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.3
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.9
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
100.0
|
Primary Th1 act
0.1
Lung Microvascular EC
58.2
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
72.2
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
48.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
3.4
|
alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.7
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
5.2
Coronery artery SMC rest
39.5
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
40.6
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
12.1
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-
27.5
|
lymphocyte rest
1beta
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
1.7
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
1.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.6
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.5
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.8
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
1.2
|
LAK cells
0.0
NCI-H292 IL-13
0.3
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.6
|
Two Way MLR 3 day
0.0
HPAEC none
3.4
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
7.5
|
Two Way MLR 7 day
0.2
Lung fibroblast none
0.3
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.1
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.6
|
Ramos (B cell) ionomycin
0.4
Lung fibroblast IFN gamma
0.9
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
14.3
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
9.9
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.9
Dermal fibroblast CCD1070 IL-
11.2
|
1beta
|
EOL-1 dbcAMP
1.8
Dermal fibroblast IFN gamma
0.9
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.2
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.3
Neutrophils TNF a + LPS
0.3
|
Monocytes rest
0.0
Neutrophils rest
0.3
|
Monocytes LPS
0.0
Colon
2.0
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
0.0
Thymus
4.5
|
HUVEC none
3.5
Kidney
29.1
|
HUVEC starved
1.5
|
|
[1099] CNS_neurodegeneration_v1.0 Summary: Ag3979 Expression of this gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1100] General_screening_panel_v1.4 Summary: Ag3979 Expression of this gene is highest in prostate cancer cell line PC-3 and a melanoma cell line (CT=28). Thus, the expression of this gene could be used to distinguish the cells from the other samples in the panel. In addition, there is substantial expression associated with kidney cancer cell lines and colon cancer cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be of benefit in the treatment of kidney cancer, prostate cancer, colon cancer or melanoma. Finally, expression of this gene is much higher in fetal liver (CT=29) than adult liver (CT=40), as well as in fetal kidney (CT=30) than adult kidney (CT=40). This observation suggests that expression of this gene may be used to distinguish fetal from adult liver or kidney.
[1101] This gene encodes a protein with homology to kinases and is expressed at very low levels in the fetal brain, hippocampus, and cerebellum. This gene is predominantly expressed in fetal tissues and in cancer cell lines, suggesting that it plays a role in cell division or differentiation. Thus, this gene may therefore be of use in regulation of the cell cycle in stem cell research or therapy.
[1102] Panel 2.1 Summary: Ag3979 Expression of this gene is highest in a sample derived from a metastatic breast cancer (CT=30.9). Thus, the expression of this gene could be used to distinguish this metastatic breast cancer specimen from other samples in the panel. In addition, there appears to be substantial expression associated with a number of normal kidney tissue samples adjacent to malignant kidney. Therefore, therapeutic modulation of the activity of this gene, through the use of small molecule drugs, protein therapeutics or antibodies, might be of benefit in the treatment of breast and kidney cancer.
[1103] Panel 4.1D Summary: Ag3979 Expression of this gene is highest in lung microvascular endothelial cells (CT=29.7). The transcript is also expressed by fibroblasts, endothelium and smooth muscle cells. This gene encodes a putative protien kinase that localizes to the nucleus based on PSORT analysis. The protein encoded for by this transcript may be important in the normal function of the fibroblasts, endothelial cells and smooth muscle cells. Therefore, therapies designed with the protein encoded for by this transcript could be used to regulate fibroblast, endothelium and smooth muscle cell function and may be important in the treatment of asthma, emphysema, arthritis, and inflammatory bowel disease.
[1104] AA. CG90635-01: Nuclear Body Associated Kinase 2B
[1105] Expression of gene CG90635-01 was assessed using the primer-probe sets Ag3709 and Ag3768, described in Tables AAA and AAB. Results of the RTQ-PCR runs are shown in Tables AAC, AAD and AAE.
522TABLE AAA
|
|
Probe Name Ag3709
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-caaagctgtgtgctcaacct-3′201067309
ProbeTET-5′-261105310
acagagctcctgaaattatccttgga-
3′-TAMRA
Reverse5′-tgaccacatgtcaatagcttca-3′221142311
|
[1106]
523
TABLE AAB
|
|
|
Probe Name Ag3768
|
Start
SEQ ID
|
Primers
Sequences
Length
Position
No:
|
|
Forward
5′-ccagatttgcactcagacaga-3′
21
1952
312
|
Probe
TET-5′-
30
1978
313
|
tccaacagacatttatagtatgtccacctg-3′-
|
TAMRA
|
Reverse
5′-gcttgtagtccactttgaaacg-3′
22
2008
314
|
|
[1107]
524
TABLE AAC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3768, Run
Ag3768, Run
|
Tissue Name
211176319
Tissue Name
211176319
|
|
AD 1 Hippo
20.2
Control (Path) 3
16.2
|
Temporal Ctx
|
AD 2 Hippo
32.5
Control (Path) 4
28.5
|
Temporal Ctx
|
AD 3 Hippo
19.5
AD 1 Occipital Ctx
27.4
|
AD 4 Hippo
7.9
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
15.4
|
AD 6 Hippo
76.3
AD 4 Occipital Ctx
15.5
|
Control 2 Hippo
24.0
AD 5 Occipital Ctx
36.1
|
Control 4 Hippo
26.1
AD 6 Occipital Ctx
28.7
|
Control (Path) 3 Hippo
15.0
Control 1 Occipital Ctx
9.5
|
AD 1 Temporal Ctx
35.4
Control 2 Occipital Ctx
46.3
|
AD 2 Temporal Ctx
22.5
Control 3 Occipital Ctx
24.7
|
AD 3 Temporal Ctx
9.4
Control 4 Occipital Ctx
11.3
|
AD 4 Temporal Ctx
28.1
Control (Path) 1
71.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
73.2
Control (Path) 2
17.1
|
Occipital Ctx
|
AD 5 Sup Temporal
63.3
Control (Path) 3
13.0
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
64.2
Control (Path) 4
7.9
|
Occipital Ctx
|
AD 6 Sup Temporal
64.2
Control 1 Parietal Ctx
15.7
|
Ctx
|
Control 1 Temporal Ctx
10.3
Control 2 Parietal Ctx
49.7
|
Control 2 Temporal Ctx
30.6
Control 3 Parietal Ctx
16.8
|
Control 3 Temporal Ctx
20.0
Control (Path) 1
11.6
|
Parietal Ctx
|
Control 3 Temporal Ctx
5.4
Control (Path) 2
19.2
|
Parietal Ctx
|
Control (Path) 1
57.4
Control (Path) 3
12.9
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
39.2
Control (Path) 4
16.6
|
Temporal Ctx
Parietal Ctx
|
|
[1108]
525
TABLE AAD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3768, Run
Ag3768, Run
|
Tissue Name
218981616
Tissue Name
218981616
|
|
Adipose
6.8
Renal ca. TK-10
26.6
|
Melanoma* Hs688(A).T
17.6
Bladder
14.1
|
Melanoma* Hs688(B).T
15.6
Gastric ca. (liver met.)
36.6
|
NCI-N87
|
Melanoma* M14
20.0
Gastric ca. KATO III
26.8
|
Melanoma* LOXIMVI
14.7
Colon ca. SW-948
5.7
|
Melanoma* SK-MEL-5
11.3
Colon ca. SW480
20.6
|
Squamous cell
14.7
Colon ca.* (SW480 met)
14.9
|
carcinoma SCC-4
SW620
|
Testis Pool
26.1
Colon ca. HT29
11.1
|
Prostate ca.* (bone met)
20.7
Colon ca. HCT-116
23.5
|
PC-3
|
Prostate Pool
4.1
Colon ca. CaCo-2
19.3
|
Placenta
8.2
Colon cancer tissue
18.7
|
Uterus Pool
3.7
Colon ca. SW1116
4.2
|
Ovarian ca. OVCAR-3
12.0
Colon ca. Colo-205
7.7
|
Ovarian ca. SK-OV-3
66.0
Colon ca. SW-48
7.1
|
Ovarian ca. OVCAR-4
8.3
Colon Pool
16.6
|
Ovarian ca. OVCAR-5
28.1
Small Intestine Pool
10.4
|
Ovarian ca. IGROV-1
14.8
Stomach Pool
12.7
|
Ovarian ca. OVCAR-8
17.3
Bone Marrow Pool
5.2
|
Ovary
9.4
Fetal Heart
13.7
|
Breast ca. MCF-7
100.0
Heart Pool
6.1
|
Breast ca. MDA-MB-
25.5
Lymph Node Pool
16.2
|
231
|
Breast ca. BT 549
39.2
Fetal Skeletal Muscle
7.2
|
Breast ca. T47D
47.3
Skeletal Muscle Pool
8.5
|
Breast ca. MDA-N
6.1
Spleen Pool
10.1
|
Breast Pool
18.0
Thymus pool
20.6
|
Trachea
20.4
CNS cancer (glio/astro)
28.1
|
U87-MG
|
Lung
4.6
CNS cancer (glio/astro) U-
36.9
|
118-MG
|
Fetal Lung
51.1
CNS cancer (neuro; met)
18.0
|
SK-N-AS
|
Lung ca. NCI-N417
6.8
CNS cancer (astro) SF-539
23.2
|
Lung ca. LX-1
14.2
CNS cancer (astro) SNB-75
43.8
|
Lung ca. NCI-H146
4.1
CNS cancer (glio) SNB-19
14.4
|
Lung ca. SHP-77
14.0
CNS cancer (glio) SF-295
37.4
|
Lung ca. A549
15.4
Brain (Amygdala) Pool
8.1
|
Lung ca. NCI-H526
9.5
Brain (cerebellum)
37.6
|
Lung ca. NCI-H23
33.0
Brain (fetal)
13.5
|
Lung ca. NCI-H460
12.3
Brain (Hippocampus) Pool
11.3
|
Lung ca. HOP-62
7.4
Cerebral Cortex Pool
13.6
|
Lung ca. NCI-H522
16.8
Brain (Substantia nigra)
12.0
|
Pool
|
Liver
1.6
Brain (Thalamus) Pool
15.9
|
Fetal Liver
34.4
Brain (whole)
29.1
|
Liver ca. HepG2
8.5
Spinal Cord Pool
17.9
|
Kidney Pool
18.6
Adrenal Gland
21.5
|
Fetal Kidney
7.0
Pituitary gland Pool
7.1
|
Renal ca. 786-0
18.9
Salivary Gland
5.7
|
Renal ca. A498
7.7
Thyroid (female)
6.0
|
Renal ca. ACHN
9.1
Pancreatic ca. CAPAN2
10.7
|
Renal ca. UO-31
15.7
Pancreas Pool
16.3
|
|
[1109]
526
TABLE AAE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3768, Run
Ag3768, Run
|
Tissue Name
170069115
Tissue Name
170069115
|
|
Secondary Th1 act
30.8
HUVEC IL-1beta
22.4
|
Secondary Th2 act
44.1
HUVEC IFN gamma
11.8
|
Secondary Tr1 act
51.1
HUVEC TNF alpha + IFN
15.9
|
gamma
|
Secondary Th1 rest
13.3
HUVEC TNF alpha + IL4
17.7
|
Secondary Th2 rest
16.5
HUVEC IL-11
12.3
|
Secondary Tr1 rest
19.6
Lung Microvascular EC none
22.7
|
Primary Th1 act
16.7
Lung Microvascular EC
19.1
|
TNF alpha + IL-1beta
|
Primary Th2 act
32.5
Microvascular Dermal EC none
16.7
|
Primary Tr1 act
26.6
Microsvasular Dermal EC
19.3
|
TNF alpha + IL-1beta
|
Primary Th1 rest
20.3
Bronchial epithelium TNF
12.3
|
alpha + IL1beta
|
Primary Th2 rest
14.8
Small airway epithelium none
4.1
|
Primary Tr1 rest
19.8
Small airway epithelium
14.6
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
21.5
Coronery artery SMC rest
8.7
|
act
|
CD45RO CD4 lymphocyte
25.9
Coronery artery SMC TNF
8.8
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
31.6
Astrocytes rest
11.5
|
Secondary CD8
32.3
Astrocytes TNF alpha + IL-
6.8
|
lymphocyte rest
1beta
|
Secondary CD8
25.5
KU-812 (Basophil) rest
30.8
|
lymphocyte act
|
CD4 lymphocyte none
14.9
KU-812 (Basophil)
56.3
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
22.1
CCD1106 (Keratinocytes) none
8.4
|
CD95 CH11
|
LAK cells rest
35.6
CCD1106 (Keratinocytes)
15.2
|
TNF alpha + IL-1beta
|
LAK cells IL-2
26.4
Liver cirrhosis
6.6
|
LAK cells IL-2 + IL-12
30.8
NCI-H292 none
7.8
|
LAK cells IL-2 + IFN
31.4
NCI-H292 IL-4
16.5
|
gamma
|
LAK cells IL-2 + IL-18
31.2
NCI-H292 IL-9
19.6
|
LAK cells
19.1
NCI-H292 IL-13
11.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
63.3
NCI-H292 IFN gamma
17.9
|
Two Way MLR 3 day
46.3
HPAEC none
12.9
|
Two Way MLR 5 day
26.8
HPAEC TNF alpha + IL-1beta
28.9
|
Two Way MLR 7 day
25.9
Lung fibroblast none
7.0
|
PBMC rest
27.7
Lung fibroblast TNF alpha + IL-
7.4
|
1beta
|
PBMC PWM
33.2
Lung fibroblast IL-4
17.1
|
PBMC PHA-L
19.2
Lung fibroblast IL-9
12.9
|
Ramos (B cell) none
34.4
Lung fibroblast IL-13
9.6
|
Ramos (B cell) ionomycin
31.0
Lung fibroblast IFN gamma
15.3
|
B lymphocytes PWM
21.9
Dermal fibroblast CCD1070 rest
17.2
|
B lymphocytes CD40L
41.5
Dermal fibroblast CCD1070
48.6
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
17.1
Dermal fibroblast CCD1070 IL-
9.2
|
1beta
|
EOL-1 dbcAMP
17.0
Dermal fibroblast IFN gamma
7.9
|
PMA/ionomycin
|
Dendritic cells none
26.8
Dermal fibroblast IL-4
15.7
|
Dendritic cells LPS
18.9
Dermal Fibroblasts rest
6.0
|
Dendritic cells anti-CD40
22.4
Neutrophils TNF a + LPS
7.1
|
Monocytes rest
34.6
Neutrophils rest
35.4
|
Monocytes LPS
48.0
Colon
10.5
|
Macrophages rest
22.7
Lung
18.3
|
Macrophages LPS
18.0
Thymus
100.0
|
HUVEC none
15.8
Kidney
15.5
|
HUVEC starved
16.2
|
|
[1110] CNS13 neurodegeneration_v1.0 Summary: Ag3768 The CG90853-01 gene appears to be slightly upregulated in the temporal cortex of Alzheimer's disease patients and also in patient not demented but showing severe AD-like pathology as compared to non-demented patient with no neuropathology. The temporal cortex is a region that shows degeneration at the mid-stages of this disease. These results suggest that this gene may be a marker of Alzheimer's-like neurodegeneration, and may also be involved in the process of neurodegeneration.
[1111] General_screening13 panel_v1.4 Summary: Ag3768 Expression of the CG90635-01 gene is ubiquitous in this panel, with highest expression in a breast cancer cell line (CT=28.6). Significant expression is also seen in a cluster of breast and ovarian cancer cell lines. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of these cancers.
[1112] In addition, this gene is expressed at much higher levels in fetal lung and liver tissue (CTs=30) when compared to expression in the adult counterpart (CTs=33-34). Thus, expression of this gene may be used to differentiate between the fetal and adult source of these tissues.
[1113] Among tissues with metabolic function, this gene is expressed at moderate to low levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1114] This gene is also expressed at moderate levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useffil in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1115] A second experiment with the probe/primer set Ag3709 shows undetectable levels of expression in all samples on this panel (CTs=40). The data suggest that there is a possibility of an experimental failure.
[1116] Panel 4.1D Summary: Ag3678 Expression of the CG90635-01 gene is ubiquitous in this panel, with highest expression in the thymus (CT=29.6). This gene also is expressed at moderate to low levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1117] Two additional experiments with the probe/primer set Ag3709 show undetectable levels of expression in all samples on this panel (CTs=40). The data suggest that there is a possibility of an experimental failure.
[1118] AB. CG90729-01: Proline-Rich Inositol Polyphosphate 5-phosphatase
[1119] Expression of gene CG90729-01 was assessed using the primer-probe set Ag3713, described in Table ABA. Results of the RTQ-PCR runs are shown in Tables ABB, ABC and ABD.
527TABLE ABA
|
|
Probe Name Ag3713
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ggatgggaatacctaccaggta-3′222457315
ProbeTET-5′-262480316
cattcagtgaggaatcactgcccaag-
3′-TAMRA
Reverse5′-aggatgctgtggttgtgactat-3′222534317
|
[1120]
528
TABLE ABB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3713, Run
Ag3713, Run
|
Tissue Name
211295018
Tissue Name
211295018
|
|
AD 1 Hippo
13.1
Control (Path) 3
3.9
|
Temporal Ctx
|
AD 2 Hippo
37.4
Control (Path) 4
6.5
|
Temporal Ctx
|
AD 3 Hippo
3.0
AD 1 Occipital Ctx
12.5
|
AD 4 Hippo
2.0
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
100.0
AD 3 Occipital Ctx
2.4
|
AD 6 Hippo
73.7
AD 4 Occipital Ctx
4.2
|
Control 2 Hippo
7.5
AD 5 Occipital Ctx
23.0
|
Control 4 Hippo
8.9
AD 6 Occipital Ctx
19.5
|
Control (Path) 3 Hippo
1.8
Control 1 Occipital Ctx
1.1
|
AD 1 Temporal Ctx
11.3
Control 2 Occipital Ctx
25.5
|
AD 2 Temporal Ctx
30.4
Control 3 Occipital Ctx
18.4
|
AD 3 Temporal Ctx
3.7
Control 4 Occipital Ctx
1.6
|
AD 4 Temporal Ctx
3.4
Control (Path) 1
62.9
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
63.3
Control (Path) 2
21.3
|
Occipital Ctx
|
AD 5 Sup Temporal Ctx
39.8
Control (Path) 3
1.7
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
29.3
Control (Path) 4
6.7
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
42.3
Control 1 Parietal Ctx
6.5
|
Control 1 Temporal Ctx
4.1
Control 2 Parietal Ctx
29.9
|
Control 2 Temporal Ctx
10.7
Control 3 Parietal Ctx
23.7
|
Control 3 Temporal Ctx
19.5
Control (Path) 1
45.7
|
Parietal Ctx
|
Control 4 Temporal Ctx
6.1
Control (Path) 2
31.6
|
Parietal Ctx
|
Control (Path) 1
36.1
Control (Path) 3
3.3
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
45.4
Control (Path) 4
17.1
|
Temporal Ctx
Parietal Ctx
|
|
[1121]
529
TABLE ABC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3713, Run
Rel. Exp. (%) Ag3713, Run
|
Tissue Name
218267257
Tissue Name
218267257
|
|
Adipose
0.2
Renal ca. TK-10
0.6
|
Melanoma* Hs688(A).T
0.1
Bladder
1.1
|
Melanoma* Hs688(B).T
0.2
Gastric ca. (liver met.)
4.1
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
2.4
|
Melanoma* LOXIMVI
0.5
Colon ca. SW-948
1.7
|
Melanoma* SK-MEL-5
0.8
Colon ca. SW480
3.6
|
Squamous cell
0.0
Colon ca.* (SW480 met)
4.6
|
carcinoma SCC-4
SW620
|
Testis Pool
0.7
Colon ca. HT29
0.1
|
Prostate ca.* (bone met)
3.2
Colon ca. HCT-116
4.0
|
PC-3
|
Prostate Pool
0.9
Colon ca. CaCo-2
2.6
|
Placenta
0.1
Colon cancer tissue
3.9
|
Uterus Pool
0.3
Colon ca. SW1116
1.0
|
Ovarian ca. OVCAR-3
1.6
Colon ca. Colo-205
0.2
|
Ovarian ca. SK-OV-3
0.2
Colon ca. SW-48
0.3
|
Ovarian ca. OVCAR-4
0.6
Colon Pool
1.9
|
Ovarian ca. OVCAR-5
38.2
Small Intestine Pool
1.1
|
Ovarian ca. IGROV-1
2.6
Stomach Pool
0.6
|
Ovarian ca. OVCAR-8
4.9
Bone Marrow Pool
0.5
|
Ovary
1.7
Fetal Heart
3.2
|
Breast ca. MCF-7
8.0
Heart Pool
2.5
|
Breast ca. MDA-MB-
1.0
Lymph Node Pool
1.2
|
231
|
Breast ca. BT 549
1.3
Fetal Skeletal Muscle
0.1
|
Breast ca. T47D
100.0
Skeletal Muscle Pool
1.3
|
Breast ca. MDA-N
0.0
Spleen Pool
0.1
|
Breast Pool
1.7
Thymus Pool
0.6
|
Trachea
5.5
CNS cancer (glio/astro)
1.4
|
U87-MG
|
Lung
0.1
CNS cancer (glio/astro) U-
0.4
|
118-MG
|
Fetal Lung
1.2
CNS cancer (neuro; met)
3.1
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.2
|
Lung ca. LX-1
4.0
CNS cancer (astro) SNB-75
0.4
|
Lung ca. NCI-H146
0.5
CNS cancer (glio) SNB-19
1.9
|
Lung ca. SHP-77
2.0
CNS cancer (glio) SF-295
2.3
|
Lung ca. A549
1.2
Brain (Amygdala) Pool
1.9
|
Lung ca. NCI-H526
0.3
Brain (cerebellum)
52.5
|
Lung ca. NCI-H23
3.0
Brain (fetal)
7.2
|
Lung ca. NCI-H460
2.9
Brain (Hippocampus) Pool
2.0
|
Lung ca. HOP-62
1.2
Cerebral Cortex Pool
3.1
|
Lung ca. NCI-H522
1.7
Brain (Substantia nigra)
4.2
|
Pool
|
Liver
0.1
Brain (Thalamus) Pool
3.0
|
Fetal Liver
0.3
Brain (whole)
12.3
|
Liver ca. HepG2
0.0
Spinal Cord Pool
0.7
|
Kidney Pool
2.0
Adrenal Gland
0.5
|
Fetal Kidney
1.9
Pituitary gland Pool
0.9
|
Renal ca. 786-0
1.0
Salivary Gland
9.6
|
Renal ca. A498
0.0
Thyroid (female)
69.3
|
Renal ca. ACHN
3.1
Pancreatic ca. CAPAN2
0.6
|
Renal ca. UO-31
2.4
Pancreas Pool
2.0
|
|
[1122]
530
TABLE ABD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3713,
Rel. Exp. (%) Ag3713,
|
Tissue Name
Run 169987477
Tissue Name
Run 169987477
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.8
HUVEC IFN gamma
0.3
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.1
|
gamma
|
Secondary Th1 rest
0.6
HUVEC TNF alpha + IL4
0.2
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
1.2
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
1.0
Microvascular Dermal EC none
0.1
|
Primary Tr1 act
1.3
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
1.5
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.8
|
Primary Tr1 rest
0.0
Small airway epithelium
0.8
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.8
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.3
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
6.4
|
Secondary CD8
0.1
Astrocytes TNF alpha + IL-1beta
1.6
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
4.6
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
4.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.8
Liver cirrhosis
1.8
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
1.5
|
LAK cells IL-2 + IFN
0.4
NCI-H292 IL-4
1.1
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
3.0
|
LAK cells
0.4
NCI-H292 IL-13
2.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
1.2
|
Two Way MLR 3 day
0.0
HPAEC none
1.1
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
2.4
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
1.4
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.3
Lung fibroblast IL-9
0.8
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.7
|
B lymphocytes PWM
0.6
Dermal fibroblast CCD1070 rest
0.1
|
B lymphocytes CD40L
0.9
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.5
|
Dendritic cells LPS
1.0
Dermal Fibroblast rest
1.2
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.1
|
Monocytes LPS
0.2
Colon
75.3
|
Macrophages rest
0.0
Lung
9.0
|
Macrophages LPS
0.0
Thymus
1.4
|
HUVEC none
2.3
Kidney
100.0
|
HUVEC starved
0.4
|
|
[1123] CNS_neurodegeneration_v1.0 Summary: Ag3713 This panel confirms the expression of the CG90729-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1124] General_screening_panel_v1.4 Summary: Ag3713 Highest expression of the CG90729-01 gene is detected in breast cancer T47D cell line (CT=25.4). In addition, significant expression of this gene is associated with number of cancer (CNS, colon, gastric, lung, renal, breast, ovarian and prostate) cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.
[1125] Among tissues with metabolic or endocrine function, this gene is expressed at low to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1126] Interestingly, this gene is expressed at much higher levels in adult (CT=31.7) when compared to fetal skeletal muscle (CT=35). This observation suggests that expression of this gene can be used to distinguish fetal from adult skeletal muscle.
[1127] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1128] Panel 4.1D Summary: Ag3713 Highest expression of the CG90729-01 gene is detected in kidney (CT=30). In addition, significant expression is also seen in colon sample (CT=30.5). Thus, expression of this gene can be used to distinguish these two tissue samples from other samples in the panel. Low expression of this gene is also observed in lung, keratinocytes and astrocytes. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis, inflammatory bowel disease, allergy, asthma, psoriasis and wound healing.
[1129] AC. CG90760-01: TRANSCRIPTION FACTOR 20
[1130] Expression of gene CG90760-01 was assessed using the primer-probe set Ag3715, described in Table ACA. Results of the RTQ-PCR runs are shown in Tables ACB, ACC and ACD.
531TABLE ACA
|
|
Probe Name Ag3715
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-cttcatctgggcctctgatt-3′2018318
ProbeTET-5′-tctttattccctccatcatctagacttga-2940319
3′-TAMRA
Reverse5′-cgcatctccttggtacaaataa-3′2271320
|
[1131]
532
TABLE ACB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3715, Run
Rel. Exp. (%) Ag3715, Run
|
Tissue Name
211295020
Tissue Name
211295020
|
|
AD 1 Hippo
17.3
Control (Path) 3
3.4
|
Temporal Ctx
|
AD 2 Hippo
27.7
Control (Path) 4
32.1
|
Temporal Ctx
|
AD 3 Hippo
12.9
AD 1 Occipital Ctx
27.0
|
AD 4 Hippo
4.6
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
66.4
AD 3 Occipital Ctx
5.9
|
AD 6 Hippo
31.2
AD 4 Occipital Ctx
8.9
|
Control 2 Hippo
4.6
AD 5 Occipital Ctx
10.5
|
Control 4 Hippo
4.0
AD 6 Occipital Ctx
18.3
|
Control (Path) 3 Hippo
0.0
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
22.5
Control 2 Occipital Ctx
14.9
|
AD 2 Temporal Ctx
40.1
Control 3 Occipital Ctx
10.4
|
AD 3 Temporal Ctx
9.0
Control 4 Occipital Ctx
6.8
|
AD 4 Temporal Ctx
17.1
Control (Path) 1
73.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
13.8
|
Occipital Ctx
|
AD 5 Sup Temporal Ctx
34.2
Control (Path) 3
6.0
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
51.4
Control (Path) 4
24.0
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
42.9
Control 1 Parietal Ctx
9.3
|
Control 1 Temporal Ctx
3.3
Control 2 Parietal Ctx
66.0
|
Control 2 Temporal Ctx
0.0
Control 3 Parietal Ctx
13.4
|
Control 3 Temporal Ctx
15.9
Control (Path) 1
41.2
|
Parietal Ctx
|
Control 4 Temporal Ctx
3.6
Control (Path) 2
39.5
|
Parietal Ctx
|
Control (Path) 1
76.8
Control (Path) 3
1.9
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
37.4
Control (Path) 4
39.5
|
Temporal Ctx
Parietal Ctx
|
|
[1132]
533
TABLE ACC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3715, Run
Rel. Exp. (%) Ag3715, Run
|
Tissue Name
218267374
Tissue Name
218267374
|
|
Adipose
3.9
Renal ca. TK-10
16.4
|
Melanoma* Hs688(A).T
1.9
Bladder
23.5
|
Melanoma* Hs688(B).T
2.5
Gastric ca. (liver met.)
40.1
|
NCI-N87
|
Melanoma* M14
7.5
Gastric ca. KATO III
29.9
|
Melanoma* LOXIMVI
4.5
Colon ca. SW-948
7.1
|
Melanoma* SK-MEL-5
27.7
Colon ca. SW480
19.6
|
Squamous cell
8.5
Colon ca.* (SW480 met)
33.4
|
carcinoma SCC-4
SW620
|
Testis Pool
11.0
Colon ca. HT29
18.9
|
Prostate ca.* (bone met)
11.3
Colon ca. HCT-116
19.1
|
PC-3
|
Prostate Pool
6.8
Colon ca. CaCo-2
32.8
|
Placenta
6.4
Colon cancer tissue
7.3
|
Uterus Pool
0.9
Colon ca. SW1116
3.2
|
Ovarian ca. OVCAR-3
14.3
Colon ca. Colo-205
2.0
|
Ovarian ca. SK-OV-3
16.8
Colon ca. SW-48
0.8
|
Ovarian ca. OVCAR-4
7.2
Colon Pool
8.3
|
Ovarian ca. OVCAR-5
100.0
Small Intestine Pool
6.5
|
Ovarian ca. IGROV-1
2.3
Stomach Pool
6.2
|
Ovarian ca. OVCAR-8
0.5
Bone Marrow Pool
7.3
|
Ovary
6.7
Fetal Heart
5.6
|
Breast ca. MCF-7
10.5
Heart Pool
3.0
|
Breast ca. MDA-MB-
4.9
Lymph Node Pool
14.3
|
231
|
Breast ca. BT 549
13.9
Fetal Skeletal Muscle
6.7
|
Breast ca. T47D
30.8
Skeletal Muscle Pool
4.3
|
Breast ca. MDA-N
3.9
Spleen Pool
10.7
|
Breast Pool
14.1
Thymus Pool
13.3
|
Trachea
13.5
CNS cancer (glio/astro)
15.1
|
U87-MG
|
Lung
3.5
CNS cancer (glio/astro) U-
13.2
|
118-MG
|
Fetal Lung
23.0
CNS cancer (neuro; met)
4.2
|
SK-N-AS
|
Lung ca. NCI-N417
2.1
CNS cancer (astro) SF-539
2.8
|
Lung ca. LX-1
38.7
CNS cancer (astro) SNB-75
23.7
|
Lung ca. NCI-H146
4.7
CNS cancer (glio) SNB-19
2.0
|
Lung ca. SHP-77
18.2
CNS cancer (glio) SF-295
32.8
|
Lung ca. A549
15.5
Brain (Amygdala) Pool
4.0
|
Lung ca. NCI-H526
0.5
Brain (cerebellum)
22.8
|
Lung ca. NCI-H23
41.8
Brain (fetal)
57.4
|
Lung ca. NCI-H460
12.3
Brain (Hippocampus) Pool
8.7
|
Lung ca. HOP-62
24.7
Cerebral Cortex Pool
2.1
|
Lung ca. NCI-H522
16.3
Brain (Substantia nigra)
5.9
|
Pool
|
Liver
0.5
Brain (Thalamus) Pool
8.4
|
Fetal Liver
8.5
Brain (whole)
6.7
|
Liver ca. HepG2
5.8
Spinal Cord Pool
7.8
|
Kidney Pool
23.0
Adrenal Gland
13.0
|
Fetal Kidney
34.9
Pituitary gland Pool
5.7
|
Renal ca. 786-0
9.5
Salivary Gland
4.9
|
Renal ca. A498
1.4
Thyroid (female)
0.8
|
Renal ca. ACHN
14.0
Pancreatic ca. CAPAN2
56.6
|
Renal ca. UO-31
6.5
Pancreas Pool
17.8
|
|
[1133]
534
TABLE ACD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3715,
Rel. Exp. (%) Ag3715,
|
Tissue Name
Run 169992420
Tissue Name
Run 169992420
|
|
Secondary Th1 act
55.5
HUVEC IL-1beta
24.3
|
Secondary Th2 act
82.4
HUVEC IFN gamma
38.4
|
Secondary Tr1 act
68.3
HUVEC TNF alpha + IFN
27.0
|
gamma
|
Secondary Th1 rest
40.9
HUVEC TNF alpha + IL4
15.2
|
Secondary Th2 rest
52.1
HUVEC IL-11
12.9
|
Secondary Tr1 rest
36.1
Lung Microvascular EC none
27.7
|
Primary Th1 act
81.2
Lung Microvascular EC
28.5
|
TNF alpha + IL-1beta
|
Primary Th2 act
97.3
Microvascular Dermal EC none
28.1
|
Primary Tr1 act
60.3
Microsvasular Dermal EC
21.3
|
TNF alpha + IL-1beta
|
Primary Th1 rest
75.3
Bronchial epithelium TNF alpha +
23.5
|
IL1beta
|
Primary Th2 rest
59.9
Small airway epithelium none
16.8
|
Primary Tr1 rest
52.5
Small airway epithelium
34.2
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
43.8
Coronery artery SMC rest
9.6
|
act
|
CD45RO CD4 lymphocyte
100.0
Coronery artery SMC TNF alpha +
7.9
|
act
IL-1beta
|
CD8 lymphocyte act
54.3
Astrocytes rest
11.5
|
Secondary CD8
69.3
Astrocytes TNF alpha + IL-1beta
6.8
|
lymphocyte rest
|
Secondary CD8
25.5
KU-812 (Basophil) rest
27.7
|
lymphocyte act
|
CD4 lymphocyte none
27.9
KU-812 (Basophil)
13.5
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
61.1
CCD1106 (Keratinocytes) none
85.9
|
CD95 CH11
|
LAK cells rest
41.8
CCD1106 (Keratinocytes)
50.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
71.7
Liver cirrhosis
5.8
|
LAK cells IL-2 + IL-12
45.7
NCI-H292 none
18.2
|
LAK cells IL-2 + IFN
70.7
NCI-H292 IL-4
53.2
|
gamma
|
LAK cells IL-2 + IL-18
72.7
NCI-H292 IL-9
40.9
|
LAK cells
88.9
NCI-H292 IL-13
25.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
30.4
NCI-H292 IFN gamma
15.9
|
Two Way MLR 3 day
86.5
HPAEC none
13.4
|
Two Way MLR 5 day
53.6
HPAEC TNF alpha + IL-1beta
34.4
|
Two Way MLR 7 day
32.5
Lung fibroblast none
12.9
|
PBMC rest
39.8
Lung fibroblast TNF alpha + IL-
12.4
|
1beta
|
PBMC PWM
32.8
Lung fibroblast IL-4
9.2
|
PBMC PHA-L
47.3
Lung fibroblast IL-9
18.7
|
Ramos (B cell) none
70.2
Lung fibroblast IL-13
15.3
|
Ramos (B cell) ionomycin
55.5
Lung fibroblast IFN gamma
1.5
|
B lymphocytes PWM
41.8
Dermal fibroblast CCD1070 rest
15.9
|
B lymphocytes CD40L
48.3
Dermal fibroblast CCD1070
67.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
50.7
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
56.6
Dermal fibroblast IFN gamma
13.4
|
PMA/ionomycin
|
Dendritic cells none
29.9
Dermal fibroblast IL-4
24.5
|
Dendritic cells LPS
6.7
Dermal Fibroblast rest
11.1
|
Dendritic cells anti-CD40
12.8
Neutrophils TNFa + LPS
11.9
|
Monocytes rest
15.0
Neutrophils rest
40.3
|
Monocytes LPS
75.8
Colon
20.3
|
Macrophages rest
17.3
Lung
8.4
|
Macrophages LPS
9.9
Thymus
84.1
|
HUVEC none
7.3
Kidney
56.6
|
HUVEC starved
29.3
|
|
[1134] CNS_neurodegeneration_v1.0 Summary: Ag3715 This panel does not show differential expression of the CG90760-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the temporal cortex of an Alzheimer's patient (CT=32.6). Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1135] General_screening_panel_v1.4 Summary: Ag3715 Expression of the CG90760-01 gene is widespread in this panel, with highest expression in an ovarian cancer cell line (CT=30.5). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel.
[1136] In addition, the widespread expression of this gene suggests a broader role for this gene product in cell survival and growth.
[1137] Among tissues with metabolic function, this gene is expressed at low levels in pituitary, adrenal gland, pancreas, fetal heart and liver and adult and fetal skeletal muscle. This expression suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1138] This gene is also expressed at low levels in the CNS, including the hippocampus, thalamus, substantia nigra, and cerebellum. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1139] Panel 4.1D Summary: Ag3715 Results from one experiment with the CG90760-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1140] AC. CG90760-01: TRANSCRIPTION FACTOR 20
[1141] Expression of gene CG90760-01 was assessed using the primer-probe set Ag3715, described in Table ACA. Results of the RTQ-PCR runs are shown in Tables ACB, ACC and ACD.
535TABLE ACA
|
|
Probe Name Ag3715
StartSEQ
StartID
PrimersSequencesLengthPositionNo
|
Forward5′-cttcatctgggcctctgatt-3′2018
ProbeTET-5′-tctttattccctccatcatctagacttga-2940
3′-TAMRA
Reverse5′-cgcatctccttggtacaaataa-3′2271
|
[1142]
536
TABLE ACB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3715, Run
Rel. Exp. (%) Ag3715, Run
|
Tissue Name
211295020
Tissue Name
211295020
|
|
AD 1 Hippo
17.3
Control (Path) 3
3.4
|
Temporal Ctx
|
AD 2 Hippo
27.7
Control (Path) 4
32.1
|
Temporal Ctx
|
AD 3 Hippo
12.9
AD 1 Occipital Ctx
27.0
|
AD 4 Hippo
4.6
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
66.4
AD 3 Occipital Ctx
5.9
|
AD 6 Hippo
31.2
AD 4 Occipital Ctx
8.9
|
Control 2 Hippo
4.6
AD 5 Occipital Ctx
10.5
|
Control 4 Hippo
4.0
AD 6 Occipital Ctx
18.3
|
Control (Path) 3 Hippo
0.0
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
22.5
Control 2 Occipital Ctx
14.9
|
AD 2 Temporal Ctx
40.1
Control 3 Occipital Ctx
10.4
|
AD 3 Temporal Ctx
9.0
Control 4 Occipital Ctx
6.8
|
AD 4 Temporal Ctx
17.1
Control (Path) 1
73.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
13.8
|
Occipital Ctx
|
AD 5 Sup Temporal Ctx
34.2
Control (Path) 3
6.0
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
51.4
Control (Path) 4
24.0
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
42.9
Control 1 Parietal Ctx
9.3
|
Control 1 Temporal Ctx
3.3
Control 2 Parietal Ctx
66.0
|
Control 2 Temporal Ctx
0.0
Control 3 Parietal Ctx
13.4
|
Control 3 Temporal Ctx
15.9
Control (Path) 1
41.2
|
Parietal Ctx
|
Control 4 Temporal Ctx
3.6
Control (Path) 2
39.5
|
Parietal Ctx
|
Control (Path) 1
76.8
Control (Path) 3
1.9
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
37.4
Control (Path) 4
39.5
|
Temporal Ctx
Parietal Ctx
|
|
[1143]
537
TABLE ACC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3715, Run
Rel. Exp. (%) Ag3715, Run
|
Tissue Name
218267374
Tissue Name
218267374
|
|
Adipose
3.9
Renal ca. TK-10
16.4
|
Melanoma* Hs688(A).T
1.9
Bladder
23.5
|
Melanoma* Hs688(B).T
2.5
Gastric ca. (liver met.)
40.1
|
NCI-N87
|
Melanoma* M14
7.5
Gastric ca. KATO III
29.9
|
Melanoma* LOXIMVI
4.5
Colon ca. SW-948
7.1
|
Melanoma* SK-MEL-5
27.7
Colon ca. SW480
19.6
|
Squamous cell
8.5
Colon ca.* (SW480 met)
33.4
|
carcinoma SCC-4
SW620
|
Testis Pool
11.0
Colon ca. HT29
18.9
|
Prostate ca.* (bone met)
11.3
Colon ca. HCT-116
19.1
|
PC-3
|
Prostate Pool
6.8
Colon ca. CaCo-2
32.8
|
Placenta
6.4
Colon cancer tissue
7.3
|
Uterus Pool
0.9
Colon ca. SW1116
3.2
|
Ovarian ca. OVCAR-3
14.3
Colon ca. Colo-205
2.0
|
Ovarian ca. SK-OV-3
16.8
Colon ca. SW-48
0.8
|
Ovarian ca. OVCAR-4
7.2
Colon Pool
8.3
|
Ovarian ca. OVCAR-5
100.0
Small Intestine Pool
6.5
|
Ovarian ca. IGROV-1
2.3
Stomach Pool
6.2
|
Ovarian ca. OVCAR-8
0.5
Bone Marrow Pool
7.3
|
Ovary
6.7
Fetal Heart
5.6
|
Breast ca. MCF-7
10.5
Heart Pool
3.0
|
Breast ca. MDA-MB-
4.9
Lymph Node Pool
14.3
|
231
|
Breast ca. BT 549
13.9
Fetal Skeletal Muscle
6.7
|
Breast ca. T47D
30.8
Skeletal Muscle Pool
4.3
|
Breast ca. MDA-N
3.9
Spleen Pool
10.7
|
Breast Pool
14.1
Thymus Pool
13.3
|
Trachea
13.5
CNS cancer (glio/astro)
15.1
|
U87-MG
|
Lung
3.5
CNS cancer (glio/astro) U-
13.2
|
118-MG
|
Fetal Lung
23.0
CNS cancer (neuro; met)
4.2
|
SK-N-AS
|
Lung ca. NCI-N417
2.1
CNS cancer (astro) SF-539
2.8
|
Lung ca. LX-1
38.7
CNS cancer (astro) SNB-75
23.7
|
Lung ca. NCI-H146
4.7
CNS cancer (glio) SNB-19
2.0
|
Lung ca. SHP-77
18.2
CNS cancer (glio) SF-295
32.8
|
Lung ca. A549
15.5
Brain (Amygdala) Pool
4.0
|
Lung ca. NCI-H526
0.5
Brain (cerebellum)
22.8
|
Lung ca. NCI-H23
41.8
Brain (fetal)
57.4
|
Lung ca. NCI-H460
12.3
Brain (Hippocampus) Pool
8.7
|
Lung ca. HOP-62
24.7
Cerebral Cortex Pool
2.1
|
Lung ca. NCI-H522
16.3
Brain (Substantia nigra)
5.9
|
Pool
|
Liver
0.5
Brain (Thalamus) Pool
8.4
|
Fetal Liver
8.5
Brain (whole)
6.7
|
Liver ca. HepG2
5.8
Spinal Cord Pool
7.8
|
Kidney Pool
23.0
Adrenal Gland
13.0
|
Fetal Kidney
34.9
Pituitary gland Pool
5.7
|
Renal ca. 786-0
9.5
Salivary Gland
4.9
|
Renal ca. A498
1.4
Thyroid (female)
0.8
|
Renal ca. ACHN
14.0
Pancreatic ca. CAPAN2
56.6
|
Renal ca. UO-31
6.5
Pancreas Pool
17.8
|
|
[1144]
538
TABLE ACD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3715,
Rel. Exp. (%) Ag3715,
|
Tissue Name
Run 169992420
Tissue Name
Run 169992420
|
|
Secondary Th1 act
55.5
HUVEC IL-1beta
24.3
|
Secondary Th2 act
82.4
HUVEC IFN gamma
38.4
|
Secondary Tr1 act
68.3
HUVEC TNF alpha + IFN
27.0
|
gamma
|
Secondary Th1 rest
40.9
HUVEC TNF alpha + IL4
15.2
|
Secondary Th2 rest
52.1
HUVEC IL-11
12.9
|
Secondary Tr1 rest
36.1
Lung Microvascular EC none
27.7
|
Primary Th1 act
81.2
Lung Microvascular EC
28.5
|
TNF alpha + IL-1beta
|
Primary Th2 act
97.3
Microvascular Dermal EC none
28.1
|
Primary Tr1 act
60.3
Microsvasular Dermal EC
21.3
|
TNF alpha + IL-1beta
|
Primary Th1 rest
75.3
Bronchial epithelium TNF alpha +
23.5
|
IL1beta
|
Primary Th2 rest
59.9
Small airway epithelium none
16.8
|
Primary Tr1 rest
52.5
Small airway epithelium
34.2
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
43.8
Coronery artery SMC rest
9.6
|
act
|
CD45RO CD4 lymphocyte
100.0
Coronery artery SMC TNF alpha +
7.9
|
act
IL-1beta
|
CD8 lymphocyte act
54.3
Astrocytes rest
11.5
|
Secondary CD8
69.3
Astrocytes TNF alpha + IL-1beta
6.8
|
lymphocyte rest
|
Secondary CD8
25.5
KU-812 (Basophil) rest
27.7
|
lymphocyte act
|
CD4 lymphocyte none
27.9
KU-812 (Basophil)
13.5
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
61.1
CCD1106 (Keratinocytes) none
85.9
|
CD95 CH11
|
LAK cells rest
41.8
CCD1106 (Keratinocytes)
50.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
71.7
Liver cirrhosis
5.8
|
LAK cells IL-2 + IL-12
45.7
NCI-H292 none
18.2
|
LAK cells IL-2 + IFN
70.7
NCI-H292 IL-4
53.2
|
gamma
|
LAK cells IL-2 + IL-18
72.7
NCI-H292 IL-9
40.9
|
LAK cells
88.9
NCI-H292 IL-13
25.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
30.4
NCI-H292 IFN gamma
15.9
|
Two Way MLR 3 day
86.5
HPAEC none
13.4
|
Two Way MLR 5 day
53.6
HPAEC TNF alpha + IL-1beta
34.4
|
Two Way MLR 7 day
32.5
Lung fibroblast none
12.9
|
PBMC rest
39.8
Lung fibroblast TNF alpha + IL-
12.4
|
1beta
|
PBMC PWM
32.8
Lung fibroblast IL-4
9.2
|
PBMC PHA-L
47.3
Lung fibroblast IL-9
18.7
|
Ramos (B cell) none
70.2
Lung fibroblast IL-13
15.3
|
Ramos (B cell) ionomycin
55.5
Lung fibroblast IFN gamma
1.5
|
B lymphocytes PWM
41.8
Dermal fibroblast CCD1070 rest
15.9
|
B lymphocytes CD40L
48.3
Dermal fibroblast CCD1070
67.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
50.7
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
56.6
Dermal fibroblast IFN gamma
13.4
|
PMA/ionomycin
|
Dendritic cells none
29.9
Dermal fibroblast IL-4
24.5
|
Dendritic cells LPS
6.7
Dermal Fibroblasts rest
11.1
|
Dendritic cells anti-CD40
12.8
Neutrophils TNFa + LPS
11.9
|
Monocytes rest
15.0
Neutrophils rest
40.3
|
Monocytes LPS
75.8
Colon
20.3
|
Macrophages rest
17.3
Lung
8.4
|
Macrophages LPS
9.9
Thymus
84.1
|
HUVEC none
7.3
Kidney
56.6
|
HUVEC starved
29.3
|
|
[1145] CNS_neurodegenerafion_v1.0 Summary: Ag3715 This panel does not show differential expression of the CG90760-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the temporal cortex of an Alzheimer's patient (CT=32.6). Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1146] General_screening_panel_v1.4 Summary: Ag3715 Expression of the CG90760-01 gene is widespread in this panel, with highest expression in an ovarian cancer cell line (CT=30.5). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel.
[1147] In addition, the widespread expression of this gene suggests a broader role for this gene product in cell survival and growth.
[1148] Among tissues with metabolic function, this gene is expressed at low levels in pituitary, adrenal gland, pancreas, fetal heart and liver and adult and fetal skeletal muscle. This expression suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1149] This gene is also expressed at low levels in the CNS, including the hippocampus, thalamus, substantia nigra, and cerebellum. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1150] Panel 4.1D Summary: Ag3715 Results from one experiment with the CG90760-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1151] AD. CG90770-01: 9530058B02RIK Protein
[1152] Expression of gene CG90770-01 was assessed using the primer-probe set ag3668, described in Table ADA. Results of the RTQ-PCR runs are shown in Table ADB.
539TABLE ADA
|
|
Probe Name ag3668
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-cattgacttggacgagtggt-3′20453321
ProbeTET-5′-gcacttcctggctagaatcaccagct-26480322
3′-TAMRA
Reverse5′-aggtagaccatagcagcgct-3′20529323
|
[1153]
540
TABLE ADB
|
|
|
Panel 4.1D
|
Rel. Exp. (%) ag3668,
Rel. Exp. (%) ag3668,
|
Tissue Name
Run 169990916
Tissue Name
Run 169990916
|
|
Secondary Th1 act
16.5
HUVEC IL-1beta
1.7
|
Secondary Th2 act
3.6
HUVEC IFN gamma
0.3
|
Secondary Tr1 act
8.3
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.6
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
1.6
HUVEC IL-11
1.3
|
Secondary Tr1 rest
0.7
Lung Microvascular EC none
0.0
|
Primary Th1 act
14.2
Lung Microvascular EC
2.2
|
TNF alpha + IL-1beta
|
Primary Th2 act
9.5
Microvascular Dermal EC none
1.5
|
Primary Tr1 act
4.4
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
4.0
Bronchial epithelium TNF alpha +
4.4
|
IL1beta
|
Primary Th2 rest
5.8
Small airway epithelium none
0.0
|
Primary Tr1 rest
15.7
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.3
Coronery artery SMC rest
1.1
|
act
|
CD45RO CD4 lymphocyte
8.9
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
8.8
Astrocytes rest
0.0
|
Secondary CD8
9.9
Astrocytes TNF alpha + IL-1beta
1.7
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
63.3
|
lymphocyte act
|
CD4 lymphocyte none
5.7
KU-812 (Basophil)
31.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
1.7
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
11.9
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
11.4
Liver cirrhosis
1.1
|
LAK cells IL-2 + IL-12
8.8
NCI-H292 none
4.3
|
LAK cells IL-2 + IFN
4.9
NCI-H292 IL-4
4.5
|
gamma
|
LAK cells IL-2 + IL-18
4.3
NCI-H292 IL-9
5.0
|
LAK cells
0.0
NCI-H292 IL-13
3.8
|
PMA/ionomycin
|
NK Cells IL-2 rest
3.8
NCI-H292 IFN gamma
9.1
|
Two Way MLR 3 day
3.6
HPAEC none
2.5
|
Two Way MLR 5 day
3.8
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
5.3
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
13.3
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
13.9
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
100.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
55.1
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
13.1
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
3.2
Dermal fibroblast CCD1070
1.7
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
19.9
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
5.2
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
10.7
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
16.7
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
1.1
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
2.7
Neutrophils rest
0.0
|
Monocytes LPS
13.3
Colon
0.0
|
Macrophages rest
1.2
Lung
0.0
|
Macrophages LPS
32.5
Thymus
19.3
|
HUVEC none
0.0
Kidney
2.6
|
HUVEC starved
4.1
|
|
[1154] CNS_neurodegeneration_v1.0 Summary: Ag3668 Expression of the CG90770-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1155] General_screening_panel_v1.5 Summary: Ag3668 Results from one experiment with the CG90770-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1156] Panel 4.1D Summary: Ag3668 Highest expression of the CG90770-01 gene is seen in an untreated sample derived from the B cell line Ramos (CT=31.5). Lower but still significant levels of expression are seen in Ramos B cells stimulated with ionomycin. B cells represent a principle component of immunity and contribute to the immune response in a number of important functional roles, including antibody production. Production of antibodies against self-antigens is a major component in autoimmune disorders. Since B cells play an important role in autoimmunity, inflammatory processes and inflammatory cascades, therapeutic modulation of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, allergies, chronic obstructive pulmonary disease, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriasis, osteoarthritis, systemic lupus erythematosus and other autoimmune disorders.
[1157] This transcript is also expressed at low but significant levels in induced in the basophil cell line KU-812. Basophils release histamines and other biological modifiers in reponse to allergens and play an important role in the pathology of asthma and hypersensitivity reactions. Therefore, therapeutics designed against the putative protein encoded by this gene may reduce or inhibit inflammation by blocking basophil function in these diseases.
[1158] AE. CG91002-01: 3beta-hydroxy-Delta5-steroid Dehydrogenase
[1159] Expression of gene CG91002-01 was assessed using the primer-probe set Ag3721, described in Table AEA. Results of the RTQ-PCR runs are shown in Tables AEB and AEC.
541TABLE AEA
|
|
Probe Name Ag3721
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-tagaaccgtggagtgggttt-3′201101324
ProbeTET-5′-accgtgaagtccaagactcagcgatt-261144325
3′-TAMRA
Reverse5′-atacccacatgcacatctctgt-3′221178326
|
[1160]
542
TABLE AEB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3721, Run
Rel. Exp. (%) Ag3721, Run
|
Tissue Name
211295023
Tissue Name
211295023
|
|
AD 1 Hippo
10.6
Control (Path) 3
2.8
|
Temporal Ctx
|
AD 2 Hippo
12.9
Control (Path) 4
26.4
|
Temporal Ctx
|
AD 3 Hippo
3.6
AD 1 Occipital Ctx
10.4
|
AD 4 Hippo
1.7
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
6.7
|
AD 6 Hippo
10.7
AD 4 Occipital Ctx
7.8
|
Control 2 Hippo
2.2
AD 5 Occipital Ctx
18.8
|
Control 4 Hippo
1.4
AD 6 Occipital Ctx
10.3
|
Control (Path) 3 Hippo
1.9
Control 1 Occipital Ctx
2.2
|
AD 1 Temporal Ctx
6.7
Control 2 Occipital Ctx
31.9
|
AD 2 Temporal Ctx
20.6
Control 3 Occipital Ctx
13.2
|
AD 3 Temporal Ctx
4.2
Control 4 Occipital Ctx
3.3
|
AD 4 Temporal Ctx
9.5
Control (Path) 1
50.3
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
79.6
Control (Path) 2
8.4
|
Occipital Ctx
|
AD 5 Sup Temporal
23.8
Control (Path) 3
2.9
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
28.1
Control (Path) 4
8.6
|
Occipital Ctx
|
AD 6 Sup Temporal
26.8
Control 1 Parietal Ctx
2.5
|
Ctx
|
Control 1 Temporal Ctx
2.2
Control 2 Parietal Ctx
33.9
|
Control 2 Temporal Ctx
11.5
Control 3 Parietal Ctx
16.0
|
Control 3 Temporal Ctx
8.7
Control (Path) 1
26.2
|
Parietal Ctx
|
Control 3 Temporal Ctx
6.3
Control (Path) 2
17.2
|
Parietal Ctx
|
Control (Path) 1
34.9
Control (Path) 3
1.2
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
33.9
Control (Path) 4
30.4
|
Temporal Ctx
Parietal Ctx
|
|
[1161]
543
TABLE AEC
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3721,
Rel. Exp. (%) Ag3721,
|
Tissue Name
Run 169993119
Tissue Name
Run 169993119
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
17.4
|
Secondary Th2 act
0.0
HUVEC IFN gamma
19.1
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
16.5
|
gamma
|
Secondary Th1 rest
0.8
HUVEC TNF alpha + IL4
26.2
|
Secondary Th2 rest
0.0
HUVEC IL-11
10.4
|
Secondary Tr1 rest
0.7
Lung Microvascular EC none
32.3
|
Primary Th1 act
0.0
Lung Microvascular EC
39.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
17.9
|
Primary Tr1 act
0.5
Microsvasular Dermal EC
13.4
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
21.8
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
2.8
|
Primary Tr1 rest
0.0
Small airway epithelium
16.8
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
5.4
Coronery artery SMC rest
6.6
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
7.9
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.4
|
Secondary CD8
1.0
Astrocytes TNF alpha + IL-1beta
1.7
|
lymphocyte rest
|
Secondary CD8
0.6
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.5
CCD1106 (Keratinocytes) none
6.4
|
CD95 CH11
|
LAK cells rest
60.7
CCD1106 (Keratinocytes)
8.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.4
|
LAK cells IL-2 + IL-12
1.1
NCI-H292 none
0.8
|
LAK cells IL-2 + IFN
0.8
NCI-H292 IL-4
20.2
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
3.9
|
LAK cells
23.8
NCI-H292 IL-13
20.3
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
3.8
|
Two Way MLR 3 day
23.2
HPAEC none
6.0
|
Two Way MLR 5 day
9.8
HPAEC TNF alpha + IL-1beta
16.8
|
Two Way MLR 7 day
1.4
Lung fibroblast none
6.9
|
PBMC rest
0.3
Lung fibroblast TNF alpha + IL-
11.0
|
1beta
|
PBMC PWM
2.7
Lung fibroblast IL-4
10.9
|
PBMC PHA-L
3.5
Lung fibroblast IL-9
5.8
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
8.7
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
6.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
1.8
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
13.9
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.8
Dermal fibroblast CCD1070 IL-
8.6
|
1beta
|
EOL-1 dbcAMP
63.7
Dermal fibroblast IFN gamma
0.5
|
PMA/ionomycin
|
Dendritic cells none
66.9
Dermal fibroblast IL-4
2.4
|
Dendritic cells LPS
42.9
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
91.4
Neutrophils TNFa + LPS
2.4
|
Monocytes rest
18.4
Neutrophils rest
1.1
|
Monocytes LPS
100.0
Colon
1.6
|
Macrophages rest
64.6
Lung
10.3
|
Macrophages LPS
11.7
Thymus
1.0
|
HUVEC none
8.5
Kidney
62.9
|
HUVEC starved
11.3
|
|
[1162] CNS_neurodegeneration_v1.0 Summary: Ag3721 This panel does not show differential expression of the CG91002-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the hippocampus of an Alzheimer's patient (CT=29.2). This gene encodes a homolog of a steroid dehydrogenase, an enzyme that is known to function in the processing of hormones. Brain hormone regulation mediates numerous clinically significant conditions, including psychiatric disorders such as anxiety, overeating and memory disorders. Therefore, agents that modulate the activity of this gene product have potential utility in the treatment of these disorders. In addition, steroid treatment is used in a number of clinical conditions including Alzheimer's disease (estrogen), menopause associated symptoms (estrogen), multiple sclerosis (glucocorticoids), and spinal cord injury (methylprednisolone). Treatment with an antagonist of this gene product, or reduction of the levels of this gene product could slow steroid degradation and lower the necessary amount given for therapeutic effect, thus reducing peripheral side effects (Biswas M G, Russell D W. Expression cloning and characterization of oxidative 17beta- and 3alpha-hydroxysteroid dehydrogenases from rat and human prostate. J Biol Chem Jun. 20, 1997;272(25): 15959-66; Matsumoto T, Tamaki T, Kawakami M, Yoshida M, Ando M, Yamada H. Early complications of high-dose methylprednisolone sodium succinate treatment in the follow-up of acute cervical spinal cord injury. Spine Feb. 15, 2001;26(4):426-30; Holinka CF.Design and conduct of clinical trials in hormone replacement therapy. Ann NY Acad Sci 2001 Sep;943:89-108; Burklman R T, Collins J A, Greene R A. Current perspectives on benefits and risks of hormone replacement therapy. Am J Obstet Gynecol 2001 August; 185(2 Suppl):S 13-23; Gaillard P J, van Der Meide P H, de Boer A G, Breimer DD. Glucocorticoid and type I interferon interactions at the blood-brain barrier: relevance for drug therapies for multiple sclerosis. Neuroreport Jul. 20, 2001;12(10):2189-93; Penning T M, Burczynski M E, Jez J M, Hung C F, Lin H K, Ma H, Moore M, Palackal N, Ratnam K. Human 3alpha-hydroxysteroid dehydrogenase isoforms (AKR1C1-AKRLC4) of the aldo-keto reductase superfamily: functional plasticity and tissue distribution reveals roles in the inactivation and formation of male and female sex hormones. Biochem J Oct. 1, 2000;351(Pt 1):67-77).
[1163] General_screening_panel_v1.4 Summary: Ag3721 Results from one experiment with the CG91002-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1164] Panel 4.1D Summary: Ag3721 Highest expression of the CG91002-01 gene is seen in LPS treated monocytes. Expression in monocytes, macrophages, and dendritic cells suggests that small molecule drugs that antagonize the function of this gene product may reduce or eliminate the symptoms of autoimmune and inflammatory diseases, such as, but not limited to, Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive pulmonary disease, asthma, emphysema, rheumatoid arthritis, lupus erythematosus, or psoriasis.
[1165] AF. CG91002-02: 3beta-hydroxy-Delta5-steroid Dehydrogenase
[1166] Expression of gene CG91002-02 was assessed using the primer-probe set Ag3722, described in Table AFA. Results of the RTQ-PCR runs are shown in Tables AFB, AFC and AFD.
544TABLE AFA
|
|
Probe Name Ag3722
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-tgcatgtgggtattgttagga-3′211172327
ProbeTET-5′-aagctccatcctcctggcttcataca-261202328
3′-TAMRA
Reverse5′-ctggacttttgcccttgtc-3′191231329
|
[1167]
545
TABLE AFB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3722, Run
Rel. Exp. (%) Ag3722, Run
|
Tissue Name
211295024
Tissue Name
211295024
|
|
AD 1 Hippo
7.4
Control (Path) 3
7.4
|
Temporal Ctx
|
AD 2 Hippo
14.7
Control (Path) 4
35.4
|
Temporal Ctx
|
AD 3 Hippo
8.2
AD 1 Occipital Ctx
29.7
|
AD 4 Hippo
6.3
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
100.0
AD 3 Occipital Ctx
9.8
|
AD 6 Hippo
13.7
AD 4 Occipital Ctx
26.1
|
Control 2 Hippo
6.5
AD 5 Occipital Ctx
12.9
|
Control 4 Hippo
3.1
AD 6 Occipital Ctx
21.2
|
Control (Path) 3 Hippo
3.9
Control 1 Occipital Ctx
3.6
|
AD 1 Temporal Ctx
16.7
Control 2 Occipital Ctx
24.7
|
AD 2 Temporal Ctx
34.2
Control 3 Occipital Ctx
31.6
|
AD 3 Temporal Ctx
10.7
Control 4 Occipital Ctx
6.3
|
AD 4 Temporal Ctx
29.7
Control (Path) 1
48.6
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
63.3
Control (Path) 2
13.9
|
Occipital Ctx
|
AD 5 Sup Temporal Ctx
22.5
Control (Path) 3
1.5
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
41.5
Control (Path) 4
22.5
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
53.6
Control 1 Parietal Ctx
12.4
|
Control 1 Temporal Ctx
7.4
Control 2 Parietal Ctx
51.1
|
Control 2 Temporal Ctx
12.2
Control 3 Parietal Ctx
17.6
|
Control 3 Temporal Ctx
22.4
Control (Path) 1
56.3
|
Parietal Ctx
|
Control 4 Temporal Ctx
13.5
Control (Path) 2
28.3
|
Parietal Ctx
|
Control (Path) 1
30.8
Control (Path) 3
4.6
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
49.3
Control (Path) 4
52.9
|
Temporal Ctx
Parietal Ctx
|
|
[1168]
546
TABLE AFC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3722, Run
Rel. Exp. (%) Ag3722, Run
|
Tissue Name
218296850
Tissue Name
218296850
|
|
Adipose
47.6
Renal ca. TK-10
35.1
|
Melanoma* Hs688(A).T
1.5
Bladder
26.8
|
Melanoma* Hs688(B).T
6.5
Gastric ca. (liver met.)
50.3
|
NCI-N87
|
Melanoma* M14
25.7
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
73.7
Colon ca. SW480
7.4
|
Squamous cell
0.0
Colon ca.* (SW480 met)
13.3
|
carcinoma SCC-4
SW620
|
Testis Pool
6.8
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
1.5
Colon ca. HCT-116
1.8
|
PC-3
|
Prostate Pool
22.4
Colon ca. CaCo-2
5.4
|
Placenta
78.5
Colon cancer tissue
9.3
|
Uterus Pool
4.4
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
10.7
Colon ca. Colo-205
1.7
|
Ovarian ca. SK-OV-3
0.0
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
3.6
Colon Pool
19.1
|
Ovarian ca. OVCAR-5
3.3
Small Intestine Pool
36.3
|
Ovarian ca. IGROV-1
12.4
Stomach Pool
31.6
|
Ovarian ca. OVCAR-8
2.7
Bone Marrow Pool
9.3
|
Ovary
95.3
Fetal Heart
63.3
|
Breast ca. MCF-7
0.0
Heart Pool
14.9
|
Breast ca. MDA-MB-
3.7
Lymph Node Pool
23.0
|
231
|
Breast ca. BT 549
4.8
Fetal Skeletal Muscle
14.4
|
Breast ca. T47D
3.2
Skeletal Muscle Pool
22.7
|
Breast ca. MDA-N
24.5
Spleen Pool
13.4
|
Breast Pool
33.7
Thymus Pool
26.4
|
Trachea
2.6
CNS cancer (glio/astro)
10.4
|
U87-MG
|
Lung
17.1
CNS cancer (glio/astro) U-
3.1
|
118-MG
|
Fetal Lung
92.7
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0
|
Lung ca. LX-1
43.5
CNS cancer (astro) SNB-75
26.2
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
7.7
|
Lung ca. SHP-77
44.4
CNS cancer (glio) SF-295
16.3
|
Lung ca. A549
0.0
Brain (Amygdala) Pool
31.4
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
16.0
|
Lung ca. NCI-H23
4.3
Brain (fetal)
51.4
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
37.1
|
Lung ca. HOP-62
6.6
Cerebral Cortex Pool
99.3
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
99.3
|
Pool
|
Liver
1.6
Brain (Thalamus) Pool
76.8
|
Fetal Liver
12.6
Brain (whole)
100.0
|
Liver ca. HepG2
0.0
Spinal Cord Pool
29.5
|
Kidney Pool
83.5
Adrenal Gland
17.9
|
Fetal Kidney
99.3
Pituitary gland Pool
11.3
|
Renal ca. 786-0
0.0
Salivary Gland
4.2
|
Renal ca. A498
6.7
Thyroid (female)
2.3
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
1.3
|
Renal ca. UO-31
0.0
Pancreas Pool
13.4
|
|
[1169]
547
TABLE AFD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3722,
Rel. Exp (%) Ag3722,
|
Tissue Name
Run 169993268
Tissue Name
Run 169993268
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
2.9
|
Secondary Th2 act
0.0
HUVEC IFN gamma
4.6
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
6.3
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
42.6
|
Primary Th1 act
0.0
Lung Microvascular EC
1.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
6.4
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
6.8
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
9.5
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
1.5
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
5.4
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
3.2
|
CD95 CH11
|
LAK cells rest
71.2
CCD1106 (Keratinocytes)
4.1
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
1.0
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
1.7
NCI-H292 IL-4
31.4
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
3.0
|
LAK cells
17.9
NCI-H292 IL-13
16.8
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
1.2
|
Two Way MLR 3 day
20.0
HPAEC none
0.0
|
Two Way MLR 5 day
3.6
HPAEC TNF alpha + IL-1beta
7.1
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
1.7
Lung fibroblast TNF alpha + IL-
4.1
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.9
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
1.8
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
9.5
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
3.2
|
1beta
|
EOL-1 dbcAMP
24.1
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
72.2
Dermal fibroblast IL-4
0.9
|
Dendritic cells LPS
64.6
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
100.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
2.1
Neutrophils rest
0.0
|
Monocytes LPS
84.7
Colon
0.0
|
Macrophages rest
66.9
Lung
6.7
|
Macrophages LPS
2.9
Thymus
0.0
|
HUVEC none
0.9
Kidney
84.1
|
HUVEC starved
0.0
|
|
[1170] CNS_neurodegeneration_v1.0 Summary: Ag3722 This panel does not show differential expression of the CG91002-02 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the hippocampus of an Alzheimer's patient (CT=29.4). This gene encodes a homolog of a steroid dehydrogenase, an enzyme that is known to function in the processing of hormones. Brain hormone regulation mediates numerous clinically significant conditions, including psychiatric disorders such as anxiety, overeating and memory disorders. Therefore, agents that modulate the activity of this gene product have potential utility in the treatment of these disorders. In addition, steroid treatment is used in a number of clinical conditions including Alzheimer's disease (estrogen), menopause associated symptoms (estrogen), multiple sclerosis (glucocorticoids), and spinal cord injury (methylprednisolone). Treatment with an antagonist of this gene product, or reduction of the levels of this gene product could slow steroid degredation and lower the necessary amount given for therapeutic effect, thus reducing peripheral side effects (Biswas M G, Russell D W. Expression cloning and characterization of oxidative 17beta- and 3alpha-hydroxysteroid dehydrogenases from rat and human prostate. J Biol Chem Jun. 20, 1997;272(25):15959-66; Matsumoto T, Tamaki T, Kawakami M, Yoshida M, Ando M, Yamada H. Early complications of high-dose methylprednisolone sodium succinate treatment in the follow-up of acute cervical spinal cord injury. Spine 2001 Feb 15;26(4):426-30; Holinka CF.Design and conduct of clinical trials in hormone replacement therapy. Ann NY Acad Sci 2001 Sep;943:89-108; Burkman R T, Collins J A, Greene R A. Current perspectives on benefits and risks of hormone replacement therapy. Am J Obstet Gynecol 2001 August;185(2 Suppl):S13-23; Gaillard P J, van Der Meide P H, de Boer A G, Breimer DD. Glucocorticoid and type 1 interferon interactions at the blood-brain barrier: relevance for drug therapies for multiple sclerosis. Neuroreport Jul. 20, 2001;12(10):2189-93; Penning TM, Burczynski M E, Jez J M, Hung C F, Lin H K, Ma H, Moore M, Palackal N, Ratnam K. Human 3alpha-hydroxysteroid dehydrogenase isoforms (AKR1 C1-AKR1 C4) of the aldo-keto reductase superfamily: functional plasticity and tissue distribution reveals roles in the inactivation and formation of male and female sex hormones. Biochem J Oct. 1, 2000;351(Pt 1):67-77).
[1171] General_screening_panel_v1.4 Summary: Ag3722 Highest expression of the CG91002-02 gene is seen in the brain (CT=31.6). Prominent expression is seen at low but significant levels throughout the CNS. Please see CNS_neurodegeneration_v1.0 for discussion of utility of this gene in the central nervous system.
[1172] Among tissues with metabolic function, this gene is expressed at moderate to low levels in adipose, adrenal gland, pancreas, thyroid, fetal liver and adult and fetal skeletal muscle, and heart. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1173] Low but significant levels of expression are also seen in lung cancer and melanoma cell lines and may suggest involvement of this gene product in those cancers.
[1174] Panel 4.1D Summary: Ag3722 Highest expression of the CG91002-02 gene is seen in anti CD40 dendritic cells (CT=33.1). Expression in monocytes, macrophages, and dendritic cells suggests that small molecule drugs that antagonize the function of this gene product may reduce or eliminate the symptoms of autoimmune and inflammatory diseases, such as, but not limited to, Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive pulmonary disease, asthma, emphysema, rheumatoid arthritis, lupus erythematosus, or psoriasis.
[1175] AG. CG91298-01: PHOSPHATIDYLGLYCEROPHOSPHATE SYNTHASE
[1176] Expression of gene CG91298-01 was assessed using the primer-probe set Ag3727, described in Table AGA. Results of the RTQ-PCR runs are shown in Tables AGB and AGC.
548TABLE AGA
|
|
Probe Name Ag3727
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-acctggatttatccgctgatt-3′21904330
ProbeTET-5′-aagcccttcgagattcaaatcgatga-26931331
3′-TAMRA
Reverse5′-agtcaacagggtctcagtgaca-3′22960332
|
[1177]
549
TABLE AGB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3727, Run
Ag3727,
|
Tissue Name
212143355
Tissue Name
Run 212143355
|
|
AD 1 Hippo
18.8
Control (Path) 3
5.7
|
Temporal Ctx
|
AD 2 Hippo
19.3
Control (Path) 4
20.4
|
Temporal Ctx
|
AD 3 Hippo
10.7
AD 1 Occipital Ctx
27.5
|
AD 4 Hippo
9.5
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
100.0
AD 3 Occipital Ctx
12.1
|
AD 6 Hippo
62.4
AD 4 Occipital Ctx
15.9
|
Control 2 Hippo
40.9
AD 5 Occipital Ctx
20.0
|
Control 4 Hippo
10.2
AD 6 Occipital Ctx
61.6
|
Control (Path) 3 Hippo
6.9
Control 1 Occipital Ctx
5.3
|
AD 1 Temporal Ctx
45.7
Control 2 Occipital Ctx
63.3
|
AD 2 Temporal Ctx
23.5
Control 3 Occipital Ctx
20.4
|
AD 3 Temporal Ctx
9.7
Control 4 Occipital Ctx
7.7
|
AD 4 Temporal Ctx
20.4
Control (Path) 1
61.6
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
84.1
Control (Path) 2
11.6
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
53.6
Control (Path) 3
5.9
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
71.7
Control (Path) 4
17.2
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
78.5
Control 1 Parietal Ctx
10.6
|
Control 1 Temporal Ctx
5.8
Control 2 Parietal Ctx
89.5
|
Control 2 Temporal Ctx
33.4
Control 3 Parietal Ctx
19.3
|
Control 3 Temporal Ctx
34.2
Control (Path) 1
50.0
|
Parietal Ctx
|
Control 4 Temporal Ctx
8.2
Control (Path) 2
16.2
|
Parietal Ctx
|
Control (Path) 1
38.4
Control (Path) 3
5.4
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
34.9
Control (Path) 4
48.0
|
Temporal Ctx
Parietal Ctx
|
|
[1178]
550
TABLE AGC
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3727, Run
Ag3727, Run
Ag3727, Run
Ag3727, Run
|
Tissue Name
170047715
170128711
Tissue Name
170047715
170128711
|
|
Secondary Th1 act
29.1
0.3
HUVEC IL-1beta
9.2
13.9
|
Secondary Th2 act
34.4
100.0
HUVEC IFN gamma
19.6
11.5
|
Secondary Tr1 act
33.7
5.4
HUVEC TNF alpha +
12.8
6.9
|
IFN gamma
|
Secondary Th1 rest
14.5
14.2
HUVEC TNF alpha +
13.5
5.1
|
IL4
|
Secondary Th2 rest
28.7
6.6
HUVEC IL-11
9.5
6.7
|
Secondary Tr1 rest
15.5
2.5
Lung Microvascular
19.1
7.6
|
EC none
|
Primary Th1 act
18.9
6.0
Lung Microvascular
16.6
48.6
|
EC TNF alpha + IL-
|
1beta
|
Primary Th2 act
25.5
8.5
Microvascular
13.2
21.9
|
Dermal EC none
|
Primary Tr1 act
17.6
20.2
Microsvasular Dermal
12.8
9.6
|
EC TNF alpha + IL-
|
1beta
|
Primary Th1 rest
20.4
5.8
Bronchial epithelium
11.1
9.3
|
TNF alpha + IL1beta
|
Primary Th2 rest
21.6
11.4
Small airway
6.6
9.6
|
epithelium none
|
Primary Tr1 rest
27.5
6.2
Small airway
8.3
12.9
|
epithelium TNF alpha +
|
IL-1beta
|
CD45RA CD4
20.6
12.6
Coronery artery SMC
7.9
10.7
|
lymphocyte act
rest
|
CD45RO CD4
27.4
6.9
Coronery artery SMC
6.4
22.4
|
lymphocyte act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
20.9
6.3
Astrocytes rest
5.1
8.8
|
Secondary CD8
32.3
5.4
Astrocytes TNF alpha
4.4
8.5
|
lymphocyte rest
+ IL-1beta
|
Secondary CD8
16.4
6.9
KU-812 (Basophil)
21.6
9.6
|
lymphocyte act
rest
|
CD4 lymphocyte
8.9
6.0
KU-812 (Basophil)
26.6
12.7
|
none
PMA/ionomycin
|
2ry
18.9
7.0
CCD1106
14.8
14.0
|
Th1/Th2/Tr1_anti-
(Keratinocytes) none
|
CD95 CH11
|
LAK cells rest
17.1
6.4
CCD1106
12.6
5.0
|
(Keratinocytes)
|
TNF alpha + IL-1beta
|
LAK cells IL-2
20.6
11.4
Liver cirrhosis
6.3
7.2
|
LAK cells IL-2 + IL-
25.5
8.7
NCI-H292 none
8.6
15.3
|
12
|
LAK cells IL-2 + IFN
23.7
8.7
NCI-H292 IL-4
15.5
10.7
|
gamma
|
LAK cells IL-2 + IL-
27.9
7.2
NCI-H292 IL-9
18.7
19.5
|
18
|
LAK cells
33.7
9.2
NCI-H292 IL-13
13.0
12.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
23.2
6.7
NCI-H292 IFN
13.4
11.5
|
gamma
|
Two Way MLR 3
27.2
4.3
HPAEC none
12.9
11.7
|
day
|
Two Way MLR 5
16.8
31.0
HPAEC TNF alpha +
19.3
12.6
|
day
IL-1beta
|
Two Way MLR 7
15.4
5.0
Lung fibroblast none
9.3
9.1
|
day
|
PBMC rest
11.3
7.5
Lung fibroblast TNF
12.3
11.8
|
alpha + IL-1beta
|
PBMC PWM
30.8
10.3
Lung fibroblast IL-4
9.9
4.3
|
PBMC PHA-L
27.5
11.6
Lung fibroblast IL-9
9.6
6.5
|
Ramos (B cell) none
17.2
11.9
Lung fibroblast IL-13
8.5
18.4
|
Ramos (B cell)
15.1
4.3
Lung fibroblast IFN
14.5
11.8
|
ionomycin
gamma
|
B lymphocytes PWM
16.4
3.3
Dermal fibroblast
13.5
13.9
|
CCD1070 rest
|
B lymphocytes
36.6
4.9
Dermal fibroblast
20.0
9.2
|
CD40L and IL-4
CCD1070 TNF alpha
|
EOL-1 dbcAMP
18.3
3.5
Dermal fibroblast
10.4
13.7
|
CCD1070 IL-1beta
|
EOL-1 dbcAMP
20.9
3.8
Dermal fibroblast IFN
13.0
12.9
|
PMA/ionomycin
gamma
|
Dendritic cells none
18.3
2.9
Dermal fibroblast IL-4
12.5
10.5
|
Dendritic cells LPS
14.7
4.7
Dermal Fibroblasts
10.2
8.1
|
rest
|
Dendritic cells anti-
20.9
6.5
Neutrophils
16.5
10.6
|
CD40
TNFa + LPS
|
Monocytes rest
27.5
6.9
Neutrophils rest
32.3
9.5
|
Monocytes LPS
100.0
8.8
Colon
5.8
10.4
|
Macrophages rest
12.8
12.1
Lung
10.1
17.8
|
Macrophages LPS
14.3
8.1
Thymus
29.5
6.9
|
HUVEC none
11.1
7.1
Kidney
9.4
17.2
|
HUVEC starved
15.4
5.7
|
|
[1179] CNS_neurodegeneration_v1.0 Summary: Ag3727 This panel does not show differential expression of the CG91298-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the hippocampus of an Alzheimer's patient. Moderate to low levels of expression in the CNS suggests that therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1180] General_screening_panel_v1.4 Summary: Ag3727 Results from one experiment with the CG91298-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1181] Panel 4.1D Summary: Ag3727 Two experiments with the same probe and primer both show ubiquitous expression of the CG91298-01 gene on this panel. Highest expression is seen in LPS stimulated monocytes and chronically activated Th2 cells (CTs=26-28). In addition, this gene is expressed at moderate to low levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This gene encodes a homolog of phosphatidylglycerophosphate synthase, an enzyme that has been shown to be necessary for cell growth. The expression profile in this experiment supports a role for this gene product in cell growth and survival, as well. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis (Ohtsuka T, Nishijima M, Akamatsu Y. A somatic cell mutant defective in phosphatidylglycerophosphate synthase, with impaired phosphatidylglycerol and cardiolipin biosynthesis. J Biol Chem Oct. 25, 1993;268(30):22908-13).
[1182] AH. CG91383-01: ALDEHYDE DEHYDROGENASE
[1183] Expression of gene CG91383-01 was assessed using the primer-probe set Ag3719, described in Table AHA.
551TABLE AHA
|
|
Probe Name Ag3719
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-acctgagcaaaagtgaactcaa-3′22151333
ProbeTET-5′-cagtcatgaagtcattaccatccttgga-28179334
3′-TAMRA
Reverse5′-aggaagattccccaacataaaa-3′22215335
|
[1184] CNS_neurodegeneration_v1.0 Summary: Ag3719 Expression of the CG91383-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1185] General_screening_panel_v1.4 Summary: Ag3719 Results from one experiment with the CG91383-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1186] Panel 2.2 Summary: Ag3719 Expression of the CG91383-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1187] Panel 4.1D Summary: Ag3719 Expression of the CG91383-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown). Panel 5 Islet Summary: Ag3719 Expression of the CG91383-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1188] AI. CG91403-01: PROLINE RICH SYNAPSE ASSOCIATED PROTEIN 2
[1189] Expression of gene CG91403-01 was assessed using the primer-probe set Ag3728, described in Table AIA. Results of the RTQ-PCR runs are shown in Tables AIB, AIC, AID and AIE.
552TABLE AIA
|
|
Probe Name Ag3728
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-agaacctcatcgatgataagca-3′22590336
ProbeTET-5′-aaagcttcacacaaaggcgaacctga-26618337
3′-TAMRA
Reverse5′-tggacgtagtccatgaacttct-3′22644338
|
[1190]
553
TABLE AIB
|
|
|
AI_comprehensive panel_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3728,
Ag3728,
|
Tissue Name
Run 248445826
Tissue Name
Run 248445826
|
|
110967 COPD-F
1.1
112427 Match Control
4.2
|
Psoriasis-F
|
110980 COPD-F
1.1
112418 Psoriasis-M
0.6
|
110968 COPD-M
0.9
112723 Match Control
0.1
|
Psoriasis-M
|
110977 COPD-M
2.5
112419 Psoriasis-M
0.8
|
110989 Emphysema-F
2.2
112424 Match Control
0.6
|
Psoriasis-M
|
110992 Emphysema-F
0.8
112420 Psoriasis-M
0.8
|
110993 Emphysema-F
1.2
112425 Match Control
1.7
|
Psoriasis-M
|
110994 Emphysema-F
0.8
104689 (MF) OA Bone-
6.6
|
Backus
|
110995 Emphysema-F
1.2
104690 (MF) Adj “Normal”
3.7
|
Bone-Backus
|
110996 Emphysema-F
0.1
104691 (MF) OA
1.6
|
Synovium-Backus
|
110997 Asthma-M
0.4
104692 (BA) OA Cartilage-
0.0
|
Backus
|
111001 Asthma-F
1.4
104694 (BA) OA Bone-
2.4
|
Backus
|
111002 Asthma-F
1.8
104695 (BA) Adj “Normal”
3.7
|
Bone-Backus
|
111003 Atopic Asthma-F
1.6
104696 (BA) OA Synovium-
1.3
|
Backus
|
111004 Atopic Asthma-F
0.6
104700 (SS) OA Bone-
2.4
|
Backus
|
111005 Atopic Asthma-F
0.8
104701 (SS) Adj “Normal”
4.0
|
Bone-Backus
|
111006 Atopic Asthma-F
0.2
104702 (SS) OA Synovium-
5.3
|
Backus
|
111417 Allergy-M
0.7
117093 OA Cartilage Rep7
0.8
|
112347 Allergy-M
0.0
112672 OA Bone5
1.2
|
112349 Normal Lung-F
0.0
112673 OA Synovium5
0.7
|
112357 Normal Lung-F
0.8
112674 OA Synovial Fluid
0.6
|
cells5
|
112354 Normal Lung-M
0.4
117100 OA Cartilage Rep14
0.4
|
112374 Crohns-F
0.5
112756 OA Bone9
0.1
|
112389 Match Control
0.6
112757 OA Synovium9
0.6
|
Crohns-F
|
112375 Crohns-F
0.6
112758 OA Synovial Fluid
0.9
|
Cells9
|
112732 Match Control
0.4
117125 RA Cartilage Rep2
1.4
|
Crohns-F
|
112725 Crohns-M
0.1
113492 Bone2 RA
1.1
|
112387 Match Control
1.0
113493 Synovium2 RA
0.4
|
Crohns-M
|
112378 Crohns-M
0.0
113494 Syn Fluid Cells RA
0.8
|
112390 Match Control
1.6
113499 Cartilage4 RA
0.9
|
Crohns-M
|
112726 Crohns-M
0.8
113500 Bone4 RA
1.4
|
112731 Match Control
0.9
113501 Synovium4 RA
0.8
|
Crohns-M
|
112380 Ulcer Col-F
0.5
113502 Syn Fluid Cells4 RA
0.5
|
112734 Match Control
0.8
113495 Cartilage3 RA
1.1
|
Ulcer Col-F
|
112384 Ulcer Col-F
1.4
113496 Bone3 RA
1.0
|
112737 Match Control
0.4
113497 Synovium3 RA
0.3
|
Ulcer Col-F
|
112386 Ulcer Col-F
0.4
113498 Syn Fluid Cells3 RA
1.4
|
112738 Match Control
100.0
117106 Normal Cartilage
1.2
|
Ulcer Col-F
Rep20
|
112381 Ulcer Col-M
0.0
113663 Bone3 Normal
0.0
|
112735 Match Control
0.4
113664 Synovium3 Normal
0.0
|
Ulcer Col-M
|
112382 Ulcer Col-M
0.4
113665 Syn Fluid Cells3
0.0
|
Normal
|
112394 Match Control
0.2
117107 Normal Cartilage
0.3
|
Ulcer Col-M
Rep22
|
112383 Ulcer Col-M
1.1
113667 Bone4 Normal
0.4
|
112736 Match Control
0.4
113668 Synovium4 Normal
0.3
|
Ulcer Col-M
|
112423 Psoriasis-F
0.8
113669 Syn Fluid Cells4
0.9
|
Normal
|
|
[1191]
554
TABLE AIC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3728, Run
Ag3728,
|
Tissue Name
212143476
Tissue Name
Run 212143476
|
|
AD 1 Hippo
16.3
Control (Path) 3
14.0
|
Temporal Ctx
|
AD 2 Hippo
31.4
Control (Path) 4
56.6
|
Temporal Ctx
|
AD 3 Hippo
17.3
AD 1 Occipital Ctx
19.1
|
AD 4 Hippo
10.0
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
95.3
AD 3 Occipital Ctx
13.3
|
AD 6 Hippo
52.1
AD 4 Occipital Ctx
17.3
|
Control 2 Hippo
44.8
AD 5 Occipital Ctx
39.2
|
Control 4 Hippo
8.6
AD 6 Occipital Ctx
12.8
|
Control (Path) 3 Hippo
10.4
Control 1 Occipital Ctx
7.5
|
AD 1 Temporal Ctx
51.1
Control 2 Occipital Ctx
66.9
|
AD 2 Temporal Ctx
34.9
Control 3 Occipital Ctx
28.7
|
AD 3 Temporal Ctx
12.4
Control 4 Occipital Ctx
7.2
|
AD 4 Temporal Ctx
26.4
Control (Path) 1
84.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
92.0
Control (Path) 2
15.8
|
Occipital Ctx
|
AD 5 Sup Temporal
41.5
Control (Path) 3
10.0
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
42.0
Control (Path) 4
22.2
|
Occipital Ctx
|
AD 6 Sup Temporal
55.5
Control 1 Parietal Ctx
11.0
|
Ctx
|
Control 1 Temporal Ctx
10.4
Control 2 Parietal Ctx
54.7
|
Control 2 Temporal Ctx
48.0
Control 3 Parietal Ctx
18.6
|
Control 3 Temporal Ctx
24.7
Control (Path) 1
100.0
|
Parietal Ctx
|
Control 3 Temporal Ctx
13.9
Control (Path) 2
27.7
|
Parietal Ctx
|
Control (Path) 1
93.3
Control (Path) 3
8.3
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
55.1
Control (Path) 4
58.2
|
Temporal Ctx
Parietal Ctx
|
|
[1192]
555
TABLE AID
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3728, Run
Ag3728, Run
|
Tissue Name
218297645
Tissue Name
218297645
|
|
Adipose
18.8
Renal ca. TK-10
1.6
|
Melanoma* Hs688(A).T
1.0
Bladder
21.6
|
Melanoma* Hs688(B).T
0.3
Gastric ca. (liver met.)
10.8
|
NCI-N87
|
Melanoma* M14
1.4
Gastric ca. KATO III
1.3
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.1
|
Melanoma* SK-MEL-5
10.7
Colon ca. SW480
2.8
|
Squamous cell
6.1
Colon ca.* (SW480 met)
0.3
|
carcinoma SCC-4
SW620
|
Testis Pool
3.6
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
2.5
Colon ca. HCT-116
20.2
|
PC-3
|
Prostate Pool
5.0
Colon ca. CaCo-2
3.3
|
Placenta
23.0
Colon cancer tissue
14.1
|
Uterus Pool
10.1
Colon ca. SW1116
1.3
|
Ovarian ca. OVCAR-3
2.4
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
70.2
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
2.8
Colon Pool
36.9
|
Ovarian ca. OVCAR-5
2.3
Small Intestine Pool
15.7
|
Ovarian ca. IGROV-1
1.1
Stomach Pool
9.4
|
Ovarian ca. OVCAR-8
1.3
Bone Marrow Pool
9.0
|
Ovary
6.7
Fetal Heart
23.3
|
Breast ca. MCF-7
0.9
Heart Pool
16.2
|
Breast ca. MDA-MB-
10.2
Lymph Node Pool
22.8
|
231
|
Breast ca. BT 549
4.5
Fetal Skeletal Muscle
14.0
|
Breast ca. T47D
3.9
Skeletal Muscle Pool
15.3
|
Breast ca. MDA-N
0.0
Spleen Pool
75.8
|
Breast Pool
25.2
Thymus Pool
12.1
|
Trachea
10.5
CNS cancer (glio/astro)
0.0
|
U87-MG
|
Lung
1.8
CNS cancer (glio/astro) U-
0.1
|
118-MG
|
Fetal Lung
100.0
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
1.1
|
Lung ca. LX-1
0.8
CNS cancer (astro) SNB-75
1.5
|
Lung ca. NCI-H146
1.6
CNS cancer (glio) SNB-19
1.1
|
Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
1.4
|
Lung ca. A549
0.6
Brain (Amygdala) Pool
26.4
|
Lung ca. NCI-H526
0.2
Brain (cerebellum)
11.6
|
Lung ca. NCI-H23
3.7
Brain (fetal)
32.8
|
Lung ca. NCI-H460
0.2
Brain (Hippocampus) Pool
27.4
|
Lung ca. HOP-62
0.7
Cerebral Cortex Pool
44.8
|
Lung ca. NCI-H522
3.2
Brain (Substantia nigra)
31.4
|
Pool
|
Liver
2.6
Brain (Thalamus) Pool
42.9
|
Fetal Liver
11.3
Brain (whole)
55.5
|
Liver ca. HepG2
3.3
Spinal Cord Pool
3.7
|
Kidney Pool
42.9
Adrenal Gland
12.4
|
Fetal Kidney
15.0
Pituitary gland Pool
1.3
|
Renal ca. 786-0
0.0
Salivary Gland
1.6
|
Renal ca. A498
0.2
Thyroid (female)
9.2
|
Renal ca. ACHN
0.4
Pancreatic ca. CAPAN2
15.7
|
Renal ca. UO-31
1.0
Pancreas Pool
26.1
|
|
[1193]
556
TABLE AIE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3728,
Ag3728,
|
Tissue Name
Run 170222884
Tissue Name
Run 170222884
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
52.9
|
Secondary Th2 act
0.2
HUVEC IFN gamma
57.8
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
13.8
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
12.9
|
Secondary Th2 rest
0.0
HUVEC IL-11
42.9
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
100.0
|
Primary Th1 act
0.1
Lung Microvascular EC
55.1
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
62.9
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
38.4
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium
2.4
|
TNF alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.8
|
Primary Tr1 rest
0.0
Small airway epithelium
4.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.2
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.1
Coronery artery SMC
0.0
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
0.1
Astrocytes rest
0.3
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.2
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
6.8
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
6.3
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
2.8
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
2.9
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
2.2
|
LAK cells IL-2 + IL-12
0.1
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.3
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.2
|
LAK cells
0.5
NCI-H292 IL-13
0.4
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.6
|
Two Way MLR 3 day
0.1
HPAEC none
57.4
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
54.0
|
Two Way MLR 7 day
0.1
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.1
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.2
Lung fibroblast IL-9
0.2
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.2
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.1
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.2
|
PMA/ionomycin
|
Dendritic cells none
0.3
Dermal fibroblast IL-4
0.2
|
Dendritic cells LPS
0.5
Dermal Fibroblasts rest
0.3
|
Dendritic cells anti-CD40
1.6
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
2.2
|
Macrophages rest
3.4
Lung
19.3
|
Macrophages LPS
0.1
Thymus
3.5
|
HUVEC none
35.8
Kidney
12.4
|
HUVEC starved
46.0
|
|
[1194] AI_comprehensive panel_v1.0 Summary: Ag3728 Highest expression of the CG91403-01 gene is detected in 112738 match control ulcerative colitis sample (CT=24). Interestingly, expression of this gene is lower in 2 samples derived from ulcerative colitis patients and 2 samples derived from Crohn's patient as compared to their matched control sample. Therefore, therapeutic modulation of the activity of the protein encoded by this gene may be useful in the treatment of inflammatory bowel disease. In addition, this gene shows low to moderate expression in all the samples used in this panel. Taken together with the expression of this gene in activated endothelial, as shown on Panel 4.1D, therapeutic modulation of the activity of the protein encoded by this gene with small molecule antagonists may reduce or eliminate the symptoms of autoimmune and inflammatory diseases that involve activated endothelial cells, such as, but not limited to, Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive pulmonary disease, asthma, emphysema, rheumatoid arthritis, lupus erythematosus, or psoriasis.
[1195] CNS_neurodegeneration_v1.0 Summary: Ag3728 This panel confirms the expression of the CG91403-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1196] General_screening_panel_v1.4 Summary: Ag3728 Highest expression of the CG91403-01 gene is detected in fetal lung (CT=27). Interestingly, this gene is expressed at much higher levels in fetal when compared to adult lung (CT=33). This observation suggests that expression of this gene can be used to distinguish fetal from adult lung. In addition, the relative overexpression of this gene in fetal lung suggests that the protein product may be required for the lung development in the fetus and thus may also act in a regenerative capacity in the adult.
[1197] High to moderate expression is seen in some of the cancer (ovarian, colon, pancreatic, breast, squamous cell carcinoma, prostate) cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs might be beneficial in the treatment of these cancers.
[1198] Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1199] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1200] The CG91403-01 gene codes for a homolog of rat proline rich synapse associated protein 2 (ProSAP2). Recently, ProSAP2 haploinsufficiency has been shown to be associated with the 22q13.3 deletion syndrome (Bonaglia MC, Giorda R. Borgatti R. Felisari G. Gagliardi C, Selicomni A, Zuffardi O. (2001) Disruption of the ProSAP2 gene in a t(12;22)(q24.1;q13.3) is associated with the 22q13.3 deletion syndrome. Am J Hum Genet 69(2):261-8). Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs might be beneficial in the treatment of the 22q13.3 deletion syndrome.
[1201] Panel 4.1D Summary: Ag3728 Highest expression of the CG91403-01 gene is w detected in lung microvascular EC (CT=28.6). High to moderate expression of this gene is also seen in HUVFC, HPAEC, lung microvascular EC, microvascular dermal EC, TNFalpha+ IL-1beta treated small airway epithelium, basophils and keratinocytes, lung, kidney, thymus, and lung. Therefore, modulation of the expression or activity of the protein encoded by this gene through the application of small molecule therapeutics may be useful in the treatment of asthma, COPD, emphysema, psoriasis, lupus erythematosus, inflammatory bowel diseases, such as Crohn's and ulcerative colitis, rheumatoid arthritis, osteoarthritis and wound healing.
[1202] AJ. CG91434-01: ALDEHYDE DEHYDROGENASE
[1203] Expression of gene CG91434-01 was assessed using the primer-probe sets Ag3729 and Ag5999, described in Tables AJA and AJB. Results of the RTQ-PCR runs are shown in Tables AJC and AJD.
557TABLE AJA
|
|
Probe Name Ag3729
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ttcaagaaaacaagcagcttct-3′22122339
ProbeTET-5′-cccaggacctgcataagccagct-23158340
3′-TAMRA
Reverse5′-ctcagatatgtctgcctcgaa-3′21181341
|
[1204]
558
TABLE AJB
|
|
|
Probe Name Ag5999
|
Start
SEQ ID
|
Primers
Sequences
Length
Position
NO:
|
|
Forward
5′-cctgtacgccttctccaaca-3′
20
1101
342
|
Probe
TET-5′-agccaggtggtcaagcgggtgctg-
24
1123
343
|
3′-TAMRA
|
Reverse
5′-actcctccaaaaggcaggct-3′
20
1207
344
|
|
[1205]
559
TABLE AJC
|
|
|
Panel 2.2
|
Rel.Exp.(%)
Rel.Exp.(%)
Rel.Exp.(%)
Rel.Exp.(%)
|
Ag3729, Run
Ag3729, Run
Ag3729, Run
Ag3729, Run
|
Tissue Name
174441818
259034396
Tissue Name
174441818
259034396
|
|
Normal Colon
0.4
0.3
Kidney Margin
0.0
0.0
|
(OD04348)
|
Colon Cancer
1.4
1.0
Kidney malignant
0.0
0.0
|
(OD06064)
cancer
|
(OD06204B)
|
Colon Margin
0.0
0.0
Kidney normal
0.0
0.0
|
(OD06064)
adjacent tissue
|
Colon cancer
0.2
0.1
Kidney Cancer
0.0
0.0
|
(OD06159)
(OD04450-01)
|
Colon Margin
0.0
0.0
Kidney Margin
1.3
0.9
|
(OD06159)
(OD04450-03)
|
Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0
|
(OD06297-04)
8120613
|
Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0
|
(OD06297-05)
8120614
|
CC Gr.2 ascend
1.1
0.8
Kidney Cancer
0.5
0.3
|
colon (ODO3921)
9010320
|
CC Margin
0.0
0.0
Kidney Margin
1.8
1.4
|
(ODO3921)
9010321
|
Colon cancer
0.2
0.1
Kidney Cancer
0.0
0.0
|
metastasis
Kidney Cancer
|
(OD06104)
8120607
|
Lung Margin
0.0
0.0
Kidney Margin
1.0
0.8
|
(OD06104)
8120608
|
Colon mets to lung
0.2
0.2
Normal Uterus
0.0
0.0
|
(OD04451-01)
|
Lung Margin
0.0
0.0
Uterine Cancer
1.8
1.2
|
(OD04451-02)
064011
|
Normal Prostate
2.3
1.8
Normal Thyroid
0.0
0.0
|
Prostate Cancer
2.2
1.6
Thyroid Cancer
0.0
0.0
|
(OD04410)
064010
|
Prostate Margin
5.1
3.8
Thyroid Cancer
0.0
0.0
|
(OD04410)
A302152
|
Normal Ovary
0.7
0.3
Thyroid Margin
0.0
0.0
|
A302153
|
Ovarian cancer
2.5
1.7
Normal Breast
9.2
6.5
|
(OD06283-03)
|
Ovarian Margin
0.0
0.0
Breast Cancer
17.4
12.9
|
(OD06283-07)
(OD04566)
|
Ovarian Cancer
1.0
0.6
Breast Cancer 1024
100.0
100.0
|
064008
|
Ovarian cancer
0.4
0.3
Breast Cancer
3.9
2.5
|
(OD06145)
(OD04590-01)
|
Ovarian Margin
0.5
0.3
Breast Cancer Mets
1.2
0.9
|
(OD06145)
(OD04590-03)
|
Ovarian cancer
0.9
0.5
Breast Cancer
48.6
34.4
|
(OD06455-03)
Metastasis
|
(OD04655-05)
|
Ovarian Margin
0.0
0.0
Breast Cancer
2.4
2.1
|
(OD06455-07)
064006
|
Normal Lung
0.0
0.0
Breast Cancer
55.1
43.8
|
9100266
|
Invasive poor diff.
9.2
7.5
Breast Margin
14.7
10.8
|
lung adeno
9100265
|
(ODO4945-01
|
Lung Margin
0.0
0.0
Breast Cancer
32.1
24.5
|
(ODO4945-03)
A209073
|
Lung Malignant
0.5
0.4
Breast Margin
9.1
6.4
|
Cancer (OD03126)
A2090734
|
Lung Margin
0.4
0.3
Breast cancer
69.7
61.6
|
(OD03126)
(OD06083)
|
Lung Cancer
0.0
0.0
Breast cancer node
28.5
23.3
|
(OD05014A)
metastasis
|
Lung Margin
0.8
0.6
Normal Liver
0.0
0.0
|
(OD05014B)
|
Lung cancer
44.8
0.3
Liver Cancer 1026
0.0
0.0
|
(OD06081)
|
Lung Margin
0.0
0.0
Liver Cancer 1025
0.8
0.6
|
(OD06081)
|
Lung Cancer
3.1
2.6
Liver Cancer 6004-
0.2
0.1
|
(OD04237-01)
|
Lung Margin
0.4
0.3
Liver Tissue 6004-
0.4
0.3
|
(OD04237-02)
N
|
Ocular Melanoma
0.0
0.0
Liver Cancer 6005-
0.0
0.0
|
Metastasis
T
|
Ocular Melanoma
0.0
0.0
Liver Tissue 6005-
0.0
0.0
|
Margin (Liver)
N
|
Melanoma
0.0
0.0
Liver Cancer
0.0
0.0
|
Metastatis
064003
|
Melanoma Margin
0.3
0.2
Normal Bladder
0.0
0.0
|
(Lung)
|
Normal Kidney
0.0
0.0
Bladder Cancer
3.2
2.3
|
1023
|
Kidney Ca, Nuclear
1.5
1.2
Bladder Cancer
4.5
3.2
|
grade (OD04338)
A302173
|
Kidney Margin
0.4
0.3
Normal Stomach
0.0
0.0
|
(OD04338)
|
Kidney Ca Nuclear
0.0
0.0
Gastric Cancer
0.5
0.3
|
grade 1/2
9060397
|
(OD04339)
|
Kidney Margin
0.0
0.0
Stomach Margin
2.1
1.4
|
(OD04339)
9060396
|
Kidney Ca, Clear
0.0
0.0
Gastric Cancer
2.5
1.7
|
cell type
9060395
|
(OD04340)
|
Kidney Margin
0.4
0.3
Stomach Margin
1.8
1.1
|
(OD04340)
9060394
|
Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0
|
grade (OD04348)
064005
|
|
[1206]
560
TABLE AJD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3729,
Rel. Exp. (%) Ag3729,
|
Tissue Name
Run 170222887
Tissue Name
Run 170222887
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
26.8
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
25.5
|
Primary Tr1 rest
0.0
Small airway epithelium
46.7
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
6.7
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
100.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
55.9
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
82.9
|
LAK cells
0.0
NCI-H292 IL-13
58.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
60.3
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
7.4
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
3.1
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monycytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
0.0
|
Macrophages rest
0.0
Lung
6.3
|
Macrophages LPS
0.0
Thymus
7.8
|
HUVEC none
0.0
Kidney
2.6
|
HUVEC starved
0.0
|
|
[1207] CNS_neurodegeneration_v1.0 Summary: Ag3729 Expression of the CG91434-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1208] General_screening_panel_v1.4 Summary: Ag3729 Results from one experiment with this probe and primer and the CG91434-01 gene are not included. The data suggests that there were experimental difficulties with this run.
[1209] General_screening_panel_v1.5 Summary: Ag5999 Results from one experiment with this probe and primer and the CG91434-01 gene are not included. The data suggests that there were experimental difficulties with this run.
[1210] Panel 2.2 Summary: Ag3729 Highest expression of the CG91434-01 gene is seen in breast cancer (CT=29). In addition, this gene is more highly expressed in breast cancer than in the normal adjacent tissue. Thus, expression of this gene could be used to differentiate between the breast cancer samples and other samples on this panel and as a marker for breast cancer. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of breast cancer (Rekha G K, Devaraj V R, Sreerama L, Lee M J, Nagasawa H T, Sladek N E. Inhibition of human class 3 aldehyde dehydrogenase, and sensitization of tumor cells that express significant amounts of this enzyme to oxazaphosphorines, by chlorpropamide analogues. Biochem Pharmacol Feb. 15, 1998;55(4):465-74).
[1211] Panel 4.1D Summary: Ag3729 Expression of the CG91434-01 gene is restricted to a few samples, with highest expression is seen in untreated NCI-H292 cells (CT=31.4). The gene is also expressed in a cluster of treated and untreated samples derived from the NCI-H292 cell line, a human airway epithelial cell line that produces mucins. Mucus overproduction is an important feature of bronchial asthma and chronic obstructive pulmonary disease samples. Interestingly, the transcript is also expressed at lower but still significant levels in small airway and bronchial epithelium treated with IL-1 beta and TNF-alpha and untreated small airway epithelium. The expression of the transcript in this mucoepidermoid cell line that is often used as a model for airway epithelium (NCI-H292 cells) suggests that this transcript may be important in the proliferation or activation of airway epithelium. Therefore, therapeutics designed with the protein encoded by the transcript may reduce or eliminate symptoms caused by inflammation in lung epithelia in chronic obstructive pulmonary disease, asthma, allergy, and emphysema.
[1212] AK CG91484-01: GPCR
[1213] Expression of gene CG91484-01 was assessed using the primer-probe set Ag3732, described in Table AKA.
561TABLE AKA
|
|
Probe Name Ag3732
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-acaatgattctgccatttatgg-3′22238345
ProbeTET-5′-cctcccaacttcaagacaatcactgg-26263346
3′-TAMRA
Reverse5′-ccattgaagaccttgcacaa-3′20304347
|
[1214] CNS_neurodegeneration_v1.0 Summary: Ag3732 Expression of the CG91484-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1215] General_screening_panel_v1.4 Summary: Ag3732 Expression of the CG91484-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1216] Panel 4.1D Summary: Ag3732 Expression of the CG91484-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1217] AL. CG91514-01: Telokin
[1218] Expression of gene CG91514-01 was assessed using the primer-probe sets Ag3772 and Ag3795, described in Tables ALA and ALB. Results of the RTQ-PCR runs are shown in Tables ALC, ALD, ALE, ALF and ALG.
562TABLE ALA
|
|
Probe Name Ag3772
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-actccctttggaatgcaataaa-3′22602348
ProbeTET-5′-attgtgtgccctgcttgccctcat-3′-24630349
TAMRA
Reverse5′-ctccacagacctccaacca-3′19660350
|
[1219]
563
TABLE ALB
|
|
|
Probe Name Ag3795
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-cctgcatcgtcacaggatac-3′
20
312
351
|
Probe
TET-
26
334
352
|
5′-agagccagaggtgacctggtacaagg-3′-
|
TAMRA
|
Reverse
5′-catattttggcaagccacag-3′
20
382
353
|
|
[1220]
564
TABLE ALC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3772, Run
Rel. Exp. (%) Ag3772, Run
|
Tissue Name
211176609
Tissue Name
211176609
|
|
AD 1 Hippo
11.5
Control (Path) 3
5.4
|
Temporal Ctx
|
AD 2 Hippo
30.1
Control (Path) 4
17.7
|
Temporal Ctx
|
AD 3 Hippo
5.5
AD 1 Occipital Ctx
4.3
|
AD 4 Hippo
6.9
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
3.5
|
AD 6 Hippo
75.8
AD 4 Occipital Ctx
16.2
|
Control 2 Hippo
14.0
AD 5 Occipital Ctx
22.5
|
Control 4 Hippo
25.7
AD 6 Occipital Ctx
33.0
|
Control (Path) 3 Hippo
6.0
Control 1 Occipital Ctx
7.0
|
AD 1 Temporal Ctx
19.5
Control 2 Occipital Ctx
55.5
|
AD 2 Temporal Ctx
39.2
Control 3 Occipital Ctx
6.1
|
AD 3 Temporal Ctx
2.9
Control 4 Occipital Ctx
7.3
|
AD 4 Temporal Ctx
13.9
Control (Path) 1
76.3
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
42.0
Control (Path) 2
14.3
|
Occipital Ctx
|
AD 5 Sup Temporal
36.3
Control (Path) 3
2.7
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
71.2
Control (Path) 4
14.1
|
Occipital Ctx
|
AD 6 Sup Temporal
93.3
Control 1 Parietal Ctx
3.3
|
Ctx
|
Control 1 Temporal Ctx
5.9
Control 2 Parietal Ctx
34.9
|
Control 2 Temporal Ctx
18.0
Control 3 Parietal Ctx
12.3
|
Control 3 Temporal Ctx
14.9
Control (Path) 1
0.0
|
Parietal Ctx
|
Control 3 Temporal Ctx
12.4
Control (Path) 2
21.6
|
Parietal Ctx
|
Control (Path) 1
48.0
Control (Path) 3
3.4
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
20.0
Control (Path) 4
24.3
|
Temporal Ctx
Parietal Ctx
|
|
[1221]
565
TABLE ALD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3772, Run
Rel. Exp. (%) Ag3772, Run
|
Tissue Name
219514529
Tissue Name
219514529
|
|
Adipose
2.3
Renal ca. TK-10
18.4
|
Melanoma* Hs688(A).T
0.1
Bladder
4.0
|
Melanoma* Hs688(B).T
0.1
Gastric ca. (liver met.)
0.9
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.1
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
3.0
|
Melanoma* SK-MEL-5
0.1
Colon ca. SW480
0.1
|
Squamous cell
1.9
Colon ca.* (SW480 met)
0.4
|
carcinoma SCC-4
SW620
|
Testis Pool
2.0
Colon ca. HT29
22.1
|
Prostate ca.* (bone met)
0.1
Colon ca. HCT-116
0.4
|
PC-3
|
Prostate Pool
0.8
Colon ca. CaCo-2
0.4
|
Placenta
7.0
Colon cancer tissue
1.1
|
Uterus Pool
0.4
Colon ca. SW1116
0.2
|
Ovarian ca. OVCAR-3
0.1
Colon ca. Colo-205
0.2
|
Ovarian ca. SK-OV-3
0.5
Colon ca. SW-48
0.1
|
Ovarian ca. OVCAR-4
0.2
Colon Pool
1.7
|
Ovarian ca. OVCAR-5
2.5
Small Intestine Pool
1.6
|
Ovarian ca. IGROV-1
1.0
Stomach Pool
1.0
|
Ovarian ca. OVCAR-8
0.4
Bone Marrow Pool
1.1
|
Ovary
0.5
Fetal Heart
59.5
|
Breast ca. MCF-7
0.8
Heart Pool
4.8
|
Breast ca. MDA-MB-
0.1
Lymph Node Pool
2.3
|
231
|
Breast ca. BT 549
2.0
Fetal Skeletal Muscle
32.8
|
Breast ca. T47D
2.2
Skeletal Muscle Pool
100.0
|
Breast ca. MDA-N
0.0
Spleen Pool
0.9
|
Breast Pool
3.2
Thymus Pool
1.7
|
Trachea
1.7
CNS cancer (glio/astro)
0.1
|
U87-MG
|
Lung
0.4
CNS cancer (glio/astro) U-
0.3
|
118-MG
|
Fetal Lung
18.0
CNS cancer (neuro; met)
0.2
|
SK-N-AS
|
Lung ca. NCI-N417
2.0
CNS cancer (astro) SF-539
0.2
|
Lung ca. LX-1
0.3
CNS cancer (astro) SNB-75
0.2
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.9
|
Lung ca. SHP-77
17.7
CNS cancer (glio) SF-295
0.8
|
Lung ca. A549
0.3
Brain (Amygdala) Pool
0.6
|
Lung ca. NCI-H526
0.9
Brain (cerebellum)
0.8
|
Lung ca. NCI-H23
0.3
Brain (fetal)
1.5
|
Lung ca. NCI-H460
0.1
Brain (Hippocampus) Pool
1.0
|
Lung ca. HOP-62
0.2
Cerebral Cortex Pool
0.5
|
Lung ca. NCI-H522
0.1
Brain (Substantia nigra)
0.5
|
Pool
|
Liver
0.4
Brain (Thalamus) Pool
1.0
|
Fetal Liver
4.9
Brain (whole)
1.1
|
Liver ca. HepG2
0.0
Spinal Cord Pool
0.8
|
Kidney Pool
2.4
Adrenal Gland
7.3
|
Fetal Kidney
8.7
Pituitary gland Pool
4.8
|
Renal ca. 786-0
7.7
Salivary Gland
0.5
|
Renal ca. A498
0.6
Thyroid (female)
0.8
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
8.6
|
Renal ca. UO-31
5.7
Pancreas Pool
2.3
|
|
[1222]
566
TABLE ALE
|
|
|
General_screening_panel_v1.5
|
Rel. Exp. (%) Ag3795, Run
Rel. Exp. (%) Ag3795, Run
|
Tissue Name
258001456
Tissue Name
258001456
|
|
Adipose
0.9
Renal ca. TK-10
12.6
|
Melanoma* Hs688(A).T
0.0
Bladder
2.2
|
Melanoma* Hs688(B).T
0.1
Gastric ca. (liver met.)
0.2
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
1.3
|
Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.1
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.1
|
carcinoma SCC-4
SW620
|
Testis Pool
0.5
Colon ca. HT29
13.1
|
Prostate ca.* (bone met)
0.0
Colon ca. HCT-116
0.1
|
PC-3
|
Prostate Pool
0.6
Colon ca. CaCo-2
0.2
|
Placenta
0.3
Colon cancer tissue
0.7
|
Uterus Pool
0.2
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
0.1
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
0.4
|
Ovarian ca. OVCAR-5
0.3
Small Intestine Pool
0.6
|
Ovarian ca. IGROV-1
0.3
Stomach Pool
0.5
|
Ovarian ca. OVCAR-8
0.2
Bone Marrow Pool
0.1
|
Ovary
0.1
Fetal Heart
13.1
|
Breast ca. MCF-7
0.3
Heart Pool
2.6
|
Breast ca. MDA-MB-
0.1
Lymph Node Pool
0.0
|
231
|
Breast ca. BT 549
0.1
Fetal Skeletal Muscle
6.7
|
Breast ca. T47D
0.0
Skeletal Muscle Pool
100.0
|
Breast ca. MDA-N
0.0
Spleen Pool
0.5
|
Breast Pool
0.6
Thymus Pool
0.5
|
Trachea
0.4
CNS cancer (glio/astro)
0.0
|
U87-MG
|
Lung
0.0
CNS cancer (glio/astro) U-
0.1
|
118-MG
|
Fetal Lung
4.2
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.1
|
Lung ca. LX-1
0.1
CNS cancer (astro) SNB-75
0.1
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.4
|
Lung ca. SHP-77
12.8
CNS cancer (glio) SF-295
0.4
|
Lung ca. A549
0.0
Brain (Amygdala) Pool
0.4
|
Lung ca. NCI-H526
0.4
Brain (cerebellum)
0.3
|
Lung ca. NCI-H23
0.1
Brain (fetal)
0.5
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
0.5
|
Lung ca. HOP-62
0.1
Cerebral Cortex Pool
0.3
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
0.3
|
Pool
|
Liver
0.2
Brain (Thalamus) Pool
0.5
|
Fetal Liver
1.8
Brain (whole)
0.8
|
Liver ca. HepG2
0.0
Spinal Cord Pool
0.4
|
Kidney Pool
0.8
Adrenal Gland
3.5
|
Fetal Kidney
1.7
Pituitary gland Pool
2.5
|
Renal ca. 786-0
6.4
Salivary Gland
0.3
|
Renal ca. A498
0.1
Thyroid (female)
0.3
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
5.2
|
Renal ca. UO-31
2.2
Pancreas Pool
0.7
|
|
[1223]
567
TABLE ALF
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3795,
Rel. Exp. (%) Ag3795,
|
Tissue Name
Run 172208559
Tissue Name
Run 172208559
|
|
Secondary Th1 act
0.4
HUVEC IL-1beta
97.3
|
Secondary Th2 act
0.5
HUVEC IFN gamma
92.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
23.7
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
88.3
|
Secondary Th2 rest
1.3
HUVEC IL-11
66.4
|
Secondary Tr1 rest
0.9
Lung Microvascular EC none
100.0
|
Primary Th1 act
0.0
Lung Microvascular EC
68.3
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.9
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
76.3
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
1.5
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
1.3
|
Primary Tr1 rest
0.5
Small airway epithelium
0.6
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.5
|
act
|
CD45RO CD4 lymphocyte
3.3
Coronery artery SMC TNF alpha +
2.1
|
act
IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
2.9
|
Secondary CD8
0.4
Astrocytes TNF alpha + IL-1beta
4.8
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.8
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.5
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
7.2
|
LAK cells IL-2 + IL-12
0.9
NCI-H292 none
1.4
|
LAK cells IL-2 + IFN
1.5
NCI-H292 IL-4
1.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
1.0
|
LAK cells
0.3
NCI-H292 IL-13
0.6
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.6
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
46.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
6.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
1.2
|
PBMC rest
0.2
Lung fibroblast TNF alpha + IL-
0.9
|
1beta
|
PBMC PWM
1.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
1.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.5
Dermal fibroblast CCD1070 rest
1.8
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
1.5
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.3
Dermal fibroblast CCD1070 IL-
0.6
|
1beta
|
EOL-1 dbcAMP
0.4
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
2.3
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
1.1
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
1.6
Neutrophils TNFa + LPS
0.5
|
Monocytes rest
0.0
Neutrophils rest
2.6
|
Monocytes LPS
0.0
Colon
3.6
|
Macrophages rest
11.0
Lung
17.0
|
Macrophages LPS
1.1
Thymus
6.1
|
HUVEC none
66.9
Kidney
30.8
|
HUVEC starved
75.8
|
|
[1224]
568
TABLE ALG
|
|
|
Panel 5 Islet
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3795, Run
Ag3795, Run
|
Tissue Name
256791465
Tissue Name
256791465
|
|
97457_Patient-
0.3
94709_Donor 2 AM - A_adipose
0.5
|
02go_adipose
|
97476_Patient-07sk_skeletal
5.1
94710_Donor 2 AM - B_adipose
0.0
|
muscle
|
97477_Patient-07ut_uterus
0.0
94711_Donor 2 AM - C_adipose
0.0
|
97478_Patient-
0.4
94712_Donor 2 AD - A_adipose
0.0
|
07pl_placenta
|
99167_Bayer Patient 1
42.9
94713_Donor 2 AD - B_adipose
0.1
|
97482_Patient-08ut_uterus
0.5
94714_Donor 2 AD - C_adipose
0.4
|
97483_Patient-
0.7
94742_Donor 3 U - A_Mesenchymal
0.1
|
08pl_placenta
Stem Cells
|
97486_Patient-09sk_skeletal
8.1
94743_Donor 3 U - B_Mesenchymal
0.0
|
muscle
Stem Cells
|
97487_Patient-09ut_uterus
0.2
94730_Donor 3 AM - A_adipose
0.3
|
97488_Patient-
0.4
94731_Donor 3 AM - B_adipose
0.3
|
09pl_placenta
|
97492_Patient-10ut_uterus
0.5
94732_Donor 3 AM - C_adipose
0.4
|
97493_Patient-
2.6
94733_Donor 3 AD - A_adipose
0.1
|
10pl_placenta
|
97495_Patient-
0.5
94734_Donor 3 AD - B_adipose
0.0
|
11go_adipose
|
97496_Patient-11sk_skeletal
48.0
94735_Donor 3 AD - C_adipose
0.0
|
muscle
|
97497_Patient-11ut_uterus
0.8
77138_Liver_HepG2untreated
0.3
|
97498_Patient-
0.6
73556_Heart_Cardiac stromal cells
5.8
|
11pl_placenta
(primary)
|
97500_Patient-
0.8
81735_Small Intestine
2.5
|
12go_adipose
|
97501_Patient-12sk_skeletal
100.0
72409_Kidney_Proximal Convoluted
1.0
|
muscle
Tubule
|
97502_Patient-12ut_uterus
0.0
82685_Small intestine_Duodenum
0.0
|
97503_Patient-
0.3
90650_Adrenal_Adrenocortical
2.4
|
12pl_placenta
adenoma
|
94721_Donor 2 U -
0.0
72410_Kidney_HRCE
5.2
|
A_Mesenchymal Stem Cells
|
94722_Donor 2 U -
0.1
72411_Kidney_HRE
16.0
|
B_Mesenchymal Stem Cells
|
94723_Donor 2 U -
0.1
73139_Uterus_Uterine smooth
0.0
|
C_Mesenchymal Stem Cells
muscle cells
|
|
[1225] CNS_neurodegeneration_v1.0 Summary: Ag3772 This panel does not show differential expression of the CG91514-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the hippocampus of an Alzheimer's patient (CT=30.3). Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1226] General_screening_panel_v1.4 Summary: Ag3772 Highest expression of the CG91514-01, a telokin homolog, is seen in skeletal muscle (CT=24.6). This gene is ubiquitously expressed in this panel, with high levels of expression also seen in fetal heart, skeletal muscle and lung and renal, lung and colon cancer cell lines. Thus, expression of this gene could be used to differentiate between skeletal muscle and other samples on this panel and as a marker of skeletal muscle. Telokin is the name for the carboxy-terminal 154 codons of muscle light chain kinase that are expressed as an independent protein in smooth muscle. This expression profile supports the identification of this gene product as a telokin homolog. This gene is also expressed in other metabolic tissues including pancreas, thyroid, pituitary, adrenal, heart, and adult and fetal liver. This expression profile suggests that this gene product may be involved in the pathogenesis and/or treatment of metabolic disorders, including obesity and type 2 diabetes.
[1227] In addition, this gene is expressed at much higher levels in fetal lung and heart tissue (CTs=25-27) when compared to expression in the adult counterpart (CTs=29-32). Thus, expression of this gene may be used to differentiate between the fetal and adult source of these tissues.
[1228] This gene is also expressed at moderate to low levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1229] Results from a second experiment with the probe/primer set Ag3795 are not included. The amp plot indicates there were experimental difficulties with this run.
[1230] General_screening_panel_v1.5 Summary: Ag3795 Highest expression of the CG91514-01 gene is seen in skeletal muscle (CT=26.6). Expression in this panel is in excellent agreement with expression in Panel 1.4. Please see that Panel 1.4 for discussion of utility of this gene in the central nervous system and metabolic disorders.
[1231] Panel 4.1D Summary: Ag3795 Highest expression of the CG91514-01 gene is seen in untreated lung microvacular EC (CT=31). Significant expression of this gene appears to be limited to a cluster of cells derived from treated and untreated HUVEC and lung microvasculature and untreated dermal EC. Endothelial cells are known to play important roles in inflammatory responses by altering the expression of surface proteins that are involved in activation and recruitment of effector inflammatory cells. The expression of this gene in dermal microvascular endothelial cells suggests that this protein product may be involved in inflammatory responses to skin disorders, including psoriasis. Expression in lung microvascular endothelial cells suggests that the protein encoded by this transcript may also be involved in lung disorders including asthma, allergies, chronic obstructive pulmonary disease, and emphysema. Therefore, therapeutic modulation of the protein encoded by this gene may lead to amelioration of symptoms associated with psoriasis, asthma, allergies, chronic obstructive pulmonary disease, and emphysema.
[1232] Expression of this transcript in the microvasculature of the lung and the dermis also suggests a role for this gene in the maintenance of the integrity of the microvasculature. Therefore, therapeutics designed for this putative protein could be beneficial for the treatment of diseases associated with damaged microvasculature including heart diseases or inflammatory diseases, such as psoriasis, asthma, and chronic obstructive pulmonary diseases.
[1233] Results from a second experiment with the probe and primer set Ag3772 are not included. The amp plot indicates that there were experimental difficulties with this run.
[1234] Panel 5 Islet Summary: Ag3795 Expression of the CG91514-05 gene confirms the expression seen in Panels 1.4 and 1.5, with prominent expression in skeletal muscle (CTs=29-33). Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1235] AM. CG91587-01: Tripartite Motif Protein TRIM4 Isoform Alpha
[1236] Expression of gene CG91587-01 was assessed using the primer-probe sets Agl 115, Agl 166 and Ag759, described in Tables AMA, AMB and AMC. Results of the RTQ-PCR runs are shown in Table AMD.
569TABLE AMA
|
|
Probe Name Ag1115
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-gccaactgcttttgtagagaga-3′221150354
ProbeTET-5′-tcagcacttaccctgtgttctgggaa-261174355
3′-TAMRA
Reverse5′-aatgtttccctgaggtgaaaac-3′221205356
|
[1237]
570
TABLE AMB
|
|
|
Probe Name Ag1166
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-gccaactgcttttgtagagaga-3′
22
1150
357
|
Probe
TET-5′-tcagcacttaccctgtgttctgggaa-
26
1174
358
|
3′-TAMRA
|
Reverse
5′-aatgtttccctgaggtgaaaac-3′
22
1205
359
|
|
[1238]
571
TABLE AMC
|
|
|
Probe Name Ag759
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-agtgcacctgcacaccttt-3′
19
1480
360
|
Probe
TET-5′-cttgttcttctgtctcacgcctccg-
25
1500
361
|
3′-TAMRA
|
Reverse
5′-atcagtcactggtggaatgact-3′
22
1558
362
|
|
[1239]
572
TABLE AMD
|
|
|
Panel 4D
|
Rel. Exp. (%) Ag1166,
Rel. Exp. (%) Ag1166,
|
Tissue Name
Run 140171235
Tissue Name
Run 140171235
|
|
Secondary Th1 act
39.5
HUVEC IL-1beta
8.1
|
Secondary Th2 act
38.4
HUVEC IFN gamma
31.0
|
Secondary Tr1 act
41.5
HUVEC TNF alpha + IFN
11.5
|
gamma
|
Secondary Th1 rest
20.9
HUVEC TNF alpha + IL4
18.4
|
Secondary Th2 rest
26.1
HUVEC IL-11
10.7
|
Secondary Tr1 rest
24.3
Lung Microvascular EC none
31.6
|
Primary Th1 act
29.5
Lung Microvascular EC
20.6
|
TNF alpha + IL-1beta
|
Primary Th2 act
40.9
Microvascular Dermal EC none
73.7
|
Primary Tr1 act
52.1
Microsvasular Dermal EC
45.7
|
TNF alpha + IL-1beta
|
Primary Th1 rest
82.4
Bronchial epithelium TNF
44.4
|
alpha + IL1beta
|
Primary Th2 rest
44.8
Small airway epithelium none
13.5
|
Primary Tr1 rest
54.3
Small airway epithelium
67.8
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
15.6
Coronery artery SMC rest
19.9
|
act
|
CD45RO CD4 lymphocyte
33.0
Coronery artery SMC TNF
24.3
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
28.9
Astrocytes rest
9.9
|
Secondary CD8
33.7
Astrocytes TNF alpha + IL-1beta
21.3
|
lymphocyte rest
|
Secondary CD8
18.3
KU-812 (Basophil) rest
27.0
|
lymphocyte act
|
CD4 lymphocyte none
18.3
KU-812 (Basophil)
58.2
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
42.9
CCD1106 (Keratinocytes) none
5.4
|
CD95 CH11
|
LAK cells rest
43.2
CCD1106 (Keratinocytes)
17.9
|
TNF alpha + IL-1beta
|
LAK cells IL-2
42.3
Liver cirrhosis
7.5
|
LAK cells IL-2 + IL-12
42.3
Lupus kidney
16.4
|
LAK cells IL-2 + IFN
41.8
NCI-H292 none
50.3
|
gamma
|
LAK cells IL-2 + IL-18
36.6
NCI-H292 IL-4
40.1
|
LAK cells
13.0
NCI-H292 IL-9
42.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
21.3
NCI-H292 IL-13
34.9
|
Two Way MLR 3 day
41.2
NCI-H292 IFN gamma
19.3
|
Two Way MLR 5 day
30.4
HPAEC none
22.1
|
Two Way MLR 7 day
19.5
HPAEC TNF alpha + IL-1beta
17.1
|
PBMC rest
12.8
Lung fibroblast none
17.8
|
PBMC PWM
100.0
Lung fibroblast TNF alpha + IL-
11.8
|
1beta
|
PBMC PHA-L
77.4
Lung fibroblast IL-4
33.4
|
Ramos (B cell) none
59.9
Lung fibroblast IL-9
19.1
|
Ramos (B cell) ionomycin
88.9
Lung fibroblast IL-13
51.8
|
B lymphocytes PWM
34.4
Lung fibroblast IFN gamma
40.3
|
B lymphocytes CD40L
17.7
Dermal fibroblast CCD1070 rest
29.9
|
and IL-4
|
EOL-1 dbcAMP
20.9
Dermal fibroblast CCD1070
46.7
|
TNF alpha
|
EOL-1 dbcAMP
24.3
Dermal fibroblast CCD1070 IL-
23.2
|
PMA/ionomycin
1beta
|
Dendritic cells none
25.2
Dermal fibroblast IFN gamma
20.2
|
Dendritic cells LPS
19.2
Dermal fibroblast IL-4
38.4
|
Dendritic cells anti-CD40
32.5
IBD Colitis 2
3.4
|
Monocytes rest
26.4
IBD Crohn's
3.0
|
Monocytes LPS
31.6
Colon
17.1
|
Macrophages rest
35.4
Lung
23.7
|
Macrophages LPS
30.4
Thymus
87.1
|
HUVEC none
21.5
Kidney
43.2
|
HUVEC starved
33.4
|
|
[1240] Panel 4D Summary: Ag1166 Highest expression of the CG91587-01 gene is detected in PWM treated PBMC cells (CT=28.8). This gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1241] AN. CG91911-01: Mitogen-Activated Protein Kinase Kinase Kinase 8 (MAP3K8)
[1242] Expression of gene CG91911-01 was assessed using the primer-probe sets Ag3116, Ag3551 and Ag4828, described in Tables ANA, ANB and ANC. Results of the RTQ-PCR runs are shown in Tables AND, ANE, ANF, ANG, ANH and ANI.
573TABLE ANA
|
|
Probe Name Ag3116
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-catgttctcaagggacttgatt-3′22802363
ProbeTET-5′-26829364
cactcaaagaaagtgatccatcatga-
3′-TAMRA
Reverse5′-ttttgtggacatgaaaacaatg-3′22870365
|
[1243]
574
TABLE ANB
|
|
|
Probe Name Ag3551
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-catgttctcaagggacttgatt-3′
22
802
366
|
Probe
TET-5′-
26
829
367
|
cactcaaagaaagtgatccatcatga-
|
3′-TAMRA
|
Reverse
5′-ttttgtggacatgaaaacaatg-3′
22
870
368
|
|
[1244]
575
TABLE ANC
|
|
|
Probe Name Ag4828
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-gaggaatctgagatgctcaaga-3′
22
1393
369
|
Probe
TET-5′-caacgctctctctacatcgacctcgg-
26
1417
370
|
3′-TAMRA
|
Reverse
5′-tccccgaacaagattgaagt-3′
20
1457
371
|
|
[1245]
576
TABLE AND
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3551,
Rel. Exp. (%) Ag3551,
|
Tissue Name
Run 209990366
Tissue Name
Run 209990366
|
|
AD 1 Hippo
20.0
Control (Path) 3
14.6
|
Temporal Ctx
|
AD 2 Hippo
44.1
Control (Path) 4
18.8
|
Temporal Ctx
|
AD 3 Hippo
7.1
AD 1 Occipital Ctx
13.5
|
AD 4 Hippo
5.6
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
100.0
AD 3 Occipital Ctx
4.0
|
AD 6 Hippo
57.0
AD 4 Occipital Ctx
15.8
|
Control 2 Hippo
24.7
AD 5 Occipital Ctx
34.6
|
Control 4 Hippo
51.4
AD 6 Occipital Ctx
46.0
|
Control (Path) 3 Hippo
48.6
Control 1 Occipital Ctx
21.0
|
AD 1 Temporal Ctx
21.3
Control 2 Occipital Ctx
41.5
|
AD 2 Temporal Ctx
39.5
Control 3 Occipital Ctx
16.3
|
AD 3 Temporal Ctx
6.1
Control 4 Occipital Ctx
13.0
|
AD 4 Temporal Ctx
16.8
Control (Path) 1
95.3
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
10.2
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
91.4
Control (Path) 3
21.5
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
58.2
Control (Path) 4
24.0
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
65.5
Control 1 Parietal Ctx
17.2
|
Control 1 Temporal Ctx
20.3
Control 2 Parietal Ctx
57.4
|
Control 2 Temporal Ctx
21.2
Control 3 Parietal Ctx
16.5
|
Control 3 Temporal Ctx
10.8
Control (Path) 1
28.3
|
Parietal Ctx
|
Control 4 Temporal Ctx
6.9
Control (Path) 2
15.8
|
Parietal Ctx
|
Control (Path) 1
42.0
Control (Path) 3
19.6
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
26.4
Control (Path) 4
61.1
|
Temporal Ctx
Parietal Ctx
|
|
[1246]
577
TABLE ANE
|
|
|
General_screening_panel_v1.4
|
Rel.
Rel.
Rel.
Rel.
Rel.
Rel.
|
Exp. (%)
Exp. (%)
Exp. (%)
Exp. (%)
Exp. (%)
Exp. (%)
|
Ag3116,
Ag3551,
Ag4828,
Ag3116,
Ag3551,
Ag4828,
|
Tissue
Run
Run
Run
Run
Run
Run
|
Name
219923407
218328114
217081802
Tissue Name
219923407
218328114
217081802
|
|
Adipose
100.0
58.2
53.6
Renal ca. TK-10
6.4
8.2
10.6
|
Melanoma*
18.8
9.0
15.5
Bladder
32.5
24.1
31.9
|
Hs688(A).T
|
Melanoma*
21.3
10.7
17.4
Gastric ca. (liver
26.8
23.5
36.3
|
Hs688(B).T
met.) NCI-N87
|
Melanoma*
1.0
0.9
3.5
Gastric ca.
8.7
8.0
12.2
|
M14
KATO III
|
Melanoma*
2.9
1.5
3.2
Colon ca. SW-
2.6
2.6
5.4
|
LOXIMVI
948
|
Melanoma*
0.8
0.8
0.9
Colon ca.
13.5
12.3
25.0
|
SK-MEL-5
SW480
|
Squamous
1.0
2.2
7.0
Colon ca.*
1.6
1.4
2.5
|
cell
(SW480 met)
|
carcinoma
SW620
|
SCC-4
|
Testis Pool
3.5
3.3
4.7
Colon ca. HT29
7.2
5.7
14.3
|
Prostate ca.*
6.4
1.8
6.3
Colon ca. HCT-
2.1
1.7
2.1
|
(bone met)
116
|
PC-3
|
Prostate Pool
2.1
2.0
3.9
Colon ca. CaCo-2
13.5
15.7
15.9
|
Placenta
30.8
25.9
39.0
Colon cancer
34.9
42.3
39.8
|
tissue
|
Uters Pool
7.7
4.7
9.0
Colon ca.
0.1
0.3
3.4
|
SW1116
|
Ovarian ca.
4.4
6.1
15.7
Colon ca. Colo-
2.7
2.6
8.8
|
OVCAR-3
205
|
Ovarian ca.
9.7
18.2
46.3
Colon ca. SW-
3.3
4.7
5.4
|
SK-OV-3
48
|
Ovarian ca.
3.7
5.4
7.1
Colon Pool
16.6
9.8
16.2
|
OVCAR-4
|
Ovarian ca.
19.2
19.9
30.6
Small Intestine
7.3
5.5
9.3
|
OVCAR-5
Pool
|
Ovarian ca.
7.0
9.1
14.1
Stomach Pool
6.6
8.0
17.3
|
IGROV-1
|
Ovarian ca.
1.8
1.9
2.7
Bone Marrow
5.2
3.3
7.0
|
OVCAR-8
Pool
|
Ovary
2.7
2.5
4.5
Fetal Heart
4.5
4.6
2.9
|
Breast ca.
64.6
81.8
100.0
Heart Pool
9.2
6.8
7.9
|
MCF-7
|
Breast ca.
3.1
2.1
9.2
Lymph Node
10.4
9.9
15.2
|
MDA-MB-
Pool
|
231
|
Breast ca.
24.5
36.3
73.2
Fetal Skeletal
2.4
2.9
1.7
|
BT 549
Muscle
|
Breast ca.
37.4
60.3
66.0
Skeletal Muscle
7.7
8.5
9.8
|
T47D
Pool
|
Breast ca.
0.3
0.5
0.9
Spleen Pool
16.0
22.8
45.7
|
MDA-N
|
Breast Pool
33.2
9.8
24.1
Thymus Pool
7.5
6.9
15.9
|
Trachea
14.5
15.5
18.0
CNS cancer
2.1
2.4
7.6
|
(glio/astro)
|
U87-MG
|
Lung
4.2
3.4
6.7
CNS cancer
5.4
2.7
7.9
|
(glio/astro) U-
|
118-MG
|
Fetal Lung
83.5
100.0
68.3
CNS cancer
0.7
1.2
2.6
|
(neuro; met) SK-
|
N-AS
|
Lung ca.
0.0
0.0
0.2
CNS cancer
1.4
1.8
2.3
|
NCI-N417
(astro) SF-539
|
Lung ca.
8.0
6.0
11.8
CNS cancer
4.7
5.9
14.1
|
LX-1
(astro) SNB-75
|
Lung ca.
0.0
0.0
0.0
CNS cancer
6.2
10.7
11.1
|
NCI-H146
(glio) SNB-19
|
Lung ca.
0.0
0.0
0.1
CNS cancer
16.0
18.8
31.9
|
SHP-77
(glio) SF-295
|
Lung ca.
35.4
0.0
36.6
Brain
1.6
0.7
2.7
|
A549
(Amygdala)
|
Pool
|
Lung ca.
0.0
0.0
0.0
Brain
1.1
0.3
1.4
|
NCI-H526
(cerebellum)
|
Lung ca.
10.9
13.0
13.4
Brain (fetal)
6.0
4.1
4.9
|
NCI-H23
|
Lung ca.
7.4
5.8
17.6
Brain
3.6
1.5
3.7
|
NCI-H460
(Hippocampus)
|
Pool
|
Lung ca.
11.4
4.3
13.2
Cerebral Cortex
2.1
2.0
3.5
|
HOP-62
Pool
|
Lung ca.
1.6
1.5
2.1
Brain
2.4
2.0
2.7
|
NCI-H522
(Substantia
|
nigra) Pool
|
Liver
0.6
0.2
1.0
Brain
2.6
2.2
4.5
|
(Thalamus) Pool
|
Fetal Liver
5.0
4.0
2.8
Brain (whole)
2.7
2.5
4.5
|
Liver ca.
4.5
5.4
8.1
Spinal Cord
2.1
3.2
3.8
|
HepG2
Pool
|
Kidney Pool
26.6
21.0
31.4
Adrenal Gland
11.7
3.8
9.5
|
Fetal Kidney
9.0
10.7
7.7
Pituitary gland
0.7
0.7
1.4
|
Pool
|
Renal ca.
6.0
7.9
10.9
Salivary Gland
1.9
1.5
2.5
|
786-0
|
Renal ca.
1.2
2.3
5.2
Thyroid
3.3
3.6
7.7
|
A498
(female)
|
Renal ca.
1.9
0.8
2.5
Pancreatic ca.
14.9
21.9
34.4
|
ACHN
CAPAN2
|
Renal ca.
11.1
10.7
14.9
Pancreas Pool
15.0
17.8
19.6
|
UO-31
|
|
[1247]
578
TABLE ANF
|
|
|
Panel 1.3D
|
Rel. Exp. (%) Ag3116,
Rel. Exp. (%) Ag3116,
|
Tissue Name
Run 167617379
Tissue Name
Run 167617379
|
|
Liver adenocarcinoma
24.8
Kidney (fetal)
34.2
|
Pancreas
3.4
Renal ca. 786-0
3.7
|
Pancreatic ca. CAPAN 2
12.1
Renal ca. A498
3.3
|
Adrenal gland
2.6
Renal ca. RXF 393
17.1
|
Thyroid
1.3
Renal ca. ACHN
1.7
|
Salivary gland
0.0
Renal ca. UO-31
0.8
|
Pituitary gland
2.1
Renal ca. TK-10
4.4
|
Brain (fetal)
3.1
Liver
2.4
|
Brain (whole)
3.1
Liver (fetal)
4.5
|
Brain (amygdala)
1.0
Liver ca. (hepatoblast)
4.4
|
HepG2
|
Brain (cerebellum)
1.0
Lung
25.0
|
Brain (hippocampus)
3.0
Lung (fetal)
29.7
|
Brain (substantia nigra)
3.7
Lung ca. (small cell) LX-1
5.5
|
Brain (thalamus)
1.2
Lung ca. (small cell)
0.0
|
NCI-H69
|
Cerebral Cortex
2.5
Lung ca. (s.cell var.)
0.0
|
SHP-77
|
Spinal cord
3.0
Lung ca. (large cell)NCI-
2.3
|
H460
|
glio/astro U87-MG
1.5
Lung ca. (non-sm. cell)
14.3
|
A549
|
glio/astro U-118-MG
2.8
Lung ca. (non-s.cell)
5.0
|
NCI-H23
|
astrocytoma SW1783
2.0
Lung ca. (non-s.cell)
5.7
|
HOP-62
|
neuro*; met SK-N-AS
1.5
Lung ca. (non-s.cl) NCI-
1.2
|
H522
|
astrocytoma SF-539
2.4
Lung ca. (squam.) SW
24.1
|
900
|
astrocytoma SNB-75
14.5
Lung ca. (squam.) NCI-
0.0
|
H596
|
glioma SNB-19
0.0
Mammary gland
7.7
|
glioma U251
0.7
Breast ca.* (pl.ef) MCF-7
57.8
|
glioma SF-295
6.9
Breast ca.* (pl.ef) MDA-
0.8
|
MB-231
|
Heart (fetal)
5.8
Breast ca.* (pl.ef) T47D
3.5
|
Heart
3.2
Breast ca. BT-549
4.8
|
Skeletal muscle (fetal)
4.6
Breast ca. MDA-N
0.0
|
Skeletal muscle
2.1
Ovary
6.1
|
Bone marrow
4.0
Ovarian ca. OVCAR-3
3.0
|
Thymus
3.4
Ovarian ca. OVCAR-4
26.1
|
Spleen
10.6
Ovarian ca. OVCAR-5
44.8
|
Lymph node
10.3
Ovarian ca. OVCAR-8
1.4
|
Colorectal
6.4
Ovarian ca. IGROV-1
6.4
|
Stomach
1.8
Ovarian ca.* (ascites)
33.2
|
SK-OV-3
|
Small intestine
3.0
Uterus
4.4
|
Colon ca. SW480
6.0
Placenta
6.8
|
Colon ca.* SW620(SW480
6.1
Prostate
0.0
|
met)
|
Colon ca. HT29
6.6
Prostate ca.* (bone
2.1
|
met)PC-3
|
Colon ca. HCT-116
0.0
Testis
0.0
|
Colon ca. CaCo-2
11.3
Melanoma Hs688(A).T
1.0
|
Colon ca.
13.1
Melanoma* (met)
3.5
|
tissue(ODO3866)
Hs688(B).T
|
Colon ca. HCC-2998
17.6
Melanoma UACC-62
0.0
|
Gastric ca.* (liver met)
11.0
Melanoma M14
1.1
|
NCI-N87
|
Bladder
10.2
Melanoma LOX IMVI
1.2
|
Trachea
3.9
Melanoma* (met) SK-
0.0
|
MEL-5
|
Kidney
5.0
Adipose
100.0
|
|
[1248]
579
TABLE ANG
|
|
|
Panel 2D
|
Rel. Exp. (%) Ag3116,
Rel. Exp. (%) Ag3116,
|
Tissue Name
Run 169556216
Tissue Name
Run 169556216
|
|
Normal Colon
58.2
Kidney Margin 8120608
2.0
|
CC Well to Mod Diff
22.7
Kidney Cancer 8120613
3.5
|
(ODO3866)
|
CC Margin (ODO3866)
14.4
Kidney Margin 8120614
2.9
|
CC Gr.2 rectosigmoid
7.5
Kidney Cancer 9010320
42.0
|
(ODO3868)
|
CC Margin (ODO3868)
3.4
Kidney Margin 9010321
7.7
|
CC Mod Diff (ODO3920)
7.0
Normal Uterus
7.0
|
CC Margin (ODO3920)
6.9
Uterus Cancer 064011
18.8
|
CC Gr.2 ascend colon
27.7
Normal Thyroid
5.8
|
(ODO3921)
|
CC Margin (ODO3921)
8.4
Thyroid Cancer 064010
6.9
|
CC from Partial Hepatectomy
34.9
Thyroid Cancer A302152
3.0
|
(ODO4309) Mets
|
Liver Margin (ODO4309)
8.5
Thyroid Margin A302153
12.1
|
Colon mets to lung (OD04451-
12.2
Normal Breast
28.9
|
01)
|
Lung Margin (OD04451-02)
21.8
Breast Cancer (OD04566)
6.3
|
Normal Prostate 6546-1
2.9
Breast Cancer (OD04590-
44.4
|
01)
|
Prostate Cancer (OD04410)
7.4
Breast Cancer Mets
43.5
|
(OD04590-03)
|
Prostate Margin (OD04410)
8.2
Breast Cancer Metastasis
6.9
|
(OD04655-05)
|
Prostate Cancer (OD04720-01)
6.6
Breast Cancer 064006
12.0
|
Prostate Margin (OD04720-02)
21.8
Breast Cancer 1024
12.9
|
Normal Lung 061010
42.6
Breast Cancer 9100266
6.9
|
Lung Met to Muscle
15.0
Breast Margin 9100265
6.9
|
(ODO4286)
|
Muscle Margin (ODO4286)
9.5
Breast Cancer A209073
7.2
|
Lung Malignant Cancer
17.4
Breast Margin A209073
4.3
|
(OD03126)
|
Lung Margin (OD03126)
59.5
Normal Liver
2.3
|
Lung Cancer (OD04404)
53.6
Liver Cancer 064003
2.1
|
Lung Margin (OD04404)
45.1
Liver Cancer 1025
5.8
|
Lung Cancer (OD04565)
10.4
Liver Cancer 1026
4.2
|
Lung Margin (OD04565)
10.8
Liver Cancer 6004-T
6.1
|
Lung Cancer (OD04237-01)
39.8
Liver Tissue 6004-N
6.4
|
Lung Margin (OD04237-02)
65.5
Liver Cancer 6005-T
7.4
|
Ocular Mel Met to Liver
1.6
Liver Tissue 6005-N
3.9
|
(ODO4310)
|
Liver Margin (ODO4310)
9.9
Normal Bladder
37.1
|
Melanoma Mets to Lung
2.0
Bladder Cancer 1023
6.5
|
(OD04321)
|
Lung Margin (OD04321)
50.7
Bladder Cancer A302173
14.8
|
Normal Kidney
13.0
Bladder Cancer
27.9
|
(OD04718-01)
|
Kidney Ca, Nuclear grade 2
16.4
Bladder Normal Adjacent
100.0
|
(OD04338)
(OD04718-03)
|
Kidney Margin (OD04338)
18.4
Normal Ovary
6.3
|
Kidney Ca Nuclear grade 1/2
10.3
Ovarian Cancer 064008
31.9
|
(OD04339)
|
Kidney Margin (OD04339)
6.5
Ovarian Cancer
21.9
|
(OD04768-07)
|
Kidney Ca, Clear cell type
28.7
Ovary Margin (OD04768-
32.5
|
(OD04340)
08)
|
Kidney Margin (OD04340)
22.7
Normal Stomach
18.8
|
Kidney Ca, Nuclear grade 3
4.5
Gastric Cancer 9060358
14.6
|
(OD04348)
|
Kidney Margin (OD04348)
6.7
Stomach Margin 9060359
16.2
|
Kidney Cancer (OD04622-01)
12.2
Gastric Cancer 9060395
33.2
|
Kidney Margin (OD04622-03)
1.8
Stomach Margin 9060394
24.8
|
Kidney Cancer (OD04450-01)
4.0
Gastric Cancer 9060397
26.8
|
Kidney Margin (OD04450-03)
7.1
Stomach Margin 9060396
7.4
|
Kidney Cancer 8120607
3.3
Gastric Cancer 064005
27.4
|
|
[1249]
580
TABLE ANH
|
|
|
Panel 4D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3116, Run
Ag3551, Run
Ag3116, Run
Ag3551, Run
|
Tissue Name
164526105
166453851
Tissue Name
164526105
166453851
|
|
Secondary Th1 act
15.6
38.4
HUVEC IL-1 beta
0.8
8.2
|
Secondary Th2 act
23.0
56.3
HUVEC IFN gamma
1.4
1.2
|
Secondary Tr1 act
23.2
78.5
HUVEC TNF alpha +
3.0
3.1
|
IFN gamma
|
Secondary Th1 rest
2.9
22.8
HUVEC TNF alpha +
2.5
2.6
|
IL4
|
Secondary Th2 rest
2.5
4.5
HUVEC IL-11
0.5
0.5
|
Secondary Tr1 rest
2.0
7.0
Lung Microvascular
0.0
0.1
|
EC none
|
Primary Th1 act
13.5
18.3
Lung Microvascular
4.2
2.8
|
EC TNF alpha + IL-
|
1 beta
|
Primary Th2 act
6.6
15.5
Microvascular
0.1
0.1
|
Dermal EC none
|
Primary Tr1 act
17.7
33.2
Microsvascular Dermal
5.7
7.3
|
EC TNF alpha + IL-
|
1 beta
|
Primary Th1 rest
9.2
32.1
Bronchial epithelium
2.4
1.5
|
TNF alpha + IL1 beta
|
Primary Th2 rest
1.2
2.9
Small airway
0.6
1.1
|
epithelium none
|
Primary Tr1 rest
1.7
3.8
Small airway
5.5
5.0
|
epithelium TNF alpha +
|
IL-1 beta
|
CD45RA CD4
4.9
6.7
Coronery artery SMC
1.0
0.8
|
lymphocyte act
rest
|
CD45RO CD4
11.1
44.8
Coronery artery SMC
0.7
0.6
|
lymphocyte act
TNF alpha + IL-1 beta
|
CD8 lymphocyte act
5.3
12.2
Astrocytes rest
0.5
1.0
|
Secondary CD8
4.9
16.0
Astrocytes TNF alpha +
14.9
61.1
|
lymphocyte rest
IL-1 beta
|
Secondary CD8
7.6
25.5
KU-812 (Basophil)
0.2
0.2
|
lymphocyte act
rest
|
CD4 lymphocyte
0.8
1.1
KU-812 (Basophil)
1.0
1.5
|
none
PMA/ionomycin
|
2ry
3.0
11.0
CCD1106
0.4
0.5
|
Th1/Th2/Tr1_anti-
(Keratinocytes) none
|
CD95 CH11
|
LAK cells rest
6.8
5.3
CCD1106
0.8
12.4
|
(Keratinocytes)
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
6.4
23.2
Liver cirrhosis
1.1
5.3
|
LAK cells IL-2 + IL-
22.4
73.7
Lupus kidney
1.1
4.8
|
12
|
LAK cells IL-2 + IFN
17.4
44.1
NCI-H292 none
8.4
9.7
|
gamma
|
LAK cells IL-2 + IL-
12.2
25.0
NCI-H292 IL-4
17.6
18.4
|
18
|
LAK cells
12.3
20.7
NCI-H292 IL-9
6.5
5.3
|
PMA/ionomycin
|
NK Cells IL-2 rest
12.9
23.0
NCI-H292 IL-13
9.2
12.0
|
Two Way MLR 3
12.5
24.0
NCI-H292 IFN
4.3
3.5
|
day
gamma
|
Two Way MLR 5
6.0
17.1
HPAEC none
0.5
0.5
|
day
|
Two Way MLR 7
3.0
6.3
HPAEC TNF alpha +
8.2
11.0
|
day
IL-1 beta
|
PBMC rest
4.0
5.4
Lung fibroblast none
0.2
1.0
|
PBMC PWM
100.0
49.3
Lung fibroblast TNF
1.7
9.8
|
alpha + IL-1 beta
|
PBMC PHA-L
11.8
5.6
Lung fibroblast IL-4
3.3
3.2
|
Ramos (B cell) none
0.8
2.0
Lung fibroblast IL-9
0.9
0.5
|
Ramos (B cell)
16.7
6.5
Lung fibroblast IL-13
1.4
1.8
|
ionomycin
|
B lymphocytes PWM
53.2
25.3
Lung fibroblast IFN
3.4
4.0
|
gamma
|
B lymphocytes
61.1
81.8
Dermal fibroblast
1.9
1.1
|
CD40L and IL-4
CCD1070 rest
|
EOL-1 dbcAMP
0.7
0.4
Dermal fibroblast
11.9
13.7
|
CCD1070 TNF alpha
|
EOL-1 dbcAMP
2.2
3.0
Dermal fibroblast
6.1
6.3
|
PMA/ionomycin
CCD1070 IL-1 beta
|
Dendritic cells none
4.8
8.7
Dermal fibroblast IFN
0.6
0.9
|
gamma
|
Dendritic cells LPS
12.3
25.2
Dermal fibroblast IL-4
4.2
6.7
|
Dendritic cells anti-
3.2
6.8
IBD Colitis 2
1.1
4.1
|
CD40
|
Monocytes rest
5.0
7.3
IBD Crohn's
1.8
6.0
|
Monocytes LPS
43.8
100.0
Colon
2.6
15.7
|
Macrophages rest
8.2
11.7
Lung
8.2
7.5
|
Macrophages LPS
26.8
57.4
Thymus
2.3
3.5
|
HUVEC none
0.2
0.5
Kidney
4.2
3.8
|
HUVEC starved
0.6
1.5
|
|
[1250]
581
TABLE ANI
|
|
|
Panel 5D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3116, Run
Ag4828, Run
Ag3116, Run
Ag4828, Run
|
Tissue Name
170863008
219436967
Tissue Name
170863008
219436967
|
|
97457_Patient-
33.4
33.9
94709_Donor 2 AM - A_adipose
5.1
10.8
|
02go_adipose
|
97476_Patient-
31.2
33.4
94710_Donor 2 AM - BN—adipode
3.2
9.3
|
07sk_skeletal
|
muscle
|
97477_Patient-
7.7
59.5
94711_Donor 2 AM - C_adipose
0.0
3.0
|
07ut_uterus
|
97478_Patient-
62.0
39.8
94712_Donor 2 AD - A_adipose
12.9
13.7
|
07pl_placenta
|
97481_Patient-
20.0
25.9
94713_Donor 2 AD - B_adipose
12.9
10.0
|
08sk_skeletal
|
muscle
|
97482_Patient-
33.4
19.8
94714_Donor 2 AD - C_adipose
8.8
6.7
|
08ut_uterus
|
97483_Patient-
58.6
41.5
94742_Donor 3 U -
1.6
4.7
|
08pl_placenta
A_Mesenchymal Stem Cells
|
97486_Patient-
3.7
6.5
94743_Donor 3 U -
4.8
2.8
|
09sk_skeletal
B_Mesenchymal Stem Cells
|
muscle
|
97487_Patient-
13.6
8.1
94730_Donor 3 AM - A_adipose
6.8
6.3
|
09ut_uterus
|
97488_Patient-
41.2
38.4
94731_Donor 3 AM - B_adipose
5.3
2.4
|
09pl_placenta
|
97492_Patient-
31.9
30.6
94732_Donor 3 AM - C_adipose
1.9
2.2
|
10ut_uterus
|
97493_Patient-
74.7
72.7
94733_Donor 3 AD - A_adipose
2.5
10.2
|
10pl_placenta
|
97495_Patient-
67.4
100.0
94734_Donor 3 AD - B_adipose
2.9
5.5
|
11go_adipose
|
97496_Patient-
9.0
5.8
94735_Donor 3 AD - C_adipose
6.7
4.7
|
11sk_skeletal
|
muscle
|
97497_Patient-
35.4
20.6
77138_Liver_HepG2untreated
13.0
14.4
|
11ut_uterus
|
97498_Patient-
52.1
50.0
73556_Heart_Cardiac stromal
9.1
1.9
|
11pl_placenta
cells (primary)
|
97500_Patient-
100.0
82.4
81735_Small Intestine
20.0
17.2
|
12go_adipose
|
97501_Patient-
14.2
19.2
72409_Kidney_Proximal
0.0
0.9
|
12sk_skeletal
Convoluted Tubule
|
muscle
|
97502_Patient-
51.8
23.7
82685_Small
13.5
19.1
|
12ut_uterus
intestine_Duodenum
|
97503_Patient-
39.5
57.0
90650_Adrenal_Adrenocortical
7.3
8.8
|
12pl_placenta
adenoma
|
94721_Donor 2 U -
2.1
1.6
72410_Kidney_HRCE
9.9
7.6
|
A_Mesenchymal
|
Stem Cells
|
94722_Donor 2 U -
0.0
3.0
72411_Kidney_HRE
5.9
13.5
|
B_Mesenchymal
|
Stem Cells
|
94723_Donor 2 U -
1.8
2.1
73139_Uterus_Uterine smooth
2.5
2.0
|
C_Mesenchymal
muscle cells
|
Stem Cells
|
|
[1251] CNS_neurodegeneration_v1.0 Summary: Ag3551 This panel confirms the expression of the CG91911-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1252] General_screening_panel_v1.4 Summary: Ag3116/Ag3551/Ag4828 Results of three experiments with two different probes and primer sets are in excellent agreement. Highest expression of the CG91911-01 gene is detected in adipose (29.6), fetal lung (CT=30) and breast cancer MCF-7 cell line(CT=27.6). Interestingly, this gene is expressed at much higher levels in fetal (CT=28-30) when compared to adult lung (CT=31-35). This observation suggests that expression of this gene can be used to distinguish fetal from adult lung. In addition, the relative overexpression of this gene in fetal lung suggests that the protein product may enhance lung growth or development in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of lung related diseases.
[1253] In addition significant expression of this gene is found in number of cancer (pancreatic, CNS, colon, lung, breast, ovary, prostate, melanoma) cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, might be beneficial in the treatment of these cancers.
[1254] Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, skeletal muscle, heart, fetal liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1255] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. The CG91911-01 gene codes for mitogen-activated protein kinase kinase kinase 8 (MAP3K8). Recently, MKK6, a related protein has been shown to associated with Alzheimer's disease (Zhu X, Rottkamp C A, Hartzler A, Sun Z, Takeda A, Boux H, Shimohama S, Perry G, Smith M A. (2001) Activation of MKK6, an upstream activator of p38, in Alzheimer's disease. J Neurochem 79(2):311-8). Therefore, in analogy to MKK6 and the expression in brain, we predict that MAP3K8 may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1256] Ag3551 Results from one experiment with the CG91911-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1257] Panel 1.3D Summary: Ag3116 Highest expression of the CG91911-01 gene is detected in adipose (32.75). Low to moderate expression of this gene is also seen in number of ovarian cancer cell lines, liver adenocarcinoma and breast cancer MCF-7 cell line. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, might be beneficial in the treatment of these cancers.
[1258] In addition, low expression of this gene is also seen in fetal kidney and lung. Interestingly, this gene is expressed at much higher levels in fetal (CT=34.3) when compared to adult kidney (CT=37). This observation suggests that expression of this gene can be used to distinguish fetal from adult kidney. In addition, the relative overexpression of this gene in fetal lung suggests that the protein product may enhance lung growth or development in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of lung related diseases.
[1259] Panel 2D Summary: Ag3116 Highest expression of the CG91911-01 gene is detected in normal bladder (OD04718-03) sample (CT=31.4). Low to moderate expression of this gene is seen in large number of normal and cancer samples. Please see Panel 1.4 for a discussion of the potential utility of this gene.
[1260] Panel 4D Summary: Ag3116/Ag3551 Results from two experiments with same primer and probe set are in excellent agreement. Highest expression of the CG91911-01 gene is detected in PWM treated PBMC and LPS treated monocytes (CTs=28-29). Interestingly, expression of this gene is stimulated in activated primary Th2 and Tr1, activated secondary Th1, Th2, Tr1, PWM treated PBMC, LPS treated monocytes, TNFalpha +IL-1beta treated astrocytes and keratinocytes. Thus, expression of this gene can be used to distinguish between these activated or treated cells from the corresponding untreated or resting cells.
[1261] In addition low expression of this gene is seen in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1262] Panel 5D Summary: Ag3116/Ag4828 Results from two experiments with different primer and probe set are in excellent agreement. Highest expression of the CG91911-01 gene is detected in adipose tissue (CTs=29-33). Low to moderate expression of this gene is seen in wide range of samples used in this panel including adipose, skeletal muscle, uterus, and placenta. This wide spread expression of this gene in tissues with metabolic or endocrine function, suggests that this gene plays a role in endocrine/metabolically related diseases, such as obesity and diabetes.
[1263] The CG91911-01 gene codes for mitogen-activated protein kinase kinase kinase 8 (MAP3K8). Recently, activation of MAP kinase, ERK, a related protein, by modified LDL in vascular smooth muscle cells has been implicated in the development of atherosclerosis in diabetes (Velarde V, Jenkins A J, Christopher J, Lyons T J, Jaffa A A. (2001) Activation of MAPK by modified low-density lipoproteins in vascular smooth muscle cells. J Appl Physiol 91(3):1412-20). Therefore, MAP3K8 may also play a role in the development of this disease and therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, might be beneficial in the treatment of artherosclerosis and diabetes.
[1264] AO. CG91941-01: SERINE/THREONINE-PROTEIN KINASE
[1265] Expression of gene CG91941-01 was assessed using the primer-probe sets Ag3744 and Ag3746, described in Tables AOA and AOB. Results of the RTQ-PCR runs are shown in Tables AOC and AOD.
582TABLE AOA
|
|
Probe Name Ag3744
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ggctgcttgctgtatgagttat-3′22631372
ProbeTET-5′-tgcattaatgcctccatttacagctt-26654373
3′-TAMRA
Reverse5′-tttcccagcgagttctttct-3′20686374
|
[1266]
583
TABLE AOB
|
|
|
Probe Name Ag3746
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-ggctgcttgctgtatgagttat-3′
22
631
375
|
Probe
TET-5′-tgcattaatgcctccatttacagctt-
26
654
376
|
3′-TAMRA
|
Reverse
5′-tttcccagcgagttctttct-3′
20
686
377
|
|
[1267]
584
TABLE AOC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3744, Run
Ag3746, Run
Ag3744, Run
Ag3746, Run
|
Tissue Name
218296857
218297971
Tissue Name
218296857
218297971
|
|
Adipose
0.3
0.1
Renal ca. TK-10
41.2
37.9
|
Melanoma*
1.2
1.5
Bladder
3.6
3.7
|
Hs688(A).T
|
Melanoma*
1.9
1.9
Gastric ca. (liver
12.2
13.2
|
Hs688(B).T
met.) NCI-N87
|
Melanoma*
32.1
38.7
Gastric ca. KATO
76.8
88.9
|
M14
III
|
Melanoma*
30.8
28.5
Colon ca. SW-948
15.4
12.9
|
LOXIMVI
|
Melanoma* SK-
52.1
48.0
Colon ca. SW480
48.0
44.1
|
MEL-5
|
Squamous cell
40.9
42.6
Colon ca.* (SW480
47.0
44.1
|
carcinoma SCC-4
met) SW620
|
Testis Pool
40.9
37.9
Colon ca. HT29
39.8
31.9
|
Prostate ca.*
17.2
18.2
Colon ca. HCT-116
59.5
81.2
|
(bone met) PC-3
|
Prostate Pool
0.1
0.0
Colon ca. CaCo-2
46.0
56.3
|
Placenta
0.3
0.2
Colon cancer tissue
8.9
9.3
|
Uterus Pool
0.1
0.0
Colon ca. SW1116
11.7
11.7
|
Ovarian ca.
43.2
40.9
Colon ca. Colo-205
27.9
23.5
|
OVCAR-3
|
Ovarian ca. SK-
52.1
57.0
Colon ca. SW-48
19.3
17.2
|
OV-3
|
Ovarian ca.
11.3
7.7
Colon Pool
0.1
0.3
|
OVCAR-4
|
Ovarian ca.
33.0
28.9
Small Intestine Pool
0.2
0.2
|
OVCAR-5
|
Ovarian ca.
9.7
9.3
Stomach Pool
0.3
0.2
|
IGROV-1
|
Ovarian ca.
4.6
4.9
Bone Marrow Pool
0.2
0.1
|
OVCAR-8
|
Ovary
0.3
0.3
Fetal Heart
2.8
2.7
|
Breast ca. MCF-7
16.2
19.3
Heart Pool
0.0
0.0
|
Breast ca. MDA-
92.7
86.5
Lymph Node Pool
0.5
0.7
|
MB-231
|
Breast ca. BT
100.0
100.0
Fetal Skeletal
0.3
0.4
|
549
Muscle
|
Breast ca. T47D
44.1
57.8
Skeletal Muscle
0.0
0.0
|
Pool
|
Breast ca. MDA-N
19.9
21.6
Spleen Pool
1.0
1.4
|
Breast Pool
0.4
0.1
Thymus Pool
4.8
4.3
|
Trachea
0.6
0.8
CNS cancer
13.6
13.7
|
(glio/astro) U87-
|
MG
|
Lung
0.0
0.0
CNS cancer
35.8
36.9
|
(glio/astro) U-118-
|
MG
|
Fetal Lung
5.1
3.7
CNS cancer
45.7
48.3
|
(neuro; met) SK-N-
|
AS
|
Lung ca. NCI-
5.6
5.7
CNS cancer (astro)
25.9
30.4
|
N417
SF-539
|
Lung ca. LX-1
43.8
42.9
CNS cancer (astro)
43.5
47.0
|
SNB-75
|
Lung ca. NCI-
19.1
18.4
CNS cancer (glio)
5.7
7.5
|
H146
SNB-19
|
Lung ca. SHP-
44.1
42.0
CNS cancer (glio)
6.0
6.0
|
77
SF-295
|
Lung ca. A549
60.3
48.0
Brain (Amygdala)
0.0
0.1
|
Pool
|
Lung ca. NCI-
9.0
9.5
Brain (cerebellum)
0.0
0.0
|
H526
|
Lung ca. NCI-
41.8
38.7
Brain (fetal)
0.9
0.9
|
H23
|
Lung ca. NCI-
0.3
0.2
Brain
0.0
0.1
|
H460
(Hippocampus)
|
Pool
|
Lung ca. HOP-
5.9
6.3
Cerebral Cortex
0.2
0.2
|
62
Pool
|
Lung ca. NCI-
32.8
28.5
Brain (Substantia
0.1
0.0
|
H522
nigra) Pool
|
Liver
0.0
0.0
Brain (Thalamus)
0.1
0.2
|
Pool
|
Fetal Liver
12.2
14.6
Brain (whole)
0.2
0.2
|
Liver ca. HepG2
14.7
13.8
Spinal Cord Pool
0.0
0.1
|
Kidney Pool
0.0
0.1
Adrenal Gland
0.1
0.1
|
Fetal Kidney
8.0
5.4
Pituitary gland Pool
0.0
0.1
|
Renal ca. 786-0
43.5
43.8
Salivary Gland
0.0
0.0
|
Renal ca. A498
6.5
6.2
Thyroid (female)
0.0
0.1
|
Renal ca. ACHN
17.2
14.9
Pancreatic ca.
70.7
72.2
|
CAPAN2
|
Renal ca. UO-31
13.7
11.9
Pancreas Pool
0.5
0.3
|
|
[1268]
585
TABLE AOD
|
|
|
Panel 4.1D
|
Rel.
Rel.
Rel.
Rel.
Rel.
Rel.
|
Exp. (%)
Exp. (%)
Exp. (%)
Exp. (%)
Exp. (%)
Exp. (%)
|
Ag3744,
Ag3746,
Ag3746,
Ag3744,
Ag3746,
Ag3746,
|
Run
Run
Run
Run
Run
Run
|
Tissue Name
170068295
170068664
197450453
Tissue Name
170068295
170068664
197450453
|
|
Secondary Th1 act
60.7
42.9
59.0
HUVEC IL-
32.8
29.3
33.0
|
1 beta
|
Secondary Th2 act
40.9
30.8
47.3
HUVEC IFN
27.7
25.2
39.2
|
gamma
|
Secondary Tr1 act
62.4
41.8
41.5
HUVEC TNF
24.3
13.5
20.3
|
alpha + IFN
|
gamma
|
Secondary Th1
10.4
8.4
10.8
HUVEC TNF
22.7
16.0
26.1
|
rest
alpha + IL4
|
Secondary Th2
13.8
11.3
13.5
HUVEC IL-11
14.2
10.9
18.3
|
rest
|
Secondary Tr1 rest
7.7
9.6
5.0
Lung
15.3
10.8
21.0
|
Microvascular
|
EC none
|
Primary Th1 act
19.6
13.8
12.9
Lung
12.4
10.4
18.8
|
Microvascular
|
EC TNF alpha +
|
IL-1 beta
|
Primary Th2 act
29.3
20.6
26.4
Microvascular
28.9
15.3
32.3
|
Dermal EC none
|
Primary Tr1 act
28.9
20.2
29.3
Microsvasular
16.4
11.4
21.6
|
Dermal EC
|
TNF alpha + IL-
|
1 beta
|
Primary Th1 rest
42.6
40.6
26.1
Bronchial
2.0
2.8
3.3
|
epithelium
|
TNF alpha +
|
IL1 beta
|
Primary Th2 rest
31.2
20.2
13.4
Small airway
1.5
0.9
1.0
|
epithelium none
|
Primary Tr1 rest
35.8
25.7
33.4
Small airway
4.2
2.8
3.9
|
epithelium
|
TNF alpha + IL-
|
1 beta
|
CD45RA CD4
44.1
36.1
69.3
Coronery artery
7.1
4.5
7.0
|
lymphocyte act
SMC rest
|
CD45RO CD4
36.1
26.8
38.4
Coronery artery
6.8
3.8
4.6
|
lymphocyte act
SMC TNF alpha +
|
IL-1 beta
|
CD8 lymphocyte
51.1
33.9
36.1
Astrocytes rest
1.7
2.2
2.0
|
act
|
Secondary CD8
25.5
18.4
27.2
Astrocytes
0.9
0.4
1.0
|
lymphocyte rest
TNF alpha + IL-
|
1 beta
|
Secondary CD8
50.7
24.1
27.2
KU-812
51.1
38.4
46.0
|
lymphocyte act
(Basophil) rest
|
CD4 lymphocyte
0.0
0.3
0.0
KU-812
40.1
32.3
52.1
|
none
(Basophil)
|
PMA/ionomycin
|
2ry
33.4
17.8
36.3
CCD1106
39.2
19.8
35.8
|
Th1/Th2/Tr1_anti-
(Keratinocytes)
|
CD95 CH11
none
|
LAK cells rest
4.5
1.9
2.6
CCD1106
16.6
11.3
12.8
|
(Keratinocytes)
|
TNF alpha + IL-
|
1 beta
|
LAK cells IL-2
50.7
31.4
44.4
Liver cirrhosis
1.0
1.2
2.5
|
LAK cells IL-
27.5
20.3
25.9
NCI-H292 none
34.2
23.3
33.9
|
2 + IL-12
|
LAK cells IL-
40.6
23.5
22.8
NCI-H292 IL-4
61.1
42.0
45.4
|
2 + IFN gamma
|
LAK cells IL-2 +
35.1
21.5
24.7
NCI-H292 IL-9
88.9
51.1
66.0
|
IL-18
|
LAK cells
1.1
1.5
1.2
NCI-H292 IL-13
56.3
40.1
53.6
|
PMA/ionomycin
|
NK Cells IL-2 rest
44.4
47.3
62.0
NCI-H292 IFN
50.3
37.9
37.1
|
gamma
|
Two Way MLR 3
3.8
5.2
3.6
HPAEC none
20.0
11.7
10.5
|
day
|
Two Way MLR 5
17.3
18.7
23.8
HPAEC TNF
13.7
9.0
11.2
|
day
alpha + IL-1 beta
|
Two Way MLR 7
24.1
22.7
23.3
Lung fibroblast
8.2
4.5
7.3
|
day
none
|
PBMC rest
0.4
0.2
0.2
Lung fibroblast
12.2
12.2
13.9
|
TNF alpha + IL-
|
1 beta
|
PBMC PWM
25.5
18.6
23.2
Lung fibroblast
2.1
0.5
0.9
|
IL-4
|
PBMC PHA-L
41.5
27.9
48.3
Lung fibroblast
4.4
3.0
3.1
|
IL-9
|
Ramos (B cell)
65.1
42.9
48.0
Lung fibroblast
1.2
0.4
0.6
|
none
IL-13
|
Ramos (B cell)
45.7
32.1
54.0
Lung fibroblast
0.3
1.1
1.1
|
ionomycin
IFN gamma
|
B lymphocytes
24.7
19.5
38.4
Dermal
87.1
69.3
83.5
|
PWM
fibroblast
|
CCD1070 rest
|
B lymphocytes
27.9
20.7
25.3
Dermal
100.0
100.0
100.0
|
CD40L and IL-4
fibroblast
|
CCD1070 TNF
|
alpha
|
EOL-1 dbcAMP
20.7
12.2
20.4
Dermal
52.5
37.1
52.9
|
fibroblast
|
CCD1070 IL-1
|
beta
|
EOL-1 dbcAMP
21.5
14.0
15.8
Dermal
20.2
14.0
23.3
|
PMA/ionomycin
fibroblast IFN
|
gamma
|
Dendritic cells
1.7
0.4
1.2
Dermal
32.8
20.6
29.3
|
none
fibroblast IL-4
|
Dendritic cells
0.2
0.1
0.0
Dermal
23.8
12.5
23.0
|
LPS
Fibroblasts rest
|
Dendritic cells
0.0
0.0
0.2
Neutrophils
0.0
0.0
0.0
|
anti-CD40
TNFa + LPS
|
Monocytes rest
0.0
0.0
0.0
Neutrophils rest
0.0
0.0
0.0
|
Monocytes LPS
0.0
0.0
0.0
Colon
6.3
1.9
1.9
|
Macrophages rest
4.4
2.3
2.9
Lung
0.4
0.4
1.8
|
Macrophages LPS
1.0
0.4
0.9
Thymus
41.2
23.0
27.2
|
HUVEC none
30.4
23.7
28.1
Kidney
0.2
0.0
0.2
|
HUVEC starved
45.1
33.9
51.8
|
|
[1269] CNS_neurodegeneration_v1.0 Summary: Ag3744/Ag3746 Expression of the CG91941-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1270] General_screening13 panel_v1.4 Summary: Ag3744/Ag3746 Two experiments with same primer and probe sets are in excellent agreement with highest expression of the CG91941-01 gene in breast cancer BT 549 sample (CT=28). Interestingly, high expression of this gene is seen mainly in all the cancer cell lines (pancreatic, CNS, colon, renal, liver, lung, breast, ovarian, prostate, squamous cell carcinoma and melanoma). Thus, expression of this gene can be used as diagnostic marker for detection of these cancers and therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.
[1271] Interestingly, this gene is expressed at much higher levels in fetal (CTs=31-34.8) when compared to adult liver, lung and heart (CTs=37-40). This observation suggests that expression of this gene can be used to distinguish fetal from adult liver, lung and hear. In addition, the relative overexpression of this gene in fetal liver, lung and heart suggests that the protein product may enhance growth or development of these tissues in the fetus and thus may also act in a regenerative capacity in the adult.
[1272] Panel 4.1D Summary: Ag3744/Ag3746 Three experiments with same primer and probe sets are in excellent agreement with highest expression of the CG91941-01 gene in in TNF alpha treated dermal fibroblast CCD1070 (CT=29-30). High to moderate expression of this gene is also seen in dermal fibroblasts, NCI-H292, basophils, keratinocytes, endothelial cells, EOL-1 dbcAMP, members of B and T cells, and thymus. Interestingly, expression of this gene is stimulated in PWM/PHA treated PBMC cells, and cytokine treated LAK cells. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1273] AP. CG91951-01:/THREONINE-PROTEIN KINASE
[1274] Expression of gene CG91951-01 was assessed using the primer-probe set Ag3747, described in Table APA. Results of the RTQ-PCR runs are shown in Tables APB and APC.
586TABLE APA
|
|
Probe Name Ag3747
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ttggggctagccagaatattaa-3′22502378
ProbeTET-5′-accacgacacgagttttgcaaaa-3′-23524379
TAMRA
Reverse5′-aggacatgtgattcgtttgttc-3′22580380
|
[1275]
587
TABLE APB
|
|
|
General_screening_panel_V1.4
|
Rel. Exp. (%) Ag3747, Run
Rel. Exp. (%) Ag3747, Run
|
Tissue Name
218298221
Tissue Name
218298221
|
|
Adipose
0.0
Renal ca. TK-10
1.6
|
Melanoma* Hs688(A).T
0.4
Bladder
2.4
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.9
|
NCI-N87
|
Melanoma* M14
2.2
Gastric ca. KATO III
16.2
|
Melanoma* LOXIMVI
1.7
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
100.0
Colon ca. SW480
1.3
|
Squamous cell
0.4
Colon ca.* (SW480 met)
3.2
|
carcinoma SCC-4
SW620
|
Testis Pool
33.7
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
0.9
Colon ca. HCT-116
7.9
|
PC-3
|
Prostate Pool
46.3
Colon ca. CaCo-2
0.6
|
Placenta
0.0
Colon cancer tissue
0.0
|
Uterus Pool
0.0
Colon ca. SW1116
0.6
|
Ovarian ca. OVCAR-3
33.0
Colon ca. Colo-205
0.3
|
Ovarian ca. SK-OV-3
3.5
Colon ca. SW-48
1.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
2.5
|
Ovarian ca. OVCAR-5
0.9
Small Intestine Pool
0.0
|
Ovarian ca. IGROV-1
6.7
Stomach Pool
0.0
|
Ovarian ca. OVCAR-8
5.0
Bone Marrow Pool
0.0
|
Ovary
0.0
Fetal Heart
0.0
|
Breast ca. MCF-7
1.3
Heart Pool
3.8
|
Breast ca. MDA-MB-
1.2
Lymph Node Pool
0.7
|
231
|
Breast ca. BT 549
6.9
Fetal Skeletal Muscle
0.0
|
Breast ca. T47D
0.0
Skeletal Muscle Pool
0.0
|
Breast ca. MDA-N
12.2
Spleen Pool
0.0
|
Breast Pool
0.0
Thymus Pool
1.0
|
Trachea
0.0
CNS cancer (glio/astro)
2.5
|
U87-MG
|
Lung
0.4
CNS cancer (glio/astro) U-
13.0
|
118-MG
|
Fetal Lung
1.8
CNS cancer (neuro; met)
3.4
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
2.2
|
Lung ca. LX-1
2.5
CNS cancer (astro) SNB-75
1.9
|
Lung ca. NCI-H146
1.7
CNS cancer (glio) SNB-19
6.6
|
Lung ca. SHP-77
52.5
CNS cancer (glio) SF-295
31.6
|
Lung ca. A549
7.6
Brain (Amygdala) Pool
0.7
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0
|
Lung ca. NCI-H23
15.9
Brain (fetal)
6.0
|
Lung ca. NCI-H460
0.9
Brain (Hippocampus) Pool
1.8
|
Lung ca. HOP-62
0.7
Cerebral Cortex Pool
1.9
|
Lung ca. NCI-H522
1.0
Brain (Substantia nigra)
1.7
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
0.3
|
Fetal Liver
23.7
Brain (whole)
0.0
|
Liver ca. HepG2
0.5
Spinal Cord Pool
0.9
|
Kidney Pool
0.5
Adrenal Gland
1.0
|
Fetal Kidney
0.0
Pituitary gland Pool
0.0
|
Renal ca. 786-0
0.6
Salivary Gland
0.0
|
Renal ca. A498
0.0
Thyroid (female)
0.0
|
Renal ca. ACHN
2.0
Pancreatic ca. CAPAN2
0.0
|
Renal ca. UO-31
3.6
Pancreas Pool
1.2
|
|
[1276]
588
TABLE APC
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3747,
Rel. Exp. (%) Ag3747,
|
Tissue Name
Run 198383569
Tissue Name
Run 198383569
|
|
Secondary Th1 act
0.1
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.1
HUVEC IFN gamma
0.1
|
Secondary Tr1 act
0.1
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.1
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.2
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.2
Microsvasular Dermal EC
0.1
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.3
Bronchial epithelium TNF alpha +
0.0
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.1
Astrocytes rest
0.1
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.1
|
lymphocyte act
|
CD4 lymphocyte none
0.1
KU-812 (Basophil)
0.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.1
CCD1106 (Keratinocytes) none
0.2
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.2
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.1
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.3
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.1
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.2
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.1
Lung fibroblast IL-13
0.4
|
Ramos (B cell) ionomycin
0.1
Lung fibroblast IFN gamma
0.3
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.6
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.3
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.2
Dermal fibroblast IFN gamma
0.2
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.2
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.8
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.5
|
Monocytes rest
0.0
Neutrophils rest
0.3
|
Monocytes LPS
0.1
Colon
0.6
|
Macrophages rest
0.0
Lung
2.0
|
Macrophages LPS
0.0
Thymus
13.3
|
HUVEC none
0.0
Kidney
100.0
|
HUVEC starved
0.1
|
|
[1277] CNS_neurodegeneration_v1.0 Summary: Ag3747 Expression of the CG91951-01 gene is low/undetectable (CTs>34.5) across all of the samples on this panel (data not shown).
[1278] General_screening_panel_v1.4 Summary: Ag3747 Highest expression of the CG91951-01 gene is detected in melanoma SK-MEL-5 cell line (CT=31.4). In addition, low to moderated expression is also seen in 2 CNS cancer, gastric cancer KATO III, two lung cancer breast cancer MDA-N and Ovarian cancer OVCAR-3 cell lines. Therefore, expression of this gene can be used as diagnostic marker to detect these cancers and therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs might be beneficial in the treatment of these cancers.
[1279] Moderate expressiion of this gene is also seen in testis, prostate, and fetal liver. Interestingly, this gene is expressed at much higher levels in fetal (CT=33.5) when compared to adult liver (CT=40). This observation suggests that expression of this gene can be used to distinguish fetal from adult liver. In addition, the relative overexpression of this gene in fetal liver suggests that the protein product may enhance growth or development of liver in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of liver related diseases.
[1280] Panel 4.1D Summary: Ag3747 High expression of the CG91951-01 gene is detected in exclusively in kidney (CT=28.7). Therefore, small molecule therapies designed with the protein encoded for by this gene could modulate kidney function and be important in the treatment of inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis.
[1281] Results from one experiment, Run 170068966, with this gene using same primer and probe set are not included. The amp plot indicates that there were experimental difficulties with this run.
[1282] Panel 5 Islet Summary: Ag3747 Expression of the CG91951-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1283] AQ. CG92025-01: GAMMA-GLUTAMYLTRANSFERASE
[1284] Expression of gene CG92025-01 was assessed using the primer-probe set Ag3766, described in Table AQA. Results of the RTQ-PCR runs are shown in Tables AQB, AQC and AQD.
589TABLE AQA
|
|
Probe Name Ag3766
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ctggctgacacctatgagatg-3′21692381
ProbeTET-5′-aggccttctacaacggcagcctcat-25732382
3′-TAMRA
Reverse5′-ctggatgtccttcacaatctg-3′21761383
|
[1285]
590
TABLE AQB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3766, Run
Rel. Exp. (%) Ag3766, Run
|
Tissue Name
211176203
Tissue Name
211176203
|
|
AD 1 Hippo
15.3
Control (Path) 3
16.0
|
Temporal Ctx
|
AD 2 Hippo
14.7
Control (Path) 4
22.2
|
Temporal Ctx
|
AD 3 Hippo
11.3
AD 1 Occipital Ctx
16.4
|
AD 4 Hippo
8.2
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
37.6
AD 3 Occipital Ctx
18.0
|
AD 6 Hippo
35.4
AD 4 Occipital Ctx
86.5
|
Control 2 Hippo
29.9
AD 5 Occipital Ctx
44.8
|
Control 4 Hippo
14.1
AD 6 Occipital Ctx
10.7
|
Control (Path) 3 Hippo
5.1
Control 1 Occipital Ctx
13.3
|
AD 1 Temporal Ctx
8.7
Control 2 Occipital Ctx
35.4
|
AD 2 Temporal Ctx
25.2
Control 3 Occipital Ctx
32.1
|
AD 3 Temporal Ctx
7.2
Control 4 Occipital Ctx
14.0
|
AD 4 Temporal Ctx
10.3
Control (Path) 1
38.7
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
34.4
Control (Path) 2
7.5
|
Occipital Ctx
|
AD 5 Sup Temporal
0.0
Control (Path) 3
7.2
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
100.0
Control (Path) 4
21.3
|
Occipital Ctx
|
AD 6 Sup Temporal
79.6
Control 1 Parietal Ctx
29.1
|
Ctx
|
Control 1 Temporal Ctx
17.2
Control 2 Parietal Ctx
73.2
|
Control 2 Temporal Ctx
41.2
Control 3 Parietal Ctx
10.2
|
Control 3 Temporal Ctx
17.0
Control (Path) 1
44.4
|
Parietal Ctx
|
Control 3 Temporal Ctx
24.0
Control (Path) 2
28.1
|
Parietal Ctx
|
Control (Path) 1
28.9
Control (Path) 3
10.3
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
24.3
Control (Path) 4
43.8
|
Temporal Ctx
Parietal Ctx
|
|
[1286]
591
TABLE AQC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3766, Run
Rel. Exp. (%) Ag3766, Run
|
Tissue Name
218902548
Tissue Name
218902548
|
|
Adipose
0.5
Renal ca. TK-10
24.0
|
Melanoma* Hs688(A).T
0.1
Bladder
8.0
|
Melanoma* Hs688(B).T
0.7
Gastric ca. (liver met.)
25.9
|
NCI-N87
|
Melanoma* M14
0.1
Gastric ca. KATO III
2.2
|
Melanoma* LOXIMVI
2.6
Colon ca. SW-948
5.3
|
Melanoma* SK-MEL-5
3.4
Colon ca. SW480
0.5
|
Squamous cell
5.6
Colon ca.* (SW480 met)
1.7
|
carcinoma SCC-4
SW620
|
Testis Pool
2.9
Colon ca. HT29
3.2
|
Prostate ca.* (bone met)
0.2
Colon ca. HCT-116
5.5
|
PC-3
|
Prostate Pool
6.6
Colon ca. CaCo-2
9.3
|
Placenta
0.6
Colon cancer tissue
39.0
|
Uterus Pool
0.1
Colon ca. SW1116
0.3
|
Ovarian ca. OVCAR-3
0.9
Colon ca. Colo-205
1.8
|
Ovarian ca. SK-OV-3
0.4
Colon ca. SW-48
3.4
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
0.7
|
Ovarian ca. OVCAR-5
27.9
Small Intestine Pool
1.3
|
Ovarian ca. IGROV-1
7.6
Stomach Pool
1.1
|
Ovarian ca. OVCAR-8
3.0
Bone Marrow Pool
0.2
|
Ovary
0.3
Fetal Heart
0.3
|
Breast ca. MCF-7
1.2
Heart Pool
0.2
|
Breast ca. MDA-MB-
2.5
Lymph Node Pool
0.2
|
231
|
Breast ca. BT 549
0.6
Fetal Skeletal Muscle
0.3
|
Breast ca. T47D
50.0
Skeletal Muscle Pool
0.3
|
Breast ca. MDA-N
0.2
Spleen Pool
1.1
|
Breast Pool
0.6
Thymus pool
2.1
|
Trachea
1.8
CNS cancer (glio/astro)
41.2
|
U87-MG
|
Lung
0.1
CNS cancer (glio/astro) U-
10.9
|
118-MG
|
Fetal Lung
1.4
CNS cancer (neuro; met)
0.6
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
1.0
|
Lung ca. LX-1
1.7
CNS cancer (astro) SNB-75
0.8
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
6.8
|
Lung ca. SHP-77
3.4
CNS cancer (glio) SF-295
6.8
|
Lung ca. A549
32.3
Brain (Amygdala) Pool
1.0
|
Lung ca. NCI-H526
0.1
Brain (cerebellum)
0.8
|
Lung ca. NCI-H23
1.2
Brain (fetal)
0.6
|
Lung ca. NCI-H460
12.2
Brain (Hippocampus) Pool
0.9
|
Lung ca. HOP-62
5.1
Cerebral Cortex Pool
0.6
|
Lung ca. NCI-H522
0.5
Brain (Substantia nigra)
1.0
|
Pool
|
Liver
2.5
Brain (Thalamus) Pool
1.1
|
Fetal Liver
25.9
Brain (whole)
0.8
|
Liver ca. HepG2
7.3
Spinal Cord Pool
3.1
|
Kidney Pool
1.1
Adrenal Gland
0.2
|
Fetal Kidney
4.2
Pituitary gland Pool
0.3
|
Renal ca. 786-0
100.0
Salivary Gland
0.8
|
Renal ca. A498
29.5
Thyroid (female)
2.8
|
Renal ca. ACHN
17.8
Pancreatic ca. CAPAN2
1.3
|
Renal ca. UO-31
8.0
Pancreas Pool
12.1
|
|
[1287]
592
TABLE AQD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3766,
Rel. Exp. (%) Ag3766,
|
Tissue Name
Run 170069662
Tissue Name
Run 170069662
|
|
Secondary Th1 act
37.1
HUVEC IL-1beta
1.5
|
Secondary Th2 act
34.6
HUVEC IFN gamma
0.5
|
Secondary Tr1 act
54.7
HUVEC TNF alpha + IFN
0.6
|
gamma
|
Secondary Th1 rest
26.2
HUVEC TNF alpha + IL4
1.1
|
Secondary Th2 rest
18.2
HUVEC IL-11
1.0
|
Secondary Tr1 rest
14.1
Lung Microvascular EC none
3.4
|
Primary Th1 act
15.1
Lung Microvascular EC
3.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
21.3
Microvascular Dermal EC none
2.0
|
Primary Tr1 act
25.0
Microsvasular Dermal EC
1.2
|
TNF alpha + IL-1beta
|
Primary Th1 rest
8.7
Bronchial epithelium TNF alpha +
8.2
|
IL1beta
|
Primary Th2 rest
7.3
Small airway epithelium none
4.8
|
Primary Tr1 rest
13.9
Small airway epithelium
2.8
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
2.8
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
12.3
Coronery artery SMC TNF alpha +
0.7
|
act
IL-1beta
|
CD8 lymphocyte act
11.0
Astrocytes rest
1.2
|
Secondary CD8
14.5
Astrocytes TNF alpha + IL-1beta
3.5
|
lymphocyte rest
|
Secondary CD8
8.7
KU-812 (Basophil) rest
6.8
|
lymphocyte act
|
CD4 lymphocyte none
2.7
KU-812 (Basophil)
6.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
15.3
CCD1106 (Keratinocytes) none
2.3
|
CD95 CH11
|
LAK cells rest
9.5
CCD1106 (Keratinocytes)
4.5
|
TNF alpha + IL-1beta
|
LAK cells IL-2
6.0
Liver cirrhosis
10.7
|
LAK cells IL-2 + IL-12
7.7
NCI-H292 none
0.9
|
LAK cells IL-2 + IFN
10.7
NCI-H292 IL-4
1.0
|
gamma
|
LAK cells IL-2 + IL-18
7.3
NCI-H292 IL-9
0.4
|
LAK cells
18.9
NCI-H292 IL-13
1.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
4.9
NCI-H292 IFN gamma
2.0
|
Two Way MLR 3 day
12.4
HPAEC none
1.3
|
Two Way MLR 5 day
10.3
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
11.2
Lung fibroblast none
1.1
|
PBMC rest
6.1
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
7.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
9.3
Lung fibroblast IL-9
0.3
|
Ramos (B cell) none
3.2
Lung fibroblast IL-13
0.4
|
Ramos (B cell) ionomycin
7.1
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
4.2
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
7.8
Dermal fibroblast CCD1070
4.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
11.5
Dermal fibroblast CCD1070 IL-
0 .6
|
1beta
|
EOL-1 dbcAMP
2.1
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
20.7
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
14.0
Dermal Fibroblasts rest
0.9
|
Dendritic cells anti-CD40
29.3
Neutrophils TNFa + LPS
7.5
|
Monocytes rest
9.7
Neutrophils rest
4.9
|
Monocytes LPS
67.8
Colon
18.0
|
Macrophages rest
13.8
Lung
1.0
|
Macrophages LPS
13.0
Thymus
2.8
|
HUVEC none
1.8
Kidney
100.0
|
HUVEC starved
2.1
|
|
[1288] CNS_neurodegeneration_v1.0 Summary: Ag3766 This panel does not show differential expression of the CG92025-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1289] General_screening_panel_v1.4 Summary: Ag3766 Highest expression of the CG92025-01 gene is seen in a renal cancer cell line (CT=27.7). Moderate levels of expression are also seen in samples derived from ovarian, breast, brain, lung and colon cancer cell lines.
[1290] Thus, expression of this gene could be used to differentiate these samples from other samples on this panel and as a marker of these types of cancers. This gene encodes a protein that is homologous to gamma-glutamyl transferase (GGT), an enzyme that acts as a glutathionase and is found in tissues involved in absorption and secretion. Elevated levels of this enzyme have been in found in renal cancers. Therefore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of ovarian, breast, brain, lung and colon cancers.
[1291] Among metabolic tissues, this gene is expressed at moderate to low levels in pancreas, thyroid and adult and fetal liver. Elevated level of GGT have been linked to type 2 diabetes.
[1292] Thus, the expression of this gene suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1293] This gene is also expressed at low levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, and cerebellum. GGT is important in repletion of intercellular glutathione and is upregulated in the face of oxidative stress. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders that result from oxidative stress, including Alzheimer's disease and schizophrenia (Perry U, Wannamethee S G, Shaper A G. Prospective study of serum gamma-glutamyltransferase and risk of NIDDM. Diabetes Care 1998 May;21(5):732-7; Lusini L, Tripodi S A, Rossi R, Giannerini F, Giustarini D, del Vecchio M T, Barbanti G, Cintorino M, Tosi P, Di Simplicio P. Altered glutathione anti-oxidant metabolism during tumor progression in human renal-cell carcinoma. Int J Cancer Jan. 1, 2001;91(1):55-9; Gupta A, Gupta A, Datta M, Shukla G S. Cerebral antioxidant status and free radical generation following glutathione depletion and subsequent recovery. Mol Cell Biochem 2000 June;209(1-2):55-61).
[1294] Panel 4.1D Summary: Ag3766 Highest expression of the CG92025-01 gene is seen in kidney (CT30.7). Moderate expression is also seen in LPS stimulated monocytes. Upon activation with pathogens such as LPS, monocytes contribute to the innate and specific immunity by migrating to the site of tissue injury and releasing inflammatory cytokines. This release contributes to the inflammation process. Therefore, modulation of the expression of the protein encoded by this transcript may prevent the recruitment of monocytes and the initiation it of the inflammatory process, and reduce the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, or rheumatoid arthritis.
[1295] Low but significant levels of expression are seen in chronically stimulated T cells. Thus, therapeutics designed with the transcript may be important in the treatment of diseases such as asthma, IBD, psoriasis and arthritis in which T cells are chronically stimulated.
[1296] AR. CG92025-02: GAMMA-GLUTAMYLTRANSPEPTIDASE 2.
[1297] Expression of gene CG92025-02 was assessed using the primer-probe set Ag4127, described in Table ARA. Results of the RTQ-PCR runs are shown in Tables ARB and ARC.
593TABLE ARA
|
|
Probe Name Ag4127
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ccaggctggacagttcagt-3′1927384
ProbeTET-2660385
5′-cccacagcagagttcaactggagaca-3′-
TAMRA
Reverse5′-cctctgcctctagctggttt-3′2089386
|
[1298]
594
TABLE ARB
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag4127, Run
Rel. Exp. (%) Ag4127, Run
|
Tissue Name
220361624
Tissue Name
220361624
|
|
Adipose
0.4
Renal ca TK-10
16.0
|
Melanoma* Hs688(A).T
0.1
Bladder
7.0
|
Melanoma* Hs688(B).T
0.2
Gastric ca. (liver met.)
6.6
|
NCI-N87
|
Melanoma* M14
0.1
Gastric ca. KATO III
1.9
|
Melanoma* LOXIMVI
0.8
Colon ca. SW-948
5.9
|
Melanoma* SK-MEL-5
4.4
Colon ca. SW480
0.6
|
Squamous cell
1.6
Colon ca.* (SW480 met)
1.6
|
carcinoma SCC-4
SW620
|
Testis Pool
2.1
Colon ca. HT29
1.2
|
Prostate ca.* (bone met)
0.1
Colon ca. HCT-116
7.2
|
PC-3
|
Prostate Pool
4.2
Colon ca. CaCo-2
5.6
|
Placenta
1.0
Colon cancer tissue
55.5
|
Uterus Pool
0.1
Colon ca. SW1116
0.1
|
Ovarian ca. OVCAR-3
0.9
Colon ca. Colo-205
2.2
|
Ovarian ca. SK-OV-3
1.6
Colon ca. SW-48
2.6
|
Ovarian ca. OVCAR-4
0.3
Colon Pool
0.8
|
Ovarian ca. OVCAR-5
23.5
Small Intestine Pool
0.5
|
Ovarian ca. IGROV-1
4.8
Stomach Pool
0.8
|
Ovarian ca. OVCAR-8
2.3
Bone Marrow Pool
0.3
|
Ovary
0.5
Fetal Heart
0.2
|
Breast ca. MCF-7
1.4
Heart Pool
0.2
|
Breast ca. MDA-MB-
1.4
Lymph Node Pool
0.8
|
231
|
Breast ca. BT 549
0.1
Fetal Skeletal Muscle
0.6
|
Breast ca. T47D
50.0
Skeletal Muscle Pool
0.3
|
Breast ca. MDA-N
0.0
Spleen Pool
0.5
|
Breast Pool
1.0
Thymus Pool
1.4
|
Trachea
0.8
CNS cancer (glio/astro)
26.6
|
U87-MG
|
Lung
0.0
CNS cancer (glio/astro) U-
8.2
|
118-MG
|
Fetal Lung
1.7
CNS cancer (neuro; met)
0.8
|
SK-N-AS
|
Lung ca. NCI-N417
0.1
CNS cancer (astro) SF-539
0.9
|
Lung ca. LX-1
2.7
CNS cancer (astro) SNB-75
1.4
|
Lung ca. NCI-H146
0.3
CNS cancer (glio) SNB-19
4.0
|
Lung ca. SHP-77
2.3
CNS cancer (glio) SF-295
3.3
|
Lung ca. A549
31.6
Brain (Amygdala) Pool
0.6
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
1.4
|
Lung ca. NCI-H23
0.6
Brain (fetal)
1.0
|
Lung ca. NCI-H460
15.7
Brain (Hippocampus) Pool
0.5
|
Lung ca. HOP-62
0.8
Cerebral Cortex Pool
0.5
|
Lung ca. NCI-H522
0.6
Brain (Substantia nigra)
0.7
|
Pool
|
Liver
7.1
Brain (Thalamus) Pool
0.6
|
Fetal Liver
24.3
Brain (whole)
1.1
|
Liver ca. HepG2
4.4
Spinal Cord Pool
0.9
|
Kidney Pool
0.4
Adrenal Gland
0.3
|
Fetal Kidney
2.3
Pituitary gland Pool
0.5
|
Renal ca. 786-0
100.0
Salivary Gland
0.6
|
Renal ca. A498
15.5
Thyroid (female)
2.4
|
Renal ca. ACHN
17.6
Pancreatic ca. CAPAN2
1.1
|
Renal ca. UO-31
5.0
Pancreas Pool
8.1
|
|
[1299]
595
TABLE ARC
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag4127,
Rel. Exp. (%) Ag4127,
|
Tissue Name
Run 172859319
Tissue Name
Run 172859319
|
|
Secondary Th1 act
44.4
HUVEC IL-1beta
0.9
|
Secondary Th2 act
37.4
HUVEC IFN gamma
0.8
|
Secondary Tr1 act
30.8
HUVEC TNF alpha + IFN
0.4
|
gamma
|
Secondary Th1 rest
28.7
HUVEC TNF alpha + IL4
0.7
|
Secondary Th2 rest
28.7
HUVEC IL-11
0.4
|
Secondary Tr1 rest
34.4
Lung Microvascular EC none
3.4
|
Primary Th1 act
30.1
Lung Microvascular EC
1.2
|
TNF alpha + IL-1beta
|
Primary Th2 act
45.4
Microvascular Dermal EC none
0.4
|
Primary Tr1 act
40.3
Microsvasular Dermal EC
0.4
|
TNF alpha + IL-1beta
|
Primary Th1 rest
13.0
Bronchial epithelium TNF alpha +
8.1
|
IL1beta
|
Primary Th2 rest
13.2
Small airway epithelium none
4.9
|
Primary Tr1 rest
18.6
Small airway epithelium
4.7
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
3.4
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
11.6
Coronery artery SMC TNF alpha +
3.0
|
act
IL-1beta
|
CD8 lymphocyte act
12.4
Astrocytes rest
0.0
|
Secondary CD8
19.3
Astrocytes TNF alpha + IL-1beta
0.2
|
lymphocyte rest
|
Secondary CD8
8.2
KU-812 (Basophil) rest
3.2
|
lymphocyte act
|
CD4 lymphocyte none
3.5
KU-812 (Basophil)
7.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
26.1
CCD1106 (Keratinocytes) none
3.9
|
CD95 CH11
|
LAK cells rest
7.3
CCD1106 (Keratinocytes)
6.6
|
TNF alpha + IL-1beta
|
LAK cells IL-2
4.1
Liver cirrhosis
10.9
|
LAK cells IL-2 + IL-12
12.9
NCI-H292 none
1.2
|
LAK cells IL-2 + IFN
17.8
NCI-H292 IL-4
1.3
|
gamma
|
LAK cells IL-2 + IL-18
26.8
NCI-H292 IL-9
2.4
|
LAK cells
10.7
NCI-H292 IL-13
2.6
|
PMA/ionomycin
|
NK Cells IL-2 rest
9.2
NCI-H292 IFN gamma
0.5
|
Two Way MLR 3 day
8.3
HPAEC none
1.2
|
Two Way MLR 5 day
4.7
HPAEC TNF alpha + IL-1beta
0.6
|
Two Way MLR 7 day
9.2
Lung fibroblast none
2.8
|
PBMC rest
0.9
Lung fibroblast TNF alpha + IL-
1.4
|
1beta
|
PBMC PWM
9.5
Lung fibroblast IL-4
0.4
|
PBMC PHA-L
7.5
Lung fibroblast IL-9
0.4
|
Ramos (B cell) none
4.2
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
12.3
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
15.1
Dermal fibroblast CCD1070 rest
1.2
|
B lymphocytes CD40L
16.5
Dermal fibroblast CCD1070
7.3
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
9.5
Dermal fibroblast CCD1070 IL-
0.5
|
1beta
|
EOL-1 dbcAMP
6.3
Dermal fibroblast IFN gamma
1.7
|
PMA/ionomycin
|
Dendritic cells none
17.4
Dermal fibroblast IL-4
1.9
|
Dendritic cells LPS
5.5
Dermal Fibroblasts rest
1.9
|
Dendritic cells anti-CD40
5.1
Neutrophils TNFa + LPS
6.6
|
Monocytes rest
9.3
Neutrophils rest
6.8
|
Monocytes LPS
39.2
Colon
13.8
|
Macrophages rest
8.7
Lung
4.6
|
Macrophages LPS
10.1
Thymus
13.7
|
HUVEC none
0.4
Kidney
100.0
|
HUVEC starved
0.2
|
|
[1300] CNS_neurodegeneration_v1.0 Summary: Ag4127 Results from one experiment with this probe and primer set and the CG92025-02 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1301] General_screening_panel_v1.4 Summary: Ag4127 Highest expression of the CG92025-02 gene is seen in a renal cancer cell line (CT=26.6). High to moderate levels of expression are also seen in samples derived from colon, brain, breast, ovarian, and lung cancers. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker for these cancers. This gene encodes a protein that is homologous to gamma-glutamyl transferase (GGT), an enzyme that acts as a glutathionase and is found in tissues involved in absorption and secretion. Elevated levels of this enzyme have been in found in renal cancers. Therefore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of renal, colon, brain, breast, ovarian and lung cancers.
[1302] Among metabolic tissues, this gene is expressed at moderate to low levels in pancreas, thyroid, pituitary, adipose, fetal skeletal muscle and fetal and adult liver. Elevated level of GGT have been linked to type 2 diabetes. Thus, the expression of this genen suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1303] This gene is also expressed at low levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. GGT is important in repletion of intercellular glutathione and is upregulated in the face of oxidative stress. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders that result from oxidative stress, including Alzheimer's disease and schizophrenia (Perry I J, Wannamethee S G, Shaper A G. Prospective study of serum gamma-glutamyltransferase and risk of NIDDM. Diabetes Care 1998 May;21(5):732-7; Lusini L, Tripodi S A, Rossi R, Giannerini F, Giustarini D, del Vecchio M T, Barbanti G, Cintorino M, Tosi P, Di Simplicio P. Altered glutathione anti-oxidant metabolism during tumor progression in human renal-cell carcinoma. Int J Cancer Jan. 1, 2001;91(1):55-9; Gupta A, Gupta A, Datta M, Shukla GS. Cerebral antioxidant status and free radical generation following glutathione depletion and subsequent recovery. Mol Cell Biochem 2000 June;209(1-2):55-61).
[1304] Panel 4.1D Summary: Ag4127 Highest expression of the CG92025-02 gene is seen in kidney (CT=30.6). Moderate to low levels of expression are also seen in LPS stimulated monocytes, primary activated T cells, and resting and activated secondary T cells. Thus, expression of this gene could be used as to differentiate the kidney sample from other samples on this panel and as a marker of kidney tissue. In addition, expression stimulated monocytes and T cells suggests that modulation of the expression of the protein encoded by this transcript may prevent the recruitment of monocytes, the initiation of the inflammatory process, and reduce the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, and rheumatoid arthritis.
[1305] AS. CG92078-01: PERMEASE
[1306] Expression of gene CG92078-01 was assessed using the primer-probe set Ag3769, described in Table ASA. Results of the RTQ-PCR runs are shown in Tables ASB and ASC.
596TABLE ASA
|
|
Probe Name Ag3769
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ttaggtccagaaatgaccagaa-3′221854387
ProbeTET-5′-cgcgtacttctgccctggttaattta-261876388
3′-TAMRA
Reverse5′-tgactctccagcagatgagagt-3′221908389
|
[1307]
597
TABLE ASB
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3769, Run
Rel. Exp. (%) Ag3769, Run
|
Tissue Name
218982365
Tissue Name
218982365
|
|
Adipose
2.3
Renal ca. TK-10
27.0
|
Melanoma* Hs688(A).T
6.8
Bladder
13.2
|
Melanoma* Hs688(B).T
3.7
Gastric ca. (liver met.)
62.4
|
NCI-N87
|
Melanoma* M14
7.9
Gastric ca. KATO III
3.9
|
Melanoma* LOXIMVI
1.0
Colon ca. SW-948
0.8
|
Melanoma* SK-MEL-5
4.5
Colon ca. SW480
7.4
|
Squamous cell
1.0
Colon ca.* (SW480 met)
11.3
|
carcinoma SCC-4
SW620
|
Testis Pool
2.5
Colon ca. HT29
22.4
|
Prostate ca.* (bone met)
22.7
Colon ca. HCT-116
11.3
|
PC-3
|
Prostate Pool
4.0
Colon ca. CaCo-2
100.0
|
Placenta
3.4
Colon cancer tissue
5.0
|
Uterus Pool
1.4
Colon ca. SW1116
2.2
|
Ovarian ca. OVCAR-3
14.2
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
9.4
Colon ca. SW-48
2.2
|
Ovarian ca. OVCAR-4
2.5
Colon Pool
8.8
|
Ovarian ca. OVCAR-5
16.2
Small Intestine Pool
8.7
|
Ovarian ca. IGROV-1
6.0
Stomach Pool
7.0
|
Ovarian ca. OVCAR-8
3.5
Bone Marrow Pool
4.0
|
Ovary
5.8
Fetal Heart
2.0
|
Breast ca. MCF-7
4.4
Heart Pool
3.2
|
Breast ca. MDA-MB-
12.3
Lymph Node Pool
17.7
|
231
|
Breast ca. BT 549
4.9
Fetal Skeletal Muscle
2.9
|
Breast ca. T47D
20.6
Skeletal Muscle Pool
1.9
|
Breast ca. MDA-N
15.4
Spleen Pool
5.8
|
Breast Pool
12.2
Thymus Pool
8.2
|
Trachea
0.0
CNS cancer (glio/astro)
6.0
|
U87-MG
|
Lung
5.0
CNS cancer (glio/astro) U-
34.6
|
118-MG
|
Fetal Lung
25.9
CNS cancer (neuro; met)
5.9
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
1.0
|
Lung ca. LX-1
20.7
CNS cancer (astro) SNB-75
9.5
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
4.5
|
Lung ca. SHP-77
5.1
CNS cancer (glio) SF-295
71.7
|
Lung ca. A549
5.0
Brain (Amygdala) Pool
1.0
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
3.1
|
Lung ca. NCI-H23
25.7
Brain (fetal)
1.1
|
Lung ca. NCI-H460
13.2
Brain (Hippocampus) Pool
0.0
|
Lung ca. HOP-62
6.3
Cerebral Cortex Pool
0.0
|
Lung ca. NCI-H522
12.5
Brain (Substantia nigra)
0.0
|
Pool
|
Liver
1.3
Brain (Thalamus) Pool
1.9
|
Fetal Liver
5.0
Brain (whole)
1.2
|
Liver ca. HepG2
6.5
Spinal Cord Pool
2.2
|
Kidney Pool
21.2
Adrenal Gland
1.6
|
Fetal Kidney
43.8
Pituitary gland Pool
0.0
|
Renal ca. 786-0
3.8
Salivary Gland
1.4
|
Renal ca. A498
2.3
Thyroid (female)
1.3
|
Renal ca. ACHN
15.0
Pancreatic ca. CAPAN2
11.1
|
Renal ca. UO-31
1.2
Pancreas Pool
15.8
|
|
[1308]
598
TABLE ASC
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3769,
Rel. Exp. (%) Ag3769,
|
Tissue Name
Run 170069130
Tissue Name
Run 170069130
|
|
Secondary Th1 act
0.6
HUVEC IL-1beta
0.5
|
Secondary Th2 act
1.2
HUVEC IFN gamma
1.0
|
Secondary Tr1 act
0.7
HUVEC TNF alpha + IFN
1.1
|
gamma
|
Secondary Th1 rest
0.9
HUVEC TNF alpha + IL4
0.8
|
Secondary Th2 rest
1.7
HUVEC IL-11
0.4
|
Secondary Tr1 rest
1.3
Lung Microvascular EC none
4.9
|
Primary Th1 act
1.0
Lung Microvascular EC
2.8
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.8
Microvascular Dermal EC none
1.6
|
Primary Tr1 act
1.1
Microsvasular Dermal EC
0.7
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.7
Bronchial epithelium TNF alpha +
1.9
|
IL1beta
|
Primary Th2 rest
1.5
Small airway epithelium none
0.4
|
Primary Tr1 rest
2.3
Small airway epithelium
1.1
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.6
Coronery artery SMC rest
0.3
|
act
|
CD45RO CD4 lymphocyte
1.3
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
0.9
Astrocytes rest
0.7
|
Secondary CD8
1.1
Astrocytes TNF alpha + IL-1beta
0.2
|
lymphocyte rest
|
Secondary CD8
0.3
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.4
KU-812 (Basophil)
1.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
1.0
CCD1106 (Keratinocytes) none
1.1
|
CD95 CH11
|
LAK cells rest
0.8
CCD1106 (Keratinocytes)
0.4
|
TNF alpha + IL-1beta
|
LAK cells IL-2
2.0
Liver cirrhosis
0.2
|
LAK cells IL-2 + IL-12
1.2
NCI-H292 none
0.2
|
LAK cells IL-2 + IFN
0.9
NCI-H292 IL-4
0.5
|
gamma
|
LAK cells IL-2 + IL-18
1.4
NCI-H292 IL-9
1.7
|
LAK cells
1.1
NCI-H292 IL-13
0.5
|
PMA/ionomycin
|
NK Cells IL-2 rest
1.2
NCI-H292 IFN gamma
0.6
|
Two Way MLR 3 day
1.0
HPAEC none
0.6
|
Two Way MLR 5 day
0.6
HPAEC TNF alpha + IL-1beta
1.0
|
Two Way MLR 7 day
0.5
Lung fibroblast none
0.7
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
1.5
Lung fibroblast IL-4
0.3
|
PBMC PHA-L
1.2
Lung fibroblast IL-9
0.8
|
Ramos (B cell) none
1.2
Lung fibroblast IL-13
0.5
|
Ramos (B cell) ionomycin
1.3
Lung fibroblast IFN gamma
0.5
|
B lymphocytes PWM
0.2
Dermal fibroblast CCD1070 rest
0.6
|
B lymphocytes CD40L
1.9
Dermal fibroblast CCD1070
0.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.3
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
4.3
Dermal fibroblast IFN gamma
0.3
|
PMA/ionomycin
|
Dendritic cells none
1.0
Dermal fibroblast IL-4
2.1
|
Dendritic cells LPS
0.8
Dermal Fibroblasts rest
0.3
|
Dendritic cells anti-CD40
0.8
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.8
Neutrophils rest
0.3
|
Monocytes LPS
1.0
Colon
17.7
|
Macrophages rest
0.9
Lung
48.0
|
Macrophages LPS
0.7
Thymus
1.0
|
HUVEC none
0.3
Kidney
100.0
|
HUVEC starved
1.1
|
|
[1309] CNS_neurodegeneration_v1.0 Summary: Ag3769 Expression of the CG92078-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1310] General_screening13 panel_v1.4 Summary: Ag3769 Highest expression of the CG92078-01 gene is detected in colon cancer CaCo-2 cell line (CT=33). In addition, significant expression of this gene is associated with two CNS cancer, and gastric cancer (liver metatstasis) NCI-N87 cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancer.
[1311] Panel 4.1D Summary: Ag3769 Highest expression of the CG92078-01 gene is detected in kidney (CT=30.6). In addition, moderate to low expression of this gene is also seen in lung and colon samples. Therefore, expression of this gene can be used to distinguish these samples from other samples used in this panel. In addition, therapeutic modulation of the activity of the permease encoded by this gene may be useful in the treatment of inflammatory bowel disease, inflammatory or autoimmune diseases that affect the kidney and lung, including lupus and glomerulonephritis, asthma, COPD, allergies and emphysema.
[1312] Panel 5 Islet Summary: Ag3769 Expression of the CG92078-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1313] AT. CG92088-01: IGb3 synthase
[1314] Expression of gene CG92088-01 was assessed using the primer-probe sets Ag3773 and Ag4123, described in Tables ATA and ATB. Results of the RTQ-PCR runs are shown in Tables ATC and ATD.
599TABLE ATA
|
|
Probe Name Ag3773
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-aagaggctagacagcagaacct-3′22284390
ProbeTET-5′-caccattgggctgactatctttgctg-26306391
3′-TAMRA
Reverse5′-ccaggtacttctccaggtacct-3′22337392
|
[1315]
600
TABLE ATB
|
|
|
Probe Name Ag4123
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-ttctgtatggcctccctaaatt-3′
22
86
393
|
Probe
TET-5′-atctggaagccctcatccccatg-3′-
23
113
394
|
TAMRA
|
Reverse
5′-gtctctcagctgggacattgt-3′
21
154
395
|
|
[1316]
601
TABLE ATC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag4123, Run
Rel. Exp. (%) Ag4123, Run
|
Tissue Name
214301671
Tissue Name
214301671
|
|
AD 1 Hippo
0.0
Control (Path) 3
0.0
|
Temporal Ctx
|
AD 2 Hippo
17.2
Control (Path) 4
100.0
|
Temporal Ctx
|
AD 3 Hippo
4.4
AD 1 Occipital Ctx
20.3
|
AD 4 Hippo
0.0
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
57.4
AD 3 Occipital Ctx
0.0
|
AD 6 Hippo
43.2
AD 4 Occipital Ctx
14.7
|
Control 2 Hippo
0.0
AD 5 Occipital Ctx
14.7
|
Control 4 Hippo
0.0
AD 6 Occipital Ctx
7.2
|
Control (Path) 3 Hippo
0.0
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
12.3
Control 2 Occipital Ctx
2.0
|
AD 2 Temporal Ctx
18.9
Control 3 Occipital Ctx
0.0
|
AD 3 Temporal Ctx
0.0
Control 4 Occipital Ctx
13.7
|
AD 4 Temporal Ctx
36.6
Control (Path) 1
41.2
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
88.9
Control (Path) 2
0.0
|
Occipital Ctx
|
AD 5 Sup Temporal Ctx
23.0
Control (Path) 3
0.0
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
60.7
Control (Path) 4
0.0
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
39.5
Control 1 Parietal Ctx
0.0
|
Control 1 Temporal Ctx
0.0
Control 2 Parietal Ctx
92.0
|
Control 2 Temporal Ctx
0.0
Control 3 Parietal Ctx
0.0
|
Control 3 Temporal Ctx
0.0
Control (Path) 1
0.0
|
Parietal Ctx
|
Control 4 Temporal Ctx
0.0
Control (Path) 2
19.3
|
Parietal Ctx
|
Control (Path) 1
2.8
Control (Path) 3
0.0
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
17.9
Control (Path) 4
57.4
|
Temporal Ctx
Parietal Ctx
|
|
[1317]
602
TABLE ATD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag4123,
Rel. Exp. (%) Ag4123,
|
Tissue Name
Run 172859311
Tissue Name
Run 172859311
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
11.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
18.3
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0
|
IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Th1 rest
24.3
Small airway epithelium
26.6
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
10.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1beta
|
CD8 lymphocyte act
19.9
Astrocytes rest
0.0
|
Secondary CD8
11.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
24.5
CCD1106 (Keratinocytes) none
1.2
|
CD95 CH11
|
LAK cells rest
1.3
CCD1106 (Keratinocytes)
1.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
10.1
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
1.8
|
LAK cells IL-2 + IFN
11.3
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
7.2
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
9.2
HPAEC none
0.0
|
Two Way MLR 5 day
29.3
HPAEC TNF alpha + IL-1beta
7.1
|
Two Way MLR 7 day
12.2
Lung fibroblast none
7.3
|
PBMC rest
9.7
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
26.8
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
42.9
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
55.1
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
10.5
|
B lymphocytes CD40L
9.0
Dermal fibroblast CCD1070
8.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
9.2
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
12.2
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
17.9
Dermal fibroblast IL-4
1.4
|
Dendritic cells LPS
29.9
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
100.0
|
Monocytes rest
40.1
Neutrophils rest
94.6
|
Monocytes LPS
21.2
Colon
0.0
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
1.1
Thymus
0.0
|
HUVEC none
0.0
Kidney
9.1
|
HUVEC starved
11.7
|
|
[1318] AI_comprehensive panel_v1.0 Summary: Ag3773 Expression of the CG92088-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1319] CNS_neurodegeneration_v1.0 Summary: Ag4123 Low expression of the CG92088-01 gene is exclusively seen in the temporal cortex of Alzheimer's disease patients. The CG92088-01 gene encodes a homolog of rat iGb3 synthase gene, which is a member of the ABO blood group glycosyltransferases. As seen in panel 4.1D, expression of this gene seems to be restricted to neutrophils. This suggests that this low expression could be from the infiltrating neutrophils in the diseased tissue. Therefore, blockade of this gene product may be of use in the treatment of this disease and decrease neuronal death. Ag3773 Expression of the CG92088-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1320] General_screening_panel_v1.4 Summary: Ag3773 Expression of the CG92088-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1321] Panel 2.2 Summary: Ag3773 Expression of the CG92088-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1322] Panel 4.1D Summary: Ag4123 Expression of the CG92088-01 gene is exclusively seen in neutrophils. Thus, expression of this gene can be used to distinguish this sample from other samples in this panel. In addition, modulation of the expression or activity of the protein encoded by this gene through the application of small molecule therapeutics may be useful in the treatment of Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive pulmonary disease, asthma, emphysema, rheumatoid arthritis, lupus erythematosus, or psoriasis. Ag3773 Expression of the CG92088-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1323] AU. CG92142-01: GLYCEROL-3-PHOSPHATE ACYLTRANSFERASE
[1324] Expression of gene CG92142-01 was assessed using the primer-probe set Ag3774, described in Table AUA. Results of the RTQ-PCR runs are shown in Tables AUB, AUC, AUD, AUE and AUF.
603TABLE AUA
|
|
Probe Name Ag3774
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ggtgctgctaaaactgttcaac-3′22673396
ProbeTET-5′-tggaacattcaaattcacaaaggtca-26704397
3′-TAMRA
Reverse5′-attcgtctcagttgcagcttt-3′21743398
|
[1325]
604
TABLE AUB
|
|
|
CNS_neurodegeneration_v1 .0
|
Rel. Exp. (%) Ag3774, Run
Rel. Exp. (%) Ag3774, Run
|
Tissue Name
206871268
Tissue Name
206871268
|
|
AD 1 Hippo
29.1
Control (Path) 3
29.3
|
Temporal Ctx
|
AD 2 Hippo
73.7
Control (Path) 4
50.3
|
Temporal Ctx
|
AD 3 Hippo
10.0
AD 1 Occipital Ctx
22.4
|
AD 4 Hippo
14.6
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
92.0
AD 3 Occipital Ctx
20.3
|
AD 6 Hippo
45.1
AD 4 Occipital Ctx
33.9
|
Control 2 Hippo
44.1
AD 5 Occipital Ctx
37.6
|
Control 4 Hippo
20.3
AD 6 Occipital Ctx
24.7
|
Control (Path) 3 Hippo
19.9
Control 1 Occipital Ctx
11.3
|
AD 1 Temporal Ctx
20.6
Control 2 Occipital Ctx
48.0
|
AD 2 Temporal Ctx
75.3
Control 3 Occipital Ctx
43.5
|
AD 3 Temporal Ctx
13.4
Control 4 Occipital Ctx
21.2
|
AD 4 Temporal Ctx
45.1
Control (Path) 1
81.8
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
12.9
|
Occipital Ctx
|
AD 5 Sup Temporal
78.5
Control (Path) 3
13.6
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
43.5
Control (Path) 4
45.1
|
Occipital Ctx
|
AD 6 Sup Temporal
50.7
Control 1 Parietal Ctx
25.2
|
Ctx
|
Control 1 Temporal Ctx
25.5
Control 2 Parietal Ctx
84.7
|
Control 2 Temporal Ctx
46.7
Control 3 Parietal Ctx
41.2
|
Control 3 Temporal Ctx
57.0
Control (Path) 1
91.4
|
Parietal Ctx
|
Control 3 Temporal Ctx
25.2
Control (Path) 2
38.2
|
Parietal Ctx
|
Control (Path) 1
66.4
Control (Path) 3
19.1
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
52.1
Control (Path) 4
48.0
|
Temporal Ctx
Parietal Ctx
|
|
[1326]
605
TABLE AUC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3774, Run
Rel. Exp. (%) Ag3774, Run
|
Tissue Name
213515543
Tissue Name
213515543
|
|
Adipose
63.7
Renal ca. TK-10
21.5
|
Melanoma* Hs688(A).T
16.0
Bladder
6.3
|
Melanoma* Hs688(B).T
74.7
Gastric ca. (liver met.)
9.7
|
NCI-N87
|
Melanoma* M14
10.2
Gastric ca. KATO III
16.5
|
Melanoma* LOXIMVI
76.8
Colon ca. SW-948
3.3
|
Melanoma* SK-MEL-5
23.8
Colon ca. SW480
12.9
|
Squamous cell
5.8
Colon ca.* (SW480 met)
8.6
|
carcinoma SCC-4
SW620
|
Testis Pool
12.8
Colon ca. HT29
4.1
|
Prostate ca.* (bone met)
10.3
Colon ca. HCT-116
25.3
|
PC-3
|
Prostate Pool
2.3
Colon ca. CaCo-2
52.5
|
Placenta
1.3
Colon cancer tissue
10.4
|
Uterus Pool
1.6
Colon ca. SW1116
3.0
|
Ovarian ca. OVCAR-3
10.6
Colon ca. Colo-205
2.9
|
Ovarian ca. SK-OV-3
15.6
Colon ca. SW-48
2.5
|
Ovarian ca. OVCAR-4
5.4
Colon Pool
4.5
|
Ovarian ca. OVCAR-5
6.3
Small Intestine Pool
5.9
|
Ovarian ca. IGROV-1
5.5
Stomach Pool
3.3
|
Ovarian ca. OVCAR-8
4.9
Bone Marrow Pool
2.8
|
Ovary
4.0
Fetal Heart
3.1
|
Breast ca. MCF-7
11.7
Heart Pool
4.0
|
Breast ca. MDA-MB-
8.5
Lymph Node Pool
7.2
|
231
|
Breast ca. BT 549
6.5
Fetal Skeletal Muscle
11.0
|
Breast ca. T47D
8.9
Skeletal Muscle Pool
10.9
|
Breast ca. MDA-N
10.7
Spleen Pool
5.3
|
Breast Pool
5.0
Thymus pool
7.6
|
Trachea
10.6
CNS cancer (glio/astro)
9.7
|
U87-MG
|
Lung
1.0
CNS cancer (glio/astro) U-
19.1
|
118-MG
|
Fetal Lung
6.2
CNS cancer (neuro; met)
22.1
|
SK-N-AS
|
Lung ca. NCI-N417
3.2
CNS cancer (astro) SF-539
5.9
|
Lung ca. LX-1
9.3
CNS cancer (astro) SNB-75
22.5
|
Lung ca. NCI-H146
2.9
CNS cancer (glio) SNB-19
5.0
|
Lung ca. SHP-77
16.2
CNS cancer (glio) SF-295
100.0
|
Lung ca. A549
7.6
Brain (Amygdala) Pool
2.9
|
Lung ca. NCI-H526
1.9
Brain (cerebellum)
2.4
|
Lung ca. NCI-H23
12.7
Brain (fetal)
17.9
|
Lung ca. NCI-H460
7.7
Brain (Hippocampus) Pool
5.9
|
Lung ca. HOP-62
6.0
Cerebral Cortex Pool
7.5
|
Lung ca. NCI-H522
17.6
Brain (Substantia nigra)
5.8
|
Pool
|
Liver
16.3
Brain (Thalamus) Pool
8.1
|
Fetal Liver
70.7
Brain (whole)
8.4
|
Liver ca. HepG2
42.9
Spinal Cord Pool
4.8
|
Kidney Pool
8.5
Adrenal Gland
65.5
|
Fetal Kidney
6.6
Pituitary gland Pool
1.0
|
Renal ca. 786-0
10.3
Salivary Gland
3.0
|
Renal ca. A498
2.5
Thyroid (female)
3.8
|
Renal ca. ACHN
7.3
Pancreatic ca. CAPAN2
5.4
|
Renal ca. UO-31
7.2
Pancreas Pool
5.7
|
|
[1327]
606
TABLE AUD
|
|
|
Panel 2.2
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3774,
Ag3774,
|
Tissue Name
Run 174448446
Tissue Name
Run 174448446
|
|
Normal Colon
7.9
Kidney Margin (OD04348)
8.7
|
Colon cancer (OD06064)
4.9
Kidney malignant cancer
2.2
|
(OD06204B)
|
Colon Margin (OD06064)
3.6
Kidney normal adjacent
0.4
|
tissue (OD06204E)
|
Colon cancer (OD06159)
0.2
Kidney Cancer (OD04450-
3.4
|
01)
|
Colon Margin (OD06159)
2.8
Kidney Margin (OD04450-
3.3
|
03)
|
Colon cancer (OD06297-04)
0.6
Kidney Cancer 8120613
0.8
|
Colon Margin (OD06297-05)
2.3
Kidney Margin 8120614
1.0
|
CC Gr.2 ascend colon
0.5
Kidney Cancer 9010320
1.6
|
(ODO3921)
|
CC Margin (ODO3921)
1.0
Kidney Margin 9010321
0.2
|
Colon cancer metastasis
1.6
Kidney Cancer 8120607
0.8
|
(OD06104)
|
Lung Margin (OD06104)
1.1
Kidney Margin 8120608
0.3
|
Colon mets to lung
2.2
Normal Uterus
5.0
|
(OD04451-01)
|
Lung Margin (OD04451-02)
2.3
Uterine Cancer 064011
1.1
|
Normal Prostate
0.6
Normal Thyroid
0.3
|
Prostate Cancer (OD04410)
1.2
Thyroid Cancer 064010
0.6
|
Prostate Margin (OD04410)
1.2
Thyroid Cancer A302152
2.2
|
Normal Ovary
1.0
Thyroid Margin A302153
2.9
|
Ovarian cancer (OD06283-
1.0
Normal Breast
61.6
|
03)
|
Ovarian Margin (OD06283-
10.1
Breast Cancer (OD04566)
2.7
|
07)
|
Ovarian Cancer 064008
3.3
Breast Cancer 1024
4.8
|
Ovarian cancer (OD06145)
2.1
Breast Cancer (OD04590-
4.8
|
01)
|
Ovarian Margin (OD06145)
2.4
Breast Cancer Mets
30.1
|
(OD04590-03)
|
Ovarian cancer (OD06455-
1.7
Breast Cancer Metastasis
6.0
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
1.3
Breast Cancer 064006
2.0
|
07)
|
Normal Lung
3.1
Breast Cancer 9100266
1.5
|
Invasive poor diff. lung
1.4
Breast Margin 9100265
3.6
|
adeno (ODO4945-01
|
Lung Margin (ODO4945-03)
2.2
Breast Cancer A209073
1.1
|
Lung Malignant Cancer
2.0
Breast Margin A2090734
5.8
|
(OD03126)
|
Lung Margin (OD03126)
0.7
Breast cancer (OD06083)
4.2
|
Lung Cancer (OD05014A)
1.2
Breast cancer node
12.6
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
7.1
Normal Liver
87.7
|
Lung cancer (OD06081)
0.1
Liver Cancer 1026
12.5
|
Lung Margin (OD06081)
2.0
Liver Cancer 1025
100.0
|
Lung Cancer (OD04237-01)
1.0
Liver Cancer 6004-T
63.7
|
Lung Margin (OD04237-02)
2.6
Liver Tissue 6004-N
4.8
|
Ocular Melanoma Metastasis
7.5
Liver Cancer 6005-T
28.5
|
Ocular Melanoma Margin
19.5
Liver Tissue 6005-N
67.8
|
(Liver)
|
Melanoma Metastasis
2.0
Liver Cancer 064003
12.2
|
Melanoma Margin (Lung)
3.6
Normal Bladder
2.3
|
Normal Kidney
1.6
Bladder Cancer 1023
0.3
|
Kidney Ca, Nuclear grade 2
3.3
Bladder Cancer A302173
1.4
|
(OD04338)
|
Kidney Margin (OD04338)
1.3
Normal Stomach
6.0
|
Kidney Ca Nuclear grade 1/2
2.2
Gastric Cancer 9060397
0.9
|
(OD04339)
|
Kidney Margin (OD04339)
2.2
Stomach Margin 9060396
1.7
|
Kidney Ca, Clear cell type
0.7
Gastric Cancer 9060395
1.9
|
(OD04340)
|
Kidney Margin (OD04340)
4.0
Stomach Margin 9060394
2.3
|
Kidney Ca, Nuclear grade 3
0.9
Gastric Cancer 064005
1.9
|
(OD04348)
|
|
[1328]
607
TABLE AUE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3774,
Ag3774,
|
Tissue Name
Run 170130276
Tissue Name
Run 170130276
|
|
Secondary Th1 act
39.8
HUVEC IL-1beta
38.2
|
Secondary Th2 act
44.4
HUVEC IFN gamma
39.0
|
Secondary Tr1 act
33.7
HUVEC TNF alpha + IFN
19.1
|
gamma
|
Secondary Th1 rest
9.5
HUVEC TNF alpha + IL4
28.1
|
Secondary Th2 rest
11.4
HUVEC IL-11
25.2
|
Secondary Tr1 rest
12.2
Lung Microvascular EC none
32.3
|
Primary Th1 act
36.6
Lung Microvascular EC
36.3
|
TNF alpha + IL-1beta
|
Primary Th2 act
39.8
Microvascular Dermal EC none
26.4
|
Primary Tr1 act
28.9
Microvascular Dermal EC
23.3
|
TNF alpha + IL-1beta
|
Primary Th1 rest
24.8
Bronchial epithelium
38.4
|
TNF alpha + IL1beta
|
Primary Th2 rest
11.7
Small airway epithelium none
24.1
|
Primary Tr1 rest
23.2
Small airway epithelium
28.9
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
45.1
Coronery artery SMC rest
31.4
|
act
|
CD45RO CD4 lymphocyte
45.1
Coronery artery SMC
24.5
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
49.0
Astrocytes rest
46.3
|
Secondary CD8
31.2
Astrocytes TNF alpha + IL-1beta
12.1
|
lymphocyte rest
|
Secondary CD8
22.1
KU-812 (Basophil) rest
37.9
|
lymphocyte act
|
CD4 lymphocyte none
11.0
KU-812 (Basophil)
49.3
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
15.9
CCD1106 (Keratinocytes) none
56.3
|
CD95 CH11
|
LAK cells rest
18.7
CCD1106 (Keratinocytes)
34.6
|
TNF alpha + IL-1beta
|
LAK cells IL-2
31.4
Liver cirrhosis
38.4
|
LAK cells IL-2 + IL-12
25.3
NCI-H292 none
25.2
|
LAK cells IL-2 + IFN
46.7
NCI-H292 IL-4
36.3
|
gamma
|
LAK cells IL-2 + IL-18
32.8
NCI-H292 IL-9
47.6
|
LAK cells
3.9
NCI-H292 IL-13
37.1
|
PMA/ionomycin
|
NK Cells IL-2 rest
30.8
NCI-H292 IFN gamma
49.3
|
Two Way MLR 3 day
23.3
HPAEC none
27.7
|
Two Way MLR 5 day
37.6
HPAEC TNF alpha + IL-1beta
31.9
|
Two Way MLR 7 day
17.8
Lung fibroblast none
44.1
|
PBMC rest
4.1
Lung fibroblast TNF alpha + IL-
17.0
|
1beta
|
PBMC PWM
35.4
Lung fibroblast IL-4
34.9
|
PBMC PHA-L
20.9
Lung fibroblast IL-9
62.4
|
Ramos (B cell) none
76.8
Lung fibroblast IL-13
42.0
|
Ramos (B cell) ionomycin
68.8
Lung fibroblast IFN gamma
25.2
|
B lymphocytes PWM
41.2
Dermal fibroblast CCD1070 rest
100.0
|
B lymphocytes CD40L
28.9
Dermal fibroblast CCD1070
66.4
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
17.4
Dermal fibroblast CCD1070 IL-
38.2
|
1beta
|
EOL-1 dbcAMP
20.9
Dermal fibroblast IFN gamma
17.0
|
PMA/ionomycin
|
Dendritic cells none
21.0
Dermal fibroblast IL-4
47.3
|
Dendritic cells LPS
5.7
Dermal Fibroblasts rest
29.5
|
Dendritic cells anti-CD40
22.5
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
7.9
Neutrophils rest
2.3
|
Monocytes LPS
2.6
Colon
15.4
|
Macrophages rest
22.2
Lung
23.8
|
Macrophages LPS
4.5
Thymus
68.3
|
HUVEC none
29.7
Kidney
49.3
|
HUVEC starved
34.6
|
|
[1329]
608
TABLE AUF
|
|
|
Panel 5D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3774, Run
Ag3774, Run
|
Tissue Name
223675472
Tissue Name
223675472
|
|
97457_Patient-
17.7
94709_Donor 2 AM - A_adipose
19.6
|
02go_adipose
|
97476_Patient-07sk_skeletal
3.6
94710_Donor 2 AM - B_adipose
9.3
|
muscle
|
97477_Patient-07ut_uterus
2.3
94711_Donor 2 AM - C_adipose
7.5
|
97478_Patient-
2.2
94712_Donor 2 AD - A_adipose
56.6
|
07pl_placenta
|
97481_Patient-08sk_skeletal
6.4
94713_Donor 2 AD - B_adipose
72.2
|
muscle
|
97482_Patient-08ut_uterus
1.6
94714_Donor 2 AD - C_adipose
70.2
|
97483_Patient-
0.8
94742_Donor 3 U - A_Mesenchymal
1.6
|
08pl_placenta
Stem Cells
|
97486_Patient-09sk_skeletal
0.5
94743_Donor 3 U - B_Mesenchymal
1.8
|
muscle
Stem Cells
|
97487_Patient-09ut_uterus
2.1
94730_Donor 3 AM - A_adipose
13.1
|
97488_Patient-
0.8
94731_Donor 3 AM - B_adipose
8.5
|
09pl_placenta
|
97492_Patient-10ut_uterus
1.6
94732_Donor 3 AM - C_adipose
8.7
|
97493_Patient-
1.4
94733_Donor 3 AD - A_adipose
100.0
|
10pl_placenta
|
97495_Patient-
10.4
94734_Donor 3 AD - B_adipose
62.9
|
11go_adipose
|
97496_Patient-11sk_skeletal
2.8
94735_Donor 3 AD - C_adipose
53.2
|
muscle
|
97497_Patient-11ut_uterus
2.1
77138_Liver_HepG2untreated
56.6
|
97498_Patient-
1.8
73556_Heart_Cardiac stromal cells
0.5
|
11pl_placenta
(primary)
|
97500_Patient-
13.5
81735_Small Intestine
2.3
|
12go_adipose
|
97501_Patient-12sk_skeletal
6.0
72409_Kidney_Proximal Convoluted
1.0
|
muscle
Tubule
|
97502_Patient-12ut_uterus
2.6
82685_Small intestine_Duodenum
1.6
|
97503_Patient-
0.4
90650_Adrenal_Adrenocortical
4.6
|
12pl_placenta
adenoma
|
94721_Donor 2 U -
3.5
72410_Kidney_HRCE
3.3
|
A_Mesenchymal Stem Cells
|
94722_Donor 2 U -
3.7
72411_Kidney_HRE
2.7
|
B_Mesenchymal Stem Cells
|
94723_Donor 2 U -
2.7
73139_Uterus_Uterine smooth
1.2
|
C_Mesenchymal Stem Cells
muscle cells
|
|
[1330] CNS_neurodegeneration_v1.0 Summary: Ag3774 This panel confirms the expression of the CG92142-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1331] General_screening_panel_v1.4 Summary: Ag3774 Highest expression of the CG92142-01 gene is detected in CNS cancer (glio) SF-295 cell line (CT=26). High expression of this gene is also in number of cancer cell lines (pancreatic, CNS, colon, gastric, renal, lung, breast, ovarian, squamous cell carcinoma, prostate and melanoma). Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs might be beneficial in the treatment of these cancers.
[1332] Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1333] The CG92142-01 gene encodes a mitochondrial glycerol-3-phosphate acyltransferase (GPAT). GPAT is an adipocyte determination and differentiation factor 1 (ADD1) and sterol regulatory element-binding protein-i (SREBP-1) regulated differentiation gene (Ericsson J, Jackson S M, Kim J B, Spiegelman B M, Edwards P A. (1997) Identification of glycerol-3-phosphate acyltransferase as an adipocyte determination and differentiation factor 1- and sterol regulatory element-binding protein-responsive gene. J Biol Chem 272(11):7298-305). It is up-regulated by insulin and high-carbohydrate diets (Dircks L K, Sul HS. (1997) Mammalian mitochondrial glycerol-3-phosphate acyltransferase. Biochim Biophys Acta 1348(1-2):17-26). GPAT up-regulation increases triglyceride (TG) synthesis and fat deposition. Inhibition of GPAT activiy could lead to decreased TG synthesis and fat deposition. Troglitazone, a thiazolidinedione compound used to treat non-insulin-dependent diabetes mellitus (NIDDM), was shown to decreases GPAT activity and adipogenesis in ZDF rat islets (Shimabukuro M, Zhou Y T, Lee Y, Unger R H. (1998) Troglitazone lowers islet fat and restores beta cell function of Zucker diabetic fatty rats. J Biol Chem 273(6):3547-50). Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of diabetes.
[1334] In addition, this gene is expressed at moderate levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1335] Panel 2.2 Summary: Ag3774 Highest expression of the CG92142-01 gene is detected in liver cancer 1025 sample (CT=28.7). In addition, low to moderate expression of this gene is seen in number of cancer and normal samples used in this panel. Please see Panel 1.4 for a discussion of the potential utility of this gene.
[1336] Panel 4.1D Summary: Ag3774 Highest expression of the CG92142-01 gene is detected in resting dermal fibroblast CCD1070 (CT=31). This gene is expressed at low to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1337] Interestingly, expression of this gene is stimulated in PWM treated PBMC cells (CT=32.5) as compared to resting PBMC (35.6). Therefore, expression of this gene can be used to distinguish between resting and stimulated PBMC cells.
[1338] Panel 5D Summary: Ag3774 Highest expression of the CG92142-01 gene is detected in 94733_Donor 3 AD-A_adipose sample (CT=27.6). In addition, high to moderated expression of this gene is also seen in number of adipose, small intestine, uterus, skeletal muscle, placenta and mesenchymal stem cell samples. Please see Panel 1.4 for a discussion of the potential utility of this gene.
[1339] AV. CG92152-01: PLASMINOGEN ACTIVATOR SPA
[1340] Expression of gene CG92152-01 was assessed using the primer-probe set Ag3775, described in Table AVA. Results of the RTQ-PCR runs are shown in Tables AVB, AVC and AVD.
609TABLE AVA
|
|
Probe Name Ag3775
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-gcagattcaagtgcatgtgtta-3′22517399
ProbeTET-5′-ttactattctgcccatccaggaggga-26562400
3′-TAMRA
Reverse5′-gcacagatcatcttctctgtga-3′22588401
|
[1341]
610
TABLE AVB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3775, Run
Ag3775, Run
Ag3775, Run
Ag3775, Run
|
Tissue Name
211176610
224339887
Tissue Name
211176610
224339887
|
|
AD 1 Hippo
34.2
29.1
Control (Path)
4.4
2.1
|
3 Temporal
|
Ctx
|
AD 2 Hippo
33.4
25.7
Control (Path)
37.1
21.5
|
4 Temporal
|
Ctx
|
AD 3 Hippo
25.7
9.7
AD 1 Occipital
24.7
19.9
|
Ctx
|
AD 4 Hippo
9.0
8.1
AD 2 Occipital
0.0
0.0
|
Ctx (Missing)
|
AD 5 hippo
100.0
61.6
AD 3 Occipital
8.1
9.0
|
Ctx
|
AD 6 Hippo
88.9
100.0
AD 4 Occipital
12.2
16.0
|
Ctx
|
Control 2 Hippo
10.7
20.9
AD 5 Occipital
29.5
11.3
|
Ctx
|
Control 4 Hippo
12.2
5.9
AD 6 Occipital
29.9
29.9
|
Ctx
|
Control (Path) 3
4.7
2.8
Control 1
1.8
0.9
|
Hippo
Occipital Ctx
|
AD 1 Temporal
24.0
26.8
Control 2
21.9
23.3
|
Ctx
Occipital Ctx
|
AD 2 Temporal
22.2
21.5
Control 3
8.9
3.3
|
Ctx
Occipital Ctx
|
AD 3 Temporal
15.9
5.0
Control 4
3.3
3.5
|
Ctx
Occipital Ctx
|
AD 4 Temporal
16.2
21.9
Control (Path)
46.0
36.1
|
Ctx
1 Occipital Ctx
|
AD 5 Inf
79.0
36.1
Control (Path)
3.9
3.3
|
Temporal Ctx
2 Occipital Ctx
|
AD 5
73.2
47.3
Control (Path)
4.6
1.3
|
SupTemporal Ctx
3 Occipital Ctx
|
AD 6 Inf
70.2
33.9
Control (Path)
3.3
3.2
|
Temporal Ctx
4 Occipital Ctx
|
AD 6 Sup
74.2
51.1
Control 1
3.7
3.3
|
Temporal Ctx
Parietal Ctx
|
Control 1
1.4
3.7
Control 2
26.8
34.9
|
Temporal Ctx
Parietal Ctx
|
Control 2
18.7
12.9
Control 3
9.3
3.1
|
Temporal Ctx
Parietal Ctx
|
Control 3
8.4
6.5
Control (Path)
44.4
27.7
|
Temporal Ctx
1 Parietal Ctx
|
Control 4
14.2
2.9
Control (Path)
13.1
17.7
|
Temporal Ctx
2 Parietal Ctx
|
Control (Path) 1
53.6
29.5
Control (Path)
1.4
4.5
|
Temporal Ctx
3 Parietal Ctx
|
Control (Path) 2
43.2
21.8
Control (Path)
32.8
23.3
|
Temporal Ctx
4 Parietal Ctx
|
|
[1342]
611
TABLE AVC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3775, Run
Ag3775, Run
|
Tissue Name
219514534
Tissue Name
219514534
|
|
Adipose
17.1
Renal ca. TK-10
0.0
|
Melanoma* Hs688(A).T
0.0
Bladder
3.9
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.0
|
NCI-N87
|
Melanoma* M14
21.6
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
5.2
Colon ca. SW480
0.0
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
SW620
|
Testis Pool
40.1
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
6.0
Colon ca. HCT-116
0.0
|
PC-3
|
Prostate Pool
5.7
Colon ca. CaCo-2
0.0
|
Placenta
0.0
Colon cancer tissue
12.8
|
Uterus Pool
6.1
Colon ca. SW1116
0.0
|
Ovarian ca OVCAR-3
100.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
5.4
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
13.6
|
Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
12.2
|
Ovarian ca. IGROV-1
12.5
Stomach Pool
15.6
|
Ovarian ca. OVCAR-8
11.3
Bone Marrow Pool
10.5
|
Ovary
12.1
Fetal Heart
3.7
|
Breast ca. MCF-7
10.4
Heart Pool
17.2
|
Breast ca. MDA-MB-
0.0
Lymph Node Pool
24.5
|
231
|
Breast ca. BT 549
56.3
Fetal Skeletal Muscle
14.7
|
Breast ca. T47D
0.0
Skeletal Muscle Pool
61.1
|
Breast ca. MDA-N
6.2
Spleen Pool
90.8
|
Breast Pool
9.9
Thymus Pool
40.3
|
Trachea
11.2
CNS cancer (glio/astro)
54.3
|
U87-MG
|
Lung
9.7
CNS cancer (glio/astro) U-
0.0
|
118-MG
|
Fetal Lung
3.8
CNS cancer (neuro; met)
2.3
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
3.0
|
Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
35.1
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
3.8
|
Lung ca. SHP-77
12.3
CNS cancer (glio) SF-295
39.5
|
Lung ca. A549
0.0
Brain (Amygdala) Pool
21.5
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
6.4
|
Lung ca. NCI-H23
0.0
Brain (fetal)
45.1
|
Lung ca. NCI-H460
30.4
Brain (Hippocampus) Pool
25.7
|
Lung ca. HOP-62
23.0
Cerebral Cortex Pool
34.4
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
12.9
|
Pool
|
Liver
3.8
Brain (Thalamus) Pool
36.3
|
Fetal Liver
3.1
Brain (whole)
33.7
|
Liver ca. HepG2
1.2
Spinal Cord Pool
21.5
|
Kidney Pool
27.7
Adrenal Gland
20.3
|
Fetal Kidney
49.0
Pituitary gland Pool
6.1
|
Renal ca. 786-0
0.0
Salivary Gland
6.6
|
Renal ca. A498
0.0
Thyroid (female)
0.0
|
Renal ca. ACHN
1.9
Pancreatic ca. CAPAN2
0.0
|
Renal ca. UO-31
0.0
Pancreas Pool
18.9
|
|
[1343]
612
TABLE AVD
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3775,
Ag3775,
|
Tissue Name
Run 170129781
Tissue Name
Run 170129781
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
5.1
|
Secondary Th2 act
51.4
HUVEC IFN gamma
4.8
|
Secondary Tr1 act
29.1
HUVEC TNF alpha + IFN
1.4
|
gamma
|
Secondary Th1 rest
4.0
HUVEC TNF alpha + IL4
4.7
|
Secondary Th2 rest
20.0
HUVEC IL-11
1.8
|
Secondary Tr1 rest
14.9
Lung Microvascular EC none
0.0
|
Primary Th1 act
6.7
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
10.4
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
8.8
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.9
Bronchial epithelium
4.2
|
TNF alpha + IL1beta
|
Primary Th2 rest
2.1
Small airway epithelium none
0.0
|
Primary Tr1 rest
13.4
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
7.8
Coronery artery SMC rest
2.8
|
act
|
CD45RO CD4 lymphocyte
2.8
Coronery artery SMC
2.0
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
6.6
Astrocytes rest
15.3
|
Secondary CD8
3.5
Astrocytes TNF alpha + IL-1beta
9.1
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
4.3
|
lymphocyte act
|
CD4 lymphocyte none
3.9
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
10.7
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
7.8
CCD1106 (Keratinocytes)
2.2
|
TNF alpha + IL-1beta
|
LAK cells IL-2
5.8
Liver cirrhosis
1.5
|
LAK cells IL-2 + IL-12
11.2
NCI-H292 none
0.0
|
LAK cells IL-2 +IFN
11.2
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
14.9
NCI-H292 IL-9
0.0
|
LAK cells
13.5
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
3.8
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
14.3
HPAEC none
0.0
|
Two Way MLR 5 day
2.5
HPAEC TNF alpha + IL-1beta
8.0
|
Two Way MLR 7 day
2.0
Lung fibroblast none
13.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
2.3
|
1beta
|
PBMC PWM
3.8
Lung fibroblast IL-4
32.1
|
PBMC PHA-L
17.6
Lung fibroblast IL-9
56.6
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
45.4
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
18.2
|
B lymphocytes PWM
1.9
Dermal fibroblast CCD1070 rest
9.0
|
B lymphocytes CD40L
10.1
Dermal fibroblast CCD1070
4.7
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
38.4
|
PMA/ionomycin
|
Dendritic cells none
8.3
Dermal fibroblast IL-4
100.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
5.3
|
Dendritic cells anti-CD40
15.4
Neutrophils TNFa + LPS
4.0
|
Monocytes rest
1.1
Neutrophils rest
2.1
|
Monocytes LPS
9.4
Colon
0.0
|
Macrophages rest
7.2
Lung
21.8
|
Macrophages LPS
5.6
Thymus
57.4
|
HUVEC none
0.0
Kidney
2.2
|
HUVEC starved
0.0
|
|
[1344] CNS_neurodegeneration_v1.0 Summary: Ag3775 Two experiments with two probe and primer sets produce results that are in excellent agreement. This panel does not show differential expression of the CG92152-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain, with highest expression in the hippocampus of an Alzheimer's patient (CTs=31-32). Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1345] General_screening13 panel_v1.4 Summary: Ag3775 Highest expression of the CG92152-01 gene is seen in an ovarian cancer cell line (CT=32). significant levels of expression are seen in a cluster of samples derived from breast and lung cancer cell lines. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of ovarian, breast and lung cancers.
[1346] This gene is also expressed at low levels in the CNS, including the hippocampus, thalamus, substantia nigra, amygdala, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurologic disorders, such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.
[1347] Among tissues with metabolic function, this gene is expressed at low but significant levels in adipose, adrenal gland, pancreas, heart and adult and fetal skeletal muscle. This expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1348] Panel 4.1D Summary: Ag3775 Highest expression of the CG92152-01 gene in IL4 treated with dermal fibroblasts (CTs=32.5). Low, but significant levels of expression are also seen in treated and untreated lung and dermal fibroblasts, and chronically activated Th2 cells. The expression of this gene in lung and skin derived fibroblasts suggests that this gene may be involved in normal conditions as well as pathological and inflammatory lung disorders that include chronic obstructive pulmonary disease, asthma, allergy, psoriasis, and emphysema.
[1349] AW. CG92228-01 and CG92228-O2: Transmembrane Tryptase
[1350] Expression of gene CG92228-01 and variant CG92228-02 was assessed using the primer-probe sets Ag1291 and Ag749, described in Tables AWA and AWB. Results of the RTQ-PCR runs are shown in Table AWC.
613TABLE AWA
|
|
Probe Name Ag1291
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ccatcccaagtaccaagataca-3′22511402
ProbeTET-5′-agacgtcgccttgttgaaactgtcct-26538403
3′-TAMRA
Reverse5′-gcagaagtgaaggtgacttgag-3′22564404
|
[1351]
614
TABLE AWB
|
|
|
Probe Name Ag749
|
Start
SEQ ID
|
Primers
Sequences
Length
Position
NO:
|
|
Forward
5′-aatcgtcatccatcccaagt-3′
20
502
405
|
Probe
TET-5′-tcgccttgttgaaactgtcctctcaa-
26
543
406
|
3′-TAMRA
|
Reverse
5′-ataggcaggatggcagaagt-3′
20
578
407
|
|
[1352]
615
TABLE AWC
|
|
|
Panel 4D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag1291,
Ag1291,
|
Tissue Name
Run 138673859
Tissue Name
Run 138673859
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
21.6
|
Secondary Th2 act
0.0
HUVEC IFN gamma
31.4
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
10.2
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
22.1
|
Secondary Th2 rest
0.0
HUVEC IL-11
20.7
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
39.8
|
Primary Th1 act
0.0
Lung Microvascular EC
22.5
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
100.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
44.4
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium
42.3
|
TNF alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
12.3
|
Primary Tr1 rest
0.0
Small airway epithelium
75.3
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
3.1
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC
6.1
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
13.5
|
Secondary CD8
0.6
Astrocytes TNF alpha + IL-1beta
8.8
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
22.4
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
85.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
18.8
|
LAK cells IL-2 + IL-12
0.0
Lupus kidney
21.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 none
55.9
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
52.9
|
LAK cells
0.0
NCI-H292 IL-9
53.6
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IL-13
50.0
|
Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
37.6
|
Two Way MLR 5 day
0.0
HPAEC none
31.9
|
Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
37.4
|
PBMC rest
0.0
Lung fibroblast none
2.9
|
PBMC PWM
0.4
Lung fibroblast TNF alpha + IL-
0.5
|
1beta
|
PBMC PHA-L
0.0
lung fibroblast IL-4
0.5
|
Ramos (B cell) none
0.0
Lung fibroblast IL-9
2.5
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
1.9
|
B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
1.7
|
B lymphocytes CD40L
0.6
Dermal fibroblast CCD1070 rest
14.0
|
and IL-4
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
8.4
|
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
8.5
|
PMA/ionomycin
1beta
|
Dendritic cells none
0.0
Dermal fibroblast IFN gamma
6.9
|
Dendritic cells LPS
0.0
Dermal fibroblast IL-4
16.6
|
Dendritic cells anti-CD40
0.0
IBD Colitis 2
1.7
|
Monocytes rest
0.0
IBD Crohn's
8.3
|
Monocytes LPS
0.0
Colon
45.1
|
Macrophages rest
0.4
Lung
16.4
|
Macrophages LPS
0.0
Thymus
76.8
|
HUVEC none
36.9
Kidney
5.7
|
HUVEC starved
59.0
|
|
[1353] Panel 4D Summary: Ag1291 Highest expression of the CG92228-01 gene is seen in untreated dermal microvascular endothelium (CT=29). Moderate levels of expression are also seen in other endothelial cells including treated and untreated HUVECs, lung microvascular EC, and HPAECs. Moderate levels of expression are also seen in untreated and activated keratinocytes, activated small airway and bronchial epithelium, normal thymus and colon and a cluster of samples from the mucoepidermoid NCI-H292 cell line. This expression in many cell types involved in the inflammatory and processes in the lung and skin suggest that this gene product may be involved in asthma, allergy, emphysema and psoriasis.
[1354] AX. CG92425-01 and CG92425-O2: RETINOL DEHYDROGENASE
[1355] Expression of gene CG92425-01 and full length clone CG92425-02 was assessed using the primer-probe set Ag3789, described in Table AXA. Please note that CG92425-02 represents a full-length physical clone of the CG92425-01 gene, validating the prediction of the gene sequence.
616TABLE AXA
|
|
Probe Name Ag3789
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ctacatccctctggctaagttg-3′22889408
ProbeTET-5′-tcatcctaagccggtaccttccaagg-26930409
3′-TAMRA
Reverse5′-gatcctccccagtttagacact-3′22965410
|
[1356] CNS_neurodegeneration_v1.0 Summary: Ag3789 Results from one experiment with the CG92425-01 gene are not included. (All CTs=40). The data suggest that there is the possibility of an experimental failure.
[1357] General_screening13 panel_v1.4 Summary: Ag3789 Results from one experiment with the CG92425-01 gene are not included. (All CTs=40). The data suggest that there is the possibility of an experimental failure.
[1358] Panel 2.2 Summary: Ag3789 Results from one experiment with the CG92425-01 gene are not included. (All CTs=40). The data suggest that there is the possibility of an experimental failure.
[1359] Panel 4.1D Summary: Ag3789 Results from one experiment with the CG92425-01 gene are not included. (All CTs=40). The data suggest that there is the possibility of an experimental failure.
[1360] AY. CG92499-01: PUTATIVE NOVEL SEVEN TRANSMEMBRANE DOMAIN PROTEIN
[1361] Expression of gene CG92499-01 was assessed using the primer-probe set Ag780, described in Table AYA. Results of the RTQ-PCR runs are shown in Table AYB.
617TABLE AYA
|
|
Probe Name Ag780
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-ctcagcgtatcatccctgttac-3′22568411
ProbeTET-5′-tcaaattttagcattggtcttccaagca-28607412
3′-TAMRA
Reverse5′-caaaatccatctggaaatacga-3′22643413
|
[1362]
618
TABLE AYB
|
|
|
Panel 1.2
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag780, Run
Ag780, Run
|
Tissue Name
117132955
Tissue Name
117132955
|
|
Endothelial cells
9.0
Renal ca. 786-0
1.3
|
Heart (Fetal)
0.0
Renal ca. A498
0.1
|
Pancreas
0.0
Renal ca. RXF 393
0.0
|
Pancreatic ca. CAPAN2
9.2
Renal ca. ACHN
0.1
|
Adrenal Gland
0.1
Renal ca. UO-31
0.2
|
Thyroid
0.0
Renal ca. TK-10
5.3
|
Salivary gland
0.1
Liver
16.7
|
Pituitary gland
0.0
Liver (fetal)
10.7
|
Brain (fetal)
5.2
Liver ca. (hepatoblast)
11.6
|
HepG2
|
Brain (whole)
0.0
Lung
0.0
|
Brain (amygdala)
0.3
Lung (fetal)
1.6
|
Brain (cerebellum)
0.2
Lung ca. (small cell) LX-1
0.0
|
Brain (hippocampus)
0.0
Lung ca. (small cell)
0.0
|
NCI-H69
|
Brain (thalamus)
0.0
Lung ca. (s. cell var.)
0.0
|
SHP-77
|
Cerebral Cortex
0.0
Lung ca. (large cell)NCI-
0.5
|
H460
|
Spinal cord
0.5
Lung ca. (non-sm. cell)
17.3
|
A549
|
glio/astro U87-MG
0.0
Lung ca. (non-s. cell)
1.9
|
NCI-H23
|
glio/astro U-118-MG
0.0
Lung ca. (non-s. cell)
2.4
|
HOP-62
|
astrocytoma SW1783
0.0
Lung ca. (non-s. cl) NCI-
49.7
|
H522
|
neuro*; met SK-N-AS
0.0
Lung ca. (squam.) SW
11.6
|
900
|
astrocytoma SF-539
0.0
Lung ca. (squam.) NCI-
0.0
|
H596
|
astrocytoma SNB-75
0.1
Mammary gland
0.0
|
glioma SNB-19
0.0
Breast ca.* (pl. ef) MCF-7
8.1
|
glioma U251
0.0
Breast ca.* (pl. ef) MDA-
1.8
|
MB-231
|
glioma SF-295
0.0
Breast ca.* (pl. ef) T47D
2.0
|
Heart
0.0
Breast ca. BT-549
0.0
|
Skeletal Muscle
0.0
Breast ca. MDA-N
0.2
|
Bone marrow
0.1
Ovary
0.0
|
Thymus
0.0
Ovarian ca. OVCAR-3
0.0
|
Spleen
0.0
Ovarian ca. OVCAR-4
0.0
|
Lymph node
0.0
Ovarian ca. OVCAR-5
1.7
|
Colorectal Tissue
0.0
Ovarian ca. OVCAR-8
0.5
|
Stomach
0.0
Ovarian ca. IGROV-1
1.3
|
Small intestine
0.0
Ovarian ca. (ascites) SK-
0.0
|
OV-3
|
Colon ca. SW480
0.0
Uterus
0.1
|
Colon ca.* SW620
0.0
Placenta
14.0
|
(SW480 met)
|
Colon ca. HT29
0.2
Prostate
0.0
|
Colon ca. HCT-116
5.8
Prostate ca.* (bone met)
29.7
|
PC-3
|
Colon ca. CaCo-2
14.9
Testis
0.0
|
Colon ca. Tissue
0.0
Melanoma Hs688(A).T
0.0
|
(ODO3866)
|
Colon ca. HCC-2998
0.9
Melanoma* (met)
0.0
|
Hs688(B).T
|
Gastric ca.* (liver met)
5.7
Melanoma UACC-62
26.1
|
NCI-N87
|
Bladder
18.6
Melanoma M14
9.2
|
Trachea
0.0
Melanoma LOX IMVI
3.5
|
Kidney
0.3
Melanoma* (met) SK-
100.0
|
MEL-5
|
Kidney (fetal)
0.5
|
|
[1363] Panel 1.2 Summary: Ag780 The CG92499-01 gene shows a restricted expression pattern, with highest expression in a melanoma cell line (CT=28). Moderate levels of expression are also seen in samples derived from prostate, lung, colon and liver cancer cell lines. Thus, expression of this gene could be used to differentiate between the melanoma cell line and other samples on this panel. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of these cancers.
[1364] Low but significant expression is also seen in the fetal brain (CT-32.4). Thus, expression of this gene could be used to differentiate between fetal and adult brain tissue (CT=40).
[1365] AZ. CG92541-01: MUNC13-4
[1366] Expression of gene CG92541-01 was assessed using the primer-probe set Ag3815, described in Table AZA. Results of the RTQ-PCR runs are shown in Tables AZB, AZC and AZD.
619TABLE AZA
|
|
Probe Name Ag3815
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ctggacccacacactcaca-3′192566414
ProbeTET-5′-cagctcatccctggcttccaacag-242614415
3′-TAMRA
Reverse5′-gttctgcagggcaatcttc-3′192641416
|
[1367]
620
TABLE AZB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3815, Run
Ag3815,
|
Tissue Name
211292378
Tissue Name
Run 211292378
|
|
AD 1 Hippo
45.1
Control (Path) 3
4.9
|
Temporal Ctx
|
AD 2 Hippo
50.0
Control (Path) 4
32.5
|
Temporal Ctx
|
AD 3 Hippo
26.6
AD 1 Occipital Ctx
36.1
|
AD 4 Hippo
17.9
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
46.7
AD 3 Occipital Ctx
41.2
|
AD 6 Hippo
61.6
AD 4 Occipital Ctx
3.3
|
Control 2 Hippo
59.0
AD 5 Occipital Ctx
18.7
|
Control 4 Hippo
44.4
AD 6 Occipital Ctx
23.7
|
Control (Path) 3 Hippo
6.2
Control 1 Occipital Ctx
11.0
|
AD 1 Temporal Ctx
26.2
Control 2 Occipital Ctx
32.3
|
AD 2 Temporal Ctx
49.7
Control 3 Occipital Ctx
16.8
|
AD 3 Temporal Ctx
42.0
Control 4 Occipital Ctx
14.0
|
AD 4 Temporal Ctx
29.5
Control (Path) 1
100.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
51.1
Control (Path) 2
23.0
|
Occipital Ctx
|
AD 5 SupTemporal Ctx
57.4
Control (Path) 3
2.1
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
48.6
Control (Path) 4
34.6
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
41.2
Control 1 Parietal Ctx
87.1
|
Control 1 Temporal Ctx
26.4
Control 2 Parietal Ctx
59.5
|
Control 2 Temporal Ctx
48.3
Control 3 Parietal Ctx
33.9
|
Control 3 Temporal Ctx
18.8
Control (Path) 1
38.7
|
Parietal Ctx
|
Control 4 Temporal Ctx
14.3
Control (Path) 2
43.8
|
Parietal Ctx
|
Control (Path) 1
35.8
Control (Path) 3
4.0
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
20.6
Control (Path) 4
45.4
|
Temporal Ctx
Parietal Ctx
|
|
[1368]
621
TABLE AZC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3815, Run
Ag3815, Run
|
Tissue Name
218668399
Tissue Name
218668399
|
|
Adipose
0.4
Renal ca. TK-10
8.3
|
Melanoma* Hs688(A).T
0.0
Bladder
4.8
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
9.0
|
NCI-N87
|
Melanoma* M14
0.2
Gastric ca. KATO III
15.2
|
Melanoma* LOXIMVI
0.1
Colon ca. SW-948
2.2
|
Melanoma* SK-MEL-5
0.9
Colon ca. SW480
45.1
|
Squamous cell
1.8
Colon ca.* (SW480 met)
7.7
|
carcinoma SCC-4
SW620
|
Testis Pool
0.3
Colon ca. HT29
3.9
|
Prostate ca.* (bone met)
1.2
Colon ca. HCT-116
3.9
|
PC-3
|
Prostate Pool
1.0
Colon ca. CaCo-2
0.9
|
Placenta
9.2
Colon cancer tissue
13.7
|
Uterus Pool
1.1
Colon ca. SW1116
0.1
|
Ovarian ca. OVCAR-3
3.2
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
13.7
Colon ca. SW-48
0.2
|
Ovarian ca. OVCAR-4
3.2
Colon Pool
1.8
|
Ovarian ca. OVCAR-5
37.9
Small Intestine Pool
4.4
|
Ovarian ca. IGROV-1
1.2
Stomach Pool
1.3
|
Ovarian ca. OVCAR-8
4.5
Bone Marrow Pool
0.8
|
Ovary
1.4
Fetal Heart
0.9
|
Breast ca. MCF-7
5.9
Heart Pool
0.6
|
Breast ca. MDA-MB-
92.0
Lymph Node Pool
1.3
|
231
|
Breast ca. BT 549
0.1
Fetal Skeletal Muscle
0.2
|
Breast ca. T47D
100.0
Skeletal Muscle Pool
2.2
|
Breast ca. MDA-N
0.0
Spleen Pool
7.9
|
Breast Pool
2.1
Thymus Pool
5.9
|
Trachea
2.0
CNS cancer (glio/astro)
0.5
|
U87-MG
|
Lung
0.3
CNS cancer (glio/astro) U-
0.3
|
118-MG
|
Fetal Lung
10.5
CNS cancer (neuro; met)
0.5
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.1
|
Lung ca. LX-1
11.0
CNS cancer (astro) SNB-75
0.4
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.7
|
Lung ca. SHP-77
1.6
CNS cancer (glio) SF-295
10.1
|
Lung ca. A549
1.4
Brain (Amygdala) Pool
0.2
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.9
|
Lung ca. NCI-H23
0.2
Brain (fetal)
0.1
|
Lung ca. NCI-H460
0.2
Brain (Hippocampus) Pool
0.8
|
Lung ca. HOP-62
28.9
Cerebral Cortex Pool
0.9
|
Lung ca. NCI-H522
1.0
Brain (Substantia nigra)
1.4
|
Pool
|
Liver
0.2
Brain (Thalamus) Pool
0.5
|
Fetal Liver
3.7
Brain (whole)
0.6
|
Liver ca. HepG2
15.2
Spinal Cord Pool
0.9
|
Kidney Pool
4.5
Adrenal Gland
0.6
|
Fetal Kidney
1.0
Pituitary gland Pool
0.8
|
Renal ca. 786-0
0.2
Salivary Gland
0.6
|
Renal ca. A498
0.2
Thyroid (female)
0.4
|
Renal ca. ACHN
0.2
Pancreatic ca. CAPAN2
17.2
|
Renal ca. UO-31
35.4
Pancreas Pool
3.1
|
|
[1369]
622
TABLE AZD
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3815,
Ag3815,
|
Tissue Name
Run 170129244
Tissue Name
Run 170129244
|
|
Secondary Th1 act
68.8
HUVEC IL-1beta
2.8
|
Secondary Th2 act
66.0
HUVEC IFN gamma
1.9
|
Secondary Tr1 act
87.1
HUVEC TNF alpha + IFN
1.8
|
gamma
|
Secondary Th1 rest
29.5
HUVEC TNF alpha + IL4
1.8
|
Secondary Th2 rest
50.3
HUVEC IL-11
1.8
|
Secondary Tr1 rest
41.2
Lung Microvascular EC none
44.4
|
Primary Th1 act
55.5
Lung Microvascular EC
16.5
|
TNF alpha + IL-1beta
|
Primary Th2 act
67.4
Microvascular Dermal EC none
4.9
|
Primary Tr1 act
84.1
Microsvasular Dermal EC
5.6
|
TNF alpha + IL-1beta
|
Primary Th1 rest
30.1
Bronchial epithelium
3.7
|
TNF alpha + IL1beta
|
Primary Th2 rest
28.5
Small airway epithelium none
3.7
|
Primary Tr1 rest
52.9
Small airway epithelium
9.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
15.2
Coronery artery SMC rest
0.5
|
act
|
CD45RO CD4 lymphocyte
37.6
Coronery artery SMC
0.2
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
78.5
Astrocytes rest
0.0
|
Secondary CD8
37.4
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
36.6
KU-812 (Basophil) rest
100.0
|
lymphocyte act
|
CD4 lymphocyte none
14.7
KU-812 (Basophil)
72.2
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
58.6
CCD1106 (Keratinocytes) none
13.3
|
CD95 CH11
|
LAK cells rest
37.6
CCD1106 (Keratinocytes)
10.2
|
TNF alpha + IL-1beta
|
LAK cells IL-2
34.9
Liver cirrhosis
7.2
|
LAK cells IL-2 + IL-12
33.7
NCI-H292 none
33.7
|
LAK cells IL-2 + IFN
30.1
NCI-H292 IL-4
24.0
|
gamma
|
LAK cells IL-2 + IL-18
31.4
NCI-H292 IL-9
59.0
|
LAK cells
9.9
NCI-H292 IL-13
21.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
50.7
NCI-H292 IFN gamma
28.7
|
Two Way MLR 3 day
56.3
HPAEC none
6.5
|
Two Way MLR 5 day
66.4
HPAEC TNF alpha + IL-1beta
4.9
|
Two Way MLR 7 day
43.2
Lung fibroblast none
0.6
|
PBMC rest
18.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
43.5
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
35.8
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.2
Lung fibroblast IL-13
0.8
|
Ramos (B cell) ionomycin
1.9
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
13.8
Dermal fibroblast CCD1070 rest
0.3
|
B lymphocytes CD40L
32.1
Dermal fibroblast CCD1070
15.7
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
24.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
14.8
Dermal fibroblast IFN gamma
0.2
|
PMA/ionomycin
|
Dendritic cells none
48.0
Dermal fibroblast IL-4
0.9
|
Dendritic cells LPS
31.4
Dermal Fibroblasts rest
0.4
|
Dendritic cells anti-CD40
29.9
Neutrophils TNFa + LPS
16.5
|
Monocytes rest
57.0
Neutrophils rest
57.8
|
Monocytes LPS
59.9
Colon
3.7
|
Macrophages rest
27.7
Lung
7.8
|
Macrophages LPS
30.4
Thymus
23.2
|
HUVEC none
2.6
Kidney
0.9
|
HUVEC starved
2.0
|
|
[1370] CNS_neurodegeneration_v1.0 Summary: Ag3815 This panel does not show differential expression of the CG925413-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1371] General_screening_panel_v1.4 Summary: Ag3815 Highest expression of the CG92541-01 gene is seen in a breast cancer cell line (C=27). Moderate levels of expression are also seen in samples derived from ovarian, renal, lung and colon cancers. Thus, expression of this gene could be used to differentiate the breast cancer cell lines from other samples on this panel and as a marker for these cancers. Furthermore, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of breast, ovarian, lung, colon and renal cancers.
[1372] Among metabolic tissues, low levels of expression are seen in pancreas, adrenal, pituitary, skeletal muscle and fetal and adult heart and liver. This expression suggests that this gene product may be involved in the pathogenesis and/or treatment of metabolic disordes, including obesity and diabetes.
[1373] In addition, this gene is expressed at much higher levels in fetal lung and liver (CTs=30-32) when compared to expression in the adult counterparts (CTs—35). Thus, expression of this gene may be used to differentiate between the fetal and adult source of these tissues.
[1374] This gene is also expressed at low but significant levels in cerebellum, substantia nigra, hippocampus, and cerebral cortex. This gene encodes a homolog of MUNC 13, a protein involved in neurotransmitter release at the synaptic junction. Therefore, therapeutic modulation of the expression or function of this protein may be useful in the treatment of disease where reduction in neurotransmission has been shown to ameliorate symptomology (e.g., epilepsy or other seizure disorders, schizophrenia, bipolar disorder or anxiety) (Richmond J E, Weimer R M, Jorgensen E M. An open form of syntaxin bypasses the requirement for UNC-13 in vesicle priming. Nature Jul. 19, 2001;412(6844):338-41).
[1375] Panel 4.1D Summary: Ag3815 Highest expression of the CG92541-01 gene is seen the resting basophil cell line KU-812 (CT=29.3). In addition, moderate levels of expression are seen in many of the hematopoietic samples on this panel, including macrophages, monocytes, dendritic cells, T cells, neutrophils and B cells. In addition, this transcript appears to be slightly upregulated in activated T cells when compared to expression in resting T cells. Thus, this gene product may be involved in autoimmune or inflammatory conditions such as asthma, emphysema, inflammatory bowel disease, and rheumatoid arthritis.
[1376] BA. CG92662-01: PROSTAGLANDIN-E29-REDUCTASE
[1377] Expression of gene CG92662-01 was assessed using the primer-probe set Ag3816, described in Table BAA. Results of the RTQ-PCR runs are shown in Tables BAB, BAC and BAD.
623TABLE BAA
|
|
Probe Name Ag3816
StartSEQ ID
PrimersSequencesLengthPositionNO:
|
Forward5′-ccctgtgaagagagaggacatt-3′22231417
ProbeTET-5′-caccactgagctttggacaactttct-26258418
3′-TAMRA
Reverse5′-gggcgaactaattctggtctaa-3′22284419
|
[1378]
624
TABLE BAB
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3816, Run
Ag3816, Run
|
Tissue Name
213515569
Tissue Name
213515569
|
|
Adipose
0.4
Renal ca. TK-10
0.3
|
Melanoma* Hs688(A).T
0.4
Bladder
0.0
|
Melanoma* Hs688(B).T
1.4
Gastric ca. (liver met.)
6.7
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
5.4
|
Melanoma* LOXIMVI
16.8
Colon ca. SW-948
3.6
|
Melanoma* SK-MEL-5
0.0
Colon ca. SW480
1.1
|
Squamous cell
6.0
Colon ca.* (SW480 met)
1.0
|
carcinoma SCC-4
SW620
|
Testis Pool
1.5
Colon ca. HT29
1.3
|
Prostate ca.* (bone met)
1.3
Colon ca. HCT-116
0.0
|
PC-3
|
Prostate Pool
0.0
Colon ca. CaCo-2
0.3
|
Placenta
2.8
Colon cancer tissue
2.1
|
Uterus Pool
0.4
Colon ca. SW1116
0.2
|
Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.4
|
Ovarian ca. SK-OV-3
27.0
Colon ca. SW-48
2.1
|
Ovarian ca. OVCAR-4
1.5
Colon Pool
0.4
|
Ovarian ca. OVCAR-5
3.1
Small Intestine Pool
1.4
|
Ovarian ca. IGROV-1
4.8
Stomach Pool
0.3
|
Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
0.8
|
Ovary
0.2
Fetal Heart
0.0
|
Breast ca. MCF-7
0.0
Heart Pool
0.3
|
Breast ca. MDA-MB-
14.4
Lymph Node Pool
0.8
|
231
|
Breast ca. BT 549
1.7
Fetal Skeletal Muscle
0.0
|
Breast ca. T47D
6.4
Skeletal Muscle Pool
1.8
|
Breast ca. MDA-N
0.0
Spleen Pool
3.8
|
Breast Pool
0.8
Thymus Pool
3.0
|
Trachea
9.5
CNS cancer (glio/astro)
100.0
|
U87-MG
|
Lung
0.0
CNS cancer (glio/astro) U-
0.0
|
118-MG
|
Fetal Lung
1.4
CNS cancer (neuro; met)
4.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0
|
Lung ca. LX-1
8.2
CNS cancer (astro) SNB-75
2.7
|
Lung ca. NCI-H146
4.8
CNS cancer (glio) SNB-19
3.9
|
Lung ca. SHP-77
39.2
CNS cancer (glio) SF-295
51.4
|
Lung ca. A549
71.2
Brain (Amygdala) Pool
0.3
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0
|
Lung ca. NCI-H23
0.2
Brain (fetal)
0.0
|
Lung ca. NCI-H460
26.4
Brain (Hippocampus) Pool
0.0
|
Lung ca. HOP-62
18.6
Cerebral Cortex Pool
0.3
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
0.2
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
0.0
|
Fetal Liver
1.7
Brain (whole)
0.0
|
Liver ca. HepG2
0.0
Spinal Cord Pool
0.0
|
Kidney Pool
0.6
Adrenal Gland
0.0
|
Fetal Kidney
0.3
Pituitary gland Pool
0.0
|
Renal ca.786-0
0.0
Salivary Gland
3.0
|
Renal ca. A498
0.8
Thyroid (female)
0.0
|
Renal ca. ACHN
1.1
Pancreatic ca. CAPAN2
5.1
|
Renal ca. UO-31
0.0
Pancreas Pool
2.0
|
|
[1379]
625
TABLE BAC
|
|
|
Panel 2.2
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3816,
Ag3816,
|
Tissue Name
Run 174448448
Tissue Name
Run 174448448
|
|
Normal Colon
8.8
Kidney Margin (OD04348)
14.2
|
Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0
|
(OD06204B)
|
Colon Margin (OD06064)
0.0
Kidney normal adjacent
1.5
|
tissue (OD06204E)
|
Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0
|
01)
|
Colon Margin (OD06159)
5.3
Kidney Margin (OD04450-
5.8
|
03)
|
Colon cancer (OD06297-04)
2.0
Kidney Cancer 8120613
2.7
|
Colon Margin (OD06297-05)
0.0
Kidney Margin 8120614
0.0
|
CC Gr.2 ascend colon
2.5
Kidney Cancer 9010320
0.0
|
(ODO3921)
|
CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0
|
Colon cancer metastasis
0.0
Kidney Cancer 8120607
3.0
|
(OD06104)
|
Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0
|
Colon mets to lung
10.6
Normal Uterus
2.1
|
(OD04451-01)
|
Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0
|
Normal Prostate
0.0
Normal Thyroid
0.0
|
Prostate Cancer (OD04410)
7.5
Thyroid Cancer 064010
0.0
|
Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0
|
Normal Ovary
0.0
Thyroid Margin A302153
0.0
|
Ovarian cancer (OD06283-
0.0
Normal Breast
6.0
|
03)
|
Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
0.0
|
07)
|
Ovarian Cancer 064008
0.0
Breast Cancer 1024
1.9
|
Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0
|
01)
|
Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
6.8
|
(OD04590-03)
|
Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
100.0
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
1.9
Breast Cancer 064006
17.7
|
07)
|
Normal Lung
5.4
Breast Cancer 9100266
0.0
|
Invasive poor diff. lung
72.7
Breast Margin 9100265
2.2
|
adeno (ODO4945-01
|
Lung Margin (ODO4945-03)
2.4
Breast Cancer A209073
0.0
|
Lung Malignant Cancer
0.0
Breast Margin A2090734
2.7
|
(OD03126)
|
Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
84.7
|
Lung Cancer (OD05014A)
0.0
Breast cancer node
49.0
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
4.9
Normal Liver
0.0
|
Lung cancer (OD06081)
8.3
Liver Cancer 1026
0.0
|
Lung Margin (OD06081)
8.5
Liver Cancer 1025
3.0
|
Lung Cancer (OD04237-01)
3.1
Liver Cancer 6004-T
0.0
|
Lung Margin (OD04237-02)
9.0
Liver Tissue 6004-N
0.0
|
Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
2.6
|
Ocular Melanoma Margin
0.0
Liver Tissue 6005-N
0.0
|
(Liver)
|
Melanoma Metastasis
0.0
Liver Cancer 064003
0.0
|
Melanoma Margin (Lung)
7.5
Normal Bladder
2.7
|
Normal Kidney
7.7
Bladder Cancer 1023
0.0
|
Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer A302173
0.0
|
(OD04338)
|
Kidney Margin (OD04338)
5.3
Normal Stomach
2.9
|
Kidney Ca Nuclear grade 1/2
5.6
Gastric Cancer 9060397
0.0
|
(OD04339)
|
Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0
|
Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
2.2
|
(OD04340)
|
Kidney Margin (OD04340)
7.7
Stomach Margin 9060394
0.0
|
Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0
|
(OD04348)
|
|
[1380]
626
TABLE BAD
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3816,
Ag3816,
|
Tissue Name
Run 170129269
Tissue Name
Run 170129269
|
|
Secondary Th1 act
8.0
HUVEC IL-1beta
3.7
|
Secondary Th2 act
3.8
HUVEC IFN gamma
1.9
|
Secondary Tr1 act
9.5
HUVEC TNF alpha + IFN
0.7
|
gamma
|
Secondary Th1 rest
27.7
HUVEC TNF alpha + IL4
1.8
|
Secondary Th2 rest
13.7
HUVEC IL-11
0.7
|
Secondary Tr1 rest
17.2
Lung Microvascular EC none
22.1
|
Primary Th1 act
8.8
Lung Microvascular EC
11.7
|
TNF alpha + IL-1beta
|
Primary Th2 act
1.4
Microvascular Dermal EC none
21.3
|
Primary Tr1 act
3.5
Microsvasular Dermal EC
14.1
|
TNF alpha + IL-1beta
|
Primary Th1 rest
20.6
Bronchial epithelium
8.4
|
TNF alpha + IL1beta
|
Primary Th2 rest
13.7
Small airway epithelium none
0.9
|
Primary Tr1 rest
14.1
Small airway epithelium
3.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
4.1
Coronery artery SMC rest
0.6
|
act
|
CD45RO CD4 lymphocyte
26.8
Coronery artery SMC
0.0
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
3.7
Astrocytes rest
0.0
|
Secondary CD8
20.4
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
1.0
KU-812 (Basophil) rest
16.3
|
lymphocyte act
|
CD4 lymphocyte none
31.2
KU-812 (Basophil)
30.6
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
15.6
CCD1106 (Keratinocytes) none
1.5
|
CD95 CH11
|
LAK cells rest
39.0
CCD1106 (Keratinocytes)
1.3
|
TNF alpha + IL-1beta
|
LAK cells IL-2
48.3
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
31.6
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
28.5
NCI-H292 IL-4
1.1
|
gamma
|
LAK cells IL-2 + IL-18
33.4
NCI-H292 IL-9
0.9
|
LAK cells
100.0
NCI-H292 IL-13
0.8
|
PMA/ionomycin
|
NK Cells IL-2 rest
43.5
NCI-H292 IFN gamma
2.0
|
Two Way MLR 3 day
71.2
HPAEC none
0.6
|
Two Way MLR 5 day
31.4
HPAEC TNF alpha + IL-1beta
5.4
|
Two Way MLR 7 day
27.0
Lung fibroblast none
0.0
|
PBMC rest
13.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
23.2
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
9.1
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
12.4
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
14.0
Dermal fibroblast CCD1070
3.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
1.0
Dermal fibroblast CCD1070 IL-
4.5
|
1beta
|
EOL-1 dbcAMP
4.3
Dermal fibroblast IFN gamma
1.5
|
PMA/ionomycin
|
Dendritic cells none
23.8
Dermal fibroblast IL-4
3.9
|
Dendritic cells LPS
14.9
Dermal Fibroblasts rest
2.7
|
Dendritic cells anti-CD40
16.6
Neutrophils TNFa + LPS
3.6
|
Monocytes rest
3.2
Neutrophils rest
3.2
|
Monocytes LPS
11.6
Colon
1.4
|
Macrophages rest
7.4
Lung
1.9
|
Macrophages LPS
3.8
Thymus
2.1
|
HUVEC none
0.0
Kidney
0.6
|
HUVEC starved
2.1
|
|
[1381] CNS_neurodegeneration_v1.0 Summary: Ag3816 Expression of the CG92662-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1382] General_screening13 panel_v1.4 Summary: Ag3816 Expression of the CG92662-01 gene is predominantly associated with normal tissues, with highest expression in a brain cancer cell line (CT=30). Moderate levels of expression are also seen in a cluster of lung cancer cell lines, with low but significant expression in ovarian, breast and melanoma cancer cell lines. Thus, expression of this gene could be used to differentiate these cell line samples from other samples on this panel and as a diagnostic marker for the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of brain, ovarian, breast and melanoma cancers.
[1383] Panel 2.2 Summary: Ag3816 Expression of the CG92662-01 gene is almost exclusive to breast cancer (CTs=33-34). Thus, expression of this gene could be used to differentiate between the breast cancer cell lines and other samples on this panel and as a diagnostic marker for the presence of breast cancer. This gene encodes a protein homologous to the activity of 9-ketoreductase, the enzyme which converts prostaglandin E2 (PGE2) into prostaglandin F2 alpha (PGF2 alpha) and can influence estrogen synthesis and thus potentially the rate of breast cancer growth. Therefore, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of breast cancer (Brueggemeier R W, Richards J A, Joomprabutra S, Bhat A S, Whetstone J L. Molecular pharmacology of aromatase and its regulation by endogenous and exogenous agents. J Steroid Biochem Mol Biol 2001 Dec;79(1-5):75-84; Brodie A M, Lu Q, Long B J, Fulton A, Chen T, Macpherson N, DeJong P C, Blankenstein M A, Nortier J W, Slee P H, van de Ven J, van Gorp J M, Elbers J R, Schipper M E, Blijham G H, Thijssen J H. Aromatase and COX-2 expression in human breast cancers. J Steroid Biochem Mol Biol 2001 Dec;79(1-5):41-7).
[1384] Panel 4.1D Summary: Ag3816 Expression of the CG92662-01 gene highest in LAK cells treated with PMA/ionomycin (CT=32.5). Low, but significant levels of expression are also seen in a cluster of both treated and untreated LAK cells, PMA/ionomycin treated basophils, untreated lung and dermal microvasculature, and resting TH1 cells (both primary and secondary). LAK cells are involved in tumor immunology and cell clearance of virally and bacterial infected cells as well as tumors. Therefore, modulation of the function of the protein encoded by this gene through the application of a small molecule drug or antibody may alter the functions of these cells and lead to improvement of symptoms associated with these conditions.
[1385] BB. CG92683-01: C2PA
[1386] Expression of gene CG92683-01 was assessed using the primer-probe set Ag3817, described in Table BBA.
627TABLE BBA
|
|
Probe Name Ag3817
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-aggatgatcagaagcgtctctt-3′22363420
ProbeTET-5′-25405421
ccagccagtccaggagacagagtct-
3′-TAMRA
Reverse5′-aaagctcatgcagccaatg-3′19430422
|
[1387] CNS_neurodegeneration_v1.0 Summary: Ag3817 Expression of the CG92683-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1388] General_screening_panel_v1.4 Summary: Ag3817 Expression of the CG92683-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1389] Panel 4.1D Summary: Ag3817 Expression of the CG92683-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1390] BC. CG92694-01: LONG-CHAIN 3-KETOACYL-COA THIOLASE
[1391] Expression of gene CG92694-01 was assessed using the primer-probe set Ag3818, described in Table BCA.
628TABLE BCA
|
|
Probe Name Ag3818
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ctctgagctattcctcccagtt-3′22133423
ProbeTET-5′-23165424
ccagccatccagaccgaaacgag-
3′-TAMRA
Reverse5′-attgggtttggctaatgtcttc-3′22188425
|
[1392] CNS_neurodegeneration_v1.0 Summary: Ag3818 Expression of the CG92694-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1393] General_screening_panel_v1.4 Summary: Ag3818 Expression of the CG92694-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1394] Panel 4.1D Summary: Ag3818 Expression of the CG92694-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1395] Panel 5 Islet Summary: Ag3818 Expression of the CG92694-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1396] BD. CG92896-01 and CG92896-O2: PHOSPHATIDYLINOSITOL 5-PHOSPHATE 4-KINASE GAMMA
[1397] Expression of gene CG92896-01 and full length clone CG92896-02 was assessed using the primer-probe set Ag3832, described in Table BDA. Results of the RTQ-PCR runs are shown in Tables BDB, BDC, BDD and BDE. Please note that CG92896-02 represents a full-length physical clone of the CG92896-01 gene, validating the prediction of the gene sequence.
629TABLE BDA
|
|
Probe Name Ag3832
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-aagtatccagtgaggacattgc-3′22461426
ProbeTET-5′-26483427
tgacatgcatagcaacctctccaact-
3′-TAMRA
Reverse5′-catggcacttcacaatgtactg-3′22514428
|
[1398]
630
TABLE BDB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3832, Run
Ag3832, Run
|
Tissue Name
206873102
Tissue Name
206873102
|
|
AD 1 Hippo
14.9
Control (Path) 3
5.1
|
Temporal Ctx
|
AD 2 Hippo
27.0
Control (Path) 4
34.2
|
Temporal Ctx
|
AD 3 Hippo
8.5
AD 1 Occipital Ctx
14.7
|
AD 4 Hippo
5.4
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
6.7
|
AD 6 Hippo
42.9
AD 4 Occipital Ctx
17.9
|
Control 2 Hippo
32.1
AD 5 Occipital Ctx
52.9
|
Control 4 Hippo
6.8
AD 6 Occipital Ctx
27.0
|
Control (Path) 3 Hippo
4.3
Control 1 Occipital Ctx
1.9
|
AD 1 Temporal Ctx
12.8
Control 2 Occipital Ctx
80.1
|
AD 2 Temporal Ctx
34.9
Control 3 Occipital Ctx
12.6
|
AD 3 Temporal Ctx
5.0
Control 4 Occipital Ctx
4.7
|
AD 4 Temporal Ctx
21.8
Control (Path) 1
74.2
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
7.3
|
Occipital Ctx
|
AD 5 Sup Temporal
33.9
Control (Path) 3
2.6
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
41.2
Control (Path) 4
11.0
|
Occipital Ctx
|
AD 6 Sup Temporal
51.4
Control 1 Parietal Ctx
5.1
|
Ctx
|
Control 1 Temporal Ctx
3.4
Control 2 Parietal Ctx
37.1
|
Control 2 Temporal Ctx
47.0
Control 3 Parietal Ctx
26.4
|
Control 3 Temporal Ctx
14.8
Control (Path) 1
92.7
|
Parietal Ctx
|
Control 3 Temporal Ctx
7.3
Control (Path) 2
20.4
|
Parietal Ctx
|
Control (Path) 1
55.5
Control (Path) 3
3.8
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
34.6
Control (Path) 4
41.5
|
Temporal Ctx
Parietal Ctx
|
|
[1399]
631
TABLE BDC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3832, Run
Ag3832, Run
|
Tissue Name
213603655
Tissue Name
213603655
|
|
Adipose
4.2
Renal ca. TK-10
32.3
|
Melanoma* Hs688(A).T
14.4
Bladder
15.5
|
Melanoma* Hs688(B).T
17.1
Gastric ca. (liver met.)
57.8
|
NCI-N87
|
Melanoma* M14
47.0
Gastric ca. KATO III
40.9
|
Melanoma* LOXIMVI
22.2
Colon ca. SW-948
14.6
|
Melanoma* SK-MEL-5
44.4
Colon ca. SW480
59.5
|
Squamous cell
28.3
Colon ca.* (SW480 met)
20.9
|
carcinoma SCC-4
SW620
|
Testis Pool
16.8
Colon ca. HT29
17.8
|
Prostate ca.* (bone met)
66.9
Colon ca. HCT-116
42.3
|
PC-3
|
Prostate Pool
6.6
Colon ca. CaCo-2
34.9
|
Placenta
9.9
Colon cancer tissue
22.1
|
Uterus Pool
2.9
Colon ca. SW1116
11.7
|
Ovarian ca. OVCAR-3
33.2
Colon ca. Colo-205
10.5
|
Ovarian ca. SK-OV-3
33.2
Colon ca. SW-48
9.9
|
Ovarian ca. OVCAR-4
47.0
Colon Pool
6.1
|
Ovarian ca. OVCAR-5
45.1
Small Intestine Pool
5.3
|
Ovarian ca. IGROV-1
14.1
Stomach Pool
4.8
|
Ovarian ca. OVCAR-8
16.0
Bone Marrow Pool
1.9
|
Ovary
7.4
Fetal Heart
2.7
|
Breast ca. MCF-7
87.7
Heart Pool
2.6
|
Breast ca. MDA-MB-
18.7
Lymph Node Pool
6.1
|
231
|
Breast ca. BT 549
32.1
Fetal Skeletal Muscle
2.0
|
Breast ca. T47D
100.0
Skeletal Muscle Pool
3.8
|
Breast ca. MDA-N
16.4
Spleen Pool
3.0
|
Breast Pool
8.0
Thymus Pool
3.9
|
Trachea
23.0
CNS cancer (glio/astro)
43.5
|
U87-MG
|
Lung
1.1
CNS cancer (glio/astro) U-
32.8
|
118-MG
|
Fetal Lung
10.8
CNS cancer (neuro; met)
28.5
|
SK-N-AS
|
Lung ca. NCI-N417
4.2
CNS cancer (astro) SF-539
12.1
|
Lung ca. LX-1
34.9
CNS cancer (astro) SNB-75
18.3
|
Lung ca. NCI-H146
16.7
CNS cancer (glio) SNB-19
13.2
|
Lung ca. SHP-77
45.1
CNS cancer (glio) SF-295
45.4
|
Lung ca. A549
39.0
Brain (Amygdala) Pool
11.2
|
Lung ca. NCI-H526
13.1
Brain (cerebellum)
23.0
|
Lung ca. NCI-H23
31.0
Brain (fetal)
10.9
|
Lung ca. NCI-H460
19.3
Brain (Hippocampus) Pool
11.7
|
Lung ca. HOP-62
16.0
Cerebral Cortex Pool
23.5
|
Lung ca. NCI-H522
36.1
Brain (Substantia nigra)
22.7
|
Pool
|
Liver
1.8
Brain (Thalamus) Pool
22.7
|
Fetal Liver
6.6
Brain (whole)
27.9
|
Liver ca. HepG2
19.6
Spinal Cord Pool
9.2
|
Kidney Pool
15.0
Adrenal Gland
11.5
|
Fetal Kidney
5.0
Pituitary gland Pool
2.9
|
Renal ca. 786-0
19.6
Salivary Gland
14.1
|
Renal ca. A498
7.5
Thyroid (female)
6.6
|
Renal ca. ACHN
37.4
Pancreatic ca. CAPAN2
44.1
|
Renal ca. UO-31
33.0
Pancreas Pool
6.1
|
|
[1400]
632
TABLE BDD
|
|
|
Panel 2.2
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3832, Run
Ag3832, Run
|
Tissue Name
174448449
Tissue Name
174448449
|
|
Normal Colon
21.2
Kidney Margin (OD04348)
100.0
|
Colon cancer (OD06064)
30.8
Kidney malignant cancer
16.3
|
(OD06204B)
|
Colon Margin (OD06064)
14.0
Kidney normal adjacent
18.4
|
tissue (OD06204E)
|
Colon cancer (OD06159)
14.2
Kidney Cancer (OD04450-
38.4
|
01)
|
Colon Margin (OD06159)
11.6
Kidney Margin (OD04450-
39.8
|
03)
|
Colon cancer (OD06297-04)
8.8
Kidney Cancer 8120613
11.3
|
Colon Margin (OD06297-05)
21.8
Kidney Margin 8120614
26.6
|
CC Gr.2 ascend colon
12.7
Kidney Cancer 9010320
8.1
|
(ODO3921)
|
CC Margin (ODO3921)
7.9
Kidney Margin 9010321
10.2
|
Colon cancer metastasis
9.7
Kidney Cancer 8120607
28.1
|
(OD06104)
|
Lung Margin (OD06104)
9.2
Kidney Margin 8120608
19.9
|
Colon mets to lung
19.6
Normal Uterus
11.5
|
(OD04451-01)
|
Lung Margin (OD04451-02)
27.9
Uterine Cancer 064011
5.6
|
Normal Prostate
11.5
Normal Thyroid
4.0
|
Prostate Cancer (OD04410)
6.5
Thyroid Cancer 064010
10.7
|
Prostate Margin (OD04410)
8.3
Thyroid Cancer A302152
14.5
|
Normal Ovary
21.2
Thyroid Margin A302153
3.1
|
Ovarian cancer (OD06283-
10.4
Normal Breast
16.3
|
03)
|
Ovarian Margin (OD06283-
8.4
Breast Cancer (OD04566)
16.4
|
07)
|
Ovarian Cancer 064008
6.8
Breast Cancer 1024
24.7
|
Ovarian cancer (OD06145)
3.1
Breast Cancer (OD04590-
45.1
|
01)
|
Ovarian Margin (OD06145)
22.7
Breast Cancer Mets
33.4
|
(OD04590-03)
|
Ovarian cancer (OD06455-
10.7
Breast Cancer Metastasis
49.3
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
6.1
Breast Cancer 064006
12.7
|
07)
|
Normal Lung
15.4
Breast Cancer 9100266
29.1
|
Invasive poor diff. lung
13.0
Breast Margin 9100265
14.4
|
adeno (ODO4945-01)
|
Lung Margin (ODO4945-03)
10.3
Breast Cancer A209073
14.9
|
Lung Malignant Cancer
26.1
Breast Margin A2090734
13.8
|
(OD03126)
|
Lung Margin (OD03126)
9.9
Breast cancer (OD06083)
35.4
|
Lung Cancer (OD05014A)
29.3
Breast cancer node
19.3
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
24.1
Normal Liver
12.9
|
Lung cancer (OD06081)
8.2
Liver Cancer 1026
7.3
|
Lung Margin (OD06081)
8.5
Liver Cancer 1025
16.8
|
Lung Cancer (OD04237-01)
8.4
Liver Cancer 6004-T
12.3
|
Lung Margin (OD04237-02)
22.8
Liver Tissue 6004-N
7.7
|
Ocular Melanoma Metastasis
18.6
Liver Cancer 6005-T
24.3
|
Ocular Melanoma Margin
7.8
Liver Tissue 6005-N
26.2
|
(Liver)
|
Melanoma Metastasis
12.5
Liver Cancer 064003
15.3
|
Melanoma Margin (Lung)
27.5
Normal Bladder
11.0
|
Normal Kidney
20.4
Bladder Cancer 1023
11.9
|
Kidney Ca, Nuclear grade 2
68.8
Bladder Cancer A302173
14.3
|
(OD04338)
|
Kidney Margin (OD04338)
15.1
Normal Stomach
45.1
|
Kidney Ca Nuclear grade 1/2
27.2
Gastric Cancer 9060397
14.7
|
(OD04339)
|
Kidney Margin (OD04339)
20.4
Stomach Margin 9060396
43.5
|
Kidney Ca, Clear cell type
9.9
Gastric Cancer 9060395
14.9
|
(OD04340)
|
Kidney Margin (OD04340)
28.7
Stomach Margin 9060394
48.3
|
Kidney Ca, Nuclear grade 3
8.5
Gastric Cancer 064005
11.3
|
(OD04348)
|
|
[1401]
633
TABLE BDE
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3832, Run
Ag3832, Run
|
Tissue Name
170127273
Tissue Name
170127273
|
|
Secondary Th1 act
26.2
HUVEC IL-1beta
9.5
|
Secondary Th2 act
40.1
HUVEC IFN gamma
9.2
|
Secondary Tr1 act
36.9
HUVEC TNF alpha + IFN
5.4
|
gamma
|
Secondary Th1 rest
13.6
HUVEC TNF alpha + IL4
4.8
|
Secondary Th2 rest
18.8
HUVEC IL-11
5.8
|
Secondary Tr1 rest
15.3
Lung Microvascular EC none
16.8
|
Primary Th1 act
20.7
Lung Microvascular EC
13.9
|
TNF alpha + IL-1beta
|
Primary Th2 act
22.2
Microvascular Dermal EC none
12.3
|
Primary Tr1 act
18.9
Microsvasular Dermal EC
6.1
|
TNF alpha + IL-1beta
|
Primary Th1 rest
15.3
Bronchial epithelium
62.4
|
TNF alpha + IL-1beta
|
Primary Th2 rest
13.4
Small airway epithelium none
44.8
|
Primary Tr1 rest
17.4
Small airway epithelium
90.1
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
20.6
Coronery artery SMC rest
18.6
|
act
|
CD45RO CD4 lymphocyte
39.0
Coronery artery SMC
20.2
|
act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
29.9
Astrocytes rest
36.1
|
Secondary CD8
41.8
Astrocytes TNF alpha + IL-
35.6
|
lymphocyte rest
1beta
|
Secondary CD8
16.7
KU-812 (Basophil) rest
12.5
|
lymphocyte act
|
CD4 lymphocyte none
9.2
KU-812 (Basophil)
27.4
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
16.3
CCD1106 (Keratinocytes) none
80.1
|
CD95 CH11
|
LAK cells rest
35.6
CCD1106 (Keratinocytes)
100.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
26.1
Liver cirrhosis
7.6
|
LAK cells IL-2 + IL-12
24.3
NCI-H292 none
33.0
|
LAK cells IL-2 + IFN
29.9
NCI-H292 IL-4
69.3
|
gamma
|
LAK cells IL-2 + IL-18
37.1
NCI-H292 IL-9
68.3
|
LAK cells
14.4
NCI-H292 IL-13
70.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
34.6
NCI-H292 IFN gamma
71.2
|
Two Way MLR 3 day
32.3
HPAEC none
8.4
|
Two Way MLR 5 day
27.5
HPAEC TNF alpha + IL-1beta
10.5
|
Two Way MLR 7 day
20.6
Lung fibroblast none
23.8
|
PBMC rest
9.2
Lung fibroblast TNF alpha + IL-
10.8
|
1beta
|
PBMC PWM
30.6
Lung fibroblast IL-4
15.5
|
PBMC PHA-L
15.0
Lung fibroblast IL-9
24.0
|
Ramos (B cell) none
26.8
Lung fibroblast IL-13
16.8
|
Ramos (B cell) ionomycin
15.6
Lung fibroblast IFN gamma
18.2
|
B lymphocytes PWM
15.8
Dermal fibroblast CCD1070 rest
27.4
|
B lymphocytes CD40L
15.3
Dermal fibroblast CCD1070
29.7
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
17.9
Dermal fibroblast CCD1070 IL-
8.6
|
1beta
|
EOL-1 dbcAMP
21.3
Dermal fibroblast IFN gamma
5.6
|
PMA/ionomycin
|
Dendritic cells none
28.1
Dermal fibroblast IL-4
8.5
|
Dendritic cells LPS
35.6
Dermal Fibroblasts rest
12.9
|
Dendritic cells anti-CD40
32.8
Neutrophils TNFa + LPS
3.3
|
Monocytes rest
15.1
Neutrophils rest
14.4
|
Monocytes LPS
27.9
Colon
15.5
|
Macrophages rest
37.4
Lung
18.2
|
Macrophages LPS
33.9
Thymus
12.7
|
HUVEC none
7.1
Kidney
40.9
|
HUVEC starved
10.2
|
|
[1402] CNS_neurodegeneration_v1.0 Summary: Ag3832 This panel confirms the expression of the CG92896-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1403] General_screening13 panel_v1.4 Summary: Ag3832 Highest expression of the CG92896-01 gene is detected in breast cancer cell line T47D (CT=25.9). In addition high expression of this gene is also seen in cluster of cancer (pancreatic, CNS, colon, gastric, renal, breast, ovarian, prostate, squamous cell carcinoma, and melanoma) cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs might be beneficial in the treatment of these cancers.
[1404] Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1405] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1406] High expression of this gene is also detected in fetal lung. Interestingly, this gene is expressed at much higher levels in fetal (CT=29) when compared to adult lung (CT=32.4). This observation suggests that expression of this gene can be used to distinguish fetal from adult lung. In addition, the relative overexpression of this gene in fetal lung suggests that the protein product may be required for growth or development of lung in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of lung related diseases.
[1407] Panel 2.2 Summary: Ag3832 Highest expression of the CG92896-01 gene is detected in kidney margin (OD04348) sample (CT=29.5). Interestingly, expression of this gene is much higher in control kidney margin (OD04348) than in corresponding cancer sample (CT=33). Therefore, expression of this gene can be used to distinguish the kidney cancer from control sample. In addition, expression of this gene is seen in both normal and cancer tissue samples used in this panel. Please see Panel 1.4 for a discussion of the potential utility of this gene.
[1408] Panel 4.1D Summary: Ag3832 Highest expression of the CG92896-01 gene is detected in TNFalpha +IL-1beta treated keratinocytes (CT=27.6). This gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1409] BE. CG93265-01 and CG93265-O2: L-serine dehydratase
[1410] Expression of gene CG93265-01 and full length clone CG93265-02 was assessed using the primer-probe set Ag3847, described in Table BEA. Results of the RTQ-PCR runs are shown in Tables BEB, BEC, BED and BEE. Please note that CG93265-02 represents a full-length physical clone of the CG93265-01 gene, validating the prediction of the gene sequence.
634TABLE BEA
|
|
Probe Name Ag3847
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-gccttagcagccatctactca-3′21902429
ProbeTET-5′-23960430
ccccttccctgacttcagttgtg-
3′-TAMRA
Reverse5′-tgcctccacacacgattac-3′19983431
|
[1411]
635
TABLE BEB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3847, Run
Ag3847, Run
|
Tissue Name
212186740
Tissue Name
212186740
|
|
AD 1 Hippo
7.2
Control (Path) 3
8.2
|
Temporal Ctx
|
AD 2 Hippo
64.6
Control (Path) 4
36.3
|
Temporal Ctx
|
AD 3 Hippo
9.7
AD 1 Occipital Ctx
29.1
|
AD 4 Hippo
33.7
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
72.7
AD 3 Occipital Ctx
10.8
|
AD 6 Hippo
39.0
AD 4 Occipital Ctx
38.7
|
Control 2 Hippo
34.2
AD 5 Occipital Ctx
95.9
|
Control 4 Hippo
67.4
AD 6 Occipital Ctx
27.5
|
Control (Path) 3 Hippo
1.9
Control 1 Occipital Ctx
32.3
|
AD 1 Temporal Ctx
21.8
Control 2 Occipital Ctx
100.0
|
AD 2 Temporal Ctx
63.3
Control 3 Occipital Ctx
56.3
|
AD 3 Temporal Ctx
7.0
Control 4 Occipital Ctx
36.3
|
AD 4 Temporal Ctx
25.5
Control (Path) 1
98.6
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
62.0
Control (Path) 2
24.3
|
Occipital Ctx
|
AD 5 Sup Temporal
29.7
Control (Path) 3
4.8
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
32.5
Control (Path) 4
39.2
|
Occipital Ctx
|
AD 6 Sup Temporal
37.6
Control 1 Parietal Ctx
35.8
|
Ctx
|
Control 1 Temporal Ctx
35.6
Control 2 Parietal Ctx
12.7
|
Control 2 Temporal Ctx
30.6
Control 3 Parietal Ctx
31.9
|
Control 3 Temporal Ctx
24.0
Control (Path) 1
64.2
|
Parietal Ctx
|
Control 3 Temporal Ctx
18.4
Control (Path) 2
66.9
|
Parietal Ctx
|
Control (Path) 1
62.4
Control (Path) 3
6.6
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
48.3
Control (Path) 4
73.7
|
Temporal Ctx
Parietal Ctx
|
|
[1412]
636
TABLE BEC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3847, Run
Ag3847, Run
|
Tissue Name
218906045
Tissue Name
218906045
|
|
Adipose
11.4
Renal ca. TK-10
40.6
|
Melanoma* Hs688(A).T
4.6
Bladder
30.6
|
Melanoma* Hs688(B).T
5.6
Gastric ca. (liver met.)
55.1
|
NCI-N87
|
Melanoma* M14
4.8
Gastric ca. KATO III
25.5
|
Melanoma* LOXIMVI
1.5
Colon ca. SW-948
8.7
|
Melanoma* SK-MEL-5
20.9
Colon ca. SW480
28.9
|
Squamous cell
2.9
Colon ca.* (SW480 met)
6.2
|
carcinoma SCC-4
SW620
|
Testis Pool
11.0
Colon ca. HT29
11.7
|
Prostate ca.* (bone met)
14.8
Colon ca. HCT-116
43.8
|
PC-3
|
Prostate Pool
8.5
Colon ca. CaCo-2
9.9
|
Placenta
8.8
Colon cancer tissue
19.8
|
Uterus Pool
5.2
Colon ca. SW1116
16.2
|
Ovarian ca. OVCAR-3
42.3
Colon ca. Colo-205
4.8
|
Ovarian ca. SK-OV-3
9.2
Colon ca. SW-48
5.0
|
Ovarian ca. OVCAR-4
21.3
Colon Pool
7.2
|
Ovarian ca. OVCAR-5
39.8
Small Intestine Pool
5.6
|
Ovarian ca. IGROV-1
18.7
Stomach Pool
8.7
|
Ovarian ca. OVCAR-8
22.4
Bone Marrow Pool
2.3
|
Ovary
13.2
Fetal Heart
2.2
|
Breast ca. MCF-7
28.3
Heart Pool
6.8
|
Breast ca. MDA-MB-
48.6
Lymph Node Pool
22.8
|
231
|
Breast ca. BT 549
35.4
Fetal Skeletal Muscle
2.2
|
Breast ca. T47D
100.0
Skeletal Muscle Pool
5.4
|
Breast ca. MDA-N
9.4
Spleen Pool
9.6
|
Breast Pool
9.9
Thymus Pool
10.3
|
Trachea
7.1
CNS cancer (glio/astro)
48.0
|
U87-MG
|
Lung
2.4
CNS cancer (glio/astro) U-
50.3
|
118-MG
|
Fetal Lung
5.5
CNS cancer (neuro; met)
12.9
|
SK-N-AS
|
Lung ca. NCI-N417
2.2
CNS cancer (astro) SF-539
11.9
|
Lung ca. LX-1
13.9
CNS cancer (astro) SNB-75
29.7
|
Lung ca. NCI-H146
6.6
CNS cancer (glio) SNB-19
15.5
|
Lung ca. SHP-77
23.5
CNS cancer (glio) SF-295
48.3
|
Lung ca. A549
34.2
Brain (Amygdala) Pool
15.9
|
Lung ca. NCI-H526
1.5
Brain (cerebellum)
9.2
|
Lung ca. NCI-H23
18.4
Brain (fetal)
2.3
|
Lung ca. NCI-H460
17.1
Brain (Hippocampus) Pool
11.3
|
Lung ca. HOP-62
4.1
Cerebral Cortex Pool
11.2
|
Lung ca. NCI-H522
19.9
Brain (Substantia nigra)
16.8
|
Pool
|
Liver
16.2
Brain (Thalamus) Pool
11.7
|
Fetal Liver
30.4
Brain (whole)
13.1
|
Liver ca. HepG2
11.7
Spinal Cord Pool
36.1
|
Kidney Pool
4.8
Adrenal Gland
36.9
|
Fetal Kidney
21.8
Pituitary gland Pool
23.7
|
Renal ca. 786-0
18.7
Salivary Gland
9.0
|
Renal ca. A498
11.4
Thyroid (female)
25.5
|
Renal ca. ACHN
23.7
Pancreatic ca. CAPAN2
3.7
|
Renal ca. UO-31
21.6
Pancreas Pool
12.1
|
|
[1413]
637
TABLE BED
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3847, Run
Ag3847, Run
|
Tissue Name
170126781
Tissue Name
170126781
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
8.1
|
Secondary Th2 act
0.5
HUVEC IFN gamma
5.4
|
Secondary Tr1 act
1.1
HUVEC TNF alpha + IFN
12.9
|
gamma
|
Secondary Th1 rest
2.5
HUVEC TNF alpha + IL4
6.8
|
Secondary Th2 rest
2.4
HUVEC IL-11
6.7
|
Secondary Tr1 rest
5.3
Lung Microvascular EC none
28.1
|
Primary Th1 act
0.6
Lung Microvascular EC
8.3
|
TNF alpha + IL-1beta
|
Primary Th2 act
1.2
Microvascular Dermal EC none
4.0
|
Primary Tr1 act
2.4
Microsvasular Dermal EC
5.5
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
13.4
|
alpha + IL-1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
12.2
|
Primary Tr1 rest
0.0
Small airway epithelium
15.7
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
12.8
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
6.4
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
11.6
|
Secondary CD8
0.5
Astrocytes TNF alpha + IL-
28.1
|
lymphocyte rest
1beta
|
Secondary CD8
0.0
KU-812 (Basophil) rest
2.3
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
14.7
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
1.7
CCD1106 (Keratinocytes) none
17.2
|
CD95 CH11
|
LAK cells rest
49.0
CCD1106 (Keratinocytes)
24.7
|
TNF alpha + IL-1beta
|
LAK cells IL-2
7.3
Liver cirrhosis
100.0
|
LAK cells IL-2 + IL-12
1.4
NCI-H292 none
13.0
|
LAK cells IL-2 + IFN
2.0
NCI-H292 IL-4
13.6
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
26.2
|
LAK cells
9.3
NCI-H292 IL-13
6.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
3.7
NCI-H292 IFN gamma
27.4
|
Two Way MLR 3 day
28.3
HPAEC none
9.7
|
Two Way MLR 5 day
11.3
HPAEC TNF alpha + IL-1beta
4.0
|
Two Way MLR 7 day
3.9
Lung fibroblast none
34.9
|
PBMC rest
2.6
Lung fibroblast TNF alpha + IL-
7.6
|
1beta
|
PBMC PWM
3.4
Lung fibroblast IL-4
8.6
|
PBMC PHA-L
0.7
Lung fibroblast IL-9
14.7
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
6.8
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
15.2
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
2.3
|
B lymphocytes CD40L
1.4
Dermal fibroblast CCD1070
0.8
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
4.0
Dermal fibroblast CCD1070 IL-
1.0
|
1beta
|
EOL-1 dbcAMP
0.5
Dermal fibroblast IFN gamma
1.6
|
PMA/ionomycin
|
Dendritic cells none
62.0
Dermal fibroblast IL-4
4.4
|
Dendritic cells LPS
22.1
Dermal Fibroblasts rest
4.6
|
Dendritic cells anti-CD40
44.1
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
4.4
Neutrophils rest
0.0
|
Monocytes LPS
24.7
Colon
3.8
|
Macrophages rest
84.1
Lung
14.9
|
Macrophages LPS
72.2
Thymus
3.6
|
HUVEC none
4.4
Kidney
55.5
|
HUVEC starved
4.1
|
|
[1414]
638
TABLE BEE
|
|
|
Panel 5 Islet
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3847, Run
Ag3847, Run
|
Tissue Name
226587525
Tissue Name
226587525
|
|
97457_Patient-
28.1
94709_Donor 2 AM - A_adipose
14.9
|
02go_adipose
|
97476_Patient-07sk_skeletal
3.1
94710_Donor 2 AM - B_adipose
7.3
|
muscle
|
97477_Patient-07ut_uterus
4.2
94711_Donor 2 AM - C_adipose
4.5
|
97478_Patient-
11.2
94712_Donor 2 AD - A_adipose
5.1
|
07pl_placenta
|
99167_Bayer Patient 1
100.0
94713_Donor 2 AD - B_adipose
14.7
|
97482_Patient-08ut_uterus
10.8
94714_Donor 2 AD - C_adipose
6.7
|
97483_Patient-
8.3
94742_Donor 3 U - A_Mesenchymal
8.3
|
08pl_placenta
Stem Cells
|
97486_Patient-09sk_skeletal
0.0
94743_Donor 3 U - B_Mesenchymal
0.0
|
muscle
Stem Cells
|
97487_Patient-09ut_uterus
3.6
94730_Donor 3 AM - A_adipose
18.2
|
97488_Patient-
3.9
94731_Donor 3 AM - B_adipose
0.0
|
09pl_placenta
|
97492_Patient-10ut_uterus
13.7
94732_Donor 3 AM - C_adipose
7.5
|
97493_Patient-
18.8
94733_Donor 3 AD - A_adipose
24.3
|
10pl_placenta
|
97495_Patient-
1.0
94734_Donor 3 AD - B_adipose
7.7
|
11go_adipose
|
97496_Patient-11sk_skeletal
1.7
94735_Donor 3 AD - C_adipose
8.0
|
muscle
|
97497_Patient-11ut_uterus
2.9
77138_Liver_HepG2untreated
0.0
|
97498_Patient-
2.7
73556_Heart_Cardiac stromal cells
9.3
|
11pl_placenta
(primary)
|
97500 Patient-
14.0
81735_Small Intestine
14.8
|
12go_adipose
|
97501_Patient-12sk_skeletal
7.7
72409_Kidney_Proximal Convoluted
16.8
|
muscle
Tubule
|
97502_Patient-12ut_uterus
3.4
82685_Small intestine_Duodenum
0.6
|
97503_Patient-
9.0
90650_Adrenal_Adrenocortical
15.0
|
12pl_placenta
adenoma
|
94721_Donor 2 U -
13.4
72410_Kidney_HRCE
31.0
|
A_Mesenchymal Stem Cells
|
94722_Donor 2 U -
0.0
72411_Kidney_HRE
25.2
|
B_Mesenchymal Stem Cells
|
94723_Donor 2 U -
15.8
73139_Uterus_Uterine smooth
12.3
|
C_Mesenchymal Stem Cells
muscle cells
|
|
[1415] CNS_neurodegeneration_v1.0 Summary: Ag3847 This panel confirms the expression of the CG93265-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1416] General_screening_panel_v1.4 Summary: Ag3847 Highest expression of the CG93265-01 gene is detected in breast cancer T47D cell line (CT=26). In addition, high expression of this gene is seen in cluster of cancer cell lines (pancreatic, CNS, colon, renal, liver, breast, ovarian, prostate, squamous cell carcinoma and melanoma) used in this panel. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.
[1417] This gene encodes a putative serine dehydratase, a gluconeogenic enzyme that produces pyruvate from serine. Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Inhibition of this enzyme in adipose or liver may decrease gluconeogenesis and be an adjunct therapy for the treatment of hyperglycemia and excess hepatic glucose production in Type 2 diabetes.
[1418] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1419] Panel 4.1D Summary: Ag3847 Highest expression of the CG93265-01 gene is detected in liver cirrhosis (CT=29.5). This gene encodes a homologue of rat L-serine dehydratase. Therefore, small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis.
[1420] In addition, moderate to low expression of this gene is seen in normal tissues represented by kidney, thymus, lung and colon, as well as in lung fibroblasts, endothelial cells, NCI-H292 cells, keratinocytes, basophils, astrocytes, coronary artery SMC cells, small airway epithelium, macrophage, monocytes, dendritic cells, LAK cells, PBMC cells, two way MLR. Interestingly, expression of this gene is down-regulated in cytokine treated LAK cells (CTs>33) as compared to resting LAK cells (CT=30). Furthermore, expression of this gene is higher in LPS treated monocytes (CT=31) as compared to resting monocytes (CT-34). Therefore, expression of this gene can be used to distinguish between activated LAK cells/monocytes and resting cells and modulation of the gene product with a functional therapeutic may lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1421] Panel 5 Islet Summary: Ag3847 Highest expression of the CG93265-01 gene is detected in pancrease from patient 1 (CT=30.3). Moderate expression of this gene is also seen in samples derived from adipose, uterus, small intestine, kidney and placenta. This gene encodes a protein that is homologous to a gluconeogenic enzyme. Since gluconeogenesis is an energetically-expensive (ATP-consuming) process, and because insulin secretion is dependent upon increases in the ATP/ADP ratio, inhibition of this gene product may be an effective therapy to increase insulin secretion in Type 2 diabetes.
[1422] BF. CG93464-01: 3-Kinase
[1423] Expression of gene CG93464-01 was assessed using the primer-probe set Ag3856, described in Table BFA.
639TABLE BFA
|
|
Probe Name Ag3856
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ctgcagtggtggaacagatt-3′202645432
ProbeTET-5′-262667433
caagtttgcccaggtaagttccctga-
3′-TAMRA
Reverse5′-aggactctggggcagtatacac-3′222720434
|
[1424] CNS_neurodegeneration_v1.0 Summary: Ag3856 Expression of the CG93464-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1425] General_screening_panel_v1.4 Summary: Ag3856 Expression of the CG93464-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1426] Panel 4.1D Summary: Ag3856 Expression of the CG93464-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1427] Panel 5 Islet Summary: Ag3856 Expression of the CG93464-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1428] BG. CG93495-01: MAP Kinase-Activating Death Domain Protein
[1429] Expression of gene CG93495-01 was assessed using the primer-probe set Ag3891, described in Table BGA. Results of the RTQ-PCR runs are shown in Tables BGB and BGC.
640TABLE BGA
|
|
Probe Name Ag3891
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-gggatcaacctcaaattcatg-3′214712435
ProbeTET-5′-294735436
caatcaggttttcatagagctgaatcaca-
3′TAMRA
Reverse5′-aagacgcctcgaactgtattg-3′214774437
|
[1430]
641
TABLE BGB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3891, Run
Ag3891, Run
|
Tissue Name
212195211
Tissue Name
212195211
|
|
AD 1 Hippo
29.9
Control (Path) 3
3.0
|
Temporal Ctx
|
AD 2 Hippo
31.2
Control (Path) 4
38.2
|
Temporal Ctx
|
AD 3 Hippo
9.7
AD 1 Occipital Ctx
23.5
|
AD 4 Hippo
10.8
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 hippo
57.4
AD 3 Occipital Ctx
10.5
|
AD 6 Hippo
76.3
AD 4 Occipital Ctx
20.4
|
Control 2 Hippo
14.4
AD 5 Occipital Ctx
11.9
|
Control 4 Hippo
15.6
AD 6 Occipital Ctx
57.8
|
Control (Path) 3 Hippo
11.1
Control 1 Occipital Ctx
6.4
|
AD 1 Temporal Ctx
15.3
Control 2 Occipital Ctx
54.7
|
AD 2 Temporal Ctx
46.3
Control 3 Occipital Ctx
21.2
|
AD 3 Temporal Ctx
7.9
Control 4 Occipital Ctx
7.5
|
AD 4 Temporal Ctx
23.0
Control (Path) 1
100.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
81.2
Control (Path) 2
6.0
|
Occipital Ctx
|
AD 5 Sup Temporal Ctx
33.0
Control (Path) 3
5.5
|
Occipital Ctx
|
AD 6 Inf Temporal Ctx
60.7
Control (Path) 4
6.8
|
Occipital Ctx
|
AD 6 Sup Temporal Ctx
51.1
Control 1 Parietal Ctx
8.9
|
Control 1 Temporal Ctx
7.4
Control 2 Parietal Ctx
29.1
|
Control 2 Temporal Ctx
65.5
Control 3 Parietal Ctx
24.8
|
Control 3 Temporal Ctx
11.8
Control (Path) 1
90.1
|
Parietal Ctx
|
Control 4 Temporal Ctx
11.1
Control (Path) 2
16.2
|
Parietal Ctx
|
Control (Path) 1
26.2
Control (Path) 3
6.5
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
42.0
Control (Path) 4
21.6
|
Temporal Ctx
Parietal Ctx
|
|
[1431]
642
TABLE BGC
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3891, Run
Ag3891, Run
|
Tissue Name
170130430
Tissue Name
170130430
|
|
Secondary Th1 act
41.2
HUVEC IL-1beta
20.4
|
Secondary Th2 act
55.9
HUVEC IFN gamma
25.5
|
Secondary Tr1 act
41.5
HUVEC TNF alpha + IFN
10.4
|
gamma
|
Secondary Th1 rest
13.1
HUVEC TNF alpha + IL4
9.7
|
Secondary Th2 rest
27.5
HUVEC IL-11
7.6
|
Secondary Tr1 rest
27.9
Lung Microvascular EC none
25.5
|
Primary Th1 act
17.0
Lung Microvascular EC
16.2
|
TNF alpha + IL-1beta
|
Primary Th2 act
45.4
Microvascular Dermal EC none
13.9
|
Primary Tr1 act
33.2
Microsvascular Dermal EC
9.1
|
TNF alpha + IL-1beta
|
Primary Th1 rest
14.8
Bronchial epithelium TNF
7.1
|
alpha + IL-1beta
|
Primary Th2 rest
18.4
Small airway epithelium none
3.5
|
Primary Tr1 rest
26.4
Small airway epithelium
7.5
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
24.8
Coronery artery SMC rest
5.3
|
act
|
CD45RO CD4 lymphocyte
47.6
Coronery artery SMC TNF
6.4
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
31.4
Astrocytes rest
4.4
|
Secondary CD8
31.9
Astrocytes TNF alpha + IL-
4.1
|
lymphocyte rest
1beta
|
Secondary CD8
17.7
KU-812 (Basophil) rest
13.6
|
lymphocyte act
|
CD4 lymphocyte none
15.5
KU-812 (Basophil)
31.9
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
52.1
CCD1106 (Keratinocytes) none
12.4
|
CD95 CH11
|
LAK cells rest
38.4
CCD1106 (Keratinocytes)
8.9
|
TNF alpha + IL-1beta
|
LAK cells IL-2
25.0
Liver cirrhosis
4.6
|
LAK cells IL-2 + IL-12
14.6
NCI-H292 none
14.9
|
LAK cells IL-2 + IFN
11.2
NCI-H292 IL-4
19.5
|
gamma
|
LAK cells IL-2 + IL-18
22.8
NCI-H292 IL-9
25.0
|
LAK cells
27.7
NCI-H292 IL-13
19.5
|
PMA/ionomycin
|
NK Cells IL-2 rest
61.6
NCI-H292 IFN gamma
20.2
|
Two Way MLR 3 day
39.0
HPAEC none
5.4
|
Two Way MLR 5 day
22.5
HPAEC TNF alpha + IL-1beta
17.7
|
Two Way MLR 7 day
21.3
Lung fibroblast none
11.0
|
PBMC rest
14.5
Lung fibroblast TNF alpha + IL-
23.7
|
1beta
|
PBMC PWM
26.2
Lung fibroblast IL-4
10.1
|
PBMC PHA-L
29.1
Lung fibroblast IL-9
19.6
|
Ramos (B cell) none
14.4
Lung fibroblast IL-13
13.0
|
Ramos (B cell) ionomycin
16.8
Lung fibroblast IFN gamma
15.4
|
B lymphocytes PWM
24.1
Dermal fibroblast CCD1070 rest
17.8
|
B lymphocytes CD40L
37.1
Dermal fibroblast CCD1070
56.3
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
27.9
Dermal fibroblast CCD1070 IL-
20.0
|
1beta
|
EOL-1 dbcAMP
23.8
Dermal fibroblast IFN gamma
10.8
|
PMA/ionomycin
|
Dendritic cells none
25.0
Dermal fibroblast IL-4
15.6
|
Dendritic cells LPS
28.5
Dermal Fibroblasts rest
10.7
|
Dendritic cells anti-CD40
24.7
Neutrophils TNFa + LPS
1.8
|
Monocytes rest
34.4
Neutrophils rest
5.8
|
Monocytes LPS
45.1
Colon
5.5
|
Macrophages rest
100.0
Lung
8.7
|
Macrophages LPS
51.4
Thymus
18.9
|
HUVEC none
9.1
Kidney
14.4
|
HUVEC starved
13.6
|
|
[1432] CNS_neurodegeneration_v1.0 Summary: Ag3891 This panel confirms the expression of the CG93495-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment.
[1433] The CG93495-01 gene encodes a splice variant of MAP kinase-activating death domain protein (MADD). The MADD gene is differentially expressed in neoplastic versus normal cells and the protein is a substrate for c-Jun N-terminal kinase in the human central nervous system (Zhang Y, Zhou L, Miller CA. (1998) A splicing variant of a death domain protein that is regulated by a mitogen-activated kinase is a substrate for c-Jun N-terminal kinase in the human central nervous system. Proc Natl Acad Sci USA 95(5):2586-91). MADD homolog from C. elegans, AEX-3, a GDP/GTP exchange proteins specific for the Rab3 subfamily members has been shown to regulate exocytosis of neurotransmitters (Iwasaki K, Staunton J, Saifee O, Nonet M, Thomas J H. (1997) aex-3 encodes a novel regulator of presynaptic activity in C. elegans. Neuron 18(4):613-22). Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of neurological disorders.
[1434] General_screening_panel_v1.4 Summary: Ag3891 Results from one experiment with the CG93495-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.
[1435] Panel 4.1D Summary: Ag3891 Highest expression of the CG93495-01 gene is detected in resting macrophage (CT=27). This gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1436] BH. CG93594-01: Phosphatidylinositol-Specific Phospholipase C
[1437] Expression of gene CG93594-01 was assessed using the primer-probe set Ag3861, described in Table BHA. Results of the RTQ-PCR runs are shown in Tables BHB and BHC.
643TABLE BHA
|
|
Probe Name Ag3861
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-tgcgagtccatgagtttatttt-3′22 984438
ProbeTET-5′-261006439
tcacaccaggaagttcattaccagaa-
3′-TAMRA
Reverse5′-agagtctgctcttgttgctttg-3′221039440
|
[1438]
644
TABLE BHB
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3861, Run
Ag3861, Run
|
Tissue Name
213609049
Tissue Name
213609049
|
|
Adipose
0.0
Renal ca. TK-10
0.0
|
Melanoma* Hs688(A).T
0.0
Bladder
0.2
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
1.1
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.8
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.0
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
SW620
|
Testis Pool
100.0
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
0.0
Colon ca. HCT-116
0.0
|
PC-3
|
Prostate Pool
0.0
Colon ca. CaCo-2
0.0
|
Placenta
0.0
Colon cancer tissue
0.0
|
Uterus Pool
0.0
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
0.0
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
1.9
Colon Pool
0.0
|
Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
0.0
|
Ovarian ca. IGROV-1
0.0
Stomach Pool
0.2
|
Ovarian ca. OVCAR-8
1.4
Bone Marrow Pool
0.0
|
Ovary
0.2
Fetal Heart
0.0
|
Breast ca. MCF-7
0.0
Heart Pool
0.1
|
Breast ca. MDA-MB-
8.5
Lymph Node Pool
0.6
|
231
|
Breast ca. BT 549
0.0
Fetal Skeletal Muscle
0.2
|
Breast ca. T47D
0.0
Skeletal Muscle Pool
0.0
|
Breast ca. MDA-N
0.0
Spleen Pool
0.3
|
Breast Pool
0.1
Thymus Pool
0.0
|
Trachea
0.8
CNS cancer (glio/astro)
0.1
|
U87-MG
|
Lung
0.0
CNS cancer (glio/astro) U-
0.4
|
118-MG
|
Fetal Lung
0.1
CNS cancer (neuro;met)
0.5
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0
|
Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
0.0
|
Lung ca. NCI-H146
0.3
CNS cancer (glio) SNB-19
0.0
|
Lung ca. SHP-77
0.1
CNS cancer (glio) SF-295
0.0
|
Lung ca. A549
0.0
Brain (Amygdala) Pool
0.0
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0
|
Lung ca. NCI-H23
0.5
Brain (fetal)
0.2
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
0.0
|
Lung ca. HOP-62
0.1
Cerebral Cortex Pool
0.5
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
0.0
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
0.1
|
Fetal Liver
1.5
Brain (whole)
0.3
|
Liver ca. HepG2
0.0
Spinal Cord Pool
0.2
|
Kidney Pool
0.0
Adrenal Gland
0.0
|
Fetal Kidney
3.2
Pituitary gland Pool
0.0
|
Renal ca. 786-0
0.0
Salivary Gland
0.1
|
Renal ca. A498
0.2
Thyroid (female)
0.0
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.0
|
Renal ca. UO-31
0.0
Pancreas Pool
0.1
|
|
[1439]
645
TABLE BHC
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3861, Run
Ag3861, Run
|
Tissue Name
170128790
Tissue Name
170128790
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.7
|
Secondary Th2 act
100.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
5.5
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvascular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
0.0
|
alpha + IL-1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.0
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
0.0
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.0
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-
0.0
|
lymphocyte rest
1beta
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.4
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
1.1
NCI-H292 none
0.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.0
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
2.7
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.3
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
0.4
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
0.0
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
0.3
Thymus
0.0
|
HUVEC none
0.0
Kidney
0.0
|
HUVEC starved
0.0
|
|
[1440] CNS_neurodegeneration_v1.0 Summary: Ag3861 Expression of the CG93594-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1441] General_screening_panel_v1.4 Summary: Ag3861 High expression of the CG93594-01 gene is exclusively seen in testis (CT=28.6). Therefore, expression of this gene can be used to distinguish testis from other samples used in this panel. Furthermore, therapeutic modulation of the activity of this gene may prove useful in the treatment of testis related disorders including fertility and hypogonadism.
[1442] In addition, low expression of this gene is seen in breast cancer MDA-MB-231 and ovarian cancer OVCAR-4 cell lines. Therefore, therapeutic modulation of the activity of this gene or its protein product, through the use of small molecule drugs might be beneficial in the treatment of breast and ovarian cancer.
[1443] Significant expression is also detected in fetal kidney. Interestingly, this gene is expressed at much higher levels in fetal (CT=33.5) when compared to adult kidney (CT=40). This observation suggests that expression of this gene can be used to distinguish fetal from adult kidney. In addition, the relative overexpression of this gene in fetal kidney suggests that the protein product may enhance kidney growth or development in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of kidney related diseases.
[1444] Panel 2.2 Summary: Ag3861 Expression of the CG93594-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1445] Panel 4.1D Summary: Ag3861 High expression of the CG93594-01 gene is exclusively seen in activated secondary Th2 cells (CT=29.5). Therefore, expression of this gene can be used to distinguish activated secondary Th2 cells from other samples used in this panel. Furthermore, therapeutics designed with the protein encoded by this gene could be important in the regulation of T cell function and in the treatment of diseases such as asthma, IBD, psoriasis and arthritis in which T cells are chronically stimulated.
[1446] BI. CG93669-01 and CG93669-02 and CG93669-03: Serine/Threonine-Protein Kinase NEK3
[1447] Expression of gene CG93669-01 and variants CG93669-02 and CG93669-03 was assessed using the primer-probe set Ag3874, described in Table BIA.
646TABLE BIA
|
|
Probe Name Ag3874
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-caagaaacgtgtgctacacaga-3′22602441
ProbeTET-5′-28628442
tcaagtcccagaatatcttcctcactca-
3′-TAMRA
Reverse5′-ctcccaatttcacttttccatt-3′22657443
|
[1448] CNS_neurodegeneration_v1.0 Summary: Ag3874 Expression of the CG93669-01 gene is low/undetectable (CTs>35) in all samples on this panel. (Data not shown.). The data suggest that there is a possibility of an experimental failure.
[1449] General_screening_panel_v1.4 Summary: Ag3874 Expression of the CG93669-01 gene is low/undetectable (CTs>35) in all samples on this panel. (Data not shown.). The data suggest that there is a possibility of an experimental failure.
[1450] Panel 4.1D Summary: Ag3874 Expression of the CG93669-01 gene is low/undetectable (CTs>35) in all samples on this panel. (Data not shown.). The data suggest that there is a possibility of an experimental failure.
[1451] BJ. CG93896-01 and CG93896-2: Protein Kinase (NEK like)
[1452] Expression of gene CG93896-01 and variant CG93896-02 was assessed using the primer-probe sets Ag3892 and Ag4153, described in Tables BJA and BJB. Results of the RTQ-PCR runs are shown in Tables BJC, BJD, BJE and BJF.
647TABLE BJA
|
|
Probe Name Ag3892
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-acttgctcgaacttgtattgga-3′22468444
ProbeTET-5′-acctgtccccagagatctgtcagaat-26500445
3′-TAMRA
Reverse5′-ccaagagaccaaatatccgttt-3′22542446
|
[1453]
648
TABLE BJB
|
|
|
Probe Name Ag4153
|
Start
SEQ
|
Primers
Sequences
Length
Position
ID NO:
|
|
Forward
5′-gagagcccacaaaccaagttat-3′
22
1101
447
|
Probe
TET-5′-
27
1123
448
|
caccccattcctcaagaaaatactgga-
|
3′-TAMRA
|
Reverse
5′-cctcgtttcctgaccgtaat-3′
20
1157
449
|
|
[1454]
649
TABLE BJC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3892,
Rel. Exp. (%) Ag3892,
|
Tissue Name
Run 212247893
Tissue Name
Run 212247893
|
|
AD 1 Hippo
51.8
Control (Path) 3
20.2
|
Temporal Ctx
|
AD 2 Hippo
54.0
Control (Path) 4
36.1
|
Temporal Ctx
|
AD 3 Hippo
54.7
AD 1 Occipital Ctx
24.8
|
AD 4 Hippo
6.5
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
27.4
AD 3 Occipital Ctx
27.9
|
AD 6 Hippo
81.2
AD 4 Occipital Ctx
14.1
|
Control 2 Hippo
25.5
AD 5 Occipital Ctx
25.0
|
Control 4 Hippo
60.3
AD 6 Occipital Ctx
28.1
|
Control (Path) 3 Hippo
60.3
Control 1 Occipital Ctx
8.6
|
AD 1 Temporal Ctx
61.6
Control 2 Occipital Ctx
19.3
|
AD 2 Temporal Ctx
28.5
Control 3 Occipital Ctx
9.8
|
AD 3 Temporal Ctx
27.0
Control 4 Occipital Ctx
43.8
|
AD 4 Temporal Ctx
29.5
Control (Path) 1
100.0
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
67.8
Control (Path) 2
6.8
|
Occipital Ctx
|
AD 5 Sup Temporal
97.3
Control (Path) 3
6.8
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
45.7
Control (Path) 4
33.0
|
Occipital Ctx
|
AD 6 Sup Temporal
48.0
Control 1 Parietal Ctx
11.7
|
Ctx
|
Control 1 Temporal Ctx
12.4
Control 2 Parietal Ctx
75.3
|
Control 2 Temporal Ctx
19.3
Control 3 Parietal Ctx
2.2
|
Control 3 Temporal Ctx
31.0
Control (Path) 1
49.7
|
Parietal Ctx
|
Control 3 Temporal Ctx
15.3
Control (Path) 2
48.0
|
Parietal Ctx
|
Control (Path) 1
54.0
Control (Path) 3
21.6
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
25.2
Control (Path) 4
33.2
|
Temporal Ctx
Parietal Ctx
|
|
[1455]
650
TABLE BJD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3892,
Rel. Exp. (%) Ag3892,
|
Tissue Name
Run 219166392
Tissue Name
Run 219166392
|
|
Adipose
8.0
Renal ca. TK-10
5.4
|
Melanoma* Hs688(A).T
0.8
Bladder
4.9
|
Melanoma* Hs688(B).T
2.5
Gastric ca. (liver met.)
45.1
|
NCI-N87
|
Melanoma* M14
5.2
Gastric ca. KATO III
9.3
|
Melanoma* LOXIMVI
0.5
Colon ca. SW-948
2.7
|
Melanoma* SK-MEL-5
3.9
Colon ca. SW480
33.4
|
Squamous cell
0.7
Colon ca.* (SW480 met)
2.7
|
carcinoma SSC-4
SW620
|
Testis Pool
73.2
Colon ca. HT29
3.8
|
Prostate ca.* (bone met)
3.2
Colon ca. HCT-116
18.9
|
PC-3
|
Prostate Pool
4.5
Colon ca. CaCo-2
3.4
|
Placenta
2.3
Colon cancer tissue
6.9
|
Uterus Pool
1.6
Colon ca. SW1116
3.6
|
Ovarian ca. OVCAR-3
7.2
Colon ca. Colo-205
2.5
|
Ovarian ca. SK-OV-3
7.6
Colon ca. SW-48
3.3
|
Ovarian ca. OVCAR-4
0.7
Colon Pool
8.8
|
Ovarian ca. OVCAR-5
7.7
Small Intestine Pool
4.6
|
Ovarian ca. IGROV-1
12.7
Stomach Pool
3.3
|
Ovarian ca. OVCAR-8
1.2
Bone Marrow Pool
1.4
|
Ovary
2.9
Fetal Heart
2.2
|
Breast ca. MCF-7
9.6
Heart Pool
3.1
|
Breast ca. MDA-MB-
1.6
Lymph Node Pool
2.9
|
231
|
Breast ca. BT 549
1.6
Fetal Skeletal Muscle
1.4
|
Breast ca. T47D
31.9
Skeletal Muscle Pool
0.7
|
Breast ca. MDA-N
0.1
Spleen Pool
0.5
|
Breast Pool
5.9
Thymus Pool
5.3
|
Trachea
62.4
CNS cancer (glio/astro)
0.7
|
U87-MG
|
Lung
1.8
CNS cancer (glio/astro) U-
0.8
|
118-MG
|
Fetal Lung
100.0
CNS cancer (neuro; met)
2.9
|
SK-N-AS
|
Lung ca. NCI-N417
0.2
CNS cancer (astro) SF-539
3.1
|
Lung ca. LX-1
20.0
CNS cancer (astro) SNB-75
0.6
|
Lung ca. NCI-H146
0.5
CNS cancer (glio) SNB-19
13.9
|
Lung ca. SHP-77
0.6
CNS cancer (glio) SF-295
5.0
|
Lung ca. A549
2.2
Brain (Amygdala) Pool
2.0
|
Lung ca. NCI-H526
0.2
Brain (cerebellum)
0.5
|
Lung ca. NCI-H23
6.7
Brain (fetal)
4.7
|
Lung ca. NCI-H460
1.2
Brain (Hippocampus) Pool
8.9
|
Lung ca. HOP-62
0.3
Cerebral Cortex Pool
2.9
|
Lung ca. NCI-H522
8.5
Brain (Substantia nigra)
3.3
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
7.0
|
Fetal Liver
0.9
Brain (whole)
8.5
|
Liver ca. HepG2
1.7
Spinal Cord Pool
8.8
|
Kidney Pool
5.0
Adrenal Gland
0.3
|
Fetal Kidney
24.0
Pituitary gland Pool
2.2
|
Renal ca. 786-0
4.6
Salivary Gland
0.8
|
Renal ca. A498
7.5
Thyroid (female)
2.5
|
Renal ca. ACHN
3.5
Pancreatic ca. CAPAN2
1.3
|
Renal ca. UO-31
1.0
Pancreas Pool
7.2
|
|
[1456]
651
TABLE BJE
|
|
|
Panel 2.2
|
Rel. Exp. (%) Ag3892,
Rel. Exp. (%) Ag3892,
|
Tissue Name
Run 173768795
Tissue Name
Run 173768795
|
|
Normal Colon
16.4
Kidney Margin (OD04348)
84.1
|
Colon cancer (OD06064)
7.9
Kidney malignant cancer
24.7
|
(OD06204B)
|
Colon Margin (OD06064)
6.3
Kidney normal adjacent
23.5
|
tissue (OD06204E)
|
Colon cancer (OD06159)
4.3
Kidney Cancer (OD04450-
36.6
|
01)
|
Colon Margin (OD06159)
9.6
Kidney Margin (OD04450-
36.9
|
03)
|
Colon cancer (OD06297-04)
8.4
Kidney Cancer 8120613
3.5
|
Colon Margin (OD06297-05)
15.1
Kidney Margin 8120614
51.8
|
CC Gr.2 ascend colon
11.6
Kidney Cancer 9010320
38.7
|
(ODO3921)
|
CC Margin (ODO3921)
11.3
Kidney Margin 9010321
18.6
|
Colon cancer metastasis
6.2
Kidney Cancer 8120607
52.5
|
(OD06104)
|
Lung Margin (OD06104)
22.2
Kidney Margin 8120608
34.9
|
Colon mets to lung
5.7
Normal Uterus
26.8
|
(OD04451-01)
|
Lung Margin (OD04451-02)
7.5
Uterine Cancer 064011
9.5
|
Normal Prostate
7.7
Normal Thyroid
2.3
|
Prostate Cancer (OD04410)
8.1
Thyroid Cancer 064010
8.0
|
Prostate Margin (OD04410)
11.9
Thyroid Cancer A302152
37.1
|
Normal Ovary
49.0
Thyroid Margin A302153
10.2
|
Ovarian cancer (OD06283-
16.2
Normal Breast
29.5
|
03)
|
Ovarian Margin (OD06283-
7.5
Breast Cancer (OD04566)
42.6
|
07)
|
Ovarian Cancer 064008
22.1
Breast Cancer 1024
17.1
|
Ovarian cancer (OD06145)
25.9
Breast Cancer (OD04590-
15.1
|
01)
|
Ovarian Margin (OD06145)
31.6
Breast Cancer Mets
27.7
|
(OD04590-03)
|
Ovarian cancer (OD06455-
25.9
Breast Cancer Metastasis
22.4
|
03)
(OD04655-05)
|
Ovarian Margin (OD06455-
10.7
Breast Cancer 064006
19.3
|
07)
|
Normal Lung
6.9
Breast Cancer 9100266
4.6
|
Invasive poor diff. lung
14.5
Breast Margin 9100265
7.9
|
adeno (ODO4945-01)
|
Lung Margin (ODO4945-03)
11.0
Breast Cancer A209073
10.3
|
Lung Malignant Cancer
8.1
Breast Margin A2090734
26.6
|
(OD03126)
|
Lung Margin (OD03126)
3.7
Breast cancer (OD06083)
32.8
|
Lung Cancer (OD05014A)
6.0
Breast cancer node
29.3
|
metastasis (OD06083)
|
Lung Margin (OD05014B)
8.2
Normal Liver
40.3
|
Lung cancer (OD06081)
18.4
Liver Cancer 1026
27.4
|
Lung Margin (OD06081)
5.3
Liver Cancer 1025
27.2
|
Lung Cancer (OD04237-01)
11.7
Liver Cancer 6004-T
32.8
|
Lung Margin (OD04237-02)
17.2
Liver Tissue 6004-N
21.8
|
Ocular Melanoma Metastasis
100.0
Liver Cancer 6005-T
57.0
|
Ocular Melanoma Margin
25.9
Liver Tissue 6005-N
95.9
|
(Liver)
|
Melanoma Metastasis
12.5
Liver Cancer 064003
32.3
|
Melanoma Margin (Lung)
10.2
Normal Bladder
8.1
|
Normal Kidney
16.3
Bladder Cancer 1023
17.0
|
Kidney Ca, Nuclear grade 2
57.8
Bladder Cancer A302173
24.7
|
(OD04338)
|
Kidney Margin (OD04338)
26.2
Normal Stomach
45.4
|
Kidney Ca, Nuclear grade 1/2
35.4
Gastric Cancer 9060397
3.3
|
(OD04339)
|
Kidney Margin (OD04339)
29.9
Stomach Margin 9060396
24.1
|
Kidney Ca, Clear cell type
11.7
Gastric Cancer 9060395
22.1
|
(OD04340)
|
Kidney Margin (OD04340)
24.8
Stomach Margin 9060394
29.7
|
Kidney Ca, Nuclear grade 3
6.0
Gastric Cancer 064005
17.6
|
(OD04348)
|
|
[1457]
652
TABLE BJF
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3892,
Rel. Exp. (%) Ag3892,
|
Tissue Name
Run 170130819
Tissue Name
Run 170130819
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
8.5
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
8.6
|
alpha + IL1beta
|
Primary Th2 rest
0.0
Small airway epithelium none
15.0
|
Primary Tr1 rest
0.0
Small airway epithelium
17.4
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
6.4
Coronery artery SMC TNF
0.0
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
26.6
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
16.8
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
71.2
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
100.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
9.7
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
10.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
7.2
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
25.9
|
LAK cells
0.0
NCI-H292 IL-13
21.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
16.5
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
5.5
|
PBMC PHA-L
6.4
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
12.4
Dermal fibroblast CCD1070 rest
1.4
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
6.5
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
17.1
|
Dendritic cells anti-CD40
8.7
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
4.7
|
Monocytes LPS
0.0
Colon
0.0
|
Macrophages rest
0.0
Lung
22.1
|
Macrophages LPS
0.0
Thymus
30.8
|
HUVEC none
0.0
Kidney
85.3
|
HUVEC starved
0.0
|
|
[1458] CNS_neurodegeneration_v1.0 Summary: Ag3892 This panel confirms the expression of the CG93896-01 gene at low levels in the brain in an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1459] Ag4153 Expression of the CG93896-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1460] General_screening_panel_v1.4 Summary: Ag3892 Highest expression of the CG93896-01 gene is observed in fetal lung (CT=28.7). Interestingly, this gene is expressed at much higher levels in fetal when compared to adult lung (CT=34.5). This observation suggests that expression of this gene can be used to distinguish fetal from adult lung. In addition, the relative overexpression of this gene in fetal lung suggests that the protein product may enhance growth or development of lung in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of lung related diseases.
[1461] In addition, high expression of this gene is seen in trachea and testis pool (CTs=29). Therefore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of trachea and testis related diseases including fertility and hypogonadism.
[1462] Among tissues with metabolic or endocrine function, this gene is expressed at low to moderate levels in pancreas, adipose, thyroid, pituitary gland, skeletal muscle, heart, and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1463] In addition, this gene is expressed at moderate levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression
[1464] Ag4153 Expression of the CG93896-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1465] Panel 2.2 Summary: Ag3892 Highest expression of the CG93896-01 gene is observed in ocular melenoma metastasis to liver (OD04310) (CT=31.4). Significant expression of this gene is associated with both normal and cancer tissue samples used in this panel. Interestingly, expression of this gene is higher in control kidney margin OD04348 and 8120614 and stomach margin 9060396 samples (CTs=32-33) as compared to their corresponding cancer tissue (CTs=36). Therefore, expression of this gene can be used as distinguish between normal and kidney/gastric cancer samples. Furthermore, therapeutic modulation of the protein encoded by this gene could be useful in treatment of these cancers.
[1466] Panel 4.1D Summary: Ag3892 Highest expression of the CG93896-01 gene is observed in PMA/ionomycin treated basophils (CT=33.5). Significant expression of this gene is also seen in resting basophils (CT=34). Therefore, therapeutic modulation of the protein encoded for by this gene could useful in treatment of asthma, allergies, hypersensitivity reactions, psoriasis, and viral infections.
[1467] In addition, moderate expression is also seen in kidney (CT=33.4). Therefore, expression of this gene could be used to distinguish basophils and kidney samples from other samples used in this panel. In addition, therapeutic modulation of the protein encoded for by this gene could useful in treatment of kidney related diseases such as lupus, glomerulonephritis. The CG93896-01 gene codes for NimA-related protein kinase (NEK). Mutation in one of the related protein NEK1 has recently been shown to be cause pleiotropic effects including facial dysmorphism, dwarfing, male sterility, anemia, cystic choroid plexus, and progressive polycystic kidney disease (PKD) in mice (Upadhya P, Birkenmeier E H, Birkenmeier CS, Barker J E. (2000) Mutations in a NIMA-related kinase gene, Nek1, cause pleiotropic effects including a progressive polycystic kidney disease in mice. Proc Natl Acad Sci USA 97(1):217-21). Therefore, therapeutic modulation of the NEK encoded for by this gene could useful in treatment of facial dysmorphism, dwarfing, male sterility, anemia, cystic choroid plexus, and progressive PKD.
[1468] Ag4153 Expression of the CG93896-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).
[1469] BK. CG94302-01: MUNC13-1
[1470] Expression of gene CG94302-01 was assessed using the primer-probe set Ag3911, described in Table BKA.
653TABLE BKA
|
|
Probe Name Ag3911
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ggtggtaaaggactgtgtgaaa-3′222957447
ProbeTET-5′-272999448
cgagtacatcttcaataactgccatga-
3′-TAMRA
Reverse5′-ggtactcccggctgtacagt-3′203026449
|
[1471] CNS_neurodegeneration_v1.0 Summary: Ag3911 Results from one experiment with the CG94302-01 gene are not included. (All CTs=40). The data suggest that there is a possibility of an experimental failure.
[1472] General_screening_panel_v1.4 Summary: Ag3911 Results from one experiment with the CG94302-01 gene are not included. (All CTs=40). The data suggest that there is a possibility of an experimental failure.
[1473] Panel 4.1D Summary: Ag3911 Results from one experiment with the CG94302-01 gene are not included. (All CTs=40). The data suggest that there is a possibility of an experimental failure.
[1474] BL. CG94356-01: Carboxylesterase
[1475] Expression of gene CG94356-01 was assessed using the primer-probe set Ag3915, described in Table BLA. Results of the RTQ-PCR runs are shown in Tables BLB and BLC.
654TABLE BLA
|
|
Probe Name Ag3915
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-agaagttactgagccggaagat-3′221463450
ProbeTET-5′-atactgggctacctttgctcgaaccg-261491451
3′-TAMRA
Reverse5′-gattataagctggccacagaga-3′221540452
|
[1476]
655
TABLE BLB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3915,
Rel. Exp. (%) Ag3915,
|
Tissue Name
Run 212248481
Tissue Name
Run 212248481
|
|
AD 1 Hippo
12.1
Control (Path) 3
0.0
|
Temporal Ctx
|
AD 2 Hippo
25.2
Control (Path) 4
29.5
|
Temporal Ctx
|
AD 3 Hippo
0.0
AD 1 Occipital Ctx
5.3
|
AD 4 Hippo
8.4
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
28.9
AD 3 Occipital Ctx
0.0
|
AD 6 Hippo
29.5
AD 4 Occipital Ctx
9.2
|
Control 2 Hippo
26.2
AD 5 Occipital Ctx
12.8
|
Control 4 Hippo
9.1
AD 6 Occipital Ctx
22.2
|
Control (Path) 3 Hippo
0.0
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
5.4
Control 2 Occipital Ctx
30.4
|
AD 2 Temporal Ctx
24.8
Control 3 Occipital Ctx
9.2
|
AD 3 Temporal Ctx
0.0
Control 4 Occipital Ctx
6.1
|
AD 4 Temporal Ctx
10.3
Control (Path) 1
70.2
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
36.9
Control (Path) 2
0.0
|
Occipital Ctx
|
AD 5 Sup Temporal
8.0
Control (Path) 3
11.6
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
63.3
Control (Path) 4
7.5
|
Occipital Ctx
|
AD 6 Sup Temporal
52.5
Control 1 Parietal Ctx
7.1
|
Ctx
|
Control 1 Temporal Ctx
0.0
Control 2 Parietal Ctx
22.1
|
Control 2 Temporal Ctx
65.1
Control 3 Parietal Ctx
35.4
|
Control 3 Temporal Ctx
22.7
Control (Path) 1
71.2
|
Parietal Ctx
|
Control 3 Temporal Ctx
9.5
Control (Path) 2
9.9
|
Parietal Ctx
|
Control (Path) 1
100.0
Control (Path) 3
0.0
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
35.8
Control (Path) 4
58.6
|
Temporal Ctx
Parietal Ctx
|
|
[1477]
656
TABLE BLC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3915,
Rel. Exp. (%) Ag3915,
|
Tissue Name
Run 219174535
Tissue Name
Run 219174535
|
|
Adipose
0.0
Renal ca. TK-10
0.0
|
Melanoma* Hs688(A).T
3.9
Bladder
0.0
|
Melanoma* Hs688(B).T
1.8
Gastric ca. (liver met.)
0.0
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
33.7
Colon ca. SW480
5.2
|
Squamous cell
1.4
Colon ca.* (SW480 met)
5.0
|
carcinoma SCC-4
SW620
|
Testis Pool
100.0
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
0.0
Colon ca. HCT-116
0.0
|
PC-3
|
Prostate Pool
0.0
Colon ca. CaCo-2
0.0
|
Placenta
0.0
Colon cancer tissue
0.0
|
Uterus Pool
0.0
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
0.0
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
0.0
|
Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
0.0
|
Ovarian ca. IGROV-1
0.0
Stomach Pool
0.0
|
Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
0.0
|
Ovary
1.8
Fetal Heart
0.0
|
Breast ca. MCF-7
0.0
Heart Pool
1.4
|
Breast ca. MDA-MB-
0.0
Lymph Node Pool
0.0
|
231
|
Breast ca. BT 549
0.0
Fetal Skeletal Muscle
0.0
|
Breast ca. T47D
0.0
Skeletal Muscle Pool
0.0
|
Breast ca. MDA-N
1.6
Spleen Pool
0.7
|
Breast Pool
0.0
Thymus Pool
3.6
|
Trachea
0.0
CNS cancer (glio/astro)
1.9
|
U87-MG
|
Lung
0.0
CNS cancer (glio/astro) U-
0.0
|
118-MG
|
Fetal Lung
0.0
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.7
CNS cancer (astro) SF-539
0.0
|
Lung ca. LX-1
1.2
CNS cancer (astro) SNB-75
0.0
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.0
|
Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
1.8
|
Lung ca. A549
1.8
Brain (Amygdala) Pool
18.0
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
19.6
|
Lung ca. NCI-H23
0.0
Brain (fetal)
20.6
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
14.0
|
Lung ca. HOP-62
0.0
Cerebral Cortex Pool
22.5
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
8.2
|
Pool
|
Liver
34.9
Brain (Thalamus) Pool
21.8
|
Fetal Liver
0.0
Brain (whole)
7.1
|
Liver ca. HepG2
0.0
Spinal Cord Pool
5.2
|
Kidney Pool
0.0
Adrenal Gland
0.0
|
Fetal Kidney
0.0
Pituitary gland Pool
0.0
|
Renal ca. 786-0
0.0
Salivary Gland
2.0
|
Renal ca. A498
0.0
Thyroid (female)
0.0
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.0
|
Renal ca. UO-31
0.0
Pancreas Pool
0.0
|
|
[1478] CNS_neurodegeneration_v1.0 Summary: Ag3915 The CG94356-01 gene is found to be down-regulated in the temporal cortex of Alzheimer's disease patients. Therefore, up-regulation of this gene or its protein product, or treatment with specific agonists of this enzyme may prevent, reduce or reverse the neuronal death, dementia, and memory loss associated with this disease.
[1479] General_screening_panel_v1.4 Summary: Ag3915 Highest expression of the CG94356-01 gene is seen in the testis (CT=32.4). Low, but significant levels of expression are also seen in amygdala, thalamus, cortex, cerebellum, and liver. This gene encodes a protein with homology to carboxylesterases, which catalyze the hydrolysis of a variety of compounds containing ester and amide bonds. They may also play an important role in lipid and drug metabolism by hydrolyzing endogenous long-chain fatty acid esters. Thus, expression of the CG94356-01 gene suggests that this gene product may play a similar role in the liver. In addition, the expression in the brain suggests that this gene may also play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1480] Panel 4.1D Summary: Ag3915 Expression of the CG94356-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)
[1481] BM. CG94421-01: Kelch-BTB Protein
[1482] Expression of gene CG94421-01 was assessed using the primer-probe set Ag3961, described in Table BMA. Results of the RTQ-PCR runs are shown in Tables BMB, BMC, BMD and BME.
657TABLE BMA
|
|
Probe Name Ag3961
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-aatgatgcaagatgctgattgt-3′22 945453
ProbeTET-5′-30 969454
cagacttctcgtagatgctatgaactacca-
3′-TAMRA
Reverse5′-tgttttgatgatatggaagcaa-3′221000455
|
[1483]
658
TABLE BMB
|
|
|
AI_comprehensive_panel_v1.0
|
Rel. Exp. (%) Ag3961,
Rel. Exp. (%) Ag3961,
|
Tissue Name
Run 248079817
Tissue Name
Run 248079817
|
|
110967 COPD-F
0.0
112427 Match Control
1.2
|
Psoriasis-F
|
110980 COPD-F
0.0
112418 Psoriasis-M
0.0
|
110968 COPD-M
0.3
112723 Match Control
1.7
|
Psoriasis-M
|
110977 COPD-M
0.2
112419 Psoriasis-M
0.2
|
110989 Emphysema-F
0.7
112424 Match Control
0.1
|
Psoriasis-M
|
110992 Emphysema-F
0.2
112420 Psoriasis-M
0.5
|
110993 Emphysema-F
0.1
112425 Match Control
1.5
|
Psoriasis-M
|
110994 Emphysema-F
0.1
104689 (MF) OA Bone-
0.5
|
Backus
|
110995 Emphysema-F
0.4
104690 (MF) Adj “Normal”
0.5
|
Bone-Backus
|
110996 Emphysema-F
0.3
104691 (MF) OA
3.7
|
Synovium-Backus
|
110997 Asthma-M
0.1
104692 (BA) OA Cartilage-
1.7
|
Backus
|
111001 Asthma-F
1.6
104694 (BA) OA Bone-
0.5
|
Backus
|
111002 Asthma-F
5.1
104695 (BA) Adj “Normal”
1.0
|
Bone-Backus
|
111003 Atopic Asthma-F
1.5
104696 (BA) OA Synovium-
2.6
|
Backus
|
111004 Atopic Asthma-F
4.0
104700 (SS) OA Bone-
0.2
|
Backus
|
111005 Atopic Asthma-F
2.0
104701 (SS) Adj “Normal”
1.2
|
Bone-Backus
|
111006 Atopic Asthma-F
1.2
104702 (SS) OA Synovium-
23.3
|
Backus
|
111417 Allergy-M
2.0
117093 OA Cartilage Rep7
0.5
|
112347 Allergy-M
0.0
112672 OA Bone5
0.3
|
112349 Normal Lung-F
0.0
112673 OA Synovium5
0.0
|
112357 Normal Lung-F
0.6
112674 OA Synovial Fluid
0.1
|
cells5
|
112354 Normal Lung-M
1.5
117100 OA Cartilage Rep14
0.1
|
112374 Crohns-F
0.6
112756 OA Bone9
0.0
|
112389 Match Control
0.4
112757 OA Synovium9
100.0
|
Crohns-F
|
112375 Crohns-F
0.0
112758 OA Synovial Fluid
0.0
|
Cells9
|
112732 Match Control
0.5
117125 RA Cartilage Rep2
0.1
|
Crohns-F
|
112725 Crohns-M
0.0
113492 Bone2 RA
0.5
|
112387 Match Control
0.0
113493 Synovium2 RA
0.3
|
Crohns-M
|
112378 Crohns-M
0.0
113494 Syn Fluid Cells RA
0.1
|
112390 Match Control
2.0
113499 Cartilage4 RA
0.5
|
Crohns-M
|
112726 Crohns-M
0.4
113500 Bone4 RA
0.9
|
112731 Match Control
0.2
113501 Synovium4 RA
0.7
|
Crohns-M
|
112380 Ulcer Col-F
0.6
113502 Syn Fluid Cells4 RA
0.2
|
112734 Match Control
5.7
113495 Cartilage3 RA
0.0
|
Ulcer Col-F
|
112384 Ulcer Col-F
0.1
113496 Bone3 RA
0.3
|
112737 Match Control
0.2
113497 Synovium3 RA
0.0
|
Ulcer Col-F
|
112386 Ulcer Col-F
0.0
113498 Syn Fluid Cells3 RA
0.5
|
112738 Match Control
0.2
117106 Normal Cartilage
0.3
|
Ulcer Col-F
Rep20
|
112381 Ulcer Col-M
0.0
113663 Bone3 Normal
0.0
|
112735 Match Control
0.0
113664 Synovium3 Normal
0.0
|
Ulcer Col-M
|
112382 Ulcer Col-M
0.5
113665 Syn Fluid Cells3
0.0
|
Normal
|
112394 Match Control
0.0
117107 Normal Cartilage
0.0
|
Ulcer Col-M
Rep22
|
112383 Ulcer Col-M
0.5
113667 Bone4 Normal
0.0
|
112736 Match Control
0.0
113668 Synovium4 Normal
0.1
|
Ulcer Col-M
|
112423 Psoriasis-F
0.2
113669 Syn Fluid Cells4
0.0
|
Normal
|
|
[1484]
659
TABLE BMC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3961,
Rel. Exp. (%) Ag3961,
|
Tissue Name
Run 212343353
Tissue Name
Run 212343353
|
|
AD 1 Hippo
38.7
Control (Path) 3
0.0
|
Temporal Ctx
|
AD 2 Hippo
69.3
Control (Path) 4
53.2
|
Temporal Ctx
|
AD 3 Hippo
15.6
AD 1 Occipital Ctx
20.0
|
AD 4 Hippo
19.9
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
48.3
AD 3 Occipital Ctx
19.8
|
AD 6 Hippo
68.8
AD 4 Occipital Ctx
16.7
|
Control 2 Hippo
27.5
AD 5 Occipital Ctx
30.6
|
Control 4 Hippo
52.1
AD 6 Occipital Ctx
30.8
|
Control (Path) 3 Hippo
17.8
Control 1 Occipital Ctx
0.0
|
AD 1 Temporal Ctx
3.8
Control 2 Occipital Ctx
32.3
|
AD 2 Temporal Ctx
0.0
Control 3 Occipital Ctx
20.0
|
AD 3 Temporal Ctx
8.3
Control 4 Occipital Ctx
33.9
|
AD 4 Temporal Ctx
40.3
Control (Path) 1
35.1
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
79.6
Control (Path) 2
31.6
|
Occipital Ctx
|
AD 5 Sup Temporal
100.0
Control (Path) 3
0.0
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
30.1
Control (Path) 4
7.5
|
Occipital Ctx
|
AD 6 Sup Temporal
19.5
Control 1 Parietal Ctx
9.6
|
Ctx
|
Control 1 Temporal Ctx
0.0
Control 2 Parietal Ctx
8.2
|
Control 2 Temporal Ctx
59.5
Control 3 Parietal Ctx
40.9
|
Control 3 Temporal Ctx
10.5
Control (Path) 1
21.9
|
Parietal Ctx
|
Control 3 Temporal Ctx
30.1
Control (Path) 2
0.0
|
Parietal Ctx
|
Control (Path) 1
24.3
Control (Path) 3
19.2
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
38.2
Control (Path) 4
57.4
|
Temporal Ctx
Parietal Ctx
|
|
[1485]
660
TABLE BMD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3961,
Rel. Exp. (%) Ag3961,
|
Tissue Name
Run 217332993
Tissue Name
Run 217332993
|
|
Adipose
9.0
Renal ca. TK-10
0.3
|
Melanoma* Hs688(A).T
0.0
Bladder
0.2
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.2
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.4
|
Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
0.1
Colon ca. SW480
0.2
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.1
|
carcinoma SCC-4
SW620
|
Testis Pool
0.1
Colon ca. HT29
0.0
|
Prostate ca.* (bone met)
0.1
Colon ca. HCT-116
0.3
|
PC-3
|
Prostate Pool
0.7
Colon ca. CaCo-2
0.5
|
Placenta
0.6
Colon cancer tissue
0.2
|
Uterus Pool
0.0
Colon ca. SW1116
0.2
|
Ovarian ca. OVCAR-3
0.1
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
0.0
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
0.2
|
Ovarian ca. OVCAR-5
0.5
Small Intestine Pool
0.1
|
Ovarian ca. IGROV-1
0.1
Stomach Pool
0.5
|
Ovarian ca. OVCAR-8
0.3
Bone Marrow Pool
0.0
|
Ovary
0.1
Fetal Heart
20.2
|
Breast ca. MCF-7
0.1
Heart Pool
25.3
|
Breast ca. MDA-MB-
0.0
Lymph Node Pool
0.3
|
231
|
Breast ca. BT 549
0.0
Fetal Skeletal Muscle
24.7
|
Breast ca. T47D
0.5
Skeletal Muscle Pool
100.0
|
Breast ca. MDA-N
0.0
Spleen Pool
0.1
|
Breast Pool
0.6
Thymus Pool
0.6
|
Trachea
0.6
CNS cancer (glio/astro)
0.0
|
U87-MG
|
Lung
0.1
CNS cancer (glio/astro) U-
0.1
|
118-MG
|
Fetal Lung
0.9
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.1
|
Lung ca. LX-1
0.3
CNS cancer (astro) SNB-75
0.0
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.0
|
Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
0.4
|
Lung ca. A549
0.1
Brain (Amygdala) Pool
0.0
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0
|
Lung ca. NCI-H23
0.0
Brain (fetal)
0.1
|
Lung ca. NCI-H460
0.1
Brain (Hippocampus) Pool
0.0
|
Lung ca. HOP-62
0.5
Cerebral Cortex Pool
0.0
|
Lung ca. NCI-H522
0.1
Brain (Substantia nigra)
0.0
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
0.1
|
Fetal Liver
0.7
Brain (whole)
0.0
|
Liver ca. HepG2
0.2
Spinal Cord Pool
0.4
|
Kidney Pool
0.1
Adrenal Gland
1.6
|
Fetal Kidney
1.8
Pituitary gland Pool
0.0
|
Renal ca. 786-0
0.0
Salivary Gland
0.2
|
Renal ca. A498
0.1
Thyroid (female)
0.0
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.2
|
Renal ca. UO-31
1.1
Pancreas Pool
0.4
|
|
[1486]
661
TABLE BME
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3961,
Rel. Exp. (%) Ag3961,
|
Tissue Name
Run 170739796
Tissue Name
Run 170739796
|
|
Secondary Th1 act
0.0
HUVEC IL-1beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
3.4
|
Primary Th1 act
3.2
Lung Microvascular EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF
37.6
|
alpha + IL1beta
|
Primary Th2 rest
3.3
Small airway epithelium none
3.5
|
Primary Tr1 rest
0.0
Small airway epithelium
25.2
|
TNF alpha + IL-1beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
0.0
|
act
alpha + IL-1beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
7.7
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
3.7
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1beta
|
LAK cells IL-2
0.0
Liver cirrhosis
6.5
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
6.0
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
2.8
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
14.8
|
LAK cells
0.0
NCI-H292 IL-13
6.3
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
7.5
|
Two Way MLR 3 day
0.0
HPAEC none
1.7
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.0
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
3.4
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0
|
B lymphocytes PWM
5.2
Dermal fibroblast CCD1070 rest
0.0
|
B lymphocytes CD40L
3.6
Dermal fibroblast CCD1070
0.0
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0
|
1beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.0
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
6.9
|
Macrophages rest
0.0
Lung
0.0
|
Macrophages LPS
0.0
Thymus
25.0
|
HUVEC none
0.0
Kidney
100.0
|
HUVEC starved
0.0
|
|
[1487] AI_comprehensive panel_v1.0 Summary: Ag3961 Expression of the CG94421-01 gene is highest in the synovium of an osteoarthritis (OA) patient (CT=28.4). Low, but significant levels are also seen in additional OA synovium samples and in asthma. Thus, this gene product may be involved in OA and asthma.
[1488] CNS_neurodegeneration_v1.0 Summary: Ag3961 This panel does not show differential expression of the CG94421-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. This gene encodes a homolog of Kelch, a family of actin organizing proteins in Drosophila. Thus, this gene product may play a role in organizing the actin cytoskeleton in the brain.
[1489] General_screening_panel_v1.4 Summary: Ag3961 Highest expression of the CG94421-01 gene is seen in skeletal muscle (CT=25.3). High levels of expression are also seen in fetal skeletal muscle, heart, and fetal heart tissue. This gene encodes a putative kelch protein, which in Drosophila comprise a family of actin organizing proteins. Overall, this gene appears to be preferentially expressed in normal tissue samples and is expressed at moderate to low levels in other metabolic tissues, including pancreas, adrenal, fetal liver and adipose. Thus, this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1490] Panel 4.1D Summary: Ag3961 Expression of the CG94421-01 gene is highest in the kidney (CT=32.3). Low, but significant expression is also seen in TNF-alpha/IL-1 beta treated small airway and bronchial epithelium and in thymus. This expression is in agreement with the expression seen in Panel AI. Thus, this gene product may be involved in inflammatory conditions of the lung, including asthma.
[1491] BN. CG94465-01: Protein Kinase
[1492] Expression of gene CG94465-01 was assessed using the primer-probe set Ag3959, described in Table BNA. Results of the RTQ-PCR runs are shown in Tables BNB, BNC, BND and BNE.
662TABLE BNA
|
|
Probe Name Ag3959
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ggcaaattgtatgcagtaaagg-3′22302456
ProbeTET-5′-aaatatgactcatcaggtccaagctg-26352457
3′-TAMRA
Reverse5′-cttagtgccagtgcatctctct-3′22378458
|
[1493]
663
TABLE BNB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3959,
Rel. Exp. (%) Ag3959,
|
Tissue Name
Run 212347482
Tissue Name
Run 212347482
|
|
AD 1 Hippo
6.1
Control (Path) 3
6.8
|
Temporal Ctx
|
AD 2 Hippo
24.1
Control (Path) 4
10.9
|
Temporal Ctx
|
AD 3 Hippo
9.4
AD 1 Occipital Ctx
14.9
|
AD 4 Hippo
7.3
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
6.3
|
AD 6 Hippo
52.5
AD 4 Occipital Ctx
14.1
|
Control 2 Hippo
33.9
AD 5 Occipital Ctx
50.7
|
Control 4 Hippo
8.0
AD 6 Occipital Ctx
42.9
|
Control (Path) 3 Hippo
4.1
Control 1 Occipital Ctx
2.1
|
AD 1 Temporal Ctx
17.4
Control 2 Occipital Ctx
59.0
|
AD 2 Temporal Ctx
18.3
Control 3 Occipital Ctx
11.9
|
AD 3 Temporal Ctx
4.8
Control 4 Occipital Ctx
3.1
|
AD 4 Temporal Ctx
18.4
Control (Path) 1
77.4
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
66.4
Control (Path) 2
0.0
|
Occipital Ctx
|
AD 5 Sup Temporal
37.1
Control (Path) 3
6.0
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
75.3
Control (Path) 4
29.5
|
Occipital Ctx
|
AD 6 Sup Temporal
77.4
Control 1 Parietal Ctx
4.3
|
Ctx
|
Control 1 Temporal Ctx
4.8
Control 2 Parietal Ctx
39.0
|
Control 2 Temporal Ctx
40.6
Control 3 Parietal Ctx
13.2
|
Control 3 Temporal Ctx
9.3
Control (Path) 1
58.6
|
Parietal Ctx
|
Control 3 Temporal Ctx
4.1
Control (Path) 2
17.7
|
Parietal Ctx
|
Control (Path) 1
55.5
Control (Path) 3
4.9
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
39.5
Control (Path) 4
54.0
|
Temporal Ctx
Parietal Ctx
|
|
[1494]
664
TABLE BNC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3959,
Rel. Exp. (%) Ag3959,
|
Tissue Name
Run 222691601
Tissue Name
Run 222691601
|
|
Adipose
5.9
Renal ca. TK-10
15.8
|
Melanoma* Hs688(A).T
4.9
Bladder
8.9
|
Melanoma* Hs688(B).T
4.4
Gastric ca. (liver met.)
51.8
|
NCI-N87
|
Melanoma* M14
41.8
Gastric ca. KATO III
48.0
|
Melanoma* LOXIMVI
28.3
Colon ca. SW-948
9.2
|
Melanoma* SK-MEL-5
26.2
Colon ca. SW480
51.8
|
Squamous cell
15.5
Colon ca.* (SW480 met)
26.2
|
carcinoma SCC-4
SW620
|
Testis Pool
6.8
Colon ca. HT29
14.8
|
Prostate ca.* (bone met)
5.3
Colon ca. HCT-116
49.7
|
PC-3
|
Prostate Pool
2.4
Colon ca. CaCo-2
48.0
|
Placenta
3.3
Colon cancer tissue
9.2
|
Uterus Pool
2.6
Colon ca. SW1116
2.9
|
Ovarian ca. OVCAR-3
52.9
Colon ca. Colo-205
3.9
|
Ovarian ca. SK-OV-3
40.3
Colon ca. SW-48
7.6
|
Ovarian ca. OVCAR-4
44.1
Colon Pool
4.5
|
Ovarian ca. OVCAR-5
25.2
Small Intestine Pool
3.2
|
Ovarian ca. IGROV-1
9.3
Stomach Pool
3.2
|
Ovarian ca. OVCAR-8
3.3
Bone Marrow Pool
1.9
|
Ovary
3.4
Fetal Heart
4.0
|
Breast ca. MCF-7
22.8
Heart Pool
1.1
|
Breast ca. MDA-MB-
75.8
Lymph Node Pool
4.8
|
231
|
Breast ca. BT 549
100.0
Fetal Skeletal Muscle
1.7
|
Breast ca. T47D
33.9
Skeletal Muscle Pool
1.9
|
Breast ca. MDA-N
27.5
Spleen Pool
7.7
|
Breast Pool
5.1
Thymus Pool
9.8
|
Trachea
4.5
CNS cancer (glio/astro)
26.1
|
U87-MG
|
Lung
1.0
CNS cancer (glio/astro) U-
35.6
|
118-MG
|
Fetal Lung
14.6
CNS cancer (neuro; met)
50.7
|
SK-N-AS
|
Lung ca. NCI-N417
10.2
CNS cancer (astro) SF-539
23.8
|
Lung ca. LX-1
39.0
CNS cancer (astro) SNB-75
26.1
|
Lung ca. NCI-H146
11.4
CNS cancer (glio) SNB-19
7.9
|
Lung ca. SHP-77
21.0
CNS cancer (glio) SF-295
10.8
|
Lung ca. A549
48.6
Brain (Amygdala) Pool
2.0
|
Lung ca. NCI-H526
5.7
Brain (cerebellum)
1.4
|
Lung ca. NCI-H23
29.7
Brain (fetal)
4.8
|
Lung ca. NCI-H460
3.5
Brain (Hippocampus) Pool
2.1
|
Lung ca. HOP-62
5.7
Cerebral Cortex Pool
3.1
|
Lung ca. NCI-H522
28.9
Brain (Substantia nigra)
1.9
|
Pool
|
Liver
0.2
Brain (Thalamus) Pool
3.7
|
Fetal Liver
14.0
Brain (whole)
2.5
|
Liver ca. HepG2
8.6
Spinal Cord Pool
1.8
|
Kidney Pool
6.8
Adrenal Gland
3.8
|
Fetal Kidney
8.1
Pituitary gland Pool
2.7
|
Renal ca. 786-0
40.3
Salivary Gland
1.1
|
Renal ca. A498
8.2
Thyroid (female)
1.0
|
Renal ca. ACHN
7.0
Pancreatic ca. CAPAN2
46.7
|
Renal ca. UO-31
24.5
Pancreas Pool
6.0
|
|
[1495]
665
TABLE BND
|
|
|
Panel 4.1D
|
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3959, Run
Ag3959, Run
Ag3959, Run
Ag3959, Run
|
Tissue Name
170739792
198383573
Tissue Name
170739792
198383573
|
|
Secondary Th1 act
63.7
56.3
HUVEC IL-1beta
14.3
16.6
|
Secondary Th2 act
100.0
100.0
HUVEC IFN gamma
13.5
11.1
|
Secondary Tr1 act
40.3
39.0
HUVEC TNF alpha +
14.2
16.0
|
IFN gamma
|
Secondary Th1 rest
4.6
4.2
HUVEC TNF alpha +
9.9
10.4
|
IL4
|
Secondary Th2 rest
4.9
4.4
HUVEC IL-11
7.2
6.9
|
Secondary Tr1 rest
6.2
5.6
Lung Microvascular
11.3
11.5
|
EC none
|
Primary Th1 act
16.4
17.8
Lung Microvascular
5.5
7.0
|
EC TNF alpha + IL-
|
1beta
|
Primary Th2 act
28.1
28.9
Microvascular
10.6
13.2
|
Dermal EC none
|
Primary Tr1 act
27.4
31.6
Microsvascular Dermal
8.8
7.9
|
EC TNF alpha + IL-
|
1beta
|
Primary Th1 rest
7.4
9.6
Bronchial epithelium
4.7
4.0
|
TNF alpha + IL1beta
|
Primary Th2 rest
3.2
5.3
Small airway
1.7
1.9
|
epithelium none
|
Primary Tr1 rest
7.9
10.0
Small airway
5.1
7.0
|
epithelium TNF
|
alpha + IL-1beta
|
CD45RA CD4
42.3
41.5
Coronery artery SMC
4.8
3.3
|
lymphocyte act
rest
|
CD45RO CD4
71.2
64.6
Coronery artery SMC
7.1
4.8
|
lymphocyte act
TNF alpha + IL-1beta
|
CD8 lymphocyte act
40.9
40.9
Astrocytes rest
2.1
2.7
|
Secondary CD8
47.0
43.2
Astrocytes TNF
2.2
3.3
|
lymphocyte rest
alpha + IL-1beta
|
Secondary CD8
21.3
17.3
KU-812 (Basophil)
9.5
9.5
|
lymphocyte act
rest
|
CD4 lymphocyte
4.0
3.6
KU-812 (Basophil)
21.6
21.8
|
none
PMA/ionomycin
|
2ry
13.5
13.5
CCD1106
29.7
29.7
|
Th1/Th2/Tr1_anti-
(Keratinocytes) none
|
CD95 CH11
|
LAK cells rest
12.1
13.5
CCD1106
27.0
20.3
|
(Keratinocytes)
|
TNF alpha + IL-1beta
|
LAK cells IL-2
27.4
30.4
Liver cirrhosis
3.2
2.1
|
LAK cells IL-2 + IL-
14.5
17.9
NCI-H292 none
17.4
14.8
|
12
|
LAK cells IL-2 + IFN
17.7
20.0
NCI-H292 IL-4
32.5
37.4
|
gamma
|
LAK cells IL-2 + IL-
54.7
23.3
NCI-H292 IL-9
36.3
44.8
|
18
|
LAK cells
28.1
30.1
NCI-H292 IL-13
37.4
47.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
46.7
46.3
NCI-H292 IFN
32.3
32.8
|
gamma
|
Two Way MLR 3
11.0
13.9
HPAEC none
4.6
6.8
|
day
|
Two Way MLR 5
17.6
17.1
HPAEC TNF alpha +
13.7
13.4
|
day
IL-1beta
|
Two Way MLR 7
15.5
17.3
Lung fibroblast none
5.9
4.5
|
day
|
PBMC rest
4.0
4.4
Lung fibroblast TNF
12.6
11.1
|
alpha + IL-1beta
|
PBMC PWM
26.6
20.0
Lung fibroblast IL-4
1.7
3.1
|
PBMC PHA-L
25.3
32.3
Lung fibroblast IL-9
6.9
6.6
|
Ramos (B cell) none
24.3
22.4
Lung fibroblast IL-13
3.0
3.8
|
Ramos (B cell)
26.1
26.4
Lung fibroblast IFN
4.0
3.5
|
ionomycin
gamma
|
B lymphocytes PWM
36.6
31.0
Dermal fibroblast
25.2
24.8
|
CCD1070 rest
|
B lymphocytes
20.0
24.0
Dermal fibroblast
41.8
52.1
|
CD40L and IL-4
CCD1070 TNF alpha
|
EOL-1 dbcAMP
23.8
14.6
Dermal fibroblast
23.8
31.2
|
CCD1070 IL-1beta
|
EOL-1 dbcAMP
24.8
27.7
Dermal fibroblast IFN
13.3
18.0
|
PMA/ionomycin
gamma
|
Dendritic cells none
7.1
8.9
Dermal fibroblast IL-4
16.4
18.0
|
Dendritic cells LPS
14.9
18.7
Dermal Fibroblasts
7.0
7.9
|
rest
|
Dendritic cells anti-
6.5
5.4
Neutrophils
2.7
2.4
|
CD40
TNFa + LPS
|
Monocytes rest
6.7
5.4
Neutrophils rest
3.4
4.2
|
Monocytes LPS
21.5
26.6
Colon
1.8
1.7
|
Macrophages rest
8.3
8.8
Lung
4.3
3.5
|
Macrophages LPS
20.7
23.3
Thymus
19.2
20.4
|
HUVEC none
13.5
13.7
Kidney
6.6
4.6
|
HUVEC starved
15.7
17.7
|
|
[1496]
666
TABLE BNE
|
|
|
Panel 5 Islet
|
Rel. Exp. (%)
Rel. Exp. (%)
|
Ag3959, Run
Ag3959, Run
|
Tissue Name
229431860
Tissue Name
229431860
|
|
97457_Patient-
14.5
94709_Donor 2 AM - A_adipose
29.1
|
02go_adipose
|
97476_Patient-07sk_skeletal
10.8
94710_Donor 2 AM - B_adipose
19.2
|
muscle
|
97477_Patient-07ut_uterus
7.1
94711_Donor 2 AM - C_adipose
7.7
|
97478_Patient-
14.0
94712_Donor 2 AD - A_adipose
17.1
|
07pl_placenta
|
99167_Bayer Patient 1
22.1
94713_Donor 2 AD - B_adipose
25.7
|
97482_Patient-08ut_uterus
9.6
94714_Donor 2 AD - C_adipose
17.9
|
97483_Patient-
33.0
94742_Donor 3 U - A_Mesenchymal
2.8
|
08pl_placenta
Stem Cells
|
97486_Patient-09sk_skeletal
0.8
94743_Donor 3 U—B_Mesenchymal
13.0
|
muscle
Stem Cells
|
97487_Patient-09ut_uterus
14.7
94730_Donor 3 AM - A_adipose
22.7
|
97488_Patient-
13.2
94731_Donor 3 AM - B_adipose
27.4
|
09pl_placenta
|
97492_Patient-10ut_uterus
13.9
94732_Donor 3 AM - C_adipose
24.1
|
97493_Patient-
46.3
94733_Donor 3 AD - A_adipose
24.7
|
10pl_placenta
|
97495_Patient-
13.3
94734_Donor 3 AD - B_adipose
8.7
|
11go_adipose
|
97496_Patient-11sk_skeletal
1.6
94735_Donor 3 AD - C_adipose
15.4
|
muscle
|
97497_Patient-11ut_uterus
11.7
77138_Liver_HepG2untreated
90.1
|
97498_Patient-
23.5
73556_Heart_Cardiac stromal cells
12.9
|
11pl_placenta
(primary)
|
97500_Patient-
20.2
81735_Small Intestine
20.7
|
12go_adipose
|
97501_Patient-12sk_skeletal
11.0
72409_Kidney_Proximal Convoluted
18.8
|
muscle
Tubule
|
97502_Patient-12ut_uterus
26.4
82685_Small intestine_Duodenum
24.7
|
97503_Patient-
19.1
90650_Adrenal_Adrenocortical
6.1
|
12pl_placenta
adenoma
|
94721_Donor 2 U -
11.3
72410_Kidney_HRCE
62.4
|
A_Mesenchymal Stem Cells
|
94722_Donor 2 U -
8.9
72411_Kidney_HRE
100.0
|
B_Mesenchymal Stem Cells
|
94723_Donor 2 U -
4.3
73139_Uterus_Uterine smooth
22.8
|
C_Mesenchymal Stem Cells
muscle cells
|
|
[1497] CNS_neurodegeneration_v1.0 Summary: Ag3959 This panel confirms the expression of the CG94465-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion of the potential utility of this gene in treatment of central nervous system disorders.
[1498] General_screening_panel_v1.4 Summary: Ag3959 Highest expression of the CG94465-01 gene is detected in a breast cancer BT 549 cell line (CT=27.3). In addition, high expression of this gene is associated with a number of cancer cell lines (pancreatic, CNS, colon, lung, gastric, renal, breast, ovarian, prostate, squamous cell carcinoma, and melanoma). Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of these cancers.
[1499] Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation of the activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.
[1500] In addition, this gene is expressed at high levels in all regions of the central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.
[1501] Panel 4.1D Summary: Ag3959 Two experiments with same primer and probe set are in excellent agreements with highest expression of the CG94465-01 gene in activated secondary Th2 cells (CTs=28). Expression of this gene is stimulated in activated secondary Th1 and Th2 cells (CTs=28-29) as compared to the resting cells (33). High expression of this gene is also detected in activated CD45RA CD4 lymphocyte and CD45RO CD4 lymphocyte. This expression pattern suggests that this gene may be important in T cell activation, polarization and proliferation. Thus, therapeutic regulation of the transcript or the protein encoded by this gene could be important in immune modulation and in the treatment of T cell-mediated diseases such as asthma, arthritis, psoriasis, IBD, and lupus.
[1502] In addition, this gene is expressed at low to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_v1.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1503] Panel 5 Islet Summary: Ag3959 Highest expression of the CG94465-01 gene is detected in 72411_kidney_HRE sample (CT=31). In addition, expression of this gene is ubiquitous throughout the samples used in this panel. Please see Panel 1.4 for a discussion of the potential utility of this gene in metabolic disorders.
[1504] BO. CG94511-01: Pyruvate Dehydrogenase
[1505] Expression of gene CG94511-01 was assessed using the primer-probe set Ag3922, described in Table BOA. Results of the RTQ-PCR runs are shown in Tables BOB, BOC and BOD.
667TABLE BOA
|
|
Probe Name Ag3922
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-ggctctggctttagctttactc-3′22374459
ProbeTET-5′-tgaactttctgtcccagaaattcttg-3′-26397460
TAMRA
Reverse5′-ttagcacaacctcctctttgtc-3′22435461
|
[1506]
668
TABLE BOB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3922, Run
Rel. Exp. (%) Ag3922, Run
|
Tissue Name
212343349
Tissue Name
212343349
|
|
AD 1 Hippo
7.0
Control (Path) 3
7.7
|
Temporal Ctx
|
AD 2 Hippo
11.4
Control (Path) 4
22.2
|
Temporal Ctx
|
AD 3 Hippo
10.7
AD 1 Occipital Ctx
12.2
|
AD 4 Hippo
11.5
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
5.3
|
AD 6 Hippo
66.0
AD 4 Occipital Ctx
9.7
|
Control 2 Hippo
25.7
AD 5 Occipital Ctx
20.4
|
Control 4 Hippo
5.1
AD 6 Occipital Ctx
20.3
|
Control (Path) 3 Hippo
9.9
Control 1 Occipital Ctx
10.9
|
AD 1 Temporal Ctx
26.8
Control 2 Occipital Ctx
25.5
|
AD 2 Temporal Ctx
19.1
Control 3 Occipital Ctx
11.7
|
AD 3 Temporal Ctx
7.0
Control 4 Occipital Ctx
2.0
|
AD 4 Temporal Ctx
34.6
Control (Path) 1
65.5
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
70.2
Control (Path) 2
9.3
|
Occipital Ctx
|
AD 5 SupTemporal
68.8
Control (Path) 3
1.2
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
42.6
Control (Path) 4
21.6
|
Occipital Ctx
|
AD 6 Sup Temporal
49.0
Control 1 Parietal Ctx
2.6
|
Ctx
|
Control 1 Temporal Ctx
8.5
Control 2 Parietal Ctx
61.1
|
Control 2 Temporal Ctx
12.3
Control 3 Parietal Ctx
2.9
|
Control 3 Temporal Ctx
10.2
Control (Path) 1
38.7
|
Parietal Ctx
|
Control 3 Temporal Ctx
4.9
Control (Path) 2
17.8
|
Parietal Ctx
|
Control (Path) 1
57.4
Control (Path) 3
3.8
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
23.7
Control (Path) 4
39.2
|
Temporal Ctx
Parietal Ctx
|
|
[1507]
669
TABLE BOC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3922, Run
Rel. Exp. (%) Ag3922, Run
|
Tissue Name
219515088
Tissue Name
219515088
|
|
Adipose
0.2
Renal ca. TK-10
0.4
|
Melanoma* Hs688(A).T
0.1
Bladder
1.1
|
Melanoma* Hs688(B).T
0.2
Gastric ca. (liver met.)
1.1
|
NCI-N87
|
Melanoma* M14
0.2
Gastric ca. KATO III
0.8
|
Melanoma* LOXIMVI
0.3
Colon ca. SW-948
0.1
|
Melanoma* SK-MEL-5
0.6
Colon ca. SW480
100.0
|
Squamous cell
0.2
Colon ca.* (SW480 met)
0.8
|
carcinoma SCC-4
SW620
|
Testis Pool
0.3
Colon ca. HT29
0.6
|
Prostate ca.* (bone met)
1.1
Colon ca. HCT-116
1.4
|
PC-3
|
Prostate Pool
0.3
Colon ca. CaCo-2
2.0
|
Placenta
0.1
Colon cancer tissue
0.4
|
Uterus Pool
0.1
Colon ca. SW1116
0.1
|
Ovarian ca. OVCAR-3
0.6
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
1.5
Colon ca. SW-48
0.1
|
Ovarian ca. OVCAR-4
0.2
Colon Pool
1.0
|
Ovarian ca. OVCAR-5
1.4
Small Intestine Pool
1.1
|
Ovarian ca. IGROV-1
0.3
Stomach Pool
0.6
|
Ovarian ca. OVCAR-8
0.1
Bone Marrow Pool
0.4
|
Ovary
0.5
Fetal Heart
0.9
|
Breast ca. MCF-7
0.3
Heart Pool
0.4
|
Breast ca. MDA-MB-
1.9
Lymph Node Pool
1.2
|
231
|
Breast ca. BT 549
0.6
Fetal Skeletal Muscle
0.5
|
Breast ca. T47D
0.9
Skeletal Muscle Pool
1.3
|
Breast ca. MDA-N
0.3
Spleen Pool
24.3
|
Breast Pool
1.1
Thymus Pool
1.0
|
Trachea
1.4
CNS cancer (glio/astro)
0.6
|
U87-MG
|
Lung
0.3
CNS cancer (glio/astro) U-
1.5
|
118-MG
|
Fetal Lung
2.4
CNS cancer (neuro; met)
0.5
|
SK-N-AS
|
Lung ca. NCI-N417
0.1
CNS cancer (astro) SF-539
0.2
|
Lung ca. LX-1
0.3
CNS cancer (astro) SNB-75
1.4
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.2
|
Lung ca. SHP-77
0.4
CNS cancer (glio) SF-295
80.7
|
Lung ca. A549
0.5
Brain (Amygdala) Pool
0.2
|
Lung ca. NCI-H526
0.1
Brain (cerebellum)
0.4
|
Lung ca. NCI-H23
0.9
Brain (fetal)
0.7
|
Lung ca. NCI-H460
0.6
Brain (Hippocampus) Pool
0.3
|
Lung ca. HOP-62
0.7
Cerebral Cortex Pool
0.3
|
Lung ca. NCI-H522
0.9
Brain (Substantia nigra)
0.1
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
0.5
|
Fetal Liver
0.3
Brain (whole)
0.2
|
Liver ca. HepG2
0.6
Spinal Cord Pool
0.3
|
Kidney Pool
1.4
Adrenal Gland
1.0
|
Fetal Kidney
1.2
Pituitary gland Pool
0.1
|
Renal ca. 786-0
0.7
Salivary Gland
0.1
|
Renal ca. A498
0.1
Thyroid (female)
0.1
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
1.8
|
Renal ca. UO-31
0.3
Pancreas Pool
1.3
|
|
[1508]
670
TABLE BOD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3922,
Rel. Exp. (%) Ag3922,
|
Tissue Name
Run 170552340
Tissue Name
Run 170552340
|
|
Secondary Th1 act
37.9
HUVEC IL-1 beta
8.2
|
Secondary Th2 act
49.7
HUVEC IFN gamma
14.9
|
Secondary Tr1 act
34.6
HUVEC TNF alpha + IFN
8.0
|
gamma
|
Secondary Th1 rest
14.6
HUVEC TNF alpha + IL4
9.2
|
Secondary Th2 rest
21.2
HUVEC IL-11
6.9
|
Secondary Tr1 rest
19.9
Lung Microvascular EC none
14.4
|
Primary Th1 act
33.0
Lung Microvascular EC
20.3
|
TNF alpha + IL-1 beta
|
Primary Th2 act
58.2
Microvascular Dermal EC none
9.5
|
Primary Tr1 act
35.4
Microsvasular Dermal EC
3.1
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
9.5
Bronchial epithelium TNF alpha +
11.3
|
IL1 beta
|
Primary Th2 rest
7.2
Small airway epithelium none
4.1
|
Primary Tr1 rest
31.0
Small airway epithelium
17.0
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
19.2
Coronery artery SMC rest
4.9
|
act
|
CD45RO CD4 lymphocyte
26.6
Coronery artery SMC TNF alpha +
1.8
|
act
IL-1 beta
|
CD8 lymphocyte act
29.1
Astrocytes rest
14.0
|
Secondary CD8
30.6
Astrocytes TNF alpha + IL-1 beta
6.8
|
lymphocyte rest
|
Secondary CD8
18.6
KU-812 (Basophil) rest
4.9
|
lymphocyte act
|
CD4 lymphocyte none
16.6
KU-812 (Basophil)
19.8
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
31.4
CCD1106 (Keratinocytes) none
17.4
|
CD95 CH11
|
LAK cells rest
11.7
CCD1106 (Keratinocytes)
26.8
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
33.4
Liver cirrhosis
2.8
|
LAK cells IL-2 + IL-12
14.9
NCI-H292 none
15.2
|
LAK cells IL-2 + IFN
14.0
NCI-H292 IL-4
14.4
|
gamma
|
LAK cells IL-2 + IL-18
19.1
NCI-H292 IL-9
16.3
|
LAK cells
15.2
NCI-H292 IL-13
15.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
23.3
NCI-H292 IFN gamma
18.3
|
Two Way MLR 3 day
28.3
HPAEC none
7.1
|
Two Way MLR 5 day
17.4
HPAEC TNF alpha + IL-1 beta
17.7
|
Two Way MLR 7 day
17.2
Lung fibroblast none
12.2
|
PBMC rest
8.0
Lung fibroblast TNF alpha + IL-
5.8
|
1 beta
|
PBMC PWM
24.0
Lung fibroblast IL-4
3.5
|
PBMC PHA-L
17.8
Lung fibroblast IL-9
15.5
|
Ramos (B cell) none
23.7
Lung fibroblast IL-13
7.4
|
Ramos (B cell) ionomycin
17.3
Lung fibroblast IFN gamma
15.4
|
B lymphocytes PWM
10.7
Dermal fibroblast CCD1070 rest
17.8
|
B lymphocytes CD40L
18.9
Dermal fibroblast CCD1070
37.4
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
15.4
Dermal fibroblast CCD1070 IL-
9.5
|
1 beta
|
EOL-1 dbcAMP
15.8
Dermal fibroblast IFN gamma
12.0
|
PMA/ionomycin
|
Dendritic cells none
14.8
Dermal fibroblast IL-4
16.3
|
Dendritic cells LPS
12.3
Dermal Fibroblasts rest
2.5
|
Dendritic cells anti-CD40
15.0
Neutrophils TNFa + LPS
8.5
|
Monocytes rest
33.9
Neutrophils rest
31.9
|
Monocytes LPS
40.1
Colon
7.0
|
Macrophages rest
11.5
Lung
24.5
|
Macrophages LPS
5.0
Thymus
58.6
|
HUVEC none
4.0
Kidney
100.0
|
HUVEC starved
13.6
|
|
[1509] CNS_neurodegeneration_v1.0 Summary: Ag3922 The CG94511-01 gene is found to be upregulated in the temporal cortex of Alzheimer's disease patients. Pyruvate dehydrogenase, which is a homolog of this gene product, plays a critical role in supplying the brain with glucose. The activity of pyruvate dehydrogenase may be affected in the Alzheimer's brain. Therefore, blockade of this enzyme may decrease neuronal death and be of use in the treatment of this disease. Impairment of the function of pyruvate dehydrogenase is associated with neurological deficits, neurodegeneration, and Parkinson's disease. Therefore, therapeutic modulation of this gene or gene product may therefore be of use in the treatment of these disorders (Bindoff L A, Birch-Machin M A, Farnsworth L, Gardner-Medwin D, Lindsay J G, Turnbull D M. Familial intermittent ataxia due to a defect of the El component of pyruvate dehydrogenase complex. J Neurol Sci 1989 November;93(2-3):311-8; Li H, Dryhurst G. Oxidative metabolites of 5-S-cysteinyldopamine inhibit the pyruvate dehydrogenase complex. J Neural Transm 2001;108(12):1363-74; Casley C S, Canevari L, Land J M, Clark J B, Sharpe M A. Beta-amyloid inhibits integrated mitochondrial respiration and key enzyme activities. J Neurochem 2002 January;80(1):91-100; Krugel U, Bigl V, Eschrich K, Bigl M. Deafferentation of the septo-hippocampal pathway in rats as a model of the metabolic events in Alzheimer's disease. Int J Dev Neurosci 2001 June;19(3):263-77).
[1510] General_screening13 panel_v1.4 Summary: Ag3922 The CG94511-01 gene is most highly expressed in a colon cancer cell line (CT=27). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel and as a marker to detect the presence of colon cancer. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of colon cancer.
[1511] Among tissues with metabolic function, this gene is expressed at moderate to low levels in adrenal gland, pancreas, fetal heart and adult and fetal skeletal muscle. This expression suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.
[1512] Panel 4.1D Summary: Ag3922 The CG94511-01 gene is most highly expressed in the kidney (CT=31.6). In addition, this gene appears to be slightly upregulated in activated T cells when compared to expression in resting T cells. Thus, therapeutic regulation of the transcript or the protein encoded by the transcript could be important in immune modulation and in the treatment of T cell-mediated diseases such as asthma, arthritis, psoriasis, IBD, and lupus. In addition, this gene is expressed in many samples involved in the immune response, including B cells, LAK cells, macrophages, monocytes, dendritic cells and keratinocytes. This widespread expression among cells involved in immune function suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.
[1513] BP. CG94551-01: MUNC13-3
[1514] Expression of gene CG94551-01 was assessed using the primer-probe set Ag3925, described in Table BPA. Results of the RTQ-PCR runs are shown in Tables BPB, BPC, BPD and BPE.
671TABLE BPA
|
|
Probe Name Ag3925
SEQ
StartID
PrimersSequencesLengthPositionNO:
|
Forward5′-tctggagaccggagtcattac-3′211957462
probeTET-5′-tctttacatgaggatctttctccatgg-3′-271993463
TAMRA
Reverse5′-agctccttgattccattcctt-3′212020464
|
[1515]
672
TABLE BPB
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp. (%) Ag3925, Run
Rel. Exp. (%) Ag3925, Run
|
Tissue Name
212344915
Tissue Name
212344915
|
|
AD 1 Hippo
3.5
Control (Path) 3
1.4
|
Temporal Ctx
|
AD 2 Hippo
4.2
Control (Path) 4
25.7
|
Temporal Ctx
|
AD 3 Hippo
0.8
AD 1 Occipital Ctx
5.1
|
AD 4 Hippo
1.4
AD 2 Occipital Ctx
0.0
|
(Missing)
|
AD 5 Hippo
100.0
AD 3 Occipital Ctx
2.6
|
AD 6 Hippo
12.5
AD 4 Occipital Ctx
10.7
|
Control 2 Hippo
13.0
AD 5 Occipital Ctx
22.4
|
Control 4 Hippo
3.1
AD 6 Occipital Ctx
24.1
|
Control (Path) 3 Hippo
1.2
Control 1 Occipital Ctx
1.2
|
AD 1 Temporal Ctx
4.7
Control 2 Occipital Ctx
50.3
|
AD 2 Temporal Ctx
12.1
Control 3 Occipital Ctx
6.9
|
AD 3 Temporal Ctx
1.3
Control 4 Occipital Ctx
1.9
|
AD 4 Temporal Ctx
8.1
Control (Path) 1
48.3
|
Occipital Ctx
|
AD 5 Inf Temporal Ctx
50.3
Control (Path) 2
9.3
|
Occipital Ctx
|
AD 5 Sup Temporal
16.3
Control (Path) 3
0.9
|
Ctx
Occipital Ctx
|
AD 6 Inf Temporal Ctx
23.2
Control (Path) 4
11.4
|
Occipital Ctx
|
AD 6 Sup Temporal
24.3
Control 1 Parietal Ctx
3.5
|
Ctx
|
Control 1 Temporal Ctx
2.3
Control 2 Parietal Ctx
11.7
|
Control 2 Temporal Ctx
15.5
Control 3 Parietal Ctx
8.1
|
Control 3 Temporal Ctx
8.1
Control (Path) 1
42.9
|
Parietal Ctx
|
Control 3 Temporal Ctx
2.2
Control (Path) 2
18.6
|
Parietal Ctx
|
Control (Path) 1
31.6
Control (Path) 3
1.8
|
Temporal Ctx
Parietal Ctx
|
Control (Path) 2
22.4
Control (Path) 4
45.4
|
Temporal Ctx
Parietal Ctx
|
|
[1516]
673
TABLE BPC
|
|
|
General_screening_panel_v1.4
|
Rel. Exp. (%) Ag3925, Run
Rel. Exp. (%) Ag3925, Run
|
Tissue Name
219515275
Tissue Name
219515275
|
|
Adipose
0.6
Renal ca. TK-10
0.0
|
Melanoma* Hs688(A).T
0.1
Bladder
0.2
|
Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.0
|
NCI-N87
|
Melanoma* M14
0.0
Gastric ca. KATO III
0.0
|
Melanoma* LOXIMVI
1.3
Colon ca. SW-948
0.0
|
Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.0
|
Squamous cell
0.0
Colon ca.* (SW480 met)
0.0
|
carcinoma SCC-4
SW620
|
Testis Pool
0.7
Colon ca. HT29
0.1
|
Prostate ca.* (bone met)
0.0
Colon ca. HCT-116
0.0
|
PC-3
|
Prostate Pool
0.4
Colon ca. CaCo-2
14.3
|
Placenta
0.0
Colon cancer tissue
0.0
|
Uterus Pool
0.2
Colon ca. SW1116
0.0
|
Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0
|
Ovarian ca. SK-OV-3
0.2
Colon ca. SW-48
0.0
|
Ovarian ca. OVCAR-4
0.0
Colon Pool
0.0
|
Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
3.0
|
Ovarian ca. IGROV-1
0.1
Stomach Pool
0.8
|
Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
0.1
|
Ovary
5.5
Fetal Heart
0.4
|
Breast ca. MCF-7
0.0
Heart Pool
0.2
|
Breast ca. MDA-MB-
0.0
Lymph Node Pool
0.2
|
231
|
Breast ca. BT 549
0.0
Fetal Skeletal Muscle
0.7
|
Breast ca. T47D
0.0
Skeletal Muscle Pool
18.4
|
Breast ca. MDA-N
38.4
Spleen Pool
0.1
|
Breast Pool
0.0
Thymus Pool
0.4
|
Trachea
1.4
CNS cancer (glio/astro)
0.0
|
U87-MG
|
Lung
2.3
CNS cancer (glio/astro) U-
0.0
|
118-MG
|
Fetal Lung
47.0
CNS cancer (neuro; met)
0.0
|
SK-N-AS
|
Lung ca. NCI-H417
0.0
CNS cancer (astro) SF-539
1.2
|
Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
88.9
|
Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.3
|
Lung ca. SHP-77
0.3
CNS cancer (glio) SF-295
0.0
|
Lung ca. A549
1.4
Brain (Amygdala) Pool
19.8
|
Lung ca. NCI-H526
0.0
Brain (cerebellum)
100.0
|
Lung ca. NCI-H23
0.0
Brain (fetal)
15.6
|
Lung ca. NCI-H460
0.0
Brain (Hippocampus) Pool
25.5
|
Lung ca. HOP-62
0.0
Cerebral Cortex Pool
42.3
|
Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
20.0
|
Pool
|
Liver
0.0
Brain (Thalamus) Pool
35.6
|
Fetal Liver
0.4
Brain (whole)
18.3
|
Liver ca. HepG2
0.1
Spinal Cord Pool
7.3
|
Kidney Pool
1.4
Adrenal Gland
0.0
|
Fetal Kidney
47.6
Pituitary gland Pool
6.4
|
Renal ca. 786-0
0.0
Salivary Gland
0.0
|
Renal ca. A498
0.0
Thyroid (female)
5.0
|
Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.0
|
Renal ca. UO-31
71.2
Pancreas Pool
0.1
|
|
[1517]
674
TABLE BPD
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3925,
Rel. Exp. (%) Ag3925,
|
Tissue Name
Run 170594009
Tissue Name
Run 170594009
|
|
Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0
|
Secondary Th2 act
0.0
HUVEC IFN gamma
0.0
|
Secondary Tr1 act
0.6
HUVEC TNF alpha + IFN
0.0
|
gamma
|
Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0
|
Secondary Th2 rest
0.0
HUVEC IL-11
0.0
|
Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0
|
Primary Th1 act
0.0
Lung Microvascular EC
0.0
|
TNF alpha + IL-1 beta
|
Primary Th2 act
0.0
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
2.9
|
IL1 beta
|
Primary Th2 rest
0.0
Small airway epithelium none
0.0
|
Primary Tr1 rest
0.0
Small airway epithelium
0.3
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0
|
act
|
CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0
|
act
IL-1 beta
|
CD8 lymphocyte act
0.0
Astrocytes rest
0.4
|
Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0
|
lymphocyte rest
|
Secondary CD8
0.0
KU-812 (Basophil) rest
0.0
|
lymphocyte act
|
CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0
|
CD95 CH11
|
LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
0.0
Liver cirrhosis
0.3
|
LAK cells IL-2 + IL-12
0.0
NCI-H292 none
2.5
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0
|
gamma
|
LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0
|
LAK cells
0.0
NCI-H292 IL-13
0.0
|
PMA/ionomycin
|
NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.5
|
Two Way MLR 3 day
0.0
HPAEC none
0.0
|
Two Way MLR 5 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0
|
Two Way MLR 7 day
0.0
Lung fibroblast none
0.0
|
PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0
|
1 beta
|
PBMC PWM
0.0
Lung fibroblast IL-4
0.7
|
PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
2.9
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
4.1
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
2.2
|
B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
4.6
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.5
|
1 beta
|
EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.6
|
PMA/ionomycin
|
Dendritic cells none
0.0
Dermal fibroblast IL-4
0.6
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
4.1
|
Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
0.8
|
Monocytes rest
0.0
Neutrophils rest
0.0
|
Monocytes LPS
0.0
Colon
9.3
|
Macrophages rest
0.0
Lung
10.8
|
Macrophages LPS
0.0
Thymus
39.5
|
HUVEC none
0.0
Kidney
100.0
|
HUVEC starved
0.0
|
|
[1518]
675
TABLE BPE
|
|
|
Panel CNS_1
|
Rel. Exp. (%) Ag3925, Run
Rel. Exp. (%) Ag3925, Run
|
Tissue Name
180912024
Tissue Name
180912024
|
|
BA4 Control
37.6
BA17 PSP
41.5
|
BA4 Control2
47.3
BA17 PSP2
7.9
|
BA4 Alzheimer's2
3.2
Sub Nigra Control
18.6
|
BA4 Parkinson's
34.6
Sub Nigra Control2
49.0
|
BA4 Parkinson's2
100.0
Sub Nigra Alzheimer's2
8.2
|
BA4 Huntington's
42.3
Sub Nigra Parkinson's2
24.1
|
BA4
3.3
Sub Nigra Huntington's
39.8
|
Huntington's2
|
BA4 PSP
6.8
Sub Nigra
43.8
|
Huntington's2
|
BA4 PSP2
24.8
Sub Nigra PSP2
5.6
|
BA4 Depression
13.2
Sub Nigra Depression
1.8
|
BA4 Depression2
6.2
Sub Nigra Depression2
7.4
|
BA7 Control
29.9
Glob Palladus Control
0.4
|
BA7 Control2
39.5
Glob Palladus Control2
2.2
|
BA7 Alzheimer's2
3.8
Glob Palladus
7.7
|
Alzheimer's
|
BA7 Parkinson's
12.0
Glob Palladus
0.7
|
Alzheimer's2
|
BA7 Parkinson's2
47.3
Glob Palladus
28.5
|
Parkinson's
|
BA7 Huntington's
42.9
Glob Palladus
1.8
|
Parkinson's2
|
BA7
33.7
Glob Palladus PSP
1.5
|
Huntington's2
|
BA7 PSP
75.8
Glob Palladus PSP2
2.8
|
BA7 PSP2
22.4
Glob Palladus
0.5
|
Depression
|
BA7 Depression
15.5
Temp Pole Control
19.9
|
BA9 Control
17.0
Temp Pole Control2
61.1
|
BA9 Control2
65.5
Temp Pole Alzheimer's
2.6
|
BA9 Alzheimer's
4.7
Temp Pole Alzheimer's2
2.6
|
BA9 Alzheimer's2
10.2
Temp Pole Parkinson's
34.2
|
BA9 Parkinson's
20.0
Temp Pole Parkinson's2
26.4
|
BA9 Parkinson's2
58.2
Temp Pole Huntington's
27.7
|
BA9 Huntington's
34.4
Temp Pole PSP
9.1
|
BA9
9.2
Temp Pole PSP2
2.3
|
Huntington's2
|
BA9 PSP
18.2
Temp Pole Depression2
4.3
|
BA9 PSP2
2.9
Cing Gyr Control
43.5
|
BA9 Depression
5.1
Cing Gyr Control2
28.5
|
BA9 Depression2
8.5
Cing Gyr Alzheimer's
9.5
|
BA17 Control
71.2
Cing Gyr Alzheimer's2
2.7
|
BA17 Control2
64.6
Cing Gyr Parkinson's
15.6
|
BA17
6.3
Cing Gyr Parkinson's2
25.5
|
Alzheimer's2
|
BA17 Parkinson's
29.9
Cing Gyr Huntington's
43.5
|
BA17
56.6
Cing Gyr Huntington's2
6.6
|
Parkinson's2
|
BA17
37.9
Cing Gyr PSP
13.9
|
Huntington's
|
BA17
12.9
Cing Gyr PSP2
1.6
|
Huntington's2
|
BA17 Depression
8.0
Cing Gyr Depression
4.5
|
BA17 Depression2
10.7
Cing Gyr Depression2
6.1
|
|
[1519] CNS_neurodegeneration_v1.0 Summary: Ag3925 This panel does not show differential expression of the CG9455 1-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1520] General_screening_panel_v1.4 Summary: Ag3925 The CG94551-01 gene appears to be prefrentially expressed in the brain, with highest expression in the cerebellum (CT=27). Moderate levels of expression are also seen in amygdala, hippocampus, cerebral cortex, substantia nigra, and thalamus. This gene encodes a homolog of MUNC 13, a protein involved in neurotransmitter release at the synaptic junction. Therefore, therapeutic modulation of the expression or function of this protein may be useful in the treatment of disease where reduction in neurotransmission has been shown to ameliorate symptomology (e.g., epilepsy or other seizure disorders, schizophrenia, bipolar disorder or anxiety).
[1521] Moderate levels of expression are also seen in cell lines derived from brain, colon, renal and breast cancers. Therefore, expression of this gene may be used as a diagnostic marker for the presence of these cancers.
[1522] Among metabolic tissues, moderate to low levels of expression are seen in thyroid, pituitary, fetal liver, adipose and fetal and adult skeletal muscle. This widespread expression among metabolic tissues suggests that this gene product may be involved in the pathogenesis and/or treatment of metabolic disorders, including obesity and diabetes.
[1523] In addition, this gene is expressed at much higher levels in fetal lung, kidney and skeletal muscle tissue (CTs=28-29) when compared to expression in the adult counterpart (CTs=32-34). Thus, expression of this gene may be used to differentiate between the fetal and adult source of these tissues (Richmond J E, Weimer R M, Jorgensen E M. An open form of syntaxin bypasses the requirement for UNC-13 in vesicle priming. Nature Jul. 19, 2001;412(6844):338-41).
[1524] Panel 4.1D Summary: Ag3925 Expression of the CG94551-01 gene, a MUNC 13 homolog, is highest in the kidney (CT=31). MUNC 13 has been shown to act as a diacylglycerol receptor that induces apoptosis and may contribute to diabetic nephropathy.
[1525] Moderate levels of expression are also seen in normal lung, thymus, and colon. This expression profile suggests that this gene product may be involved in the normal homeostasis of these organs. Therefore, therapeutic modulation of the expression or function of this gene may be useful in maintaining or restoring function to these organs during inflammation (Song Y, Ailenberg M, Silverman M. Human munc 13 is a diacylglycerol receptor that induces apoptosis and may contribute to renal cell injury in hyperglycemia. Mol Biol Cell 1999 May; 10(5): 1609-19).
[1526] Panel CNS—1 Summary: Ag3925 This panel confirms the presence of the CG94551-01 gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.
[1527] BQ. CG94682-O2: RENAL ORGANIC ANION TRANSPORTER 1
[1528] Expression of gene CG94682-02 was assessed using the primer-probe set Ag3948, described in Table BQA. Results of the RTQ-PCR runs are shown in Tables BQB, BQC, BQD and BQE.
676TABLE BQA
|
|
Probe Name Ag3948
StartSEQ
PrimersSequencesLengthPositionID NO:
|
Forward5′-ctctattcttggtggcttcca-3′21808465
ProbeTET-5′-ctcctgcatggcaagtcccagttag-25847466
3′-TAMRA
Reverse5′-ccaccttctgcagattctgtac-3′22874467
|
[1529]
677
TABLE BQB
|
|
|
AI_comprehensive panel_v1.0
|
Rel. Exp.(%)
Rel. Exp.(%)
|
Tissue
Ag3948,
Tissue
Ag3948,
|
Name
Run 247842315
Name
Run 247842315
|
|
110967
17.8
112427
12.1
|
COPD-F
Match Control
|
Psoriasis-F
|
110980
6.7
112418
11.2
|
COPD-F
Psoriasis-M
|
110968
9.2
112723
9.6
|
COPD-M
Match Control
|
Psoriasis-M
|
110977
20.0
112419
30.6
|
COPD-M
Psoriasis-M
|
110989
34.6
112424
18.4
|
Emphysema-F
Match Control
|
Psoriasis-M
|
110992
0.0
112420
94.6
|
Emphysema-F
Psoriasis-M
|
110993
18.6
112425
42.6
|
Emphysema-F
Match Control
|
Psoriasis-M
|
110994
9.2
104689
20.2
|
Emphysema-F
(MF)
|
OA Bone-
|
Backus
|
110995
100.0
104690
7.2
|
Emphysema-F
(MF) Adj
|
“Normal”
|
Bone-Backus
|
110996
5.2
104691
10.0
|
Emphysema-F
(MF) OA
|
Synovium-
|
Backus
|
110997
18.0
104692
0.0
|
Asthma-M
(BA) OA
|
Cartilage-
|
Backus
|
111001
0.0
104694
14.2
|
Asthma-F
(BA) OA
|
Bone-
|
Backus
|
111002
8.8
104695
7.5
|
Asthma-F
(BA) Adj
|
“Normal”
|
Bone-Backus
|
111003
5.6
104696
13.9
|
Atopic
(BA) OA
|
Asthma-F
Synovium-
|
Backus
|
111004
41.5
104700
35.6
|
Atopic
(SS) OA Bone-
|
Asthma-F
Backus
|
111005
17.6
104701
8.7
|
Atopic
(SS) Adj
|
Asthma-F
“Normal”
|
Bone-Backus
|
111006
0.0
104702
26.8
|
Atopic
(SS) OA
|
Asthma-F
Synovium-
|
Backus
|
111417
2.4
117093
18.9
|
Allergy-M
OA Cartilage
|
Rep7
|
112347
0.0
112672
13.4
|
Allergy-M
OA Bone5
|
112349
10.1
112673
0.0
|
Normal
OA Synovium5
|
Lung-F
|
112357
9.2
112674
5.8
|
Normal
OA Synovial
|
Lung-F
Fluid
|
cells5
|
112354
19.2
117100
14.8
|
Normal
OA Cartilage
|
Lung-M
Rep14
|
112374
28.3
112756
21.0
|
Crohns-F
OA Bone9
|
112389
15.2
112757
20.3
|
Match
OA Synovium9
|
Control
|
Crohns-F
|
112375
14.3
112758
7.2
|
Crohns-F
OA Synovial
|
Fluid
|
Cells9
|
112732
92.7
117125
25.9
|
Match
RA Cartilage
|
Control
Rep2
|
Crohns-F
|
112725
23.5
113492
10.9
|
Crohns-M
Bone2 RA
|
112387
13.6
113493
8.8
|
Match
Synovium2 RA
|
Control
|
Crohns-M
|
112378
14.4
113494
16.8
|
Crohns-M
Syn Fluid
|
Cells RA
|
112390
72.7
113499
12.9
|
Match
Cartilage4 RA
|
Control
|
Crohns-M
|
112726
12.8
113500
20.7
|
Crohns-M
Bone4 RA
|
112731
65.5
113501
13.8
|
Match
Synovium4 RA
|
Control
|
Crohns-M
|
112380
18.6
113502
3.1
|
Ulcer
Syn Fluid
|
Col-F
Cells4
|
RA
|
112734
89.5
113495
6.8
|
Match
Cartilage3 RA
|
Control
|
Ulcer
|
Col-F
|
112384
57.8
113496
20.7
|
Ulcer
Bone3 RA
|
Col-F
|
112737
15.6
113497
8.7
|
Match
Synovium3 RA
|
Control
|
Ulcer
|
Col-F
|
112386
2.7
113498
21.6
|
Ulcer
Syn Fluid
|
Col-F
Cells3 RA
|
112738
4.3
117106
5.7
|
Match
Normal
|
Control
Cartilage
|
Ulcer
Rep20
|
Col-F
|
112381
0.0
113663
16.2
|
Ulcer
Bone3 Normal
|
Col-M
|
112735
58.2
113664
2.6
|
Match
Synovium3
|
Control
Normal
|
Ulcer
|
Col-M
|
112382
8.4
113665
5.2
|
Ulcer
Syn Fluid
|
Col-M
Cells3
|
Normal
|
112394
17.1
117107
11.0
|
Match
Normal
|
Control
Cartilage
|
Ulcer
Rep22
|
Col-M
|
112383
47.3
113667
15.4
|
Ulcer
Bone4 Normal
|
Col-M
|
112736
1.9
113668
20.2
|
Match
Synovium4
|
Control
Normal
|
Ulcer
|
Col-M
|
112423
27.4
113669
40.9
|
Psoriasis-F
Syn Fluid
|
Cells4
|
Normal
|
|
[1530]
678
TABLE BQC
|
|
|
CNS_neurodegeneration_v1.0
|
Rel. Exp.(%)
Rel. Exp.(%)
|
Tissue
Ag3948,
Tissue
Ag3948,
|
Name
Run 212345604
Name
Run 212345604
|
|
AD 1 Hippo
0.0
Control
0.0
|
(Path) 3
|
Temporal Ctx
|
AD 2 Hippo
57.0
Control
33.4
|
(Path) 4
|
Temporal Ctx
|
AD 3 Hippo
32.5
AD 1
67.8
|
Occipital Ctx
|
AD 4 Hippo
0.0
AD 2 Occipital
5.3
|
Ctx (Missing)
|
AD 5 Hippo
47.0
AD 3 Occipital
0.0
|
Ctx
|
AD 6 Hippo
20.3
AD 4 Occipital
0.0
|
Ctx
|
Control
0.0
AD 5 Occipital
18.4
|
2 Hippo
Ctx
|
Control
0.0
AD 6 Occipital
39.2
|
4 Hippo
Ctx
|
Control
0.0
Control 1
0.0
|
(Path) 3
Occipital Ctx
|
Hippo
|
AD 1
0.0
Control 2
79.6
|
Temporal Ctx
Occipital Ctx
|
AD 2
16.7
Control 3
42.0
|
Temporal Ctx
Occipital Ctx
|
AD 3
14.9
Control 4
0.0
|
Temporal Ctx
Occipital Ctx
|
AD 4
0.0
Control
46.0
|
Temporal Ctx
(Path) 1
|
Occipital Ctx
|
AD 5 Inf
100.0
Control
0.0
|
Temporal Ctx
(Path) 2
|
Occipital Ctx
|
AD 5 Sup
0.0
Control
0.0
|
Temporal Ctx
(Path) 3
|
Occipital Ctx
|
AD 6 Inf
65.1
Control
11.7
|
Temporal Ctx
(Path) 4
|
Occipital Ctx
|
AD 6 Sup
91.4
Control 1
10.3
|
Temporal Ctx
Parietal Ctx
|
Control 1
0.0
Control 2
49.3
|
Temporal Ctx
Parietal Ctx
|
Control 2
0.0
Control 3
0.0
|
Temporal Ctx
Parietal Ctx
|
Control 3
0.0
Control
67.8
|
Temporal Ctx
(Path) 1
|
Parietal Ctx
|
Control 3
18.0
Control
14.4
|
Temporal Ctx
(Path) 2
|
Parietal Ctx
|
Control
32.1
Control
0.0
|
(Path) 1
(Path) 3
|
Temporal Ctx
Parietal Ctx
|
Control
9.7
Control
20.4
|
(Path) 2
(Path) 4
|
Temporal Ctx
Parietal Ctx
|
|
[1531]
679
TABLE BQD
|
|
|
General_screening_panel_v1.4
|
Rel. Exp.(%)
Rel. Exp.(%)
|
Tissue
Ag3948,
Tissue
Ag3948,
|
Name
Run 219279808
Name
Run 219279808
|
|
Adipose
0.3
Renal ca.
2.1
|
TK-10
|
Melanoma*
0.5
Bladder
2.8
|
Hs688 (A).T
|
Melanoma*
0.2
Gastric ca.
22.2
|
Hs688 (B).T
(liver met.)
|
NCI-N87
|
Melanoma*
0.2
Gastric ca.
4.0
|
M14
KATO III
|
Melanoma*
0.0
Colon ca.
0.1
|
LOXIMVI
SW-948
|
Melanoma*
0.1
Colon ca.
10.5
|
SK-MEL-5
SW480
|
Squamous cell
2.4
Colon ca.*
1.6
|
carcinoma
(SW480 met)
|
SCC-4
SW620
|
Testis Pool
0.3
Colon ca.
1.5
|
HT29
|
Prostate ca.*
0.9
Colon ca.
2.3
|
(bone met)
HCT-116
|
PC-3
|
Prostate Pool
0.3
Colon ca.
0.7
|
CaCo-2
|
Placenta
0.2
Colon cancer
0.2
|
tissue
|
Uterus Pool
0.1
Colon ca.
0.1
|
SW1116
|
Ovarian ca.
11.5
Colon ca.
0.6
|
OVCAR-3
Colo-205
|
Ovarian ca.
1.3
Colon ca.
0.8
|
SK-OV-3
SW-48
|
Ovarian ca.
0.4
Colon Pool
0.6
|
OVCAR-4
|
Ovarian ca.
8.0
Small Intestine
0.4
|
OVCAR-5
Pool
|
Ovarian ca.
1.9
Stomach Pool
0.1
|
IGROV-1
|
Ovarian ca.
0.5
Bone Marrow
0.4
|
OVCAR-8
Pool
|
Ovary
0.4
Fetal Heart
0.1
|
Breast ca.
3.2
Heart Pool
0.3
|
MCF-7
|
Breast ca.
2.3
Lymph Node
0.7
|
MDA-MB-231
Pool
|
Breast ca.
0.2
Fetal Skeletal
0.6
|
BT 549
Muscle
|
Breast ca.
9.6
Skeletal Muscle
0.4
|
T47D
Pool
|
Breast ca.
0.2
Spleen Pool
0.2
|
MDA-N
|
Breast Pool
0.7
Thymus Pool
0.3
|
Trachea
0.6
CNS cancer
0.5
|
(glio/astro)
|
U87-MG
|
Lung
0.2
CNS cancer
0.1
|
(glio/astro)
|
U-118-MG
|
Fetal Lung
1.7
CNS cancer
0.4
|
(neuro; met)
|
SK-N-AS
|
Lung ca.
0.1
CNS cancer
0.1
|
NCI-N417
(astro) SF-539
|
Lung ca.
100.0
CNS cancer
0.1
|
LX-1
(astro) SNB-75
|
Lung ca.
1.0
CNS cancer
0.9
|
NCI-H146
(glio) SNB-19
|
Lung ca.
0.1
CNS cancer
9.3
|
SHP-77
(glio) SF-295
|
Lung ca.
2.2
Brain (Amygdala)
0.1
|
A549
Pool
|
Lung ca.
0.1
Brain
5.1
|
NCI-H526
(cerebellum)
|
Lung ca.
0.9
Brain (fetal)
2.3
|
NCI-H23
|
Lung ca.
0.2
Brain
0.0
|
NCI-H460
(Hippocampus)
|
Pool
|
Lung ca.
0.5
Cerebral Cortex
0.0
|
HOP-62
Pool
|
Lung ca.
1.1
Brain (Substantia
0.2
|
NCI-H522
nigra) Pool
|
Liver
0.3
Brain (Thalamus)
0.0
|
Pool
|
Fetal Liver
0.8
Brain (whole)
0.8
|
Liver ca.
0.2
Spinal Cord
0.0
|
HepG2
Pool
|
Kidney Pool
0.7
Adrenal Gland
0.2
|
Fetal Kidney
0.7
Pituitary gland
0.0
|
Pool
|
Renal ca.
0.2
Salivary Gland
0.1
|
786-0
|
Renal ca.
0.2
Thyroid (female)
0.7
|
A498
|
Renal ca.
0.2
Pancreatic ca.
3.8
|
ACHN
CAPAN2
|
Renal ca.
1.6
Pancreas Pool
0.6
|
UO-31
|
|
[1532]
680
TABLE BQE
|
|
|
Panel 4.1D
|
Rel. Exp. (%) Ag3948,
Rel. Exp. (%) Ag3948,
|
Tissue Name
Run 170684837
Tissue Name
Run 170684837
|
|
Secondary Th1 act
1.6
HUVEC IL-1 beta
3.1
|
Secondary Th2 act
8.6
HUVEC IFN gamma
7.4
|
Secondary Tr1 act
4.4
HUVEC TNF alpha + IFN
2.7
|
gamma
|
Secondary Th1 rest
0.3
HUVEC TNF alpha + IL4
0.9
|
Secondary Th2 rest
3.9
HUVEC IL-11
6.0
|
Secondary Tr1 rest
1.6
Lung Microvascular EC none
7.3
|
Primary Th1 act
0.3
Lung Microvascular EC
2.7
|
TNF alpha + IL-1 beta
|
Primary Th2 act
1.6
Microvascular Dermal EC none
0.0
|
Primary Tr1 act
0.0
Microsvasular Dermal EC
0.5
|
TNF alpha + IL-1 beta
|
Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
15.5
|
IL1 beta
|
Primary Th2 rest
1.6
Small airway epithelium none
6.3
|
Primary Tr1 rest
0.4
Small airway epithelium
20.4
|
TNF alpha + IL-1 beta
|
CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
6.0
|
act
|
CD45RO CD4 lymphocyte
1.2
Coronery artery SMC TNF alpha +
1.6
|
act
IL-1 beta
|
CD8 lymphocyte act
2.8
Astrocytes rest
0.3
|
Secondary CD8
2.0
Astrocytes TNF alpha + IL-1 beta
1.5
|
lymphocyte rest
|
Secondary CD8
1.2
KU-812 (Basophil) rest
3.4
|
lymphocyte act
|
CD4 lymphocyte none
1.6
KU-812 (Basophil)
6.3
|
PMA/ionomycin
|
2ry Th1/Th2/Tr1_anti-
4.6
CCD1106 (Keratinocytes) none
14.6
|
CD95 CH11
|
LAK cells rest
1.6
CCD1106 (Keratinocytes)
20.6
|
TNF alpha + IL-1 beta
|
LAK cells IL-2
1.7
Liver cirrhosis
0.0
|
LAK cells IL-2 + IL-12
3.6
NCI-H292 none
49.7
|
LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
79.6
|
gamma
|
LAK cells IL-2 + IL-18
1.7
NCI-H292 IL-9
92.7
|
LAK cells
0.0
NCI-H292 IL-13
58.2
|
PMA/ionomycin
|
NK Cells IL-2 rest
4.9
NCI-H292 IFN gamma
50.0
|
Two Way MLR 3 day
3.1
HPAEC none
8.6
|
Two Way MLR 5 day
1.4
HPAEC TNF alpha + IL-1 beta
3.5
|
Two Way MLR 7 day
4.1
Lung fibroblast none
2.7
|
PBMC rest
2.5
Lung fibroblast TNF alpha + IL-
2.9
|
1 beta
|
PBMC PWM
2.3
Lung fibroblast IL-4
2.9
|
PBMC PHA-L
0.4
Lung fibroblast IL-9
4.0
|
Ramos (B cell) none
0.0
Lung fibroblast IL-13
5.5
|
Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
5.8
|
B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
16.2
|
B lymphocytes CD40L
7.6
Dermal fibroblast CCD1070
4.7
|
and IL-4
TNF alpha
|
EOL-1 dbcAMP
1.9
Dermal fibroblast CCD1070 IL-1
1.8
|
beta
|
EOL-1 dbcAMP
5.5
Dermal fibroblast IFN gamma
6.7
|
PMA/ionomycin
|
Dendritic cells none
1.6
Dermal fibroblast IL-4
4.1
|
Dendritic cells LPS
0.0
Dermal Fibroblasts rest
12.2
|
Dendritic cells anti-CD40
0.7
Neutrophils TNFa + LPS
0.7
|
Monocytes rest
2.8
Neutrophils rest
1.5
|
Monocytes LPS
9.3
Colon
5.5
|
Macrophages rest
0.0
Lung
11.6
|
Macrophages LPS
0.8
Thymus
40.6
|
HUVEC none
0.3
Kidney
100.0
|
HUVEC starved
9.9
|
|
[1533] AI_comprehensive panel_v1.0 Summary: Ag3948 This panel confirms expression of the CG94682-02 gene in tissue samples related to the immune and inflammatory response. Please see Panels 4 and 4.1D for discussion of utility of this gene in inflammation.
[1534] CNS_neurodegeneration_v1.0 Summary: Ag3948 This panel does not show differential expression of the CG94682-02 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel General_screening_panel_v1.4 for discussion of utility of this gene in the central nervous system.
[1535] General_screening_panel_v1.4 Summary: Ag3948 The expression of the CG94682-02 gene, an organic anion transporter homolog, is highest in a small cell lung cancer line LX-1 (CT=28.2). This gene is also expressed in some ovarian, breast, CNS, gastric, pancreatic, renal and colon cancer cell lines. Therefore, expression of this gene may be associated with these forms of cancer and therapeutic modulation of this gene might be of use in the treatment or diagnosis of these cancers.
[1536] This gene is also expressed at low levels in the cerebellum and fetal brain. The organic anion transporters are involved in transport across the blood brain barrier. This gene may therefore be of use in drug delivery to the CNS, specifically for compounds such as nerve growth factors protein therapeutics which are believed to have numerous uses in the CNS, but lack a delivery system (Sugiyama D, Kusuhara H, Shitara Y, Abe T, Meier P J, Sekine T, Endou H, Suzuki H, Sugiyama Y. Characterization of the efflux transport of 17beta-estradiol-D-17beta-glucuronide from the brain across the blood-brain barrier. J Pharmacol Exp Ther 2001 July;298(1):316-22).
[1537] Ag3532 Results from one experiment with this gene are not included. The amp plot indicates that there were instrumental difficulties with this run.
[1538] Panel 4.1D Summary: Ag3498 The highest expression of the CG94682-02 gene is found in the kidney and in the pulmonary muco-epidermoid cell line NCI-H292 (CTs=31). The expression of this gene, although constitutive in the H292 cell line, is up regulated upon treatment with IL-4, Il-9 and IL-13, cytokines that have been linked to the pathogenesis of asthma and/or COPD.
[1539] This gene is also found in small airway epithelium and keratinocytes treated with the inflammatory cytokines TNF-a and IL-1b. Therefore, modulation of the expression or activity of the protein encoded by this transcript through the application of small molecule therapeutics may be useful in the treatment of asthma, COPD, emphysema, psoriasis and wound healing.
Identification of Single Nucleotide Polymorphisms in NOVX Nucleic Acid Sequences
[1540] Variant sequences are also included in this application. A variant sequence can include a single nucleotide polymorphism (SNP). A SNP can, in some instances, be referred to as a “cSNP” to denote that the nucleotide sequence containing the SNP originates as a cDNA. A SNP can arise in several ways. For example, a SNP may be due to a substitution of one nucleotide for another at the polymorphic site. Such a substitution can be either a transition or a transversion. A SNP can also arise from a deletion of a nucleotide or an insertion of a nucleotide, relative to a reference allele. In this case, the polymorphic site is a site at which one allele bears a gap with respect to a particular nucleotide in another allele. SNPs occurring within genes may result in an alteration of the amino acid encoded by the gene at the position of the SNP. Intragenic SNPs may also be silent, when a codon including a SNP encodes the same amino acid as a result of the redundancy of the genetic code. SNPs occurring outside the region of a gene, or in an intron within a gene, do not result in changes in any amino acid sequence of a protein but may result in altered regulation of the expression pattern. Examples include alteration in temporal expression, physiological response regulation, cell type expression regulation, intensity of expression, and stability of transcribed message.
[1541] SeqCalling assemblies produced by the exon linking process were selected and extended using the following criteria. Genomic clones having regions with 98% identity to all or part of the initial or extended sequence were identified by BLASTN searches using the relevant sequence to query human genomic databases. The genomic clones that resulted were selected for further analysis because this identity indicates that these clones contain the genomic locus for these SeqCalling assemblies. These sequences were analyzed for putative coding regions as well as for similarity to the known DNA and protein sequences. Programs used for these analyses include Grail, Genscan, BLAST, HMMER, FASTA, Hybrid and other relevant programs.
[1542] Some additional genomic regions may have also been identified because selected SeqCalling assemblies map to those regions. Such SeqCalling sequences may have overlapped with regions defined by homology or exon prediction. They may also be included because the location of the fragment was in the vicinity of genomic regions identified by similarity or exon prediction that had been included in the original predicted sequence. The sequence so identified was manually assembled and then may have been extended using one or more additional sequences taken from CuraGen Corporation's human SeqCalling database. SeqCalling fragments suitable for inclusion were identified by the CuraTools™ program SeqExtend or by identifying SeqCalling fragments mapping to the appropriate regions of the genomic clones analyzed.
[1543] The regions defined by the procedures described above were then manually integrated and corrected for apparent inconsistencies that may have arisen, for example, from miscalled bases in the original fragments or from discrepancies between predicted exon junctions, EST locations and regions of sequence similarity, to derive the final sequence disclosed herein. When necessary, the process to identify and analyze SeqCalling assemblies and genomic clones was reiterated to derive the full length sequence (Alderbom et al., Determination of Single Nucleotide Polymorphisms by Real-time Pyrophosphate DNA Sequencing. Genome Research. 10 (8) 1249-1265, 2000).
[1544] Variants are reported individually but any combination of all or a select subset of variants are also included as contemplated NOVX embodiments of the invention.
[1545] NOV2a SNP Data:
[1546] NOV2a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:3 and 4, respectively. The nucleotide sequence of the NOV5a variant differs as shown in Table SNP1.
681TABLE SNP1
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377738806CT268GlyGly
|
[1547] NOV3a SNP Data:
[1548] NOV3a has six SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:7 and 8, respectively. The nucleotide sequence of the NOV3a variant differs as shown in Table SNP2.
682TABLE SNP2
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
133756991355TC388PheLeu
133756961685TC498CysArg
133756951698CT502AlaVal
133756941723GA510GlnGln
133756931727CT512ProSer
133756921769CT0
|
[1549] NOV4a SNP Data:
[1550] NOV4a has five SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:9 and 10, respectively. The nucleotide sequence of the NOV4a variant differs as shown in Table SNP3.
683TABLE SNP3
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13374823769GA242GlyGlu
13376034827TC261AlaAla
13374824835AG264HisArg
133748251089AG349LysGlu
133760331095TC351SerPro
|
[1551] NOV5a SNP Data:
[1552] NOV5a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs: 15 and 16, respectively. The nucleotide sequence of the NOV5a variant differs as shown in Table SNP4.
684TABLE SNP4
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377739596GA175ArgGln
|
[1553] NOV15a SNP Data:
[1554] NOV15a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:41 and 42, respectively. The nucleotide sequence of the NOV15a variant differs as shown in Table SNP5.
685TABLE SNP5
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377740813AG262ThrThr
133777411030TC0
|
[1555] NOV19a SNP Data:
[1556] NOV19a has eleven SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:49 and 50, respectively. The nucleotide sequence of the NOV19a variant differs as shown in Table SNP6.
686TABLE SNP6
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13375979157CT19GlyGly
13375978210TC37LeuPro
13375977219AC40LysThr
13375976269AG57ThrAla
13375975324AG75GluGly
13375974374GA92AspAsn
13375973454AG118ArgArg
13375972475TC125CysCys
13375971781TC227TyrTyr
133759701589GA497AlaThr
133777421818CT0
|
[1557] NOV20a SNP Data:
[1558] NOV20a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:53 and 54, respectively. The nucleotide sequence of the NOV20a variant differs as shown in Table SNP7.
687TABLE SNP7
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377746267CT87ProPro
13377745802AG266MetVal
133777441368CA454AsnLys
133777431671TC555TyrTyr
|
[1559] NOV21a SNP Data:
[1560] NOV21 has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:57 and 58, respectively. The nucleotide sequence of the NOV21 variant differs as shown in Table SNP8.
688TABLE SNP8
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377747360CT114LeuPhe
|
[1561] NOV23a SNP Data:
[1562] NOV23 a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:61 and 62, respectively. The nucleotide sequence of the NOV23a variant differs as shown in Table SNP9.
689TABLE SNP9
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
133777482580GA850ThrThr
133777493243TC1071AsnAsn
|
[1563] NOV24a SNP Data:
[1564] NOV24a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:63 and 64, respectively. The nucleotide sequences of the NOV24a variant differs as shown in Table SNP10.
690TABLE SNP10
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377750324GT104GluAsp
|
[1565] NOV26a SNP Data:
[1566] NOV26a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:67 and 68, respectively. The nucleotide sequence of the NOV26a variant differs as shown in Table SNP 11.
691TABLE SNP11
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377752681CT192ThrThr
13377751796AG231SerGly
|
[1567] NOV81a SNP Data:
[1568] NOV81 a has seven SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:211 and 212, respectively. The nucleotide sequence of the NOV81a variant differs as shown in Table SNP 12.
692TABLE SNP12
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13375651256TC86TyrHis
13375314392TC131LeuPro
13375429610CT204LeuPhe
13375779853GA285AspAsn
13375778908AG303AsnSer
133777531186AG396ThrAla
133756521312CT438ArgTrp
|
[1569] NOV30a SNP Data:
[1570] NOV30a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:77 and 78, respectively. The nucleotide sequence of the NOV30a variant differs as shown in Table SNP13.
693TABLE SNP13
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377756591AG197LysLys
|
[1571] NOV32a SNP Data:
[1572] NOV32a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:83 and 84 respectively. The nucleotide sequence of the NOV32a variant differs as shown in Table SNP 14.
694TABLE SNP14
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
133762982469CA801ThrThr
|
[1573] NOV38a SNP Data:
[1574] NOV38a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:97 and 98, respectively. The nucleotide sequence of the NOV38a variant differs as shown in Table SNP 15.
695TABLE SNP15
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
133777604685GA1540ArgHis
133777594763AG1566AspGly
|
[1575] NOV39a SNP Data:
[1576] NOV39a has four SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:99 and 100, respectively. The nucleotide sequence of the NOV39a variant differs as shown in Table SNP16.
696TABLE SNP16
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13375856187GA59AlaThr
13375855798CT262ProPro
133758541021GA337GlyArg
133758531337GA442ArgHis
|
[1577] NOV40a SNP Data:
[1578] NOV40a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs: 101 and 102 respectively. The nucleotide sequence of the NOV40a variant differs as shown in Table SNP 17.
697TABLE SNP17
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377761178AG58MetVal
|
[1579] NOV42A SNP Data:
[1580] NOV42A has three SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 105 and 106, respectively. The nucleotide sequence of the NOV42A variant differs as shown in Table SNP 18.
698TABLE SNP18
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377766824AT230LysEnd
13377770913TC259AlaAla
133777711042AG302LeuLeu
|
[1581] NOV44a SNP Data:
[1582] NOV44a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 109 and 110, respectively. The nucleotide sequence of the NOV44a variant differs as shown in Table SNP 19.
699TABLE SNP19
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377775450CT150IleIle
13377774551AG184AspGly
|
[1583] NOV45a NP Data:
[1584] NOV45a has eleven SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 111 and 112, respectively. The nucleotide sequence of the NOV45a variant differs as shown in Table SNP20.
700TABLE SNP20
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13376182154CT20HisTyr
13376181273TG59ArgArg
13376178977TC294MetThr
133761771052AG319HisArg
133761761111TC339TyrHis
133761751280AT395GlnLeu
133761741343AG416GluGly
133761731349AG418GluGly
133761721361AG422AsnSer
133761711427TC444LeuPro
133761701913GA0
|
[1585] NOV52a SNP Data:
[1586] NOV52a has seven SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs:127 and 128, respectively. The nucleotide sequence of the NOV52a variant differs as shown in Table SNP21.
701TABLE SNP21
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13375586430TC110SerSer
13375585492AG131GluGly
133755841713AG537IleMet
133755831756CT552AsnAsn
133755822143TA681ProPro
133775592550AG817LysArg
133777762555CT819LeuLeu
|
[1587] NOV53a SNP Data:
[1588] NOV53a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs: 129 and 130, respectively. The nucleotide sequence of the NOV53a variant differs as shown in Table SNP22.
702TABLE SNP22
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13376019384CT123SerPhe
|
[1589] NOV54a SNP Data:
[1590] NOV54a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:135 and 136, respectively. The nucleotide sequence of the NOV54a variant differs as shown in Table SNP23.
703TABLE SNP23
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13376037485AG148LysGlu
|
[1591] NOV55a SNP Data:
[1592] NOV55a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs: 135 and 136, respectively. The nucleotide sequence of the NOV55a variant differs as shown in Table SNP24.
704TABLE SNP24
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377777941CT304ArgTrp
|
[1593] NOV56a SNP Data:
[1594] NOV56a has three SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 139 and 140, respectively. The nucleotide sequence of the NOV56a variant differs as shown in Table SNP25.
705TABLE SNP25
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377780311AG102LysLys
13377779403GA133ArgHis
133777781303TC433LeuPro
|
[1595] NOV57a SNP Data:
[1596] NOV57a has nine SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 141 and 142, respectively. The nucleotide sequence of the NOV57a variant differs as shown in Table SNP26.
706TABLE SNP26
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377782140CA11SerTyr
13377783193GC29AlaPro
13377784201GA31ArgArg
13377785261CT51SerSer
13377786282TC58AsnAsn
13377787291TC61AlaAla
13377788317AT70AspVal
13377789418CT104LeuLeu
13377790461AG118AspGly
|
[1597] NOV61a SNP Data:
[1598] NOV61a has seven SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 153 and 154, respectively. The nucleotide sequence of the NOV61 a variant differs as shown in Table SNP27.
707TABLE SNP27
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13376354241GA67ArgHis
13376355283TC81IleThr
13376356420CT127LeuPhe
13376357474AG145IleVal
13376358588AG183ArgGly
13376359904TA288LeuGln
133763601384GA448GlyAsp
|
[1599] NOV62a SNP Data:
[1600] NOV62a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 155 and 156, respectively. The nucleotide sequence of the NOV62a variant differs as shown in Table SNP28.
708TABLE SNP28
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377799135CT44AlaAla
133765111228GA409AlaThr
|
[1601] NOV65a SNP Data:
[1602] NOV65a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 163 and 164, respectively. The nucleotide sequence of the NOV65a variant differs as shown in Table SNP29.
709TABLE SNP29
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
133763631001AG312AsnAsp
133763621007AG314IleVal
|
[1603] NOV67a SNP Data:
[1604] NOV67a has three SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 173 and 174, respectively. The nucleotide sequence of the NOV67a variant differs as shown in Table SNP30.
710TABLE SNP30
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377801533AT70SerCys
133762801815TC497PheSer
133778024683AG1453GluGly
|
[1605] NOV69a SNP Data:
[1606] NOV69a has one SNP variant, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs: 177 and 178, respectively. The nucleotide sequence of the NOV69a variant differs as shown in Table SNP31.
711TABLE SNP31
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13377803118GA25MetIle
|
[1607] NOV70a SNP Data:
[1608] NOV70a has eight SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 179 and 180, respectively. The nucleotide sequence of the NOV70a variant differs as shown in Table SNP32.
712TABLE SNIP32
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13376466285TC14SerPro
13376465310AT22GlnLeu
13376463502AG86AspGly
13376461603AG120LysGlu
13376457993AC250ThrPro
133764581165GA307SerAsn
133764601680AG479ThrAla
133764591711AG489AspGly
|
[1609] NOV71a SNP Data:
[1610] NOV71 a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 185 and 186, respectively. The nucleotide sequence of the NOV71 a variant differs as shown in Table SNP33.
713TABLE SNP33
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13374756535AG179AsnAsp
13374757572TC191LeuSer
|
[1611] NOV72a SNP Data:
[1612] NOV72a has three SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 189 and 190, respectively. The nucleotide sequence of the NOV72a variant differs as shown in Table SNP34.
714TABLE SNP34
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
1337780547GC13SerThr
13377806196TC63TrpArg
133778071272TC421GlyGly
|
[1613] NOV77a SNP Data:
[1614] NOV77a has four SNP variants, whose variant positions for its nucleotide and amino acid sequences are numbered according to SEQ ID NOs: 199 and 200, respectively. The nucleotide sequence of the NOV77a variant differs as shown in Table SNP35.
715TABLE SNP35
|
|
NucleotidesAmino Acids
VariantPositionInitialModifiedPositionInitialModified
|
13375891201AG22AsnSer
133758902463CT776ThrIle
133758922692CT852ProPro
133758892755TC873ThrThr
|
OTHER EMBODIMENTS
[1615] Although particular embodiments have been disclosed herein in detail, this has been done by way of example for purposes of illustration only, and is not intended to be limiting with respect to the scope of the appended claims, which follow. In particular, it is contemplated by the inventors that various substitutions, alterations, and modifications may be made to the invention without departing from the spirit and scope of the invention as defined by the claims. The choice of nucleic acid starting material, clone of interest, or library type is believed to be a matter of routine for a person of ordinary skill in the art with knowledge of the embodiments described herein. Other aspects, advantages, and modifications considered to be within the scope of the following claims. The claims presented are representative of the inventions disclosed herein. Other, unclaimed inventions are also contemplated. Applicants reserve the right to pursue such inventions in later claims.
Claims
- 1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
a) a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106; b) a variant of a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106, wherein any amino acid in the mature form is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence of the mature form are so changed; c) the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106; d) a variant of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106 wherein any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence are so changed; and e) a fragment of any of a) through d).
- 2. The polypeptide of claim 1 that is a naturally occurring allelic variant of the sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106.
- 3. The polypeptide of claim 2, wherein said allelic variant comprises an amino acid sequence that is the translation of a nucleic acid sequence differing by a single nucleotide from a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106.
- 4. The polypeptide of claim 1 that is a variant polypeptide described therein, wherein any amino acid specified in the chosen sequence is changed to provide a conservative substitution.
- 5. A method for determining the presence or amount of the polypeptide of claim 1 in a sample, the method comprising:
(a) providing said sample; (b) introducing said sample to an antibody that binds immunospecifically to the polypeptide; and (c) determining the presence or amount of antibody bound to said polypeptide, thereby determining the presence or amount of polypeptide in said sample.
- 6. A method for determining the presence of or predisposition to a disease associated with altered levels of the polypeptide of claim 1 in a first mammalian subject, the method comprising:
a) measuring the level of expression of the polypeptide in a sample from the first mammalian subject; and b) comparing the amount of said polypeptide in the sample of step (a) to the amount of the polypeptide present in a control sample from a second mammalian subject known not to have, or not to be predisposed to, said disease, wherein an alteration in the expression level of the polypeptide in the first subject as compared to the control sample indicates the presence of or predisposition to said disease.
- 7. A method of identifying an agent that binds to the polypeptide of claim 1, the method comprising:
(a) introducing said polypeptide to said agent; and (b) determining whether said agent binds to said polypeptide.
- 8. The method of claim 7 wherein the agent is a cellular receptor or a downstream effector.
- 9. A method for identifying a potential therapeutic agent for use in treatment of a pathology, wherein the pathology is related to aberrant expression or aberrant physiological interactions of the polypeptide of claim 1, the method comprising:
(a) providing a cell expressing the polypeptide of claim 1 and having a property or function ascribable to the polypeptide; (b) contacting the cell with a composition comprising a candidate substance; and (c) determining whether the substance alters the property or function ascribable to the polypeptide; whereby, if an alteration observed in the presence of the substance is not observed when the cell is contacted with a composition devoid of the substance, the substance is identified as a potential therapeutic agent.
- 10. A method for screening for a modulator of activity or of latency or predisposition to a pathology associated with the polypeptide of claim 1, said method comprising:
a) administering a test compound to a test animal at increased risk for a pathology associated with the polypeptide of claim 1, wherein said test animal recombinantly expresses the polypeptide of claim 1;b) measuring the activity of said polypeptide in said test animal after administering the compound of step (a); and c) comparing the activity of said protein in said test animal with the activity of said polypeptide in a control animal not administered said polypeptide, wherein a change in the activity of said polypeptide in said test animal relative to said control animal indicates the test compound is a modulator of latency of, or predisposition to, a pathology associated with the polypeptide of claim 1.
- 11. The method of claim 10, wherein said test animal is a recombinant test animal that expresses a test protein transgene or expresses said transgene under the control of a promoter at an increased level relative to a wild-type test animal, and wherein said promoter is not the native gene promoter of said transgene.
- 12. A method for modulating the activity of the polypeptide of claim 1, the method comprising introducing a cell sample expressing the polypeptide of said claim with a compound that binds to said polypeptide in an amount sufficient to modulate the activity of the polypeptide.
- 13. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of:
a) a mature form of the amino acid sequence given SEQ ID NO: 2n, wherein n is an integer between 1 and 106; b) a variant of a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106 wherein any amino acid in the mature form of the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence of the mature form are so changed; c) the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106; d) a variant of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106, in which any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% of the amino acid residues in the sequence are so changed; e) a nucleic acid fragment encoding at least a portion of a polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 106 or any variant of said polypeptide wherein any amino acid of the chosen sequence is changed to a different amino acid, provided that no more than 10% of the amino acid residues in the sequence are so changed; and f) the complement of any of said nucleic acid molecules.
- 14. The nucleic acid molecule of claim 13, wherein the nucleic acid molecule comprises the nucleotide sequence of a naturally occurring allelic nucleic acid variant.
- 15. The nucleic acid molecule of claim 13 that encodes a variant polypeptide, wherein the variant polypeptide has the polypeptide sequence of a naturally occurring polypeptide variant.
- 16. The nucleic acid molecule of claim 13, wherein the nucleic acid molecule differs by a single nucleotide from a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106.
- 17. The nucleic acid molecule of claim 13, wherein said nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of:
a) the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106; b) a nucleotide sequence wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% of the nucleotides are so changed; c) a nucleic acid fragment of the sequence selected from the group consisting of SEQ ID NO: 2n−1, wherein n is an integer between 1 and 106; and d) a nucleic acid fragment wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% of the nucleotides are so changed.
- 18. The nucleic acid molecule of claim 13, wherein said nucleic acid molecule hybridizes under stringent conditions to the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106, or a complement of said nucleotide sequence.
- 19. The nucleic acid molecule of claim 13, wherein the sequence is changed such that no more than 15% of the nucleotides in the coding sequence differ from the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1, wherein n is an integer between 1 and 106 or a fragment thereof.
- 20. A vector comprising the nucleic acid molecule of claim 19.
- 21. The vector of claim 20, further comprising a promoter operably linked to said nucleic acid molecule.
- 22. A cell comprising the vector of claim 20.
- 23. A method for determining the presence or amount of the nucleic acid molecule of claim 13 in a sample, the method comprising:
(a) providing said sample; (b) introducing said sample to a probe that binds to said nucleic acid molecule; and (c) determining the presence or amount of said probe bound to said nucleic acid molecule, thereby determining the presence or amount of the nucleic acid molecule in said sample.
- 24. The method of claim 23 wherein presence or amount of the nucleic acid molecule is used as a marker for cell or tissue type.
- 25. The method of claim 24 wherein the cell or tissue type is cancerous.
- 26. A method for determining the presence of or predisposition to a disease associated with altered levels of the nucleic acid molecule of claim 13 in a first mammalian subject, the method comprising:
a) measuring the amount of the nucleic acid in a sample from the first mammalian subject; and b) comparing the amount of said nucleic acid in the sample of step (a) to the amount of the nucleic acid present in a control sample from a second mammalian subject known not to have or not be predisposed to, the disease; wherein an alteration in the level of the nucleic acid in the first subject as compared to the control sample indicates the presence of or predisposition to the disease.
Provisional Applications (36)
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Number |
Date |
Country |
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60281086 |
Apr 2001 |
US |
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60281136 |
Apr 2001 |
US |
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60281863 |
Apr 2001 |
US |
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60281906 |
Apr 2001 |
US |
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60282020 |
Apr 2001 |
US |
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60282930 |
Apr 2001 |
US |
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60282934 |
Apr 2001 |
US |
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60283512 |
Apr 2001 |
US |
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60283710 |
Apr 2001 |
US |
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60284234 |
Apr 2001 |
US |
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60285325 |
Apr 2001 |
US |
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60285381 |
Apr 2001 |
US |
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60285609 |
Apr 2001 |
US |
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60285748 |
Apr 2001 |
US |
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60285890 |
Apr 2001 |
US |
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60286068 |
Apr 2001 |
US |
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60286292 |
Apr 2001 |
US |
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60287213 |
Apr 2001 |
US |
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60288257 |
May 2001 |
US |
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60294164 |
May 2001 |
US |
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60294484 |
May 2001 |
US |
|
60298952 |
Jun 2001 |
US |
|
60299237 |
Jun 2001 |
US |
|
60299276 |
Jun 2001 |
US |
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60318750 |
Sep 2001 |
US |
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60324800 |
Sep 2001 |
US |
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60324802 |
Sep 2001 |
US |
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60325684 |
Sep 2001 |
US |
|
60330143 |
Oct 2001 |
US |
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60332115 |
Nov 2001 |
US |
|
60332131 |
Nov 2001 |
US |
|
60332240 |
Nov 2001 |
US |
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60332779 |
Nov 2001 |
US |
|
60337621 |
Dec 2001 |
US |
|
60345783 |
Jan 2002 |
US |
|
60350251 |
Jan 2002 |
US |