Compounds for immunotherapy and diagnosis of tuberculosis and methods of their use

Abstract

Compounds and methods for inducing protective immunity against tuberculosis are disclosed. The compounds provided include polypeptides that contain at least one immunogenic portion of one or more M. tuberculosis proteins and DNA molecules encoding such polypeptides. Such compounds may be formulated into vaccines and/or pharmaceutical compositions for immunization against M. tuberculosis infection, or may be used for the diagnosis of tuberculosis.

Description

TECHNICAL FIELD

The present invention relates generally to detecting, treating and preventing Mycobacterium tuberculosis infection. The invention is more particularly related to polypeptides comprising a Mycobacterium tuberculosis antigen, or a portion or other variant thereof, and the use of such polypeptides for diagnosing and vaccinating against Mycobacterium tuberculosis infection.

BACKGROUND OF THE INVENTION

Tuberculosis is a chronic, infectious disease, that is generally caused by infection with Mycobacterium tuberculosis. It is a major disease in developing countries, as well as an increasing problem in developed areas of the world, with about 8 million new cases and 3 million deaths each year. Although the infection may be asymptomatic for a considerable period of time, the disease is most commonly manifested as an acute inflammation of the lungs, resulting in fever and a nonproductive cough. If left untreated, serious complications and death typically result.

Although tuberculosis can generally be controlled using extended antibiotic therapy, such treatment is not sufficient to prevent the spread of the disease. Infected individuals may be asymptomatic, but contagious, for some time. In addition, although compliance with the treatment regimen is critical, patient behavior is difficult to monitor. Some patients do not complete the course of treatment, which can lead to ineffective treatment and the development of drug resistance.

Inhibiting the spread of tuberculosis requires effective vaccination and accurate, early diagnosis of the disease. Currently, vaccination with live bacteria is the most efficient method for inducing protective immunity. The most common Mycobacterium employed for this purpose is Bacillus Calmette-Guerin (BCG), an avirulent strain of Mycobacterium bovis. However, the safety and efficacy of BCG is a source of controversy and some countries, such as the United States, do not vaccinate the general public. Diagnosis is commonly achieved using a skin test, which involves intradermal exposure to tuberculin PPD (protein-purified derivative). Antigen-specific T cell responses result in measurable induration at the injection site by 48-72 hours after injection, which indicates exposure to Mycobacterial antigens. Sensitivity and specificity have, however, been a problem with this test, and individuals vaccinated with BCG cannot be distinguished from infected individuals.

While macrophages have been shown to act as the principal effectors of M. tuberculosis immunity, T cells are the predominant inducers of such immunity. The essential role of T cells in protection against M. tuberculosis infection is illustrated by the frequent occurrence of M. tuberculosis in AIDS patients, due to the depletion of CD4 T cells associated with human immunodeficiency virus (HIV) infection. Mycobacterium-reactive CD4 T cells have been shown to be potent producers of gamma-interferon (IFN-γ), which, in turn, has been shown to trigger the anti-mycobacterial effects of macrophages in mice. While the role of IFN-γ in humans is less clear, studies have shown that 1,25-dihydroxy-vitamin D3, either alone or in combination with IFN-γ or tumor necrosis factor-alpha, activates human macrophages to inhibit M. tuberculosis infection. Furthermore, it is known that IFN-γ stimulates human macrophages to make 1,25-dihydroxy-vitamin D3. Similarly, IL-12 has been shown to play a role in stimulating resistance to M. tuberculosis infection. For a review of the immunology of M. tuberculosis infection see Chan and Kaufmann in Tuberculosis: Pathogenesis, Protection and Control, Bloom (ed.), ASM Press, Washington, D.C., 1994.

Accordingly, there is a need in the art for improved vaccines and methods for preventing, treating and detecting tuberculosis. The present invention fulfills these needs and further provides other related advantages.

SUMMARY OF THE INVENTION

Briefly stated, this invention provides compounds and methods for preventing and diagnosing tuberculosis. In one aspect, polypeptides are provided comprising an immunogenic portion of an M. tuberculosis antigen, or a variant of such an antigen that differs only in conservative substitutions and/or modifications, the antigen comprising an amino acid sequence encoded by a DNA sequence selected from the group consisting of the sequences recited in SEQ ID NO: 1, 11, 12, 83, 103-108, 125, 127, 129-137, 139 and 140, the complements of said sequences, and DNA sequences that hybridize to a sequence recited in SEQ ID NO: 1, 11, 12, 83, 103-108, 125, 127, 129-137, 139 and 140, or a complement thereof under moderately stringent conditions. In a second aspect, the present invention provides polypeptides comprising an immunogenic portion of a M. tuberculosis antigen having an amino acid sequence selected from the group consisting of sequences provided in SEQ ID NO: 16-33, 109, 126, 138, 141, 142 and variants thereof.

In related aspects, DNA sequences encoding the above polypeptides, expression vectors comprising these DNA sequences and host cells transformed or transfected with such expression vectors are also provided.

In another aspect, the present invention provides fusion proteins comprising a first and a second inventive polypeptide or, alternatively, an inventive polypeptide and a known M. tuberculosis antigen.

Within other aspects, the present invention provides pharmaceutical compositions that comprise one or more of the above polypeptides, or a DNA molecule encoding such polypeptides, and a physiologically acceptable carrier. The invention also provides vaccines comprising one or more of the polypeptides as described above and a non-specific immune response enhancer, together with vaccines comprising one or more DNA sequences encoding such polypeptides and a non-specific immune response enhancer.

In yet another aspect, methods are provided for inducing protective immunity in a patient, comprising administering to a patient an effective amount of one or more of the above polypeptides.

In further aspects of this invention, methods and diagnostic kits are provided for detecting tuberculosis in a patient. The methods comprise contacting dermal cells of a patient with one or more of the above polypeptides and detecting an immune response on the patient's skin. The diagnostic kits comprise one or more of the above polypeptides in combination with an apparatus sufficient to contact the polypeptide with the dermal cells of a patient.

In yet another aspect, methods are provided for detecting tuberculosis in a patient, such methods comprising contacting dermal cells of a patient with one or more polypeptides encoded by a DNA sequence selected from the group consisting of SEQ ID NO: 2-10, 102, 128, the complements of said sequences, and DNA sequences that hybridize to a sequence recited in SEQ ID NO: 2-10, 102, 128; and detecting an immune response on the patient's skin. Diagnostic kits for use in such methods are also provided.

These and other aspects of the present invention will become apparent upon reference to the following detailed description and attached drawings. All references disclosed herein are hereby incorporated by reference in their entirety as if each was incorporated individually.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B illustrate the stimulation of proliferation and interferon-γ production, respectively, in T cells derived from a first PPD-positive donor (referred to as D7) by recombinant ORF-2 and synthetic peptides to ORF-2.

FIGS. 2A and 2B illustrate the stimulation of proliferation and interferon-γ production, respectively, in T cells derived from a second PPD-positive donor (referred to as D160) by recombinant ORF-2 and synthetic peptides to ORF-2.

DETAILED DESCRIPTION OF THE INVENTION

As noted above, the present invention is generally directed to compositions and methods for preventing, treating and diagnosing tuberculosis. The compositions of the subject invention include polypeptides that comprise at least one immunogenic portion of a M. tuberculosis antigen, or a variant of such an antigen that differs only in conservative substitutions and/or modifications. As used herein, the term “polypeptide” encompasses amino acid chains of any length, including full length proteins (i.e., antigens), wherein the amino acid residues are linked by covalent peptide bonds. Thus, a polypeptide comprising an immunogenic portion of one of the above antigens may consist entirely of the immunogenic portion, or may contain additional sequences. The additional sequences may be derived from the native M. tuberculosis antigen or may be heterologous, and such sequences may (but need not) be immunogenic.

“Immunogenic,” as used herein, refers to the ability to elicit an immune response (e.g., cellular) in a patient, such as a human, and/or in a biological sample. In particular, antigens that are immunogenic (and immunogenic portions or other variants of such antigens) are capable of stimulating cell proliferation, interleukin-12 production and/or interferon-γ production in biological samples comprising one or more cells selected from the group of T cells, NK cells, B cells and macrophages, where the cells are derived from an M. tuberculosis -immune individual. Polypeptides comprising at least an immunogenic portion of one or more M. tuberculosis antigens may generally be used to detect tuberculosis or to induce protective immunity against tuberculosis in a patient.

The compositions and methods of this invention also encompass variants of the above polypeptides. A polypeptide “variant,” as used herein, is a polypeptide that differs from the recited polypeptide only in conservative substitutions and/or modifications, such that the therapeutic, antigenic and/or immunogenic properties of the polypeptide are retained. Polypeptide variants preferably exhibit at least about 70%, more preferably at least about 90% and most preferably at least about 95% identity to the identified polypeptides. For polypeptides with immunoreactive properties, variants may, alternatively, be identified by modifying the amino acid sequence of one of the above polypeptides, and evaluating the immunoreactivity of the modified polypeptide. For polypeptides useful for the generation of diagnostic binding agents, a variant may be identified by evaluating a modified polypeptide for the ability to generate antibodies that detect the presence or absence of tuberculosis. Alternatively, variants of the claimed antigens that may be usefully employed in the inventive diagnostic methods may be identified by evaluating modified polypeptides for their ability to detect antibodies present in the sera of tuberculosis-infected patients. Such modified sequences may be prepared and tested using, for example, the representative procedures described herein.

A “conservative substitution” is one in which an amino acid is substituted for another amino acid that has similar properties, such that one skilled in the art of peptide chemistry would expect the secondary structure and hydropathic nature of the polypeptide to be substantially unchanged. In general, the following groups of amino acids represent conservative changes: (1) ala, pro, gly, glu, asp, gln, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala, phe; (4) lys, arg, his; and (5) phe, tyr, trp, his.

Variants may also (or alternatively) be modified by, for example, the deletion or addition of amino acids that have minimal influence on the immunogenic properties, secondary structure and hydropathic nature of the polypeptide. For example, a polypeptide may be conjugated to a signal (or leader) sequence at the N-tenninal end of the protein which co-translationally or post-translationally directs transfer of the protein. The polypeptide may also be conjugated to a linker or other sequence for ease of synthesis, purification or identification of the polypeptide (e.g., poly-His), or to enhance binding of the polypeptide to a solid support. For example, a polypeptide may be conjugated to an immunoglobulin Fc region.

In general, M. tuberculosis antigens, and DNA sequences encoding such antigens, may be prepared using any of a variety of procedures. For example, genomic or cDNA libraries derived from M. tuberculosis may be screened directly using peripheral blood mononuclear cells (PBMCs) or T cell lines or clones derived from one or more M. tuberculosis -immune individuals. Direct library screens may generally be performed by assaying pools of expressed recombinant proteins for the ability of induce proliferation and/or interferon-γ production in T cells derived from an M. tuberculosis -immune individual. Potential T cell antigens may be first selected based on antibody reactivity, as described above.

Alternatively, DNA sequences encoding antigens may be identified by screening an appropriate M. tuberculosis genomic or cDNA expression library with sera obtained from patients infected with M. tuberculosis. Such screens may generally be performed using techniques well known to those of ordinary skill in the art, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y., 1989.

Purified antigens are then evaluated for their ability to elicit an appropriate immune response (e.g., cellular) using, for example, the representative methods described herein. Immunogenic antigens may then be partially sequenced using techniques such as traditional Edman chemistry. See Edman and Berg, Eur. J. Biochem. 80:116-132, 1967. Immunogenic antigens may also be produced recombinantly using a DNA sequence that encodes the antigen, which has been inserted into an expression vector and expressed in an appropriate host.

DNA sequences encoding the inventive antigens may also be obtained by screening an appropriate M. tuberculosis cDNA or genomic DNA library for DNA sequences that hybridize to degenerate oligonucleotides derived from partial amino acid sequences of isolated antigens. Degenerate oligonucleotide sequences for use in such a screen may be designed and synthesized, and the screen may be performed, as described (for example) in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y., 1989 (and references cited therein). Polymerase chain reaction (PCR) may also be employed, using the above oligonucleotides in methods well known in the art, to isolate a nucleic acid probe from a cDNA or genomic library. The library screen may then be performed using the isolated probe.

Regardless of the method of preparation, the antigens (and immunogenic portions thereof) described herein have the ability to induce an immunogenic response. More specifically, the antigens have the ability to induce proliferation and/or cytokine production (i.e., interferon-γ and/or interleukin-12 production) in T cells, NK cells, B cells and/or macrophages derived from an M. tuberculosis -immune individual. The selection of cell type for use in evaluating an immunogenic response to a antigen will, of course, depend on the desired response. For example, interleukin-12 production is most readily evaluated using preparations containing B cells and/or macrophages. An M. tuberculosis -immune individual is one who is considered to be resistant to the development of tuberculosis by virtue of having mounted an effective T cell response to M. tuberculosis (i.e., substantially free of disease symptoms). Such individuals may be identified based on a strongly positive (i.e., greater than about 10 mm diameter induration) intradermal skin test response to tuberculosis proteins (PPD) and an absence of any signs or symptoms of tuberculosis disease. T cells, NK cells, B cells and macrophages derived from M. tuberculosis -immune individuals may be prepared using methods known to those of ordinary skill in the art. For example, a preparation of PBMCs (i.e., peripheral blood mononuclear cells) may be employed without further separation of component cells. PBMCs may generally be prepared, for example, using density centrifugation through Ficoll™ (Winthrop Laboratories, NY).

T cells for use in the assays described herein may also be purified directly from PBMCs. Alternatively, an enriched T cell line reactive against mycobacterial proteins, or T cell clones reactive to individual mycobacterial proteins, may be employed. Such T cell clones may be generated by, for example, culturing PBMCs from M. tuberculosis -immune individuals with mycobacterial proteins for a period of 2-4 weeks. This allows expansion of only the mycobacterial protein-specific T cells, resulting in a line composed solely of such cells. These cells may then be cloned and tested with individual proteins, using methods known to those of ordinary skill in the art, to more accurately define individual T cell specificity. In general, antigens that test positive in assays for proliferation and/or cytokine production (i.e., interferon-γ and/or interleukin-12 production) performed using T cells, NK cells, B cells and/or macrophages derived from an M. tuberculosis -immune individual are considered immunogenic. Such assays may be performed, for example, using the representative procedures described below. Immunogenic portions of such antigens may be identified using similar assays, and may be present within the polypeptides described herein.

The ability of a polypeptide (e.g., an immunogenic antigen, or a portion or other variant thereof) to induce cell proliferation is evaluated by contacting the cells (e.g., T cells and/or NK cells) with the polypeptide and measuring the proliferation of the cells. In general, the amount of polypeptide that is sufficient for evaluation of about 10 5 cells ranges from about 10 ng/mL to about 100 μg/mL and preferably is about 10 μg/mL. The incubation of polypeptide with cells is typically performed at 37° C. for about six days. Following incubation with polypeptide, the cells are assayed for a proliferative response, which may be evaluated by methods known to those of ordinary skill in the art, such as exposing cells to a pulse of radiolabeled thymidine and measuring the incorporation of label into cellular DNA. In general, a polypeptide that results in at least a three fold increase in proliferation above background (i.e., the proliferation observed for cells cultured without polypeptide) is considered to be able to induce proliferation.

The ability of a polypeptide to stimulate the production of interferon-γ and/or interleukin-12 in cells may be evaluated by contacting the cells with the polypeptide and measuring the level of interferon-γ or interleukin-12 produced by the cells. In general, the amount of polypeptide that is sufficient for the evaluation of about 10 5 cells ranges from about 10 ng/mL to about 100 μg/mL and preferably is about 10 μg/mL. The polypeptide may, but need not, be immobilized on a solid support, such as a bead or a biodegradable microsphere, such as those described in U.S. Pat. Nos. 4,897,268 and 5,075,109. The incubation of polypeptide with the cells is typically performed at 37° C. for about six days. Following incubation with polypeptide, the cells are assayed for interferon-γ and/or interleukin-12 (or one or more subunits thereof), which may be evaluated by methods known to those of ordinary skill in the art, such as an enzyme-linked immunosorbent assay (ELISA) or, in the case of IL-12 P70 heterodimer, a bioassay such as an assay measuring proliferation of T cells. In general, a polypeptide that results in the production of at least 50 pg of interferon-γ per mL of cultured supernatant (containing 10 4 -10 5 T cells per mL) is considered able to stimulate the production of interferon-γ. A polypeptide that stimulates the production of at least 10 pg/mL of IL-12 P70 subunit, and/or at least 100 pg/mL of IL-12 P40 subunit, per 10 5 macrophages or B cells (or per 3×10 5 PBMC) is considered able to stimulate the production of IL-12.

In general, immunogenic antigens are those antigens that stimulate proliferation and/or cytokine production (i.e., interferon-γ and/or interleukin-12 production) in T cells, NK cells, B cells and/or macrophages derived from at least about 25% of M. tuberculosis -immune individuals. Among these immunogenic antigens, polypeptides having superior therapeutic properties may be distinguished based on the magnitude of the responses in the above assays and based on the percentage of individuals for which a response is observed. In addition, antigens having superior therapeutic properties will not stimulate proliferation and/or cytokine production in vitro in cells derived from more than about 25% of individuals that are not M. tuberculosis -immune, thereby eliminating responses that are not specifically due to M. tuberculosis -responsive cells. Those antigens that induce a response in a high percentage of T cell, NK cell, B cell and/or macrophage preparations from M. tuberculosis -immune individuals (with a low incidence of responses in cell preparations from other individuals) have superior therapeutic properties.

Antigens with superior therapeutic properties may also be identified based on their ability to diminish the severity of M. tuberculosis infection in experimental animals, when administered as a vaccine. Suitable vaccine preparations for use on experimental animals are described in detail below. Efficacy may be determined based on the ability of the antigen to provide at least about a 50% reduction in bacterial numbers and/or at least about a 40% decrease in mortality following experimental infection. Suitable experimental animals include mice, guinea pigs and primates.

Antigens having superior diagnostic properties may generally be identified based on the ability to elicit a response in an intradermal skin test performed on an individual with active tuberculosis, but not in a test performed on an individual who is not infected with M. tuberculosis. Skin tests may generally be performed as described below, with a response of at least 5 mm induration considered positive.

Immunogenic portions of the antigens described herein may be prepared and identified using well known techniques, such as those summarized in Paul, Fundamental Immunology, 3d ed., Raven Press, 1993, pp. 243-247 and references cited therein. Such techniques include screening polypeptide portions of the native antigen for immunogenic properties. The representative proliferation and cytokine production assays described herein may generally be employed in these screens. An immunogenic portion of a polypeptide is a portion that, within such representative assays, generates an immune response (e.g., proliferation, interferon-γ production and/or interleukin-12 production) that is substantially similar to that generated by the full length antigen. In other words, an immunogenic portion of an antigen may generate at least about 20%, and preferably about 100%, of the proliferation induced by the full length antigen in the model proliferation assay described herein. An immunogenic portion may also, or alternatively, stimulate the production of at least about 20%, and preferably about 100%, of the interferon-γ and/or interleukin-12 induced by the full length antigen in the model assay described herein.

Portions and other variants of M. tuberculosis antigens may be generated by synthetic or recombinant means. Synthetic polypeptides having fewer than about 100 amino acids, and generally fewer than about 50 amino acids, may be generated using techniques well known to those of ordinary skill in the art. For example, such polypeptides may be synthesized using any of the commercially available solid-phase techniques, such as the Merrifield solid-phase synthesis method, where amino acids are sequentially added to a growing amino acid chain. See Merrifield, J. Am. Chem. Soc. 85:2149-2146, 1963. Equipment for automated synthesis of polypeptides is commercially available from suppliers such as Perkin Elmer/Applied BioSystems Division, Foster City, Calif., and may be operated according to the manufacturer's instructions. Variants of a native antigen may generally be prepared using standard mutagenesis techniques, such as oligonucleotide-directed site-specific mutagenesis. Sections of the DNA sequence may also be removed using standard techniques to permit preparation of truncated polypeptides.

Recombinant polypeptides containing portions and/or variants of a native antigen may be readily prepared from a DNA sequence encoding the polypeptide using a variety of techniques well known to those of ordinary skill in the art. For example, supernatants from suitable host/vector systems which secrete recombinant protein into culture media may be first concentrated using a commercially available filter. Following concentration, the concentrate may be applied to a suitable purification matrix such as an affinity matrix or an ion exchange resin. Finally, one or more reverse phase HPLC steps can be employed to further purify a recombinant protein.

Any of a variety of expression vectors known to those of ordinary skill in the art may be employed to express recombinant polypeptides of this invention. Expression may be achieved in any appropriate host cell that has been transformed or transfected with an expression vector containing a DNA molecule that encodes a recombinant polypeptide. Suitable host cells include prokaryotes, yeast and higher eukaryotic cells. Preferably, the host cells employed are E. coli, yeast or a mammalian cell line such as COS or CHO. The DNA sequences expressed in this manner may encode naturally occurring antigens, portions of naturally occurring antigens, or other variants thereof.

In general, regardless of the method of preparation, the polypeptides disclosed herein are prepared in substantially pure form. Preferably, the polypeptides are at least about 80% pure, more preferably at least about 90% pure and most preferably at least about 99% pure. In certain preferred embodiments, described in detail below, the substantially pure polypeptides are incorporated into pharmaceutical compositions or vaccines for use in one or more of the methods disclosed herein.

In one embodiment, the subject invention discloses polypeptides comprising at least an immunogenic portion of an M. tuberculosis antigen (or a variant of such an antigen) that comprises one or more of the amino acid sequences encoded by (a) the DNA sequences of SEQ ID NO: 1-12, 83, 102-108, 125, 127-137, 139 and 140; (b) the complements of such DNA sequences, or (c) DNA sequences substantially homologous to a sequence of (a) or (b). In a related embodiment, the present invention provides polypeptides comprising at least an immunogenic portion of an M. tuberculosis antigen having an amino acid sequence selected from the group consisting of sequences provided in SEQ ID NO: 16-33, 109, 126, 138, 141, 142 and variants thereof.

The M. tuberculosis antigens provided herein include variants that are encoded by DNA sequences which are substantially homologous to one or more of the DNA sequences specifically recited herein. “Substantial homology,” as used herein, refers to DNA sequences that are capable of hybridizing under moderately stringent conditions. Suitable moderately stringent conditions include prewashing in a solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50° C.-65° C., 5×SSC, overnight or, in the case of cross-species homology at 45° C., 0.5×SSC; followed by washing twice at 65° C. for 20 minutes with each of 2×, 0.5× and 0.2×SSC containing 0.1% SDS). Such hybridizing DNA sequences are also within the scope of this invention, as are nucleotide sequences that, due to code degeneracy, encode an immunogenic polypeptide that is encoded by a hybridizing DNA sequence.

In a related aspect, the present invention provides fusion proteins comprising a first and a second inventive polypeptide or, alternatively, a polypeptide of the present invention and a known M. tuberculosis antigen, such as the 38 kD antigen described in Andersen and Hansen, Infect. Immun. 57:2481-2488, 1989, (Genbank Accession No. M30046), or ESAT-6 previously identified in M. bovis (Accession No. U34848) and in M. tuberculosis (Sorensen et al., Infec. Immun. 63:1710-1717, 1995). Variants of such fusion proteins are also provided. The fusion proteins of the present invention may include a linker peptide between the first and second polypeptides.

A DNA sequence encoding a fusion protein of the present invention is constructed using known recombinant DNA techniques to assemble separate DNA sequences encoding the first and second polypeptides into an appropriate expression vector. The 3′ end of a DNA sequence encoding the first polypeptide is ligated, with or without a peptide linker, to the 5′ end of a DNA sequence encoding the second polypeptide so that the reading frames of the sequences are in phase to permit mRNA translation of the two DNA sequences into a single fusion protein that retains the biological activity of both the first and the second polypeptides.

A peptide linker sequence may be employed to separate the first and the second polypeptides by a distance sufficient to ensure that each polypeptide folds into its secondary and tertiary structures. Such a peptide linker sequence is incorporated into the fusion protein using standard techniques well known in the art. Suitable peptide linker sequences may be chosen based on the following factors: (1) their ability to adopt a flexible extended conformation; (2) their inability to adopt a secondary structure that could interact with functional epitopes on the first and second polypeptides; and (3) the lack of hydrophobic or charged residues that might react with the polypeptide functional epitopes. Preferred peptide linker sequences contain Gly, Asn and Ser residues. Other near neutral amino acids, such as Thr and Ala may also be used in the linker sequence. Amino acid sequences which may be usefully employed as linkers include those disclosed in Maratea et al., Gene 40:39-46, 1985; Murphy et al., Proc. Natl. Acad Sci. USA 83:8258-8262, 1986; U.S. Pat. Nos. 4,935,233 and 4,751,180. The linker sequence may be from 1 to about 50 amino acids in length. Peptide sequences are not required when the first and second polypeptides have non-essential N-terminal amino acid regions that can be used to separate the functional domains and prevent steric interference.

The ligated DNA sequences are operably linked to suitable transcriptional or translational regulatory elements. The regulatory elements responsible for expression of DNA are located only 5′ to the DNA sequence encoding the first polypeptides. Similarly, stop codons require to end translation and transcription termination signals are only present 3′ to the DNA sequence encoding the second polypeptide.

In another aspect, the present invention provides methods for using one or more of the above polypeptides or fusion proteins (or DNA molecules encoding such polypeptides) to induce protective immunity against tuberculosis in a patient. As used herein, a “patient” refers to any warm-blooded animal, preferably a human. A patient may be afflicted with a disease, or may be free of detectable disease and/or infection. In other words, protective immunity may be induced to prevent or treat tuberculosis.

In this aspect, the polypeptide, fusion protein or DNA molecule is generally present within a pharmaceutical composition and/or a vaccine. Pharmaceutical compositions may comprise one or more polypeptides, each of which may contain one or more of the above sequences (or variants thereof), and a physiologically acceptable carrier. Vaccines may comprise one or more of the above polypeptides and a non-specific immune response enhancer, such as an adjuvant or a liposome (into which the polypeptide is incorporated). Such pharmaceutical compositions and vaccines may also contain other M. tuberculosis antigens, either incorporated into a combination polypeptide or present within a separate polypeptide.

Alternatively, a vaccine may contain DNA encoding one or more polypeptides as described above, such that the polypeptide is generated in situ. In such vaccines, the DNA may be present within any of a variety of delivery systems known to those of ordinary skill in the art, including nucleic acid expression systems, bacterial and viral expression systems. Appropriate nucleic acid expression systems contain the necessary DNA sequences for expression in the patient (such as a suitable promoter and terminating signal). Bacterial delivery systems involve the administration of a bacterium (such as Bacillus-Calmette-Guerrin) that expresses an immunogenic portion of the polypeptide on its cell surface. In a preferred embodiment, the DNA may be introduced using a viral expression system (e.g., vaccinia or other pox virus, retrovirus, or adenovirus), which may involve the use of a non-pathogenic (defective), replication competent virus. Techniques for incorporating DNA into such expression systems are well known to those of ordinary skill in the art. The DNA may also be “naked,” as described, for example, in Ulmer et al., Science 259:1745-1749, 1993 and reviewed by Cohen, Science 259:1691-1692, 1993. The uptake of naked DNA may be increased by coating the DNA onto biodegradable beads, which are efficiently transported into the cells.

In a related aspect, a DNA vaccine as described above may be administered simultaneously with or sequentially to either a polypeptide of the present invention or a known M. tuberculosis antigen, such as the 38 kD antigen described above. For example, administration of DNA encoding a polypeptide of the present invention, either “naked” or in a delivery system as described above, may be followed by administration of an antigen in order to enhance the protective immune effect of the vaccine.

Routes and frequency of administration, as well as dosage, will vary from individual to individual and may parallel those currently being employed in immunization using BCG. In general, the pharmaceutical compositions and vaccines may be administered by injection (e.g., intracutaneous, intramuscular, intravenous or subcutaneous), intranasally (e.g, by aspiration) or orally. Between 1 and 3 doses may be administered for a 1-36 week period. Preferably, 3 doses are administered, at intervals of 3-4 months, and booster vaccinations may be given periodically thereafter. Alternate protocols may be appropriate for individual patients. A suitable dose is an amount of polypeptide or DNA that, when administered as described above, is capable of raising an immune response in an immunized patient sufficient to protect the patient from M. tuberculosis infection for at least 1-2 years. In general, the amount of polypeptide present in a dose (or produced in situ by the DNA in a dose) ranges from about 1 pg to about 100 mg per kg of host, typically from about 10 pg to about 1 mg, and preferably from about 100 pg to about 1 μg. Suitable dose sizes will vary with the size of the patient, but will typically range from about 0.1 mL to about 5 mL.

While any suitable carrier known to those of ordinary skill in the art may be employed in the pharmaceutical compositions of this invention, the type of carrier will vary depending on the mode of administration. For parenteral administration, such as subcutaneous injection, the carrier preferably comprises water, saline, alcohol, lipids, a wax or a buffer. For oral administration, any of the above carriers or a solid carrier, such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, glucose, sucrose, and magnesium carbonate, may be employed. Biodegradable microspheres (e.g., polylactic galactide) may also be employed as carriers for the pharmaceutical compositions of this invention. Suitable biodegradable microspheres are disclosed, for example, in U.S. Pat. Nos. 4,897,268 and 5,075,109.

Any of a variety of adjuvants may be employed in the vaccines of this invention to nonspecifically enhance the immune response. Most adjuvants contain a substance designed to protect the antigen from rapid catabolism, such as aluminum hydroxide or mineral oil, and a nonspecific stimulator of immune responses, such as lipid A, Bortadella pertussis or Mycobacterium tuberculosis. Suitable adjuvants are commercially available as, for example, Freund's Incomplete Adjuvant and Freund's Complete Adjuvant (Difco Laboratories) and Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.). Other suitable adjuvants include alum, biodegradable microspheres, monophosphoryl lipid A and quil A.

In another aspect, this invention provides methods for using one or more of the polypeptides described above to diagnose tuberculosis using a skin test. As used herein, a “skin test” is any assay performed directly on a patient in which a delayed-type hypersensitivity (DTH) reaction (such as swelling, reddening or dermatitis) is measured following intradermal injection of one or more polypeptides as described above. Such injection may be achieved using any suitable device sufficient to contact the polypeptide or polypeptides with dermal cells of the patient, such as a tuberculin syringe or 1 mL syringe. Preferably, the reaction is measured at least 48 hours after injection, more preferably 48-72 hours.

The DTH reaction is a cell-mediated immune response, which is greater in patients that have been exposed previously to the test antigen (i.e., the immunogenic portion of the polypeptide employed, or a variant thereof). The response may be measured visually, using a ruler. In general, a response that is greater than about 0.5 cm in diameter, preferably greater than about 1.0 cm in diameter, is a positive response, indicative of tuberculosis infection, which may or may not be manifested as an active disease.

The polypeptides of this invention are preferably formulated, for use in a skin test, as pharmaceutical compositions containing a polypeptide and a physiologically acceptable carrier, as described above. Such compositions typically contain one or more of the above polypeptides in an amount ranging from about 1 μg to about 100 μg, preferably from about 10 μg to about 50 μg in a volume of 0.1 mL. Preferably, the carrier employed in such pharmaceutical compositions is a saline solution with appropriate preservatives, such as phenol and/or TWEEN 80™.

In a preferred embodiment, a polypeptide employed in a skin test is of sufficient size such that it remains at the site of injection for the duration of the reaction period. In general, a polypeptide that is at least 9 amino acids in length is sufficient. The polypeptide is also preferably broken down by macrophages within hours of injection to allow presentation to T-cells. Such polypeptides may contain repeats of one or more of the above sequences and/or other immunogenic or non-immunogenic sequences.

The following Examples are offered by way of illustration and not by way of limitation.

EXAMPLE 1

Purification and Characterization of M. Tuberculosis Polypeptide Using CD4+ T Cell Lines Generated from Human PBMC

M. tuberculosis antigens of the present invention were isolated by expression cloning of cDNA libraries of M. tuberculosis strains H37Rv and Erdman essentially as described by Sanderson et al. ( J. Exp. Med., 1995, 182:1751-1757) and were shown to induce PBMC proliferation and IFN-γ in an immunoreactive T cell line.

Two CD4+ T cell lines, referred to as DC-4 and DC-5, were generated against dendritic cells infected with M. tuberculosis. Specifically, dendritic cells were prepared from adherent PBMC from a single donor and subsequently infected with tuberculosis. Lymphocytes from the same donor were cultured under limiting dilution conditions with the infected dendritic cells to generate the CD4+ T cell lines DC-4 and DC-5. These cell lines were shown to react with crude soluble proteins from M. tuberculosis but not with Tb38-1. Limiting dilution conditions were employed to obtain a third CD4+ T cell line, referred to as DC-6, which was shown to react with both crude soluble proteins and Tb38-1.

Genomic DNA was isolated from the M. tuberculosis strains H37Rv and Erdman and used to construct expression libraries in the vector pBSK(−) using the Lambda ZAP expression system (Stratagene, La Jolla, Calif.). These libraries were transformed into E. coli, pools of induced E. coli cultures were incubated with dendritic cells, and the ability of the resulting incubated dendritic cells to stimulate cell proliferation and IFN-γ production in the CD4+ T cell line DC-6 was examined as described below in Example 2. Positive pools were fractionated and re-tested until pure M. tuberculosis clones were obtained. Nineteen clones were isolated, of which nine were found to contain the previously identified M. tuberculosis antigens TbH-9 and Tb38-1, disclosed in U.S. patent application Ser. No. 08/533,634. The determined cDNA sequences for the remaining ten clones (hereinafter referred to as Tb224, Tb636, Tb424, Tb436, Tb398, Tb508, Tb441, Tb475, Tb488 and Tb465) are provided in SEQ ID No: 1-10, respectively. The corresponding predicted amino acid sequences for Tb224 and Tb636 are provided in SEQ ID NO: 13 and 14, respectively. The open reading frames for these two antigens were found to show some homology to TbH-9, described above. Tb224 and Tb636 were also found to be overlapping clones.

Tb424, Tb436, Tb398, Tb508, Tb441, Tb475, Tb488 and Tb465 were each found to contain two small open reading frames (referred to as ORF-1 and ORF-2) or truncated forms thereof, with minor variations in ORF-1 and ORF-2 being found for each clone. The predicted amino acid sequences of ORF-1 and ORF-2 for Tb424, Th436, Tb398, Tb508, Tb441, Tb475, Tb488 and Tb465 are provided in SEQ ID NO: 16 and 17, 18 and 19, 20 and 21, 22 and 23, 24 and 25, 26 and 27, 28 and 29, and 30 and 31, respectively. In addition, clones Tb424 and Tb436 were found to contain a third apparent open reading frame, referred to as ORF-U. The predicted amino acid sequences of ORF-U for Tb424 and Tb436 are provided in SEQ ID NO: 32 and 33, respectively. Tb424 and Tb436 were found to be either overlapping clones or recently duplicated/transposed copies. Similarly Tb398, Tb508 and Tb465 were found to be either overlapping clones or recently duplicated/transposed copies, as were Tb475 and Tb488.

These sequences were compared with known sequences in the gene bank using the BLASTN system. No homologies to the antigens Tb224 and Tb431 were found. Tb636 was found to be 100% identical to a cosmid previously identified in M. tuberculosis. Similarly, Tb508, Tb488, Tb398, Tb424, Tb436, Tb441, Tb465 and Tb475 were found to show homology to known M. tuberculosis cosmids. In addition, Tb488 was found to have 100% homology to M. tuberculosis topoisomerase I.

Seventeen overlapping peptides to the open reading frame ORF-1 (referred to as 1-1-1-17; SEQ ID NO: 34-50, respectively) and thirty overlapping peptides to the open reading frame ORF-2 (referred to as 2-1-2-30, SEQ ID NO: 51-80) were synthesized using the procedure described below in Example 3.

The ability of the synthetic peptides, and of recombinant ORF-1 and ORF-2, to induce T cell proliferation and IFN-γ production in PBMC from PPD-positive donors was assayed as described below in Example 2. FIGS. 1A-B and 2 A-B illustrate stimulation of T cell proliferation and IFN-γ by recombinant ORF-2 and the synthetic peptides 2-1-2-16 for two donors, referred to as D7 and D160, respectively. Recombinant ORF-2 (referred to as MTI) stimulated T cell proliferation and IFN-γ production in PBMC from both donors. The amount of PBMC stimulation seen with the individual synthetic peptides varied with each donor, indicating that each donor recognizes different epitopes on ORF-2. The proteins encoded by ORF-1, ORF-2 and ORF-U were subsequently named MTS, MTI and MSF, respectively.

Eighteen overlapping peptides to the sequence of MSF (referred to as MSF-1-MSF-18; SEQ ID NO: 84-101, respectively) were synthesized and their ability to stimulate T cell proliferation and IFN-γ production in a CD4+ T cell line generated against M. tuberculosis culture filtrate was examined as described below. The peptides referred to as MSF-12 and MSF-13 (SEQ ID NO: 95 and 96, respectively) were found to show the highest levels of reactivity. Two overlapping peptides (SEQ ID NO:81 and 82) to the open reading frame of Tb224 were synthesized and shown to induce T cell proliferation and IFN-γ production in PBMC from PPD-positive donors.

Two CD4+ T cell lines from different donors were generated against M. tuberculosis infected dendritic cells using the above methodology. Screening of the M. tuberculosis cDNA expression library described above using this cell line, resulted in the isolation of two clones referred to as Tb867 and Tb391. The determined cDNA sequence for Tb867 (SEQ ID NO: 102) was found to be identical to the previously isolated M. tuberculosis cosmid SCY22G10, with the candidate reactive open reading frame encoding a 750 amino acid M. tuberculosis protein kinase. Comparison of the determined cDNA sequence for Tb391 (SEQ ID NO: 103) with those in the gene bank revealed no significant homologies to known sequences.

In further studies, CD4+ T cell lines were generated against M. tuberculosis culture filtrate, essentially as outlined above, and used to screen the M. tuberculosis Erdman cDNA expression library described above. Five reactive clones, referred to as Tb431, Tb472, Tb470, Tb838 and Tb962 were isolated. The determined cDNA sequences for Tb431, Tb472, Tb470, and Tb838 are provided in SEQ ID NO: 11, 12, 104 and 105, respectively, with the determined cDNA sequences for Tb962 being provided in SEQ ID NO: 106 and 107. The corresponding predicted amino acid sequence for Tb431 is provided in SEQ ID NO: 15.

Subsequent studies led to the isolation of a full-length cDNA sequence for Tb472 (SEQ ID NO: 108). Overlapping peptides were synthesized and used to identify the reactive open reading frame. The predicted amino acid sequence for the protein encoded by Tb472 (referred to as MSL) is provided in SEQ ID NO: 109. Comparison of the sequences for Tb472 and MSL with those in the gene bank, as described above, revealed no homologies to known sequences. Fifteen overlapping peptides to the sequence of MSL (referred to as MSL-1-MSL-15; SEQ ID NO: 110-124, respectively) were synthesized and their ability to stimulate T cell proliferation and IFN-γ production in a CD4+ Tcell line generated against M. tuberculosis culture filtrate was examined as described below. The peptides referred to as MSL-10 (SEQ ID NO: 119) and MSL-11 (SEQ ID NO: 120) were found to show the highest level of reactivity.

Comparison of the determined cDNA sequence for Tb838 with those in the gene bank revealed identity to the previously isolated M. tuberculosis cosmid SCY07H7. Comparison of the determined cDNA sequences for the clone Tb962 with those in the gene bank revealed some homology to two previously identified M. tuberculosis cosmids, one encoding a portion of bactoferritin. However, recombinant bactoferritin was not found to be reactive with the T cell line used to isolate Tb962.

The clone Tb470, described above, was used to recover a full-length open reading (SEQ ID NO: 125) that showed homology with TbH9 and was found to encode a 40 kDa antigen, referred to as Mtb40. The determined amino acid sequence for Mtb40 is provided in SEQ ID NO: 126. Similarly, Subsequent Studies LED to the Isolation of the Full-Length cDNA Sequence for TB43 1, Provided in SEQ ID NO: 83, which was determined to contain an open reading frame encoding Mtb40. Tb470 and Tb431 were also found to contain a potential open reading frame encoding a U-ORF-like antigen.

Screening of an M. tuberculosis Erdman cDNA expression library with multiple CD4+ Tcell lines generated against M. tuberculosis culture filtrate, resulted in the isolation of three clones, referred to as Tb366, Tb433 and Tb439. The determined EDNA sequences for Tb366, Tb433 and Tb439 are provided in SEQ ID NO: 127, 128 and 129, respectively. Comparison of these sequences with those in the gene bank revealed no significant homologies to Tb366. Tb433 was found to show some 30 homology to the previously identified M. tuberculosis antigen MPT83. Tb439 was found to show 100% identity to the previously isolated M. tuberculosis cosmid SCY02B10.

A CD4+ T cell line was generated against M. tuberculosis PPD, essentially described above, and used to screen the above M. tuberculosis Erdman cDNA expression library. One reactive clone (referred to as Tb372) was isolated, with the determined cDNA sequences being provided in SEQ ID NO: 130 and 131. Comparison of these sequences with those in the gene bank revealed no significant homologies.

In further studies, screening of an M. tuberculosis cDNA expression library with a CD4+ T cell line generated against dendritic cells that had been infected with tuberculosis for 8 days, as described above, led to the isolation of two clones referred to as Tb390R5C6 and Tb390R2C11. The determined cDNA sequence for Tb390R5C6 is provided in SEQ ID NO: 132, with the determined cDNA sequences for Tb390R2C11 being provided in SEQ ID NO: 133 and 134. Tb390R5C6 was found to show 100% identity to a previously identified M. tuberculosis cosmid.

In subsequent studies, the methodology described above was used to screen an M. tuberculosis genomic DNA library prepared as follows. Genomic DNA from M. tuberculosis Erdman strain was randomly sheared to an average size of 2 kb, and blunt ended with Klenow polymerase, followed by the addition of EcoRI adaptors. The insert was subsequently ligated into the Screen phage vector (Novagen, Madison, Wis.) and packaged in vitro using the PhageMaker extract (Novagen). The phage library (referred to as the Erd λScreen library) was amplified and a portion was converted into a plasmid expression library by an autosubcloning mechanism using the E. coli strain BM25.8 (Novagen). Plasmid DNA was purified from BM25.8 cultures containing the pSCREEN recombinants and used to transform competent cells of the expressing host strain BL21 (DE3)pLysS. Transformed cells were aliquoted into 96 well microtiter plates with each well containing a pool size of approximately 50 colonies. Replica plates of the 96 well plasmid library format were induced with IPTG to allow recombinant protein expression. Following induction, the plates were centrifuged to pellet the E. coli which was used directly in T cell expression cloning of a CD4+ T cell line prepared from a PPD-positive donor (donor 160) as described above. Pools containing E. coli expressing M. tuberculosis T cell antigens were subsequently broken down into individual colonies and reassayed in a similar fashion to identify positive hits.

Screening of the T cell line from donor 160 with one 96 well plate of the Erd λScreen library provided a total of nine positive hits. Previous experiments on the screening of the pBSK library described above with T cells from donor 160 suggested that most or all of the positive clones would be TbH-9, Tb38-1 or MTI (disclosed in U.S. patent application Ser. No. 08/533,634) or variants thereof. However, Southern analysis revealed that only three wells hybridized with a mixed probe of TbH-9, Tb38-1 and MTI. Of the remaining six positive wells, two were found to be identical. The determined 5′ cDNA sequences for two of the isolated clones (referred to as Y1-26C1 and Y1-86C11) are provided in SEQ ID NO: 135 and 136, respectively. The full length cDNA sequence for the isolated clone referred to as hTcc#1 is provided in SEQ ID NO: 137, with the corresponding predicted amino acid sequence being provided in SEQ ID NO: 138. Comparison of the sequences of hTcc#1 to those in the gene bank as described above, revealed some homology to the previously isolated M. tuberculosis cosmid MTCY07H7B.06.

EXAMPLE 2

Induction of T Cell Proliferation and Interferon-γ Production by M. Tuberculosis Antigens

The ability of recombinant M. tuberculosis antigens to induce T cell proliferation and interferon-γ production may be determined as follows.

Proteins may be induced by IPTG and purified by gel elution, as described in Skeiky et al. J. Exp. Med., 1995, 181:1527-1537. The purified polypeptides are then screened for the ability to induce T-cell proliferation in PBMC preparations. The PBMCs from donors known to be PPD skin test positive and whose T-cells are known to proliferate in response to PPD, are cultured in medium comprising RPMI 1640 supplemented with 10% pooled human serum and 50 μg/ml gentamicin. Purified polypeptides are added in duplicate at concentrations of 0.5 to 10 μg/mL. After six days of culture in 96-well round-bottom plates in a volume of 200 μl, 50 μl of medium is removed from each well for determination of IFN-γ levels, as described below. The plates are then pulsed with 1 μCi/well of tritiated thymidine for a further 18 hours, harvested and tritium uptake determined using a gas scintillation counter. Fractions that result in proliferation in both replicates three fold greater than the proliferation observed in cells cultured in medium alone are considered positive.

IFN-γ is measured using an enzyme-linked immunosorbent assay (ELISA). ELISA plates are coated with a mouse monoclonal antibody directed to human IFN-γ (PharMingen, San Diego, Calif.) in PBS for four hours at room temperature. Wells are then blocked with PBS containing 5% (W/V) non-fat dried milk for 1 hour at room temperature. The plates are washed six times in PBS/0.2% TWEEN20™ and samples diluted 1:2 in culture medium in the ELISA plates are incubated overnight at room temperature. The plates are again washed and a polyclonal rabbit anti-human IFN-γ serum diluted 1:3000 in PBS/10% normal goat serum is added to each well. The plates are then incubated for two hours at room temperature, washed and horseradish peroxidase-coupled anti-rabbit IgG (Sigma Chemical So., St. Louis, Mo.) is added at a 1:2000 dilution in PBS/5% non-fat dried milk. After a further two hour incubation at room temperature, the plates are washed and TMB substrate added. The reaction is stopped after 20 min with 1 N sulfuric acid. Optical density is determined at 450 nm using 570 nm as a reference wavelength. Fractions that result in both replicates giving an OD two fold greater than the mean OD from cells cultured in medium alone, plus 3 standard deviations, are considered positive.

EXAMPLE 3

Purification and Characterization of M. Tuberculosis Polypeptides using CD4+ T Cell Lines Generated from a Mouse M. Tuberculosis Model

Infection of C57BL/6 mice with M. tuberculosis results in the development of a progressive disease for approximately 2-3 weeks. The disease progression is then halted as a consequence of the emergence of a strong protective T cell-mediated immune response. This infection model was used to generate T cell lines capable of recognizing protective M. tuberculosis antigens.

Specifically, spleen cells were obtained from C57BL/6 mice infected with M. tuberculosis for 28 days and used to raise specific anti- M. tuberculosis T cell lines as described above. The resulting CD4+ T cell lines, in conjunction with normal antigen presenting (spleen) cells from C57BL/6 mice were used to screen the M. tuberculosis Erd λscreen library described above. One of the reactive library pools, which was found to be highly stimulatory of the T cells, was selected and the corresponding active clone (referred to as Y288C10) was isolated.

Sequencing of the clone Y288C10 revealed that it contains two potential genes, in tandem. The determined cDNA sequences for these two genes (referred to as mTCC#1 and mTCC#2) are provided in SEQ ID NO: 139 and 140, respectively, with the corresponding predicted amino acid sequences being provided in SEQ ID NO: 141 and 142, respectively. Comparison of these sequences with those in the gene bank revealed identity to unknown sequences previously found within the M. tuberculosis cosmid MTY21C12. The predicted amino acid sequences of mTCC#1 and mTCC#2 were found to show some homology to previously identified members of the TbH9 protein family, discussed above.

EXAMPLE 4

Synthesis of Synthetic Polypeptides

Polypeptides may be synthesized on a Millipore 9050 peptide synthesizer using FMOC chemistry with HPTU (O-Benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate) activation. A Gly-Cys-Gly sequence may be attached to the amino terminus of the peptide to provide a method of conjugation or labeling of the peptide. Cleavage of the peptides from the solid support may be carried out using the following cleavage mixture: trifluoroacetic acid:ethanedithiol:thioanisole:water:phenol (40:1:2:2:3). After cleaving for 2 hours, the peptides may be precipitated in cold methyl-t-butyl-ether. The peptide pellets may then be dissolved in water containing 0.1% trifluoroacetic acid (TFA) and lyophilized prior to purification by C18 reverse phase HPLC. A gradient of 0%-60% acetonitrile (containing 0.1% TFA) in water (containing 0.1% TFA) may be used to elute the peptides. Following lyophilization of the pure fractions, the peptides may be characterized using electrospray mass spectrometry and by amino acid analysis.

From the foregoing, it will be appreciated that, although specific embodiments of the invention have been described herein for the purpose of illustration, various modifications may be made without deviating from the spirit and scope of the invention.

144 1886 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 1CGCTCTGGTG ACCACCAACT TCTTCGGTGT CAACACCATC CCGATCGCCC TCAACGAGGC 60CGACTACCTG CGCATGTGGA TCCAGGCCGC CACCGTCATG AGCCACTATC AAGCCGTCGC 120GCACGAAATC TGGTGTCTCC ATGAATANGC CAGTTCGGGA AAGCCGTGGG CCAGTATCAC 180CACGGGTGCG CCGGGCTCAC CGGCCTCGAC CACTCGCAGT CGCACGCCGT TGGTATCAAC 240TAACCGTNCN GTANGTGCGC CCATCGTCTC ACCAAATCAC ACCGGGCACC GGCCTGAGAA 300GGGCTTGGGG AGCANCCAGA GGCGATTGTC GCGGGTGCTG CCGCGCATCA TTGATCGGCC 360GGCCGGACCA NTCGGGCCTC CCTTGACGTC CGGATCNCAC TTCCTGTGCA GCTGGCATGG 420CTACAGCTCA CAGTGACTGC CCCACGATTG CCGGCCAGGT CCAGTTCAAA TTCCGGTGAA 480TTCGCGGACA AAAGCAGCAG GTCAACCAAC CGCAGTCAGT CGAGGGTCCC AAACGTGAGC 540CAATCGGTGA AATGGCTTGC TGCAGTGACA CCGGTCACAG GCTTAGCCGA CAGCACCGGA 600ATAGCTCAGG CGGGCTATAG AGTCCTATAG AAACATTTGC TGATAGAATT AACCGCTGTC 660TTGGCGTGAT CTTGATACGG CTCGCCGTGC GACCGGTTGG CTCAGTAGCT GACCACCATG 720TAACCCATCC TCGGCAGGTG TCTACTAAGG CGAGACACCG CATTGGTGGG GCTGCATCGC 780AAATCGGTCC GAGCATGTAG CACTGCCGTT ATCCCGGGAT AGCAAACCAC CCGGAACCAG 840GGCTATCCCA GTCGCTCTCC GACGGAGGCC GTTTCGCTTT CCGTTGCCCG ATAACTCCCG 900AGTGGATATC GGCGTTATCA NATTCAGGCT TTTCTTCGCA AGGTACCGGT GTTCGCTATA 960TTCGGATATC TCGGACGGAT AATTACTAAA ACTTCAGTGG TTTAGATAAG GCCGCCGCAA 1020TACTTCGCCG ATCTTGCCGA GCGCAACGGA TTTCCATCGT CGGTTTTCGT CGCCTTATCA 1080AACATGATCG GAGATAATGA CAGATCGGCC TAGCTAGGTG TTTAGCGGAC GCGATTTAGG 1140ACAACCGAGA TTTGCTTTGC CTCGCAACCA TGAGAGCGCC CCGCTTCGAC GCCGAATCGG 1200GTGAGTGATG GTGGGTTAGC ACAGCCCTGA TTGCGCCACC GGCGAGGTGA TTGTGCCCGC 1260CACGAGGCCG CCGCCGGCTA GCCCCATGAG CACGNTATAT AGACTCTCCT GCAACAGATC 1320TCATACCGAT CGAAGGCGAA GCGCAGGCAT CGACGTCGGA GACACTGCCT TGGGATCGCG 1380CCGCCTACAC GGCGGTTGGC GCATTGTCGC AGCGCAGTTG CAGGAGGGCA AATGTGCGCA 1440GACGATGTAG TCGACAACAA GTGNACATGC CGTCTTCACG AACTCAAAAC TGACGATCTG 1500CTTAGCATGA AAAAAACTGT TGACATCGGC CAAGCATGAC AGCCAGACTG TAGGCCTACG 1560CGTGCAATGC AGAACCAAGG NTATGCATGG AATCGACGAC CGTTGAGATA GGCGGCAGGC 1620ATGAGCAGAG CGTTCATCAT CGATCCAACG ATCAGTGCCA TTGACGGCTT GTACGACCTT 1680CTGGGGATTG GAATACCCAA CCAAGGGGGT ATCCTTTACT CCTCACTAGA GTACTTCGAA 1740AAAGCCCTGG AGGAGCTGGC AGCAGCGTTT CCGGGTGATG GCTGGTTAGG TTCGGCCGCG 1800GACAAATACG CCGGCAAAAA CCGCAACCAC GTGAATTTTT TCCAGGAACT GGCAGACCTC 1860GATCGTCAGC TCATCAGCCT GATCCA 1886 2305 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 2GGCACGCGCT GGCCGCGCAA TACACCGAAA TTGCAACGGA ACTCGCAAGC GTGCTCGCTG 60CGGTGCAGGC AAGCTCGTGG CAGGGGCCCA GCGCCGACCG GTTCGTCGTC GCCCATCAAC 120CGTTCCGGTA TTGGCTAACC CACGCTGCCA CGGTGGCCAC CGCAGCAGCC GCCGCGCACN 180AAACGGCCGC CGCCGGGTAT ACGTCCGCAT TGGGGGGCAT GCCTACGCTA GCCGAGTTGG 240CGGCCAACCA TGCCATGCAC GGCGCTCTGG TGACCACCAA CTTCTTCGGT GTCAACACCA 300TCCCGATCGC CCTCAACGAG GCCGACTACC TGCGCATGTG GATCCAGGCC GCCACCGTCA 360TGAGCCACTA TCAAGCCGTC GCGCACGAAA GCGTGGCGGC GACCCCCAGC ACGCCGCCGG 420CGCCGCAGAT AGTGACCAGT GCGGCCAGCT CGGCGGCTAG CAGCAGCTTC CCCGACCCGA 480CCAAATTGAT CCTGCAGCTA CTCAAGGATT TCCTGGAGCT GCTGCGCTAT CTGGCTGTTG 540AGCTGCTGCC GGGGCCGCTC GGCGACCTCA TCGCCCAGGT GTTGGACTGG TTCATCTCGT 600TCGTGTCCGG TCCAGTCTTC ACGTTTCTCG CCTACCTGGT GCTGGACCCA CTGATCTATT 660TCGGACCGTT CGCCCCGCTG ACGAGTCCGG TCCTGTTGCC TGCTGTGGAG TTACGCAACC 720GCCTCAAAAC CGCCACCGGA CTGACGCTGC CACCTACCGT GATTTTCGAT CATCCCACTC 780CCACTGCGGT CGCCGAGTAT GTCGCCCAGC AAATGTCTGG CAGCCGCCCA ACGGAATCCG 840GTGATCCGAC GTCGCAGGTT GTCGAACCCG CTCGTGCCGA ATTCGGCACG AGTGCTGTTC 900ATCAAATCCC CCCGAGACCT GCGGACACCC GGCGCGCTTG CCGACATCGA GATGATGTCC 960CGCGAGATAG CAGAATTGCC CAACATCGTG ATGGTGCGGG GCTTGACCCG ACCGAACGGG 1020GAACCTCTGA AGGAGACCAA GGTCTCGTTT CAGGCTGGTG AAGTGGGCGG CAAGCTCGAC 1080GAAGCGACCA CCCTGCTCGA AGAGCACGGA GGCGAGCTGG ACCAGCTGAC CGGCGGTGCG 1140CACCAGTTGG CCGACGCCCT CGCCCAAATA CGCAACGAAA TCAATGGGGC CGTGGCCAGC 1200TCGAGCGGGA TAGTCAACAC CCTGCAGGCC ATGATGGACC TGATGGGCGG TGACAAGACC 1260ATCCGACAAC TGGAAAATGC GTCCCAATAT GTCGGGCGCA TGCGGGCTCT GGGGGACAAT 1320CTGAGCGGGA CCGTCACCGA TGCCGAACAA ATCGCCACTT GGGCCAGCCC TATGGTCAAC 1380GCCCTCAACT CCAGCCCGGT GTGTAACAGC GATCCCGCCT GTCGGACGTC GCGCGCACAG 1440TTGGCGGCGA TTGTCCAGGC GCAGGACGAC GGCCTGCTCA GGTCCATCAG AGCGCTAGCC 1500GTCACCCTGC AACAGACGCA GGAATACCAG ACACTCGCCC GGACGGTGAG CACACTGGAC 1560GGGCAACTGA AGCAAGTCGT CAGCACCCTC AAAGCGGTCG ACGGCCTACC CACCAAATTG 1620GCTCAAATGC AGCAAGGAGC CAACGCTCTC GCCGACGGCA GCGCAGCGCT GGCGGCAGGC 1680GTGCAGGAAT TGGTCGATCA GGTCAAAAAG ATGGGCTCAG GGCTCAACGA GGCCGCCGAC 1740TTCCTGTTGG GGATCAAGCG GGATGCGGAC AAGCCGTCAA TGGCGGGCTT CAACATTCCA 1800CCGCAGATTT TTTCGAGGGA CGAGTTCAAG AAGGGCGCCC AGATTTTCCT GTCGGCCGAT 1860GGTCATGCGG CGCGGTACTT CGTGCAGAGC GCGCTGAATC CGGCCACCAC CGAGGCGATG 1920GATCAGGTCA ACGATATCCT CCGTGTTGCG GATTCCGCGC GACCGAATAC CGAACTCGAG 1980GATGCCACGA TAGGTCTGGC GGGGGTTCCG ACTGCGCTGC GGGATATCCG CGACTACTAC 2040AACAGCGATA TGAAATTCAT CGTCATTGCG ACGATCGTTA TCGTATTCTT GATTCTCGTC 2100ATTCTGNTGC GCGCACTTGT GGNTCCGATA TATCTGATAG GCTCGGTGCT GATTTCTTAC 2160TTGTCGGCCC TAGGCATAGG AACTTTCGTT TTCCAATTGA TACTGGGCCA GGAAATGCAT 2220TGGAGCCTGC CGGGACTGTC CTTCATATTA TTGGTTGCCA TCGGCGCTGA CTACAACATG 2280CTGCTCATTT CACGCATCCG CGACG 2305 1742 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 3CCGCTCTCTT TCAACGTCAT AAGTTCGGTG GGCCAGTCGG CCGCGCGTGC ATATGGCACC 60AATAACGCGT GTCCCATGGA TACCCGGACC GCACGACGGT AGAGCGGATC AGCGCAGCCG 120GTGCCGAACA CTACCGCGTC CACGCTCAGC CCTGCCGCGT TGCGGAAGAT CGAGCCCAGG 180TTCTCATGGT CGTTAACGCC TTCCAACACT GCGACGGTGC GCGCCCCGGC GACCACCTGA 240GCAACGCTCG GCTCCGGCAC CCGGCGCGCG GCTGCCAACA CCCCACGATT GAGATGGAAG 300CCGATCACCC GTGCCATGAC ATCAGCCGAC GCTCGATAGT ACGGCGCGCC GACACCGGCC 360AGATCATCCT TGAGCTCGGC CAGCCGGCGG TCGGTGCCGA ACAGCGCCAG CGGCGTGAAC 420CGTGAGGCCA GCATGCGCTG CACCACCAGC ACACCCTCGG CGATCACCAA CGCCTTGCCG 480GTCGGCAGAT CGGGACNACN GTCGATGCTG TTCAGGTCAC GGAAATCGTC GAGCCGTGGG 540TCGTCGGGAT CGCAGACGTC CTGAACATCG AGGCCGTCGG GGTGCTGGGC ACAACGGCCT 600TCGGTCACGG GCTTTCGTCG ACCAGAGCCA GCATCAGATC GGCGGCGCTG CGCAGGATGT 660CACGCTCGCT GCGGTTCAGC GTCGCGAGCC GCTCAGCCAG CCACTCTTGC AGAGAGCCGT 720TGCTGGGATT AATTGGGAGA GGAAGACAGC ATGTCGTTCG TGACCACACA GCCGGAAGCC 780CTGGCAGCTG CGGCGGCGAA CCTACAGGGT ATTGGCACGA CAATGAACGC CCAGAACGCG 840GCCGCGGCTG CTCCAACCAC CGGAGTAGTG CCCGCAGCCG CCGATGAAGT ATCAGCGCTG 900ACCGCGGCTC AGTTTGCTGC GCACGCGCAG ATGTACCAAA CGGTCAGCGC CCAGGCCGCG 960GCCATTCACG AAATGTTCGT GAACACGCTG GTGGCCAGTT CTGGCTCATA CGCGGCCACC 1020GAGGCGGCCA ACGCAGCCGC TGCCGGCTGA ACGGGCTCGC ACGAACCTGC TGAAGGAGAG 1080GGGGAACATC CGGAGTTCTC GGGTCAGGGG TTGCGCCAGC GCCCAGCCGA TTCAGNTATC 1140GGCGTCCATA ACAGCAGACG ATCTAGGCAT TCAGTACTAA GGAGACAGGC AACATGGCCT 1200CACGTTTTAT GACGGATCCG CATGCGATGC GGGACATGGC GGGCCGTTTT GAGGTGCACG 1260CCCAGACGGT GGAGGACGAG GCTCGCCGGA TGTGGGCGTC CGCGCAAAAC ATTTCCGGTG 1320CGGGCTGGAG TGGCATGGCC GAGGCGACCT CGCTAGACAC CATGACCTAG ATGAATCAGG 1380CGTTTCGCAA CATCGTGAAC ATGCTGCACG GGGTGCGTGA CGGGCTGGTT CGCGACGCCA 1440ACAANTACGA ACAGCAAGAG CAGGCCTCCC AGCAGATCCT GAGCAGNTAG CGCCGAAAGC 1500CACAGCTGNG TACGNTTTCT CACATTAGGA GAACACCAAT ATGACGATTA ATTACCAGTT 1560CGGGGACGTC GACGCTCATG GCGCCATGAT CCGCGCTCAG GCGGCGTCGC TTGAGGCGGA 1620GCATCAGGCC ATCGTTCGTG ATGTGTTGGC CGCGGGTGAC TTTTGGGGCG GCGCCGGTTC 1680GGTGGCTTGC CAGGAGTTCA TTACCCAGTT GGGCCGTAAC TTCCAGGTGA TCTACGAGCA 1740GG 1742 2836 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 4GTTGATTCCG TTCGCGGCGC CGCCGAAGAC CACCAACTCC GCTGGGGTGG TCGCACAGGC 60GGTTGCGTCG GTCAGCTGGC CGAATCCCAA TGATTGGTGG CTCNGTGCGG TTGCTGGGCT 120CGATTACCCC CACGGAAAGG ACGACGATCG TTCGTTTGCT CGGTCAGTCG TACTTGGCGA 180CGGGCATGGC GCGGTTTCTT ACCTCGATCG CACAGCAGCT GACCTTCGGC CCAGGGGGCA 240CAACGGCTGG CTCCGGCGGA GCCTGGTACC CAACGCCACA ATTCGCCGGC CTGGGTGCAG 300GCCCGGCGGT GTCGGCGAGT TTGGCGCGGG CGGAGCCGGT CGGGAGGTTG TCGGTGCCGC 360CAAGTTGGGC CGTCGCGGCT CCGGCCTTCG CGGAGAAGCC TGAGGCGGGC ACGCCGATGT 420CCGTCATCGG CGAAGCGTCC AGCTGCGGTC AGGGAGGCCT GCTTCGAGGC ATACCGCTGG 480CGAGAGCGGG GCGGCGTACA GGCGCCTTCG CTCACCGATA CGGGTTCCGC CACAGCGTGA 540TTACCCGGTC TCCGTCGGCG GGATAGCTTT CGATCCGGTC TGCGCGGCCG CCGGAAATGC 600TGCAGATAGC GATCGACCGC GCCGGTCGGT AAACGCCGCA CACGGCACTA TCAATGCGCA 660CGGCGGGCGT TGATGCCAAA TTGACCGTCC CGACGGGGCT TTATCTGCGG CAAGATTTCA 720TCCCCAGCCC GGTCGGTGGG CCGATAAATA CGCTGGTCAG CGCGACTCTT CCGGCTGAAT 780TCGATGCTCT GGGCGCCCGC TCGACGCCGA GTATCTCGAG TGGGCCGCAA ACCCGGTCAA 840ACGCTGTTAC TGTGGCGTTA CCACAGGTGA ATTTGCGGTG CCAACTGGTG AACACTTGCG 900AACGGGTGGC ATCGAAATCA ACTTGTTGCG TTGCAGTGAT CTACTCTCTT GCAGAGAGCC 960GTTGCTGGGA TTAATTGGGA GAGGAAGACA GCATGTCGTT CGTGACCACA CAGCCGGAAG 1020CCCTGGCAGC TGCGGCGGCG AACCTACAGG GTATTGGCAC GACAATGAAC GCCCAGAACG 1080CGGCCGCGGC TGCTCCAACC ACCGGAGTAG TGCCCGCAGC CGCCGATGAA GTATCAGCGC 1140TGACCGCGGC TCAGTTTGCT GCGCACGCGC AGATGTACCA AACGGTCAGC GCCCAGGCCG 1200CGGCCATTCA CGAAATGTTC GTGAACACGC TGGTGGCCAG TTCTGGCTCA TACGCGGCCA 1260CCGAGGCGGC CAACGCAGCC GCTGCCGGCT GAACGGGCTC GCACGAACCT GCTGAAGGAG 1320AGGGGGAACA TCCGGAGTTC TCGGGTCAGG GGTTGCGCCA GCGCCCAGCC GATTCAGCTA 1380TCGGCGTCCA TAACAGCAGA CGATCTAGGC ATTCAGTACT AAGGAGACAG GCAACATGGC 1440CTCACGTTTT ATGACGGATC CGCATGCGAT GCGGGACATG GCGGGCCGTT TTGAGGTGCA 1500CGCCCAGACG GTGGAGGACG AGGCTCGCCG GATGTGGGCG TCCGCGCAAA ACATTTCCGG 1560TGCGGGCTGG AGTGGCATGG CCGAGGCGAC CTCGCTAGAC ACCATGACCT AGATGAATCA 1620GGCGTTTCGC AACATCGTGA ACATGCTGCA CGGGGTGCGT GACGGGCTGG TTCGCGACGC 1680CAACAACTAC GAACAGCAAG AGCAGGCCTC CCAGCAGATC CTGAGCAGCT AGCGCCGAAA 1740GCCACAGCTG CGTACGCTTT CTCACATTAG GAGAACACCA ATATGACGAT TAATTACCAG 1800TTCGGGGACG TCGACGCTCA TGGCGCCATG ATCCGCGCTC AGGCGGCGTC GCTTGAGGCG 1860GAGCATCAGG CCATCGTTCG TGATGTGTTG GCCGCGGGTG ACTTTTGGGG CGGCGCCGGT 1920TCGGTGGCTT GCCAGGAGTT CATTACCCAG TTGGGCCGTA ACTTCCAGGT GATCTACGAG 1980CAGGCCAACG CCCACGGGCA GAAGGTGCAG GCTGCCGGCA ACAACATGGC GCAAACCGAC 2040AGCGCCGTCG GCTCCAGCTG GGCCTAAAAC TGAACTTCAG TCGCGGCAGC ACACCAACCA 2100GCCGGTGTGC TGCTGTGTCC TGCAGTTAAC TAGCACTCGA CCGCTGAGGT AGCGATGGAT 2160CAACAGAGTA CCCGCACCGA CATCACCGTC AACGTCGACG GCTTCTGGAT GCTTCAGGCG 2220CTACTGGATA TCCGCCACGT TGCGCCTGAG TTACGTTGCC GGCCGTACGT CTCCACCGAT 2280TCCAATGACT GGCTAAACGA GCACCCGGGG ATGGCGGTCA TGCGCGAGCA GGGCATTGTC 2340GTCAACGACG CGGTCAACGA ACAGGTCGCT GCCCGGATGA AGGTGCTTGC CGCACCTGAT 2400CTTGAAGTCG TCGCCCTGCT GTCACGCGGC AAGTTGCTGT ACGGGGTCAT AGACGACGAG 2460AACCAGCCGC CGGGTTCGCG TGACATCCCT GACAATGAGT TCCGGGTGGT GTTGGCCCGG 2520CGAGGCCAGC ACTGGGTGTC GGCGGTACGG GTTGGCAATG ACATCACCGT CGATGACGTG 2580ACGGTCTCGG ATAGCGCCTC GATCGCCGCA CTGGTAATGG ACGGTCTGGA GTCGATTCAC 2640CACGCCGACC CAGCCGCGAT CAACGCGGTC AACGTGCCAA TGGAGGAGAT CTCGTGCCGA 2700ATTCGGCACG AGGCACGAGG CGGTGTCGGT GACGACGGGA TCGATCACGA TCATCGACCG 2760GCCGGGATCC TTGGCGATCT CGTTGAGCAC GACCCGGGCC CGCGGGAAGC TCTGCGACAT 2820CCATGGGTTC TTCCCG 2836 900 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 5AACATGCTGC ACGGGGTGCG TGACGGGCTG GTTCGCGACG CCAACAACTA CGAGCAGCAA 60GAGCAGGCCT CCCAGCAGAT CCTCAGCAGC TAACGTCAGC CGCTGCAGCA CAATACTTTT 120ACAAGCGAAG GAGAACAGGT TCGATGACCA TCAACTATCA GTTCGGTGAT GTCGACGCTC 180ACGGCGCCAT GATCCGCGCT CAGGCCGGGT TGCTGGAGGC CGAACATCAG GCCATCATTC 240GTGATGTGTT GACCGCGAGT GACTTTTGGG GCGGCGCCGG TTCGGCGGCC TGCCAGGGGT 300TCATTACCCA ATTGGGCCGT AACTTCCAGG TGATCTACGA ACAGGCCAAC GCCCACGGGC 360AGAAGGTGCA GGCTGCCGGC AACAACATGG CGCAAACCGA CAGCGCCGTC GGCTCCAGCT 420GGGCCTGACA CCAGGCCAAG GCCAGGGACG TGGTGTACGA GTGAAGGTTC CTCGCGTGAT 480CCTTCGGGTG GCAGTCTAGG TGGTCAGTGC TGGGGTGTTG GTGGTTTGCT GCTTGGCGGG 540TTCTTCGGTG CTGGTCAGTG CTGCTCGGGC TCGGGTGAGG ACCTCGAGGC CCAGGTAGCG 600CCGTCCTTCG ATCCATTCGT CGTGTTGTTC GGCGAGGACG GCTCCGACGA GGCGGATGAT 660CGAGGCGCGG TCGGGGAAGA TGCCCACGAC GTCGGTTCGG CGTCGTACCT CTCGGTTGAG 720GCGTTCCTGG GGGTTGTTGG ACCAGATTTG GCGCCAGATC TTCTTGGGGA AGGCGGTGAA 780CGCCAGCAGG TCGGTGCGGG CGGTGTCGAN GTGCTCGGCC ACCGCGGGGA GTTTGTCGGT 840CAGAGCGTCG AGTACCCGAT CATATTGGGC AACAACTGAT TCGGCGTTGG GCTGGTCGTA 900 1905 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 6GCTCGCCGGA TGTGGGCGTC CGCGCAAAAC ATTTCCGGTG CGGGCTGGAG TGGCATGGCC 60GAGGCGACCT CGCTAGACAC CATGGCCCAG ATGAATCAGG CGTTTCGCAA CATCGTGAAC 120ATGCTGCACG GGGTGCGTGA CGGGCTGGTT CGCGACGCCA ACAACTACGA GCAGCAAGAG 180CAGGCCTCCC AGCAGATCCT CAGCAGCTAA CGTCAGCCGC TGCAGCACAA TACTTTTACA 240AGCGAAGGAG AACAGGTTCG ATGACCATCA ACTATCAGTT CGGTGATGTC GACGCTCACG 300GCGCCATGAT CCGCGCTCAG GCCGGGTTGC TGGAGGCCGA GCATCAGGCC ATCATTCGTG 360ATGTGTTGAC CGCGAGTGAC TTTTGGGGCG GCGCCGGTTC GGCGGCCTGC CAGGGGTTCA 420TTACCCAGTT GGGCCGTAAC TTCCAGGTGA TCTACGAACA AGCCAACACC CACGGGCAGA 480AGGTGCAAGC TGCCGGCAAC AACATGGCGC AAACCGACAG CGCCGTCNGC TCCAGCTGGG 540CCTGACACCA GGCCAAGGCC AGGGACGTGG TGTACNAGTG AAGGTTCCTC GCGTGATCCT 600TCGGGTGGCA GTCTAGGTGG TCAGTGCTGG GGTGTTGGTG GTTTGCTGCT TGGCGGGTTC 660TTCGGTGCTG GTCAGTGCTG CTCGGGCTCG GGTGAGGACC TCGAGGCCCA GGTAGCGCCG 720TCCTTCGATC CATTCGTCGT GTTGTTCGGC GAGGACNGCT CCGACGANGC GGATGATCGA 780GGCGCGGTCG GGGAAGATGC CCACGACGTC GGTTCGGCGT CGTACCTCTC GGTTGAAGCG 840TTCCTGGGGG CCACCGCTTG GCGCCNANGC ACTCCACGCC AATTCGTCNC ACCTAACAGC 900GGTGGCCAAC GACTATGACT ACGACACCGT TTTTGCCAGG GCCCTCNAAA GGATCTGCGC 960GTCCCGGCGA CACGCTTTTT GCGATAAGTA CCTCCGGCAA TTCTATGAGT GTACTGCGGN 1020CCGCGAAAAC CGCAAGGGAG TTGGGTGTGA CGGTTNTTGC AAATGACGGG CGAATCCGGC 1080GGCCAGCTGG CAGAATTCGC AGATTTCTTG ATCAACGTCC CGTCACGCGA CACCGGGCGA 1140ATCCAGGAAT CTCACATCGT TTTTATTCAT GCGATCTCCG AACATGTCGA ACACGCGCTT 1200TTCGCGCCTC GCCAATAGGA AAGCCGATCC TTACGCGGCC ATTCGAAAGA TGGTCGCGGA 1260ACGTGCGGGA CACCAATGGT GTCTCTTCCT CGATAGAGAC GGGGTCATCA ATCGACAAGT 1320GGTCGGCGAC TACGTACGGA ACTGGCGGCA GTTTGAATGG TTGCCCGGGG CGGCGCGGGC 1380GTTGAAGAAG CTACGGGCAT GGGCTCCGTA CATCGTTGTC GTGACAAACC AGCAGGGCGT 1440GGGTGCCGGA TTGATGAGCG CCGTCGACGT GATGGTGATA CATCGGCACC TCCAAATGCA 1500GCTTGCATCC GATGGCGTGC TGATAGATGG ATTTCAGGTT TGCCCGCACC ACCGTTCGCA 1560GCGGTGTGGC TGCCGTAAGC CGAGACCGGG TCTGGTCCTC GACTGGCTCG GACGACACCC 1620CGACAGTGAG CCATTGCTGA GCATCGTGGT TGGGGACAGC CTCAGCGATC TTGACATTGG 1680CACACAACGT CGCCGCTGCT GCCGGTGCAT GTGCCAGTGT CCAGATAGGG GGCGCCAGTT 1740CTGGCGGTGT CGCTGACGCG TCATTTGACT CGCTCTGGGA GTTCGCTGTC GCAGTCGGAC 1800ATGCGCGGGG GGAGCGGGGC TAATGGCGAT CTTGCGCGGG CGAGCGCCGT NGCGGNTCGG 1860ACTNNGCGGT GGCGGGACAG ACGTGGAACC GTACTCGAGC CAGTT 1905 2921 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 7CGGGATGCCG TGGTGGTTGG TATTGCCCAA ACCCTGGCGC TGGTCCCCGG GGTATCCAGG 60TCCGGGTCGA CCATCAGCGC TGGACTGTTT CTCGGACTCG ACCGTGAACT GGCCGCCCGA 120TTCGGATTCC TGCTGGCCAT TCCAGCGGTG TTCGCCTCCG GGTTGTTCTC GTTGCCCGAC 180GCATTCCACC CGGTAACCGA GGGCATGAGC GCTACTGGCC CGCAGTTGCT GGTGGCCACC 240CTGATCGCGT TCGTCCTCGG TCTGACCGCG GTGGCCTGGC TGCTGCGGTT TCTGGTGCGA 300CACAACATGT ACTGGTTCGT CGGCTACCGG GTGCTCGTCG GGACGGGCAT GCTCGTGCTG 360CTGGCTACCG GGACGGTAGC CGCGACATGA CCGTCATCTT GCTACGCCAT GCCCGTTCCA 420CCTCGAACAC CGCGGGCGTG CTGGCCGGCC GGTCCGGCGT CGACCTCGAC GAGAAGGGGC 480GCGAGCAGGC CACCGGGTTG ATCGATCGAA TTGGTGACCT GCCGATCCGG GCGGTCGCGT 540CTTCTCCAAT GCTGCGGTGT CAACGCACCG TCGAACCGCT GGCCGAGGCG CTGTGCCTGG 600AGCCGCTCAT CGATGACCGG TTCTCCGAAG TCGACTACGG CGAATGGACT GGCAGAAAAA 660TCGGTGACCT GGTCGACGAG CCGTTGTGGC GGGTAGTCCA GGCCCACCCC AGCGCGGCGG 720TGTTTCCCGG CGGTGAGGGT TTGGCGCAGG TGCAGACGTG GTTGTCCTGA CGGATTTCCA 780TGCCGGGGAA CACCAAGACC GGATCGGCAC TGGCGGTCGC CGGCGAAAAC CCGGCCGCCA 840ATAGGGCGAC CGTCGCTGCG AATGCGCGTG GTACCAGGCG GACCACCTTG AACTCCCATC 900CGTCGGGGCC AAGCGCATCG CCCGCCGCCG GTTACGGCTA AGGCGTACCA AAACCCGACG 960GTAATACTTC GGCAATGTCG GGTCNCGACG TTACCGAGAC GTGACCAGNG AGGCNGCGGC 1020ATTGGATTTA TCGATGGTGC GCGGTTCCCA NCCCGGCGGT CCGAANACGT AGCCCAGCCG 1080ATCCCGCAGA CGTGTTGCCG ACCGCCAGTC ACGCACGATC GCCACGTACT CGCGGGTCTG 1140CAGCTTCCAG ATGTTGAACG TGTCGACCCG CTTGGTCAGG CCATAATGCG GTCGGAATAG 1200CTCCGGCTGA AAGCTACCGA ACAGGCGGTC CCAGATGATG AGGATGCCGC CATAGTTCTT 1260GTCCANATAC ACCGGGTCCA TTCCGTGGTG GACCCGGTGG TGCGACGGGG TATTGAAGAC 1320GAATTCGAAC CACCGCGGCA GCCTGTCGAT CCGCTCGGTG TGCACCCAGA ACTGGTAGAT 1380CAAGTTCAGC GACCAATTGC AGAACACCAT CCAAGGGGGA AGCCCCATCA GTGGCAGCGG 1440AACCCACATG AGAATCTCGC CGCTGTTGTT CCANTTTCTG GCGCAGCGCG GTGGCGAAGT 1500TGAAGTATTC GCTGGAGTGA TGCGCCTGGT GGGTAGCCCA GATCAGCCGA ACTCGGTGGG 1560CGATGCGGTG ATAGGAGTAG TACAGCAGAT CGACACCAAC GATCGCGATC ACCCAGGTGT 1620ACCACCGGTG GGCGGACAGC TGCCAGGGGG CAAGGTAGGC ATAGATTGCG GCATAACCGA 1680GCAGGGCAAG GGACTTCCAG CCGGCGGTGG TGGCTATCGA AACCAGCCCC ATCGAGATGC 1740TGGCCACCGA GTCGCGGGTG AGGTAAGCGC CCGAGGCGGG CCGTGGCTGC CCGGTAGCAG 1800CGGTCTCGAT GCTTTCCAGC TTGCGGGCCG CCGTCCATTC GAGAATCAGC AGCAATAGAA 1860AACATGGAAT GGCGAACAGT ACCGGGTCCC GCATTTCCTC GGGCAGCGCT GAGAAGAATC 1920CGGCGACGGC ATGGCCGAGG CGACCTCGNT AGACACCATG ACCCAGATGA ATCAGGCGTT 1980TCGCAACATC GTGAACATGC TGCACGGGGT GCGTGACGGG CTGGTTCGCG ACGCCAACAA 2040NTACGAACAG CAAGAGCAGG CCTCCCAGCA GATCCTCAGC AGCTGACCCG GCCCGACGAC 2100TCAGGAGGAC ACATGACCAT CAACTATCAA TTCGGGGACG TCGACGCTCA CGGCGCCATG 2160ATCCGCGCTC AGGCCGGGTC GCTGGAGGCC GAGCATCAGG CCATCATTTC TGATGTGTTG 2220ACCGCGAGTG ACTTTTGGGG CGGCGCCGGT TCGGCGGCCT GCCAGGGGTT CATTACCCAG 2280CTGGGCCGTA ACTTCCAGGT GATNTACGAG CAGGCCAACG CCCACGGGCA GAAGGTGCAG 2340GCTGCCGGCA ACAACATGGC ACAAACCGAC AGCGCCGTCG GCTCCAGCTG GGCATAAAGN 2400TGGCTTAAGG CCCGCGCCGT CAATTACAAC GTGGCCGCAC ACCGGTTGGT GTGTGGCCAC 2460GTTGTTATCT GAACGACTAA CTACTTCGAC CTGCTAAAGT CGGCGCGTTG ATCCCCGGTC 2520GGATGGTGCT GAACTGGGAA GATGGCCTCA ATGCCCTTGT TGCGGAAGGG ATTGAGGCCA 2580TCGTGTTTCG TACTTTAGGC GATCAGTGCT GGTTGTGGGA GTCGCTGCTG CCCGACGAGG 2640TGCGCCGACT GCCCGAGGAA CTGGCCCGGG TGGACGCATT GTTGGACGAT CCGGCGTTCT 2700TCGCCCCGTT CGTGCCGTTC TTCGACCCGC GCAGGGGCCG GCCGTCGACG CCGATGGAGG 2760TCTATCTGCA GTTGATGTTT GTGAAGTTCC GCTACCGGCT GGGCTATGAG TCGCTGTGCC 2820GGGAGGTGGC TGATTCGATC ACCTGACGGC GGTTTTGCCG CATTGCGCTG GACGGGTCGG 2880TGCCGCATCC GACCACATTG ATGAAGCTCA CCACGCGTTG C 2921 1704 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 8CGCGATCGTC GTCAACGANG TCGACCGTCA CCACGGACTG ATCAACAAGT TCGCAGGCGA 60CGCCGCCCTG GCCATCTTCG GAGCCCCGAA CCGCCTCGAC CGTCCCGAAG ACGCCGCGCT 120GGCCGCCGCC CGGGCCATAN CCGANCGGCT GGCCNACGAG ATGCCCGAGG TCCAAGCCGG 180CATCGGGGTG GCGGCAGGCC ANATCGTCGC CGGCAATGTC GGCGCCAAGC AAAGATTCNA 240ATACACAGTG GTCGGCAAGC CGGTCAACCA NGCGGCCCGA TTGTGCGAAC TGGCCAAATC 300ACACCCCGCG CGATTGGGTC TCGCCCGCTC GGCTCATGGT CACCCAATTC AAGGACTACT 360TTGGCCTGGC GCACGACCTG CCGAAGTGGG CGAGTGAAGG CGCCAAAGCC GCCGGTGAGG 420CCGCCAAGGC GTTGCCGGCC GCCGTTCCGG CCATTCCGAG TGCTGGCCTG AGCGGCGTTG 480CGGGCGCCGT CGGTCAGGCG GCGTCGGTCG GGGGATTGAA GGTTCCGGCC GTTTGGACCG 540CCACGACCCC GGCGGCGAGC CCCGCGGTGC TGGCGGCGTC CAACGGCCTC GGAGCCGCGG 600CCGCCGCTGA AGGTTCGACA CACGCGTTTG GCGGGATGCC GCTCATGGGT ANCGGTGCCG 660GACGTGCGTT TAACAACTTC GCTGCCCCTC GATACGGATT CAAGCCGACC GTGATCGCCC 720AACCGCCGGC TGGCGGATGA CCAACTACGT TCGTTGATCG AGGATCGAAT TCNACGATTC 780AAAGGGAGGA ATTCATATGA CCTCNCGTTT TATGACGGAT CCGCACGCNA TNCGGGACAT 840GGCGGGCCGT TTTGAGGTGC ACGCCCAGAC GGTGGAGGAC GAGGCTNGCN GGATGTGGGC 900GTCCGCGCAA AACATTTCCG GTGCGGGCTG GAGTGGCATG GCCGAGGCGA CCTCGNTAGA 960CACCATGGCC CAGATGAATC AGGCGTTTCN CAACATCGTG AACATGCTGC ACGGGGTGNG 1020TGACGGGCTG GTTCGCGACG CCAACAACTA CGAACAGCAA GAGCAGGCCT CCCAGCAGAT 1080CCTCAGCAGC TGACCCGGCC CGACGACTCA GGAGGACACA TGACCATCAA CTATCAATTC 1140GGGGACGTCG ACGCTCATGG CGCCATGATC CGCGCTNTGG CCGGGTTGCT GGAGGCCGAG 1200CATCAGGCCA TCATTTCTGA TGTGTTGACC GCGAGTGACT TTTGGGGCGG CGCCGGTTCG 1260GCGGCCTGCC AGGGGTTCAT TACCCAGTTG GGCCGTAACT TCCAGGTGAT TTACGAGCAG 1320GCCAACGCCC ACGGGCAGAA GGTGCAGGCT GCCGGCAACA ACATGGCACA AACCGACAGC 1380GCCGTNGGNT CCAGCTGGGC CTAACCCGGG TCNTAAGTTG GGTCCGCGCA GGGCGGGCCG 1440ATCAGCGTNG ACTTTGGCGC CCGATACACG GGCATNTTNT NGTCGGGAAC ACTGCGCCCG 1500CGTCAGNTGC CCGCTTCCCC TTGTTNGGCG ACGTGCTCGG TGATGGCTTT GACGACCGCT 1560TCGCCGGCGC GGCCAATCAA TTGGTCGCGC TTGCCTNTAG CCCATTCGTG CGACGCCCGC 1620GGCGCCGCGA GTTGTCCCTT GAAATAAGGA ATCACAGCAC GGGCGAACAG CTCATAGGAG 1680TGAAAGGTTG CCGTGGCGGG GCCC 1704 2286 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 9CCGTCTTGGC GTCTGGGCGC ATTGTGATCT GGGCCANTTG CCCCTCCACC CAGACCGCGC 60CCAGCTTGTC GATCCAGCCC GCGACCCGGA TTGCCACCGC GCGAACCGGG AACGGATTCT 120CCGCTGAATT CTGGGTCACT TCGCAGTCGC GCGGGTGATC CTGTTGGCGA NCAGCGTCTG 180GAACGGGCGT CNAACGCGTG CCGTAAGCCC AGCGTGTACG CCGTCAGCCC GACGCCGATG 240CCGAATGCCT TGCCGCCCAA GCTGAGCCGC GCGGGCTCCA CCAAGAGCGT CACGGTGAGC 300CAGCCAACCA GATGCAAGGC GACGATCACC GCGAAGTGCC GAATTCGGCA CGAGAGGTGC 360TGGAAATCCA GCAATACGCC CGCGAGCCGA TCTCGTTGGA CCAGACCATC GGCGACGANG 420GCGACAGNCA GCTTGGCGAT TTCATCGAAA ACAGCGAGGC GGTGGTGGNC GTCGACGCGG 480TGTCCTTCAC TTTGCTGCAT GATCAACTGC ANTCGGTGCT GGACACGCTC TCCGAGCGTG 540AGGCGGGCGT GGTGCGGCTA CGCTTCGGCC TTACCGACGG CCAGCCGCGC ACCCTTGACG 600AGATCGGCCA GGTCTACGGC GTGACCCGGG AACGCATCCG CCAGATCGAA TCCAAGACTA 660TGTCGAAGTT GCGCCATCCG AGCCGCTCAC AGGTCCTGCG CGACTATCGT GCCGAATTCG 720GCACGAGCCG TTTTGAGGTG CACGCCCAGA CGGTGGAGGA CGAGGCTCGC CGGATGTGGG 780CGTCCGCGCA AAACATTTCC GGTGCGGGCT GGAGTGGCAT GGCCGANGCG ACCTCGCTAG 840ACACCATGGC CCAGATGAAT CAGGCGTTTC GCAACATCGT GAACATGCTG CACGGGGTGC 900GTGACGGGCT GGTTCGCGAC GCCAACAACT ACGAACAGCA AGAGCAGGCC TCCCAGCAGA 960TCCTCAGCAG CTGACCCGGC CCGACGACTC AGGAGGACAC ATGACCATCA ACTATCAATT 1020CGGGGACGTC GACGCTCATG GCGCCATGAT CCGCGCTCTG GCCGGGTTGC TGGAGGCCGA 1080GCATCAGGCC ATCATTTCTG ATGTGTTGAC CGCGAGTGAC TTTTGGGGCG GCGCCGGTTC 1140GGCGGCCTGC CAGGGGTTCA TTACCCAGTT GGGCCGTAAC TTCCAGGTGA TCTACGAGCA 1200GGCCAACGCC CACGGGCAGA AGGTGCAGGC TGCCGGCAAC AACATGGCAC AAACCGACAG 1260CGCCGTCGGC TCCAGCTGGG CCTAACCCGG GTCCTAAGTT GGGTCCGCGC AGGGCGGGCC 1320GATCAGCGTC GACTTTGGCG CCCGATACAC GGGCATGTNG TNGTCGGGAA CACTGCGCCC 1380GCGTCAGCTG CCCGCTTCCC CTTGTTCGGC GACGTGCTCG GTGATGGCTT TGACGACCGC 1440TTCGCCGGCG CGGCCAATCA ATTGGTCGCG CTTGCCTCTA GCCTCGTGCC GAATTCGGCA 1500CGAGGGTGCT GGTGCCGCGC TATCGGCAGC ACGTGAGCTC CACGACGAAC TCATCCCAGT 1560GCTGGGTTCC GCGGAGTTCG GCATCGGCGT GTCGGCCGGA AGGGCCATCG CCGGCCACAT 1620CGGCGCTCAA GCCCGCTTCG AGTACACCGT CATCGGCGAC CCGGTCAACG AGGCCGCCCG 1680GCTCACCGAA CTGGCCAAAG TCGAGGATGG CCACGTTCTG GCGTCGGCGA TCGCGGTCAG 1740TGGCGCCCTG GACGCCGAAG CATTGTGTTG GGATGTTGGC GAGGTGGTTG AGCTCCGCGG 1800ACGTGCTGCA CCCACCCAAC TAGCCAGGCC AATGAATNTG GCNGCACCCG AAGAGGTTTC 1860CAGCGAAGTA CGCGGCTAGT CGCGCTTGGC TGCNTTCTTC GCCGGCACCT TCCGGGCAGC 1920TTTCCTGGCT GGCCGTTTTG CCGGACCCCG GGCTCGGCGA TCGGCCAACA GCTCGGCGGC 1980GCGCTCGTCG GTTATGGAAG CCACGTNGTC GCCCTTACGC AGGCTGGCAT TGGTCTCACC 2040GTCGGTGACG TACGGCCCGA ATCGGCCGTC CTTGATGACC ATTGGCTTGC CAGACGCCGG 2100ATNTGNTCCC AGCTCGCGCA GCGGCGGAGC CGAAGCGCTT TGCCGGCCAC GACNTTTCGG 2160CTCTGNGTAG ATNTTCAGGG CTTCGTCGAG CGNGATGGTG AATATATGGT CTTCGGTGAC 2220CAGTGATCGA GAATCGTTGC CGCGCTTTAG ATACGGTCNG TAGCGCCCGT TCTGCGCGGT 2280GATNTC 2286 1136 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 10GGGCATCTTC CCCGACCGCG CCTCGATCAT CCGCCTCGTC GGAGCCGTCC TCGCCGAACA 60ACACGACGAA TGGATCGAAG GACGGCGCTA CCTGGGCCTC GAGGTCCTCA CCCGAGCCCG 120AGCAGCACTG ACCAGCACCG AAGAACCGCC AAGCAGCAAA CCACCAACAC CCCAGCACTG 180ACCACCTAGA CTGCCACCCG AAGGATCACG CGAGGAACCT TCACTCGTAC ACCACGTCCC 240TGGCCTTGGC CTGGTGTCAG GCCCAGCTGG AGCCGACGGC GCTGTCGGTT TGCGCCATGT 300TGTTGCCGGC AGCCTGCACC TTCTGCCCGT GGGCGTTGGC CTGCTCGTAG ATCACCTGGA 360AGTTACGGCC CAACTGGGTA ATGAACCCCT GGCAGGCCGC CGAACCGGCG CCGCCCCAAA 420AGTCACTCGC GGTCAACACA TCACGAATGA TGGCCTGATG CTCGGCCTCC AGCAACCCGG 480CCTGAGCGCG GATCATGGCG CCGTGAGCGT CGACATCACC GAACTGATAG TTGATGGTCA 540TCGAACCTGT TCTCCTTCGC TTGTAAAAGT ATTGTGCTGC AGCGGCTGAC GTTAGCTGCT 600GAGGATCTGC TGGGAGGCCT GCTCTTGCCT CGTGCCGAAT TCGGCACGAG AGGCCGCCTT 660CGAAGAAATC CTTTGAGAAT TCGCCAAGGC CGTCGACCCA GCATGGGGTC AGCTCGCCAG 720CCGCGCCGGC TGGCAACCGT TCCCGCTCGA GAAAGACCTG GAGGAATACC AGTGACAAAC 780GACCTCCCAG ACGTCCGAGA GCGTGACGGC GGTCCACGTC CCGCTCCTCC TGCTGGCGGG 840CCACGCTTGT CAGACGTGTG GGTTTACAAC GGGCGGGCGT ACGACCTGAG TGAGTGGATT 900TCCAAGCATC CCGGCGGCGC CTTNTTCATT GGGCGGACCA AGAACCGCGA CATCACCGCA 960ATCGTCAAGT CCTACCATCG TGATCCGGCG ATTGTCGAGC GAATCCTGCA GCGGAGGTAC 1020GCGTTGGGCC GCGACGCAAC CCCTAGGGAC ATCCACCCCA AGCACAATGC ACCGGCATTT 1080CTGTTCAAAG ACGACTTCAA CAGCTGGCGG GACACCCCGA AGTATCGATT NGACGA 1136 967 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 11TGAGCGCCAA CCCTACCGTC GGTTCGTCAC ACGGACCGCA TGGCCTGCTC CGCGGACTGC 60CGCTAGGGTC GCGGATCACT CGGCGTAGCG GCGCCTTTGC CCACCGATAT GGGTTCCGTC 120ACAGTGTGGT TGCCCGCCCG CCATCGGCCG GATAACGCCA TGACCTCAGC TCGGCAGAAA 180TGACAATGCT CCCAAAGGCG TGAGCACCCG AAGACAACTA AGCAGGAGAT CGCATGCCGT 240TTGTGACTAC CCAACCAGAA GCACTGGCGG CGGCGGCCGG CAGTCTGCAG GGAATCGGCT 300CCGCATTGAA CGCCCAGAAT GCGGCTGCGG CGACTCCCAC GACGGGGGTG GTCCGGCGGC 360CGCCGATGAA NTGTCGGCGC TGACGGCGGC TCAGTTCGCG GCACACGCCC AGATCTATCA 420GGCCGTCAGC GCCCAGGCCG CGGCGATTCA CGAGATGTTC GTCAACACTC TACAGATGAG 480CTCAGGGTCG TATGCTGCTA CCGAGGCCGC CAACGCGGCC GCGGCCGGNT AGAGGAGTCA 540CTGCGATGGA TTTTGGGGCG TTGCCGCCGG AGGTCAATTC GGTGCGGATG TATGCCGTTC 600CTGGCTCGGC ACCAATGGTC GCTGCGGCGT CGGCCTGGAA CGGGTTGGCC GCGGAGCTGA 660GTTCGGCGGC CACCGGTTAT GAGACGGTGA TCACTCAGCT CAGCAGTGAG GGGTGGCTAG 720GTCCGGCGTC AGCGGCGATG GCCGAGGCAG TTGCGCCGTA TGTGGCGTGG ATGAGTGCCG 780CTGCGGCGCA AGCCGAGCAG GCGGCCACAC AGGCCAGGGC CGCCGCGGCC GCTTTTGAGG 840CGGCGTTTGC CGCGACGGTG CCTCCGCCGT TGATCGCGGC CAACCGGGCT TCGTTGATGC 900AGCTGATCTC GACGAATGTC TTTGGTCAGA ACACCTCGGC GATCGCGGCC GCCGAAGCTC 960AGTACGG 967 585 base pairs nucleic acid double linear DNA (genomic) Mycobacterium tuberculosis 12TGGATTCCGA TAGCGGTTTC GGCCCCTCGA CGGGCGACCA CGGCGCGCAG GCCTCCGAAC 60GGGGGGCCGG GACGCTGGGA TTCGCCGGGA CCGCAACCAA AGAACGCCGG GTCCGGGCGG 120TCGGGCTGAC CGCACTGGCC GGTGATGAGT TCGGCAACGG CCCCCGGATG CCGATGGTGC 180CGGGGACCTG GGAGCAGGGC AGCAACGAGC CCGAGGCGCC CGACGGATCG GGGAGAGGGG 240GAGGCGACGG CTTACCGCAC GACAGCAAGT AACCGAATTC CGAATCACGT GGACCCGTAC 300GGGTCGAAAG GAGAGATGTT ATGAGCCTTT TGGATGCTCA TATCCCACAG TTGGTGGCCT 360CCCAGTCGGC GTTTGCCGCC AAGGCGGGGC TGATGCGGCA CACGATCGGT CAGGCCGAGC 420AGGCGGCGAT GTCGGCTCAG GCGTTTCACC AGGGGGAGTC GTCGGCGGCG TTTCAGGCCG 480CCCATGCCCG GTTTGTGGCG GCGGCCGCCA AAGTCAACAC CTTGTTGGAT GTCGCGCAGG 540CGAATCTGGG TGAGGCCGCC GGTACCTATG TGGCCGCCGA TGCTG 585 144 amino acids amino acid single linear peptide Mycobacterium tuberculosis 13Ala Leu Val Thr Thr Asn Phe Phe Gly Val Asn Thr Ile Pro Ile Ala1 5 10 15Leu Asn Glu Ala Asp Tyr Leu Arg Met Trp Ile Gln Ala Ala Thr Val 20 25 30Met Ser His Tyr Gln Ala Val Ala His Glu Ile Trp Cys Leu His Glu 35 40 45Xaa Ala Ser Ser Gly Lys Pro Trp Ala Ser Ile Thr Thr Gly Ala Pro 50 55 60Gly Ser Pro Ala Ser Thr Thr Arg Ser Arg Thr Pro Leu Val Ser Thr65 70 75 80Asn Arg Xaa Val Xaa Ala Pro Ile Val Ser Pro Asn His Thr Gly His 85 90 95Arg Pro Glu Lys Gly Leu Gly Ser Xaa Gln Arg Arg Leu Ser Arg Val 100 105 110Leu Pro Arg Ile Ile Asp Arg Pro Ala Gly Pro Xaa Gly Pro Pro Leu 115 120 125Thr Ser Gly Ser His Phe Leu Cys Ser Trp His Gly Tyr Ser Ser Gln 130 135 140 352 amino acids amino acid single linear peptide Mycobacterium tuberculosis 14His Ala Leu Ala Ala Gln Tyr Thr Glu Ile Ala Thr Glu Leu Ala Ser1 5 10 15Val Leu Ala Ala Val Gln Ala Ser Ser Trp Gln Gly Pro Ser Ala Asp 20 25 30Arg Phe Val Val Ala His Gln Pro Phe Arg Tyr Trp Leu Thr His Ala 35 40 45Ala Thr Val Ala Thr Ala Ala Ala Ala Ala His Xaa Thr Ala Ala Ala 50 55 60Gly Tyr Thr Ser Ala Leu Gly Gly Met Pro Thr Leu Ala Glu Leu Ala65 70 75 80Ala Asn His Ala Met His Gly Ala Leu Val Thr Thr Asn Phe Phe Gly 85 90 95Val Asn Thr Ile Pro Ile Ala Leu Asn Glu Ala Asp Tyr Leu Arg Met 100 105 110Trp Ile Gln Ala Ala Thr Val Met Ser His Tyr Gln Ala Val Ala His 115 120 125Glu Ser Val Ala Ala Thr Pro Ser Thr Pro Pro Ala Pro Gln Ile Val 130 135 140Thr Ser Ala Ala Ser Ser Ala Ala Ser Ser Ser Phe Pro Asp Pro Thr145 150 155 160Lys Leu Ile Leu Gln Leu Leu Lys Asp Phe Leu Glu Leu Leu Arg Tyr 165 170 175Leu Ala Val Glu Leu Leu Pro Gly Pro Leu Gly Asp Leu Ile Ala Gln 180 185 190Val Leu Asp Trp Phe Ile Ser Phe Val Ser Gly Pro Val Phe Thr Phe 195 200 205Leu Ala Tyr Leu Val Leu Asp Pro Leu Ile Tyr Phe Gly Pro Phe Ala 210 215 220Pro Leu Thr Ser Pro Val Leu Leu Pro Ala Val Glu Leu Arg Asn Arg225 230 235 240Leu Lys Thr Ala Thr Gly Leu Thr Leu Pro Pro Thr Val Ile Phe Asp 245 250 255His Pro Thr Pro Thr Ala Val Ala Glu Tyr Val Ala Gln Gln Met Ser 260 265 270Gly Ser Arg Pro Thr Glu Ser Gly Asp Pro Thr Ser Gln Val Val Glu 275 280 285Pro Ala Arg Ala Glu Phe Gly Thr Ser Ala Val His Gln Ile Pro Pro 290 295 300Arg Pro Ala Asp Thr Arg Arg Ala Cys Arg His Arg Asp Asp Val Pro305 310 315 320Arg Asp Ser Arg Ile Ala Gln His Arg Asp Gly Ala Gly Leu Asp Pro 325 330 335Thr Glu Arg Gly Thr Ser Glu Gly Asp Gln Gly Leu Val Ser Gly Trp 340 345 350 141 amino acids amino acid single linear peptide Mycobacterium tuberculosis 15Met Asp Phe Gly Ala Leu Pro Pro Glu Val Asn Ser Val Arg Met Tyr1 5 10 15Ala Val Pro Gly Ser Ala Pro Met Val Ala Ala Ala Ser Ala Trp Asn 20 25 30Gly Leu Ala Ala Glu Leu Ser Ser Ala Ala Thr Gly Tyr Glu Thr Val 35 40 45Ile Thr Gln Leu Ser Ser Glu Gly Trp Leu Gly Pro Ala Ser Ala Ala 50 55 60Met Ala Glu Ala Val Ala Pro Tyr Val Ala Trp Met Ser Ala Ala Ala65 70 75 80Ala Gln Ala Glu Gln Ala Ala Thr Gln Ala Arg Ala Ala Ala Ala Ala 85 90 95Phe Glu Ala Ala Phe Ala Ala Thr Val Pro Pro Pro Leu Ile Ala Ala 100 105 110Asn Arg Ala Ser Leu Met Gln Leu Ile Ser Thr Asn Val Phe Gly Gln 115 120 125Asn Thr Ser Ala Ile Ala Ala Ala Glu Ala Gln Tyr Gly 130 135 140 58 amino acids amino acid single linear peptide Mycobacterium tuberculosis 16Met Ala Ser Arg Phe Met Thr Asp Pro His Ala Met Arg Asp Met Ala1 5 10 15Gly Arg Phe Glu Val His Ala Gln Thr Val Glu Asp Glu Ala Arg Arg 20 25 30Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly Met 35 40 45Ala Glu Ala Thr Ser Leu Asp Thr Met Thr 50 55 67 amino acids amino acid single linear peptide Mycobacterium tuberculosis 17Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Gln Ala Ala Ser Leu Glu Ala Glu His Gln Ala Ile Val 20 25 30Arg Asp Val Leu Ala Ala Gly Asp Phe Trp Gly Gly Ala Gly Ser Val 35 40 45Ala Cys Gln Glu Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln65 58 amino acids amino acid single linear peptide Mycobacterium tuberculosis 18Met Ala Ser Arg Phe Met Thr Asp Pro His Ala Met Arg Asp Met Ala1 5 10 15Gly Arg Phe Glu Val His Ala Gln Thr Val Glu Asp Glu Ala Arg Arg 20 25 30Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly Met 35 40 45Ala Glu Ala Thr Ser Leu Asp Thr Met Thr 50 55 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 19Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Gln Ala Ala Ser Leu Glu Ala Glu His Gln Ala Ile Val 20 25 30Arg Asp Val Leu Ala Ala Gly Asp Phe Trp Gly Gly Ala Gly Ser Val 35 40 45Ala Cys Gln Glu Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 30 amino acids amino acid single linear peptide Mycobacterium tuberculosis 20Asn Met Leu His Gly Val Arg Asp Gly Leu Val Arg Asp Ala Asn Asn1 5 10 15Tyr Glu Gln Gln Glu Gln Ala Ser Gln Gln Ile Leu Ser Ser 20 25 30 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 21Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Gln Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30Arg Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 69 amino acids amino acid single linear peptide Mycobacterium tuberculosis 22Ala Arg Arg Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp1 5 10 15Ser Gly Met Ala Glu Ala Thr Ser Leu Asp Thr Met Ala Gln Met Asn 20 25 30Gln Ala Phe Arg Asn Ile Val Asn Met Leu His Gly Val Arg Asp Gly 35 40 45Leu Val Arg Asp Ala Asn Asn Tyr Glu Gln Gln Glu Gln Ala Ser Gln 50 55 60Gln Ile Leu Ser Ser65 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 23Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Gln Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30Arg Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln Ala Asn Thr His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Xaa Ser Ser Trp Ala 85 90 52 amino acids amino acid single linear peptide Mycobacterium tuberculosis 24Gly Met Ala Glu Ala Thr Ser Xaa Asp Thr Met Thr Gln Met Asn Gln1 5 10 15Ala Phe Arg Asn Ile Val Asn Met Leu His Gly Val Arg Asp Gly Leu 20 25 30Val Arg Asp Ala Asn Xaa Tyr Glu Gln Gln Glu Gln Ala Ser Gln Gln 35 40 45Ile Leu Ser Ser 50 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 25Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Gln Ala Gly Ser Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30Ser Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Xaa 50 55 60Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 98 amino acids amino acid single linear peptide Mycobacterium tuberculosis 26Met Thr Ser Arg Phe Met Thr Asp Pro His Ala Met Arg Asp Met Ala1 5 10 15Gly Arg Phe Glu Val His Ala Gln Thr Val Glu Asp Glu Ala Arg Arg 20 25 30Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly Met 35 40 45Ala Glu Ala Thr Ser Leu Asp Thr Met Ala Gln Met Asn Gln Ala Phe 50 55 60Arg Asn Ile Val Asn Met Leu His Gly Val Arg Asp Gly Leu Val Arg65 70 75 80Asp Ala Asn Asn Tyr Glu Gln Gln Glu Gln Ala Ser Gln Gln Ile Leu 85 90 95Ser Ser 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 27Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Xaa Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30Ser Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 81 amino acids amino acid single linear peptide Mycobacterium tuberculosis 28Arg Phe Glu Val His Ala Gln Thr Val Glu Asp Glu Ala Arg Arg Met1 5 10 15Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly Met Ala 20 25 30Xaa Ala Thr Ser Leu Asp Thr Met Ala Gln Met Asn Gln Ala Phe Arg 35 40 45Asn Ile Val Asn Met Leu His Gly Val Arg Asp Gly Leu Val Arg Asp 50 55 60Ala Asn Asn Tyr Glu Gln Gln Glu Gln Ala Ser Gln Gln Ile Leu Ser65 70 75 80Ser 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 29Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Leu Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30Ser Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 11 amino acids amino acid single linear peptide Mycobacterium tuberculosis 30Gln Glu Gln Ala Ser Gln Gln Ile Leu Ser Ser1 5 10 94 amino acids amino acid single linear peptide Mycobacterium tuberculosis 31Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met1 5 10 15Ile Arg Ala Gln Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30Arg Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn65 70 75 80Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 99 amino acids amino acid single linear peptide Mycobacterium tuberculosis 32Met Ser Phe Val Thr Thr Gln Pro Glu Ala Leu Ala Ala Ala Ala Ala1 5 10 15Asn Leu Gln Gly Ile Gly Thr Thr Met Asn Ala Gln Asn Ala Ala Ala 20 25 30Ala Ala Pro Thr Thr Gly Val Val Pro Ala Ala Ala Asp Glu Val Ser 35 40 45Ala Leu Thr Ala Ala Gln Phe Ala Ala His Ala Gln Met Tyr Gln Thr 50 55 60Val Ser Ala Gln Ala Ala Ala Ile His Glu Met Phe Val Asn Thr Leu65 70 75 80Val Ala Ser Ser Gly Ser Tyr Ala Ala Thr Glu Ala Ala Asn Ala Ala 85 90 95Ala Ala Gly 99 amino acids amino acid single linear peptide Mycobacterium tuberculosis 33Met Ser Phe Val Thr Thr Gln Pro Glu Ala Leu Ala Ala Ala Ala Ala1 5 10 15Asn Leu Gln Gly Ile Gly Thr Thr Met Asn Ala Gln Asn Ala Ala Ala 20 25 30Ala Ala Pro Thr Thr Gly Val Val Pro Ala Ala Ala Asp Glu Val Ser 35 40 45Ala Leu Thr Ala Ala Gln Phe Ala Ala His Ala Gln Met Tyr Gln Thr 50 55 60Val Ser Ala Gln Ala Ala Ala Ile His Glu Met Phe Val Asn Thr Leu65 70 75 80Val Ala Ser Ser Gly Ser Tyr Ala Ala Thr Glu Ala Ala Asn Ala Ala 85 90 95Ala Ala Gly 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 34Asp Pro His Ala Met Arg Asp Met Ala Gly Arg Phe Glu Val His1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 35Arg Asp Met Ala Gly Arg Phe Glu Val His Ala Gln Thr Val Glu1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 36Arg Phe Glu Val His Ala Gln Thr Val Glu Asp Glu Ala Arg Arg1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 37Ala Gln Thr Val Glu Asp Glu Ala Arg Arg Met Trp Ala Ser Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 38Asp Glu Ala Arg Arg Met Trp Ala Ser Ala Gln Asn Ile Ser Gly1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 39Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 40Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly Met Ala Glu Ala Thr1 5 10 15 16 amino acids amino acid single linear peptide Mycobacterium tuberculosis 41Ala Gly Trp Ser Gly Met Ala Glu Ala Thr Ser Leu Asp Thr Met Thr1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 42Met Ala Glu Ala Thr Ser Leu Asp Thr Met Ala Gln Met Asn Gln1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 43Ser Leu Asp Thr Met Ala Gln Met Asn Gln Ala Phe Arg Asn Ile1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 44Ala Gln Met Asn Gln Ala Phe Arg Asn Ile Val Asn Met Leu His1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 45Ala Phe Arg Asn Ile Val Asn Met Leu His Gly Val Arg Asp Gly1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 46Val Asn Met Leu His Gly Val Arg Asp Gly Leu Val Arg Asp Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 47Gly Val Arg Asp Gly Leu Val Arg Asp Ala Asn Asn Tyr Glu Gln1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 48Leu Val Arg Asp Ala Asn Asn Tyr Glu Gln Gln Glu Gln Ala Ser1 5 10 15 16 amino acids amino acid single linear peptide Mycobacterium tuberculosis 49Asn Asn Tyr Glu Gln Gln Glu Gln Ala Ser Gln Gln Ile Leu Ser Ser1 5 10 15 17 amino acids amino acid single linear peptide 50Met Ala Ser Arg Phe Met Thr Asp Pro His Ala Met Arg Asp Met Ala1 5 10 15Gly 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 51Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 52Gln Phe Gly Asp Val Asp Ala His Gly Ala Met Ile Arg Ala Gln1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 53Asp Ala His Gly Ala Met Ile Arg Ala Gln Ala Ala Ser Leu Glu1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 54Met Ile Arg Ala Gln Ala Ala Ser Leu Glu Ala Glu His Gln Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 55Ala Ala Ser Leu Glu Ala Glu His Gln Ala Ile Val Arg Asp Val1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 56Ala Glu His Gln Ala Ile Val Arg Asp Val Leu Ala Ala Gly Asp1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 57Ile Val Arg Asp Val Leu Ala Ala Gly Asp Phe Trp Gly Gly Ala1 5 10 15 16 amino acids amino acid single linear peptide Mycobacterium tuberculosis 58Leu Ala Ala Gly Asp Phe Trp Gly Gly Ala Gly Ser Val Ala Cys Gln1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 59Phe Trp Gly Gly Ala Gly Ser Val Ala Cys Gln Glu Phe Ile Thr1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 60Gly Ser Val Ala Cys Gln Glu Phe Ile Thr Gln Leu Gly Arg Asn1 5 10 15 18 amino acids amino acid single linear peptide Mycobacterium tuberculosis 61Gln Glu Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile Tyr Glu1 5 10 15Gln Ala 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 62Arg Asn Phe Gln Val Ile Tyr Glu Gln Ala Asn Ala His Gly Gln1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 63Ile Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 64Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn Asn Met Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 65Lys Val Gln Ala Ala Gly Asn Asn Met Ala Gln Thr Asp Ser Ala1 5 10 15 16 amino acids amino acid single linear peptide Mycobacterium tuberculosis 66Gly Asn Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 67Asp Ala His Gly Ala Met Ile Arg Ala Leu Ala Gly Leu Leu Glu1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 68Asp Ala His Gly Ala Met Ile Arg Ala Gln Ala Gly Leu Leu Glu1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 69Met Ile Arg Ala Leu Ala Gly Leu Leu Glu Ala Glu His Gln Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 70Met Ile Arg Ala Gln Ala Gly Leu Leu Glu Ala Glu His Gln Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 71Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile Ser Asp Val1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 72Ala Gly Leu Leu Glu Ala Glu His Gln Ala Ile Ile Arg Asp Val1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 73Ala Glu His Gln Ala Ile Ile Ser Asp Val Leu Thr Ala Ser Asp1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 74Ala Glu His Gln Ala Ile Ile Arg Asp Val Leu Thr Ala Ser Asp1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 75Ile Ile Ser Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 76Ile Ile Arg Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala1 5 10 15 16 amino acids amino acid single linear peptide Mycobacterium tuberculosis 77Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala Ala Cys Gln1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 78Phe Trp Gly Gly Ala Gly Ser Ala Ala Cys Gln Gly Phe Ile Thr1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 79Gly Ser Ala Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn1 5 10 15 15 amino acids amino acid single linear peptide Mycobacterium tuberculosis 80Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile Tyr1 5 10 15 25 amino acids amino acid single linear peptide Mycobacterium tuberculosis 81Val Thr Thr Asn Phe Phe Gly Val Asn Thr Ile Pro Ile Ala Leu Asn1 5 10 15Glu Ala Asp Tyr Leu Arg Met Trp Ile 20 25 25 amino acids amino acid single linear peptide Mycobacterium tuberculosis 82Asn Glu Ala Asp Tyr Leu Arg Met Trp Ile Gln Ala Ala Thr Val Met1 5 10 15Ser His Tyr Gln Ala Val Ala His Glu 20 25 967 base pairs nucleic acid single linear cDNA 83TGAGCGCCAA CCCTACCGTC GGTTCGTCAC ACGGACCGCA TGGCCTGCTC CGCGGACTGC 60CGCTAGGGTC GCGGATCACT CGGCGTAGCG GCGCCTTTGC CCACCGATAT GGGTTCCGTC 120ACAGTGTGGT TGCCCGCCCG CCATCGGCCG GATAACGCCA TGACCTCAGC TCGGCAGAAA 180TGACAATGCT CCCAAAGGCG TGAGCACCCG AAGACAACTA AGCAGGAGAT CGCATGCCGT 240TTGTGACTAC CCAACCAGAA GCACTGGCGG CGGCGGCCGG CAGTCTGCAG GGAATCGGCT 300CCGCATTGAA CGCCCAGAAT GCGGCTGCGG CGACTCCCAC GACGGGGGTG GTCCGGCGGC 360CGCCGATGAA NTGTCGGCGC TGACGGCGGC TCAGTTCGCG GCACACGCCC AGATCTATCA 420GGCCGTCAGC GCCCAGGCCG CGGCGATTCA CGAGATGTTC GTCAACACTC TACAGATGAG 480CTCAGGGTCG TATGCTGCTA CCGAGGCCGC CAACGCGGCC GCGGCCGGNT AGAGGAGTCA 540CTGCGATGGA TTTTGGGGCG TTGCCGCCGG AGGTCAATTC GGTGCGGATG TATGCCGTTC 600CTGGCTCGGC ACCAATGGTC GCTGCGGCGT CGGCCTGGAA CGGGTTGGCC GCGGAGCTGA 660GTTCGGCGGC CACCGGTTAT GAGACGGTGA TCACTCAGCT CAGCAGTGAG GGGTGGCTAG 720GTCCGGCGTC AGCGGCGATG GCCGAGGCAG TTGCGCCGTA TGTGGCGTGG ATGAGTGCCG 780CTGCGGCGCA AGCCGAGCAG GCGGCCACAC AGGCCAGGGC CGCCGCGGCC GCTTTTGAGG 840CGGCGTTTGC CGCGACGGTG CCTCCGCCGT TGATCGCGGC CAACCGGGCT TCGTTGATGC 900AGCTGATCTC GACGAATGTC TTTGGTCAGA ACACCTCGGC GATCGCGGCC GCCGAAGCTC 960AGTACGG 967 15 amino acids amino acid single linear peptide 84Met Ser Phe Val Thr Thr Gln Pro Glu Ala Leu Ala Ala Ala Ala 1 5 10 15 15 amino acids amino acid single linear peptide 85Thr Gln Pro Glu Ala Leu Ala Ala Ala Ala Ala Asn Leu Gln Gly 1 5 10 15 15 amino acids amino acid single linear peptide 86Leu Ala Ala Ala Ala Ala Asn Leu Gln Gly Ile Gly Thr Thr Met 1 5 10 15 15 amino acids amino acid single linear peptide 87Ala Asn Leu Gln Gly Ile Gly Thr Thr Met Asn Ala Gln Asn Ala 1 5 10 15 15 amino acids amino acid single linear peptide 88Ile Gly Thr Thr Met Asn Ala Gln Asn Ala Ala Ala Ala Ala Pro 1 5 10 15 15 amino acids amino acid single linear peptide 89Asn Ala Gln Asn Ala Ala Ala Ala Ala Pro Thr Thr Gly Val Val 1 5 10 15 15 amino acids amino acid single linear peptide 90Ala Ala Ala Ala Pro Thr Thr Gly Val Val Pro Ala Ala Ala Asp 1 5 10 15 15 amino acids amino acid single linear peptide 91Thr Thr Gly Val Val Pro Ala Ala Ala Asp Glu Val Ser Ala Leu 1 5 10 15 15 amino acids amino acid single linear peptide 92Pro Ala Ala Ala Asp Glu Val Ser Ala Leu Thr Ala Ala Gln Phe 1 5 10 15 15 amino acids amino acid single linear peptide 93Glu Val Ser Ala Leu Thr Ala Ala Gln Phe Ala Ala His Ala Gln 1 5 10 15 15 amino acids amino acid single linear peptide 94Thr Ala Ala Gln Phe Ala Ala His Ala Gln Met Tyr Gln Thr Val 1 5 10 15 15 amino acids amino acid single linear peptide 95Ala Ala His Ala Gln Met Tyr Gln Thr Val Ser Ala Gln Ala Ala 1 5 10 15 16 amino acids amino acid single linear peptide 96Met Tyr Gln Thr Val Ser Ala Gln Ala Ala Ala Ile His Glu Met Phe 1 5 10 15 15 amino acids amino acid single linear peptide 97Ser Ala Gln Ala Ala Ala Ile His Glu Met Phe Val Asn Thr Leu 1 5 10 15 15 amino acids amino acid single linear peptide 98Ala Ile His Glu Met Phe Val Asn Thr Leu Val Ala Ser Ser Gly 1 5 10 15 15 amino acids amino acid single linear peptide 99Phe Val Asn Thr Leu Val Ala Ser Ser Gly Ser Tyr Ala Ala Thr 1 5 10 15 15 amino acids amino acid single linear peptide 100Val Ala Ser Ser Gly Ser Tyr Ala Ala Thr Glu Ala Ala Asn Ala 1 5 10 15 14 amino acids amino acid single linear peptide 101Ser Tyr Ala Ala Thr Glu Ala Ala Asn Ala Ala Ala Ala Gly 1 5 10 1784 base pairs nucleic acid single linear cDNA 102ATTCGTTCCT GCCGCAGCTA AATCCCGGGG ACATCGTCGC CGGCCAGTAC GAGGTCAAAG 60GCTGCATCGC GCACGGCGGA CTGGGCTGGA TCTACCTCGC TCTCGACCGC AATGTCAACG 120GCCGTCCGGT GGTGCTCAAG GGCCTGGTGC ATTCCGGTGA TGCCGAAGCG CAGGCAATGG 180CGATGGCCGA ACGCCAGTTC CTGGCCGAGG TGGTGCACCC GTCGATCGTG CAGATCTTCA 240ACTTTGTCGA GCACACCGAC AGGCACGGGG ATCCGGTCGG CTACATCGTG ATGGAATACG 300TCGGCGGGCA ATCGCTCAAA CGCAGCAAGG GTCANAAACT GCCCGTCGCG GAGGCCATCG 360CCTACCTGCT GGAGATCCTG CCGGCGCTGA GCTACCTGCA TTCCATCGGC TTGGTCTACA 420ACGACCTGAA GCCGGAAAAC ATCATGCTGA CCGAGGAACA GCTCAAGCTG ATCGACCTGG 480GCGCGGTATC GCGGATCAAC TCGTTCGGCT ACCTCTACGG GACCCCAGGC TTCCAGGCGC 540CCGAGATCGT GCGGACCGGT CCGACGGTGG CCACCGACAT CTACACCGTG GGACGCACGC 600TCGCGGCGCT CACGCTGGAC CTGCCCACCC GCAATGGCCG TTATGTGGAT GGGCTACCCG 660AAGACGACCC GGTGCTGAAA ACCTACGACT CTTACGGCCG GTTGCTGCGC AGGGCCATCG 720ACCCCGATCC GCGGCAACGG TTCACCACCG CCGAAGAGAT GTCCGCGCAA TTGACGGGCG 780TGTTGCGGGA GGTGGTCGCC CAGACACCGG GGTGCCGCGG CCAGGCTATC AACGATCTTC 840AGTCCCAGTC GGTCGACATT TGGAGTGGAC TGCTGGTGGC GCACACCGAC GTGTATCTGG 900ACGGGCAGGT GCACGCGGAG AAGCTGACCG CCAACGAGAT CGTGACCGCG CTGTCGGTGC 960CGCTGGTCGA TCCGACCGAC GTCGCAGCTT CGGTCCTGCA GGCCACGGTG CTCTCCCAGC 1020CGGTGCAGAC CCTAGACTCG NTGCGCGCGG CCCGCCACGG TGCGCTGGAC GCCGACGGCG 1080TCGATTNTCC GAGTCAGTGG AGCTGCCGCT AATGGAAGTC CGCGCGCTGC TGGATCTCGG 1140CGATGTGGCC AAGGCCACCC GAAAACTCGA CGATCTGGCC GAACGCGTTG GCTGGCGATG 1200GCGATTGGTC TGGTACCGGG CCGTCGCCGA GCTGCTCACC GGCGACTATG ACTCGGCCAC 1260CAAACATTTC ACCGAGGTGC TGGATACCTT TCCCGGCGAG CTGGCGCCCA AGCTCGCCCT 1320GGCCGCCACC GCCGAACTAG CCGGCAACAC CGACGAACAC AAGTTCTATC AGACGGTGTG 1380GAGCACCAAC GACGGCGTGA TCTCGGCGGC TTTCGGACTG GCCAGAGCCC GGTCGGCCGA 1440AGGTGATCGG GTCGGCGCCG TGCGCACGCT CGACGAGGTA CCGCCCACTT CTCGGCATTT 1500CACCACGGCA CGGCTGACCA GCGCGGTGAC TCTGTTGTCC GGCCGGTCAA CGAGTGAAGT 1560CACCGAGGAA CAGATCCGCG ACGCCGCCCG AAGAGTGGAG GCGCTGCCCC CGACCGAACC 1620ACGCGTGCTG CAGATCCGCG CCCTGGTGCT GGGTGGCGCG CTGGACTGGC TGAAGGACAA 1680CAAGGCCAGC ACCAACCACA TCCTCGGTTT CCCGTTCACC AGTCACGGGC TGCGGCTGGG 1740TGTCGAGGCG TCACTGCGCA GCCTGGCCCG GGTAGCTCCC ACTC 1784 766 base pairs nucleic acid single linear cDNA 103ACAARACACT CGGYGGCKGC CGMTCCGGCC TGATCGTCGG TGATCAGCYT CGTGCCAAAY 60TCGGCACAAG GTGCGCGCTR CCCAANGAGT TCTTCGCCGC RGTGCGMGCM KAACTGGCCT 120ATCNTGGTTG GGTGCCGTCC CGCANAACCC GCGAACTTAA ACCCATTTTA ACCGGGCAGG 180AAGTTTCCTA CATYTACCCN RGSMANCCAA CCGGGCCGCC NANAAMTCCG TCCTGGANTC 240CGANCGGTTC CCGGTGTTCG CCGCACTGCT GACCGGCACG GARTATCCGC AGGCGGCGTT 300GGCCAACGCG TGGGTGCAAC TGGCCTACGG TGCGCACCAS GACGCCATCA CCGGCTCGGA 360GTCCGACCAG GTACTCAATG CTGGCGACCA CACCAGCCAG CAGACCAAAC TGGTGCACGC 420CGATCTCCAG GCGCGCCGGC CCGGTGGCAT ACGGATTGGT CGAAACCAAT CCGAAGGAAT 480TCATCACGGA CGGTCACGGA AAACGATCGC CCCAATGGGN GGACNACCCN AGCCAGGCGN 540ATTNACCGTT NAACAAGTTG GNGTAGGTTC TTTGATATCG AKCAACCGAT ACGGAKCGGM 600CCGCGGAATG GTAGACCACC ACCAGTGCCC NCAMGTMGTG CACCAGTTTG GTCATCGCCC 660GCAGATCGGT GACCCCGCCA AGCGTTCCGG ATGCGGAGAT GASGGTGACC AGCCYGGTTG 720ACCTGTTGAT CAGGTTNTCC CAGTGCCACG TCGGCAGCTG GCCGGT 766 1231 base pairs nucleic acid single linear cDNA 104CGGCACGAGA ATGTCGCCTG TGCCTCGATA GCCACTTGCG TGTGGTCGCG CTGCCAGCGG 60GTCAGCCAGG TCGCCTGGTC CAGGCCATCG GGCCGGCGCA GGAGCGCGAT GTTGGCCAGA 120CCCGGTGTAC GAGAACCGGA CTCGACNAAG TGTCGGCGCT GACGGCGGCT CAGTTCGCGG 180CACACGCCCA GATCTATCAG GCCGTCAGCG CCCAGGCCGC GGCGATTCAC GAGATGTTCG 240TCAACACTCT ACAGATNANC TCAGGGTCGT ATGCTGCTAC CGAGGCCGCC AACGCGGCCG 300CGGCCGGCTA GAGGAGTCAC TGCGATGGAT TTTGGGGCGT TGCCGCCGGA GGTCAATTCG 360GTGCGGATGT ATGCCGGTCC TGGCTCGGCA CCAATGGTCG CTGCGGCGTC GGCCTGGAAC 420GGGTTGGCCG CGGAGCTGAG TTCGGCGGCC ACCGGTTATG AGACGGTGAT CACTCAGCTC 480AGCAGTGAGG GGTGGCTAGG TCCGGCGTCA GCGGCGATGG CCGAGGCAGT TGCGCCGTAT 540GTGGCGTGGA TGAGTGCCGC TGCGGCGCAA GCCGAGCAGG CGGCCACACA GGCCAGGGCC 600GCCGCGGCCG CTTTTGAGGC GGCGTTTGCC GCGACGGTGC CTCCGCCGTT GATCGCGGCC 660AACCGGGCTT CGTTGATGCA GCTGATCTCG ACGAATGTCT TTGGTCAGAA CACCTCGGCG 720ATCGCGGCCG CCGAAGCTCA GTACGGCGAG ATGTGGGCCC AAGACTCCGC GGCGATGTAT 780GCCTACGCGG GCAGTTCGGC GAGCGCCTCG GCGGTCACGC CGTTTAGCAC GCCGCCGCAG 840ATTGCCAACC CGACCGCTCA GGGTACGCAG GCCGCGGCCG TGGCCACCGC CGCCGGTACC 900GCCCAGTCGA CGCTGACGGA GATGATCACC GGGCTACCCA ACGCGCTGCA AAGCCTCACC 960TCACNTCTGT TGCAGTCGTC TAACGGTCCG CTGTCGTGGC TGTGGCAGAT CTTGTTCGGC 1020ACGCCCAATT TCCCCACCTC AATTTCGGCA CTGCTGACCG ACCTGCAGCC CTACGCGAGC 1080TTNTTNTATA ACACCGAGGG CCTGCCGTAC TTCAGCATCG GCATGGGCAA CAACTTCATT 1140CAGTCGGCCA AGACCCTGGG ATTGATCGGC TAGGCGGCAC CGGCTGCGGT CGCGGNTGCT 1200GGGGATNCCG CCAAGGGCTT GCCTCGTGCC G 1231 2041 base pairs nucleic acid single linear cDNA 105CGGCACGAGC TCGTGCCGAT CAGTGCCATT GACGGCTTGT ACGACCTTCT GGGGATTGGA 60ATACCCAACC AAGGGGGTAT CCTTTACTCC TCACTAGAGT ACTTCGAAAA AGCCCTGGAG 120GAGCTGGCAG CAGCGTTTCC GGGTGATGGC TGGTTAGGTT CGGCCGCGGA CAAATACGCC 180GGCAAAAACC GCAACCACGT GAATTTTTTC CAGGAACTGG CAGACCTCGA TCGTCAGCTC 240ATCAGCCTGA TCCACGACCA GGCCAACGCG GTCCAGACGA CCCGCGACAT CCTGGAGGGC 300GCCAAGAAAG GTCTCGAGTT CGTGCGCCCG GTGGCTGTGG ACCTGACCTA CATCCCGGTC 360GTCGGGCACG CCCTATCGGC CGCCTTCCAN GCGCCGTTTT GCGCGGGCGC GATGGCCGTA 420GTGGGCGGCG CGCTTGCCTA CTTGGTCGTG AAAACGCTGA TCAACGCGAC TCAACTCCTC 480AAATTGCTTG CCAAATTGGC GGAGTTGGTC GCGGCCGCCA TTGCGGACAT CATTTCGGAT 540GTGGCGGACA TCATCAAGGG CATCCTCGGA GAAGTGTGGG AGTTCATCAC AAACGCGCTC 600AACGGCCTGA AAGAGCTTTG GGACAAGCTC ACGGGGTGGG TGACCGGACT GTTCTCTCGA 660GGGTGGTCGA ACCTGGAGTC CTTCTTTGCG GGCGTCCCCG GCTTGACCGG CGCGACCAGC 720GGCTTGTCGC AAGTGACTGG CTTGTTCGGT GCGGCCGGTC TGTCCGCATC GTCGGGCTTG 780GCTCACGCGG ATAGCCTGGC GAGCTCAGCC AGCTTGCCCG CCCTGGCCGG CATTGGGGGC 840GGGTCCGGTT TTGGGGGCTT GCCGAGCCTG GCTCAGGTCC ATGCCGCCTC AACTCGGCAG 900GCGCTACGGC CCCGAGCTGA TGGCCCGGTC GGCGCCGCTG CCGAGCAGGT CGGCGGGCAG 960TCGCAGCTGG TCTCCGCGCA GGGTTCCCAA GGTATGGGCG GACCCGTAGG CATGGGCGGC 1020ATGCACCCCT CTTCGGGGGC GTCGAAAGGG ACGACGACGA AGAAGTACTC GGAAGGCGCG 1080GCGGCGGGCA CTGAAGACGC CGAGCGCGCG CCAGTCGAAG CTGACGCGGG CGGTGGGCAA 1140AAGGTGCTGG TACGAAACGT CGTCTAACGG CATGGCGAGC CAAATCCATT GCTAGCCAGC 1200GCCTAACAAC GCGCAATGCT AAACGGAAGG GACACGATCA ATGACGGAAA ACTTGACCGT 1260CCAGCCCGAG CGTCTCGGTG TACTGGCGTC GCACCATGAC AACGCGGCGG TCGATGCNTC 1320CTCGGGCGTC GAAGCTGCCG CTGGCCTAGG CGAATCTGTG GCGATCACTC ACGGTCCGTA 1380CTGCTCACAG TTCAACGACA CGTTAAATGT GTACTTGACT GCCCACAATG CCCTGGGCTC 1440GTCCTTGCAT ACGGCCGGTG TCGATCTCGC CAAAAGTCTT CGAATTGCGG CGAAGATATA 1500TAGCGAGGCC GACGAAGCGT GGCGCAAGGC TATCGACGGG TTGTTTACCT GACCACGTTT 1560GCTGCCCGCA GTGCAGGCCA CGACGTAGCG CAGGTCGTGT CCCTCGTAGG CGTGGATGCG 1620ACCGGCCAGC ACCAGCACCC GGTGCGCACC GATGGGCACG GACAGTAGCT CGCCCGCATG 1680CCCGGCTGCG GTTGGCGGCA CAAACCCGGG CAGTTCGGCC TGCGGCAGCA CGGTGGTNGG 1740GGAGCCCAAC GCCGCAACGG CCGGTAACCA TCCCGACCCG AGCACGACCG AGACGTCATG 1800TTCGCCGATC CCGGTGCGGT CAGCGATGAC CTGCGCCGCC CGCCGGGCCA GTTTGTCGGG 1860ATCGGGGCGC GGGTCAGCCA CACTGGGCGA GCTTAACTGA GCCGCTCGCC GGGGAGCGGG 1920TGCTNGTCGA TGAGATACTG CGAGCATGCC AGCAGCCAGC GCATCCGACC GCGTCGAGGA 1980ATTGGTGCGG CGCCGTGGTG GCGAGCTGGT CGAGCTGTCC CATGCCATCC ACCTCGTGCC 2040G 2041 1202 base pairs nucleic acid single linear cDNA 106GAGCTCACCG CTATCAACCA ATACTTTCTG CACTCCAAGA TGCAGGACAA CTGGGGTTTT 60ACCGAGCTGG CGGCCCACAC CCGCGCGGAG TCGTTCGACG AAATGCGGCA CGCCGAGGAA 120ATCACCGATC GCATCTTGTT GCTGGATGGT TTGCCGAACT ACCAGCGCAT CGGTTCGTTG 180CGTATCGGCC AGACGCTCCG CGAGCAATTT GAGGCCGATC TGGCGATCGA ATACGACGTG 240TTGAATCGTC TCAAGCCAGG AATCGTCATG TGCCGGGAGA AACAGGACAC CACCAGCGCC 300GTACTGCTGG AGAAAATCGT TGCCGACGAG GAAGAACACA TCGACTACTT GGAAACGCAG 360CTGGAGCTGA TGGACAAGCT AGGAGAGGAG CTTTACTCGG CGCAGTGCGT CTCTCGCCCA 420CCGACCTGAT GCCCGCTTGA GGATTCTCCG ATACCACTCC GGGCGCCGCT GACAAGCTCT 480AGCATCGACT CGAACAGCGA TGGGAGGGCG GATATGGCGG GCCCCACAGC ACCGACCACT 540GCCCCCACCG CAATCCGAGC CGGTGGCCCG CTGCTCAGTC CGGTGCGACG CAACATTATT 600TTCACCGCAC TTGTGTTCGG GGTGCTGGTC GCTGCGACCG GCCAAACCAT CGTTGTGCCC 660GCATTGCCGA CGATCGTCGC CGAGCTGGGC AGCACCGTTG ACCAGTCGTG GGCGGTCACC 720AGCTATCTGC TGGGGGGAAC ACTSKYGKKK KTGKKGKSKS KSRMRMKCTC GGTGATCTGC 780TCGGCCGCAA CAGGGTGCTG CTAGGCTCCG TCGTGGTCTT CGTCGTTGGC TCTGTGCTGT 840GCGGGTTATC GCAGACGATG ACCATGCTGG CGATCTCTCG CGCACTGCAG GGCGTCGGTG 900CCGGTGCGAT TTCCGTCACC GCCTACGCGC TGGCCGCTGA GGTGGTCCCA CTGCGGGACC 960GTGGCCGCTA CCAGGGCGTC TTANGTGCGG TGTTCGGTGT CAACACGGTC ACCGGTCCGC 1020TGCTGGGGGG CTGGCTCACC GACTATCTGA GCTGGCGGTG GGCGTTCCGA CCACCAGCCC 1080CATCACCGAC CCGATCGCGG TCATCGCGGC GAACACCGCC CTCGCGGCGT TGCGGGCAGG 1140TCCCTTGGGG AACGTGGTCC CACAGCGCCA GAACGGTCGG AAATGCGATG GCCGACCCAC 1200AC 1202 496 base pairs nucleic acid single linear cDNA 107GGCGGCGGCA GTTGGCCAGC AGTTNGGGCG GGGGAGCCGG TTCGGNGACC AAGAAATCGG 60CCTGGGCAAG CAGCCGGGAC CGCGNACCGT GATCAGTTNG GATCGCCGGG ACCGCCGCCG 120ACCAANGCCA TTCCGCCGNT GAGGAAGTCG GAANTNTGCG CAGTGATGAC GCCCTGCTGC 180AACGCNTCCC GGATTGCCGA GCGGATCGCC GCCGAACGGC GGTGCTCACC ACCGGCGAGC 240ACCCCTACNG ACAGGCCCGC ATAGCTGAAT GACGCCGGGT NACCGCCGTC CCNTCCACCG 300NGANATCGGC CCGGANGCAA AAGATCCGTC GGCGCTCCGC CTCGGCGACG ACAGCCACGT 360TCACCCGCGC GTTATCGGTG GCCGCGATCG CATACCAGGC GCCGTCAAGG TNGCCGTYGC 420GGTAGTCACG CACCGACAAG GTGATYTGGT CCATCGCCTN GACGGCGGGG GTGACGCTGG 480GGGCGATCAM GTGCAC 496 849 base pairs nucleic acid single linear cDNA 108TGGATTCCGA TAGCGGTTTC GGCCCCTCGA CGGGCGACCA CGGCGCGCAG GCCTCCGAAC 60GGGGGGCCGG GACGCTGGGA TTCGCCGGGA CCGCAACCAA AGAACGCCGG GTCCGGGCGG 120TCGGGCTGAC CGCACTGGCC GGTGATGAGT TCGGCAACGG CCCCCGGATG CCGATGGTGC 180CGGGGACCTG GGAGCAGGGC AGCAACGAGC CCGAGGCGCC CGACGGATCG GGGAGAGGGG 240GAGGCGACGG CTTACCGCAC GACAGCAAGT AACCGAATTC CGAATCACGT GGACCCGTAC 300GGGTCGAAAG GAGAGATGTT ATGAGCCTTT TGGATGCTCA TATCCCACAG TTGGTGGCCT 360CCCAGTCGGC GTTTGCCGCC AAGGCGGGGC TGATGCGGCA CACGATCGGT CAGGCCGAGC 420AGGCGGCGAT GTCGGCTCAG GCGTTTCACC AGGGGGAGTC GTCGGCGGCG TTTCAGGCCG 480CCCATGCCCG GTTTGTGGCG GCGGCCGCCA AAGTCAACAC CTTGTTGGAT GTCGCGCAGG 540CGAATCTGGG TGAGGCCGCC GGTACCTATG TGGCCGCCGA TGCTGCGGCC GCGTCGACCT 600ATACCGGGTT CTGATCGAAC CCTGCTGACC GAGAGGACTT GTGATGTCGC AAATCATGTA 660CAACTACCCC GCGATGTTGG GTCACGCCGG GGATATGGCC GGATATGCCG GCACGCTGCA 720GAGCTTGGGT GCCGAGATCG CCGTGGAGCA GGCCGCGTTG CAGAGTGCGT GGCAGGGCGA 780TACCGGGATC ACGTATCAGG CGTGGCAGGC ACANTGGTAA CCANGCCANG GAAGATTTGG 840TGCGGGCCT 849 97 amino acids amino acid single linear protein 109Met Ser Leu Leu Asp Ala His Ile Pro Gln Leu Val Ala Ser Gln Ser 1 5 10 15Ala Phe Ala Ala Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala 20 25 30Glu Gln Ala Ala Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ser 35 40 45Ala Ala Phe Gln Ala Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys 50 55 60Val Asn Thr Leu Leu Asp Val Ala Gln Ala Asn Leu Gly Glu Ala Ala65 70 75 80Gly Thr Tyr Val Ala Ala Asp Ala Ala Ala Ala Ser Thr Tyr Thr Gly 85 90 95Phe 15 amino acids amino acid single linear peptide 110Met Ser Leu Leu Asp Ala His Ile Pro Gln Leu Val Ala Ser Gln 1 5 10 15 15 amino acids amino acid single linear peptide 111Ala His Ile Pro Gln Leu Val Ala Ser Gln Ser Ala Phe Ala Ala 1 5 10 15 15 amino acids amino acid single linear peptide 112Leu Val Ala Ser Gln Ser Ala Phe Ala Ala Lys Ala Gly Leu Met 1 5 10 15 15 amino acids amino acid single linear peptide 113Ser Ala Phe Ala Ala Lys Ala Gly Leu Met Arg His Thr Ile Gly 1 5 10 15 15 amino acids amino acid single linear peptide 114Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala Glu Gln Ala 1 5 10 15 15 amino acids amino acid single linear peptide 115Arg His Thr Ile Gly Gln Ala Glu Gln Ala Ala Met Ser Ala Gln 1 5 10 15 15 amino acids amino acid single linear peptide 116Gln Ala Glu Gln Ala Ala Met Ser Ala Gln Ala Phe His Gln Gly 1 5 10 15 15 amino acids amino acid single linear peptide 117Ala Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ser Ala Ala 1 5 10 15 15 amino acids amino acid single linear peptide 118Ala Phe His Gln Gly Glu Ser Ser Ala Ala Phe Gln Ala Ala His 1 5 10 15 15 amino acids amino acid single linear peptide 119Glu Ser Ser Ala Ala Phe Gln Ala Ala His Ala Arg Phe Val Ala 1 5 10 15 15 amino acids amino acid single linear peptide 120Phe Gln Ala Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys Val 1 5 10 15 15 amino acids amino acid single linear peptide 121Ala Arg Phe Val Ala Ala Ala Ala Lys Val Asn Thr Leu Leu Asp 1 5 10 15 15 amino acids amino acid single linear peptide 122Ala Ala Ala Lys Val Asn Thr Leu Leu Asp Val Ala Gln Ala Asn 1 5 10 15 15 amino acids amino acid single linear peptide 123Asn Thr Leu Leu Asp Val Ala Gln Ala Asn Leu Gly Glu Ala Ala 1 5 10 15 18 amino acids amino acid single linear peptide 124Val Ala Gln Ala Asn Leu Gly Glu Ala Ala Gly Thr Tyr Val Ala Ala 1 5 10 15Asp Ala 1752 base pairs nucleic acid single linear cDNA 125CGGCACGAGA ATGTCGCCTG TGCCTCGATA GCCACTTGCG TGTGGTCGCG CTGCCAGCGG 60GTCAGCCAGG TCGCCTGGTC CAGGCCATCG GGCCGGCGCA GGAGCGCGAT GTTGGCCAGA 120CCCGGTGTAC GAGAACCGGA CTCGACNAAG TGTCGGCGCT GACGGCGGCT CAGTTCGCGG 180CACACGCCCA GATCTATCAG GCCGTCAGCG CCCAGGCCGC GGCGATTCAC GAGATGTTCG 240TCAACACTCT ACAGATNANC TCAGGGTCGT ATGCTGCTAC CGAGGCCGCC AACGCGGCCG 300CGGCCGGCTA GAGGAGTCAC TGCGATGGAT TTTGGGGCGT TGCCGCCGGA GGTCAATTCG 360GTGCGGATGT ATGCCGGTCC TGGCTCGGCA CCAATGGTCG CTGCGGCGTC GGCCTGGAAC 420GGGTTGGCCG CGGAGCTGAG TTCGGCGGCC ACCGGTTATG AGACGGTGAT CACTCAGCTC 480AGCAGTGAGG GGTGGCTAGG TCCGGCGTCA GCGGCGATGG CCGAGGCAGT TGCGCCGTAT 540GTGGCGTGGA TGAGTGCCGC TGCGGCGCAA GCCGAGCAGG CGGCCACACA GGCCAGGGCC 600GCCGCGGCCG CTTTTGAGGC GGCGTTTGCC GCGACGGTGC CTCCGCCGTT GATCGCGGCC 660AACCGGGCTT CGTTGATGCA GCTGATCTCG ACGAATGTCT TTGGTCAGAA CACCTCGGCG 720ATCGCGGCCG CCGAAGCTCA GTACGGCGAG ATGTGGGCCC AAGACTCCGC GGCGATGTAT 780GCCTACGCGG GCAGTTCGGC GAGCGCCTCG GCGGTCACGC CGTTTAGCAC GCCGCCGCAG 840ATTGCCAACC CGACCGCTCA GGGTACGCAG GCCGCGGCCG TGGCCACCGC CGCCGGTACC 900GCCCAGTCGA CGCTGACGGA GATGATCACC GGGCTACCCA ACGCGCTGCA AAGCCTCACC 960TCACNTCTGT TGCAGTCGTC TAACGGTCCG CTGTCGTGGC TGTGGCAGAT CTTGTTCGGC 1020ACGCCCAATT TCCCCACCTC AATTTCGGCA CTGCTGACCG ACCTGCAGCC CTACGCGAGC 1080TTNTTNTATA ACACCGAGGG CCTGCCGTAC TTCAGCATCG GCATGGGCAA CAACTTCATT 1140CAGTCGGCCA AGACCCTGGG ATTGATCGGC TAGGCGGCAC CGGCTGCGGT CGCGGCTGCT 1200GGGGATGCCG CCAAGGGCTT GCCTGGACTG GGCGGGATGC TCGGTGGCGG GCCGGTGGCG 1260GCGGGTCTGG GCAATGCGGC TTCGGTTGGC AAGCTGTCGG TGCCGCCGGT GTGGANTGGA 1320CCGTTGCCCG GGTCGGTGAC TCCGGGGGCT GCTCCGCTAC CGGTGAGTAC GGTCAGTGCC 1380GCCCCGGAGG CGGCGCCCGG AAGCCTGTTG GGCGGCCTGC CGCTANCTGG TGCGGGCGGG 1440GCCGGCGCGG GTCCACGCTA CGGATTCCRT CCCACCGTCA TGGCTCGCCC ACCCTTCGMC 1500GGGATAGTCG CTGCCGCAAC GTATTAACGC GCCGGCCTCG GCTGGTGTGG TCCGCTGCGG 1560GTGGCAATTG GTCNGCGCCG AAATCTCSGT GGGTTATTTR CGGTGGGATT TTTTCCCGAA 1620GCCGGGTTCA RCACCGGATT TCCTAACGGT CCCGCKACTC TCGTGCCGAA TTCSGCACTA 1680AGTGACGTCC GGCGGAAACC CGTTGGGTNT GAAAGCTTCA GAAAGGCCCG CTCCCAGGGG 1740TTCGGCAAAC GG 1752 400 amino acids amino acid single linear protein 126Met Asp Phe Gly Ala Leu Pro Pro Glu Val Asn Ser Val Arg Met Tyr 1 5 10 15Ala Gly Pro Gly Ser Ala Pro Met Val Ala Ala Ala Ser Ala Trp Asn 20 25 30Gly Leu Ala Ala Glu Leu Ser Ser Ala Ala Thr Gly Tyr Glu Thr Val 35 40 45Ile Thr Gln Leu Ser Ser Glu Gly Trp Leu Gly Pro Ala Ser Ala Ala 50 55 60Met Ala Glu Ala Val Ala Pro Tyr Val Ala Trp Met Ser Ala Ala Ala65 70 75 80Ala Gln Ala Glu Gln Ala Ala Thr Gln Ala Arg Ala Ala Ala Ala Ala 85 90 95Phe Glu Ala Ala Phe Ala Ala Thr Val Pro Pro Pro Leu Ile Ala Ala 100 105 110Asn Arg Ala Ser Leu Met Gln Leu Ile Ser Thr Asn Val Phe Gly Gln 115 120 125Asn Thr Ser Ala Ile Ala Ala Ala Glu Ala Gln Tyr Gly Glu Met Trp 130 135 140Ala Gln Asp Ser Ala Ala Met Tyr Ala Tyr Ala Gly Ser Ser Ala Ser145 150 155 160Ala Ser Ala Val Thr Pro Phe Ser Thr Pro Pro Gln Ile Ala Asn Pro 165 170 175Thr Ala Gln Gly Thr Gln Ala Ala Ala Val Ala Thr Ala Ala Gly Thr 180 185 190Ala Gln Ser Thr Leu Thr Glu Met Ile Thr Gly Leu Pro Asn Ala Leu 195 200 205Gln Ser Leu Thr Ser Xaa Leu Leu Gln Ser Ser Asn Gly Pro Leu Ser 210 215 220Trp Leu Trp Gln Ile Leu Phe Gly Thr Pro Asn Phe Pro Thr Ser Ile225 230 235 240Ser Ala Leu Leu Thr Asp Leu Gln Pro Tyr Ala Ser Xaa Xaa Tyr Asn 245 250 255Thr Glu Gly Leu Pro Tyr Phe Ser Ile Gly Met Gly Asn Asn Phe Ile 260 265 270Gln Ser Ala Lys Thr Leu Gly Leu Ile Gly Ser Ala Ala Pro Ala Ala 275 280 285Val Ala Ala Ala Gly Asp Ala Ala Lys Gly Leu Pro Gly Leu Gly Gly 290 295 300Met Leu Gly Gly Gly Pro Val Ala Ala Gly Leu Gly Asn Ala Ala Ser305 310 315 320Val Gly Lys Leu Ser Val Pro Pro Val Trp Xaa Gly Pro Leu Pro Gly 325 330 335Ser Val Thr Pro Gly Ala Ala Pro Leu Pro Val Ser Thr Val Ser Ala 340 345 350Ala Pro Glu Ala Ala Pro Gly Ser Leu Leu Gly Gly Leu Pro Leu Xaa 355 360 365Gly Ala Gly Gly Ala Gly Ala Gly Pro Arg Tyr Gly Phe Xaa Pro Thr 370 375 380Val Met Ala Arg Pro Pro Phe Xaa Gly Ile Val Ala Ala Ala Thr Tyr385 390 395 400 474 base pairs nucleic acid single linear cDNA 127GGCACGAGCA CCAGTTGACC CGCGAAGAAC CTGACCGCGC CACCCAGCGC CGCCCGCATC 60ACCGGCCCCG TCCCACGAAC CTTTTCGGTA AACGAGCCAC TCCAGCGGAG ATCGGTACCG 120CCCGACGCAT TTGGTGTAAG GACCACCTCG CCGAAGTAGT CCTGGACGGG TGTCCTCGCG 180CCAACCAGCT TGTAGACGTG GCGACGGTCC TGCTCATACT CGACGGTCTC TTCCTGCACG 240AACACCGGCC ACATGCCTAG TTTGCGGATG GCCCCGATGC CGCCGGGCGC GGGATCACCG 300CGTCGCGCCC AACTCGATTG AGCAACGATG GGCTTGGCCC AGGTCGCCCA GTTGCCACCG 360TCTGTCACGA GCCGAAACAA GGTTGCAGCC GGCGCGCTGC TGGTCTTGGT GACCTCGAAC 420GAAAATTTCC GACCCGACAT GCGCGACTCC CGAAACGACA ACTGAAGCTC GTGC 474 1431 base pairs nucleic acid single linear cDNA 128CTGCGCGCCG GAAAAAANTA TTACTGGCAG GACCGGCAGA ATGCATGGTG ATATTCCGGT 60GATGAGGCCG CCGAGGAACC GACTAGTGCG AGGGTCAACA CATCGGTTAT TCGTTGCCGT 120TTAGGTCTTG GATCTGCCGG GACGGCAACG AGTTGGCAGG ACCGCTCACG CGAGCGCTGT 180TGACAGAGTC GGTTCACGTC GAACTCGCCA CCCGTCAGAT GCGAATGATA GCCACATCGG 240CCACACCATC GACGGCGTCG AAGTCGCCGT CGTGGGTCAC GACCGGCACC CCTTGCGACG 300TGGCAACGGC AGCGGCCCTC ACCGGACGGG ACCGAGATCG TCGGTGGTGT CGCCAGTGAG 360CGTTGCGAGG TCGCGGGTGC AATCCCGCAT CTGCTTGCGT ATGCCGAAGC CGCCGCAGCA 420GCTCGTCTCG ACTCAACCAT CGGCGCCGTG CGGGCTGCCT GCGGTCAGCA GCGCAACGGG 480TTTGCCGTTG GCAGTGATGG TGATGTCTTC GCCGGCCTGC ACGCGCCGTA GCAGCCCGGC 540GGTGTTGTTG CGCAGTTCGC GAGACGCGAC TTCAGCAGGC ATGCTGCGGG GATCGGCTTG 600CGCTGGGCGC GGTGTCACCG TCATGCGCTT GGGATATCAC GTGATCTATC GGCACGAAGC 660CGCCGGATGA GCGAGGCAAA CCGCCTACAC GGGCTGCCTC GCCTTGACCG CGCCGAACGT 720TACTGTGCCG GGGGCATCAG CACCGTATCG ATCATGTACA CCGTCGCGTG GGCGGTGTGA 780CTCCGCCACA TACCAAACGG GCGTTGTTGA CCATGAGTCG TCGCGGGCGC CTATCACCGT 840CAGGTCGGCA CCTTGCAGGT CTGATGGGTG CCGTCGATCC TGCTCGGACT CGCCTGGCCG 900GCTATCACGT GGTAGGTCAG GATGCTGCTG AGCAGCTTGG CGTCAGTCTT GAGTTGATCG 960ATAGTGGCCG CCGGCAGCTT GTCGAATGCG GCGTTGGTGG GGGCGAAAAC GGTGTACTCG 1020CCGCCGTTGA GGGTGTCGAC CAGATTCACA TCCGGGTTCA GCTTGCCCGA CAGAGCCGAG 1080GTCAGGGTAC TGAGCATCGG GTTGTTGGAA GCCGCGGTAG CGACCGGGTC TTGCGCCATT 1140CCGGCCACCG ATCCGGGACC GGTGGGATTT TGCGCCGCGT ATTGCGCGCA CCCACGACCA 1200ATCAGGTCCG CTGCGGTCAG CCATTGCCGC CGTGGTAACG GGCGCCGCCG GGCTGGTCGC 1260CGGTTTCGGG CTGGTGTCTT GCGACACGGG TTTGGTGCTC GAACAACCCG CTAAGAACGC 1320AATCGCGATG GCTGCGAGGC TCGCTGCTGC GGCCGGTTTG GCCTGAACGT TGATCATCGC 1380TTCGATTCCT TTGCTTCTGC GGCGGCGTTG AACGCCGTCC TCCTGGGTGG A 1431 279 base pairs nucleic acid single linear cDNA 129GCACGAGAGT CGTATCTTTG CACCCAGCGC CCGTAGGAAA CCGCTGGCCT GGCTAACTCA 60GATGCGGGCG GCCGTCGATT CGAGAGGTAA CCGATCGCCC GCCGACAATG GGTTACCCAC 120CGAGACTGAT TGCCGCGCAG CCGCCTTCGA CGTGTAAGCG CCGGTTCGTG CATGCCCGGA 180ACGGCTGCAC TCACGGACCT TCTACGTAGT ACGTGACGGA CTTTTACGCA TTATCGCTGA 240CGATCTTTGC CTCCCAGGAC TCCAGAATCT ACTCGTGCC 279 1470 base pairs nucleic acid single linear cDNA 130ACCGCCACCC GCAGCCCGGA ATCACCGTCG GTAACCTGCG AATACAATTT CTTCATCGAC 60GACTTCGCGA ACAGCGAACC CGAGCCCACC GCCTGATAGC CTTCTTCCTC GATGTTCCAA 120CCGCCGGCGG CGTCGAACGA AACGATACGA CCCGCGCTCT GCGGGTCAGA CGCATGAATG 180TCGTAGCCCG CCAGCAACGG CAACGCCAGC AGACCCTGCA TCGCGGCCGC CAGATTGCCA 240CGCACCATAA TCGCCAGCCG GTTGATTTTG CCGGCAAACG TCAGCGGCAC ACCCTCGAGC 300TTCTCGTAGT GCTCAAGTTC CACGGCATAC AGCCGGGCAA ACTCAACCGC GACCGCAGCC 360GTGCCAGCGA TGCCGGTAGC GGTGTAGTCA TCGGTGATAT ACACCTTGCG CACATCACGC 420CCAGAAATCA TGTTGCCCTG CGTCGAACGC CGGTCACCCG CCATGACAAC ACCGCCGGGG 480TATTTCAGCG CGACAATGGT GGTGCCGTGC GGCAGTTGCG CATCGCCGCC TGCGAGTGGC 540GCACCGCCGC TGATGCTTGC CGGCAGCAAC TCCGGCGCCT GGCGGCGCAG GAAGTCAAGT 600GAAAGAAGAT AGGTCTACAG CGGGTGTTCC AGAGAGTGAA TTAATGGACA GGCGATCGGG 660CAACGGCCAG GTCACTGTCC GCCCTTTTGG ACGTATGCGC GGACGAAGTC CTCGGCGTTC 720TCCTCGAGGA CGTCGTCGAT TTCGTCGAGC AGATCGTCGG TCTCCTCGGT CAGCTTTTCG 780CGACGCTCCT GGCCCGCGGC GGTGCTGCCG GCGATGTCGT CATCATCGCC GCCGCCACCG 840CCACGCTTGG TCTGCTCTTG CGCCATCGCC GCCTCCTGCT TCCTCATGGC CTTTCAAAAG 900GCCGCGGGTG CGCGTCACAC GCCCGCTGTC TTTCTCTCAC CTACCGGTCA ACACCAACGT 960TTCCCGGCCT AACCAGGCTT AGCGAGGCTC AGCGGTCAGT TGCTCTACCA GCTCCACGGC 1020ACTGTCCACC GAATCCAGCA ACGCACCAAC ATGCGCCTTA CTACCCCGCA ACGGCTCCAG 1080CGTCGGGATG CGAACCAGCG AGTCGCCGCC AGGTCGAAGA TCACCGAGTC CCAGCTAGCC 1140GCGGCGATAT CAGCCCCGAA CCGGCGCAGG CATTTCGCCG CGGAAATACG CGCGGGTGTC 1200GGTCGGCGGT TCTCCACCGC ACTCAGCACC TGGTGTTTCG GTGACTAAAC GCTTTATCGA 1260GCCGCGCGCG ACCAGCCGGT TGTACAGGCC CTTGTCCAGC CGGACATCGG AGTACTGCAG 1320GTTGACGAGG TGCAGCCGGG GCGCCGACCA GCTCAGGTTC TCCCGCTGCC GGAAACCGTC 1380GAGCAGCCGC AGTTTGGCCG GCCAGTCCAG CAGCTCCGCG CAATCCATCG GGTCACGCTC 1440GAGCTGATCC AGCACGTGTG CCCAGGTTTC 1470 1059 base pairs nucleic acid single linear cDNA 131ATTCCCATCG CTCCGGCACC TATCACCAGG TAGTCGGTTT CGATGGTTTT CGCCGGCCCT 60TGCGTTGGCC TGGGCCACGG GTCGTTCATG GGCCCTCCTG TGCGGATTGG AATTTGTGAC 120AACGAAATCG GGCGATCGGT GAGCAATCGT CGCCGATGCA AGACACGCTT TCGCTGCCGC 180GGCGTCAGGT GGAGTTTAGG CCAGCGTAAC AACGTAGACC GGCCACTGAC CAAACCCCAA 240ACCCACAAAC CCTGGACGCA TGCGGGTCTC GGGCGTCAAA TTCCGGGTAG ATATCGTATA 300CCGATATCGG ATGCCGTAGC CTTATCGAGG CATGAGACGC CCGCTAGACC CACGCGATAT 360TCCAGATGAG CTGCGGCGAC GGCTGGGGCT CTTGGATGCG GTGGTGATCG GGCTTGGGTC 420CATGATCGGT GCCGGAATCT TTGCTCGTGC CGAATTCGGC ACGAGCTCGT GCCGAATTCG 480GCACGAGATT CCAATCCCCA GAAGGTCGTA CAAGCCGTCA ATGGCACTTG ATCGTTGGAT 540CGATGATGAA CGCTCTGCTC ATGCCTGCCG CCTATCTCAA CGGTCGTCGA TTCCATGCAT 600TAGCCTTGGT TCTGCATTGC ACGCGTAGGG CCTACAGTCT GGCTGTCATG CTTGGCCGAT 660GTCAACAGTT TTTTTCATGC TAAGCAGATC GTCAGTTTTG AGTTCGTGAA GACGGCATGT 720TCACTTGTTG TCGACTACAT CGTCTGCGCA CATTTGCCCT CCTGCAACTG CGCTGCGACA 780ATGCGCCAAC CGCCGTGTAG CTCGTGCCGA ATTCGGCACG AGGATCCACC GGAGATGGCC 840GACGACTACG ACGAGGCCTG GATGCTCAAC ACCGTGTTCG ACTATCACAA CGAGAACGCA 900AAAGAAGAGG TCATCCATCT CGTGCCCGAC GTGAACAAGG AGAGGGGGCC CATCGAACTC 960GTAACCAAGG TAGACAAAGA GGGACATCAG ACTCGTCTAC GATGGGGAGC CACGTTTTCA 1020TACAAGGAAC ATCCTAAGTT TTGATTCGGG AACATCCTA 1059 153 base pairs nucleic acid single linear cDNA 132GCACGAGGCA TTGGCGGGCA TCTGCATAAA CGGTGACGTA TCAGCACAAA ACAGCGGAGA 60GAACAACATG CGATCAGAAC GTCTCCGGTG GCTGGTAGCC GCAGAAGGTC CGTTCGCCTC 120GGTGTATTTC GACGACTCGC ACGACTCGTG CCG 153 387 base pairs nucleic acid single linear cDNA 133CCGCGCGGTC GATCAGCGAG CCAGGCAAAA ACTCCGTCGA GCCCGAGTCG ATGATGGTCA 60CCCGGCGCAG CATCTGGCGA ACGATCACCT CGATGTGCTT GTCGTGGATC GACACACCTT 120GGGCGCGGTA GACCTCCTGG ACCTCGCGAA CCAGGTGTAT CTGCACCTCG CGGGGGCCCT 180GCACCCGCAG CACCTCATGC GGGTCGGCCG AGCCTTCCAT CAGCTGCTGG CCCACCTCGA 240CGTGGTCGCC ATCGGAGAGC ACCCGTTCGG AACCGTCTTC GTGCTTGAAC ACCCGCAGCC 300GCTGCCGCTT GGAGATCTTG TCGTAGACCA CTTCCTCACC GCCGTCGTCA GGAACGATGG 360TGATCTTGTA GAACCGCTCG CCGTCCT 387 389 base pairs nucleic acid single linear cDNA 134GTTCAGCACG GCTATCCGAT TGTGCCGTTC GCTTCGGTGG GTGCTGAACA CGGCATCGAC 60ATCGTGCTCG ACAACGAATC CCCACTGCTG GCACCGGTCC AGTTCCTCGC CGAGAAGCTG 120CTCGGCACCA AAGACGGTCC GGCGCTGGTC CGTGGTGTCG GACTGACACC GGTACCGCGC 180CCCGAACGGC AGTATTACTG GTTCGGCGAG CCAACCGACA CCACAGAGTT TATGGGGCAG 240CAAGCCGACG ATAACGCCGC ACGCAGGGTG CGCGAGCGTG CCGCCGCCGC TATCGAACAC 300GGCATCGAGC TGATGCTGGC CGAGCGCGCA GCCGATCCAA ATCGATCCCT GGTCGGACGG 360CTCTTGCGCT CGGACGCCTA AGGCGCCCC 389 480 base pairs nucleic acid single linear cDNA 135CCCGCGGTCG GAATGATCCC CGTCTCGTCG CGCGCCCATT TGATGCTGTT GATGAGCTGT 60TTGGAGAAGC CCGGTTGGCG TACCGGTGAG CCGGAATATC TGTTGGAAGC GTCACCGGAT 120GTNCACATGA ANTNCNTTGN CCCNGTNGCG GTNTTGGNTG NGGNAAACAC GTGTTGTNTA 180AGCCTTGNTG GNCTCGNAAG NGCCGTNGAC GCCTGTGTCG CCGAAGATAA TGAGCACCTG 240ACGGTTGGCG GGATCGCCGT TATCCCAAGG AATTCCGAGG TCGGTCCCGG AGATGCCGAA 300GCGTTCCAGG GTCTTGTTGG GGCTGTCCGG TCCGGTCACC CACTCGGCGA GGGATGTGGN 360AGCCCCGGCG AGCGTGGCAC CAGGATCCGG CGCCGCCGCC GGAGCAGGGT CGGNNGCTGN 420NCTGNNTTCC TNNNGCCNAA TTNNACTCCN NCNACAANCT TGNNNCCGAC TCNNACCCGN 480 587 base pairs nucleic acid single linear cDNA 136GCACGAGGCT ACCGGCGCGT CGCCCGCCAT GCCCTGGATG CACGCGTAGC CACCCGTNCA 60TNCAGCGGGT CAGCCGCCGC GTCCGGGCTT AACGCTATAG CAGCTGCAAA CAACCCAGCG 120CCGGCAATTA CTTTGATGTT GAACCGATGA CCATNGCCTN CGNGTNCAAT CTCNTCTCTT 180NGCGCGCCNC TATTTNNGCC ATANATTTGG TTNNANNCGN AACGCTAGAC GTATCGAGTT 240CCTTTTCGAC CACCGGCTCA ATTGTCAGCA TCCTATGGGG AACATGAGCC CCGCCGCACC 300GGGCCGTTTC CAAATGGTGA CGTCACAACG GTGTCACAAG CCAGCGCAAT GTCCGCGGTA 360GGGACGCGGC GGCTGGGATC GGTGGGGTGA GCGCCCGGCT TCTCAAAGCG AGGGGAGCCC 420CGGGACTCTT ACCGGCCGAA GGCGGCGGGT GTCACTGATC TAGGCTGACG GCCAGTGGTT 480GNTNAGCCAA CAAGGATGAC NACAAATAAN CCGAGGANAG ACANGNGACG GNCCGANANG 540CTNANCCGGN NTTGNNCNAA NNNNACNCAC TTNTACCGNN CTTATGN 587 1200 base pairs nucleic acid single linear cDNA 137CAGGCATGAG CAGAGCGTTC ATCATCGATC CAACGATCAG TGCCATTGAC GGCTTGTACG 60ACCTTCTGGG GATTGGAATA CCCAACCAAG GGGGTATCCT TTACTCCTCA CTAGAGTACT 120TCGAAAAAGC CCTGGAGGAG CTGGCAGCAG CGTTTCCGGG TGATGGCTGG TTAGGTTCGG 180CCGCGGACAA ATACGCCGGC AAAAACCGCA ACCACGTGAA TTTTTTCCAG GAACTGGCAG 240ACCTCGATCG TCAGCTCATC AGCCTGATCC ACGACCAGGC CAACGCGGTC CAGACGACCC 300GCGACATCCT GGAGGGCGCC AAGAAAGGTC TCGAGTTCGT GCGCCCGGTG GCTGTGGACC 360TGACCTACAT CCCGGTCGTC GGGCACGCCC TATCGGCCGC CTTCCAGGCG CCGTTTTGCG 420CGGGCGCGAT GGCCGTAGTG GGCGGCGCGC TTGCCTACTT GGTCGTGAAA ACGCTGATCA 480ACGCGACTCA ACTCCTCAAA TTGCTTGCCA AATTGGCGGA GTTGGTCGCG GCCGCCATTG 540CGGACATCAT TTCGGATGTG GCGGACATCA TCAAGGGCAC CCTCGGAGAA GTGTGGGAGT 600TCATCACAAA CGCGCTCAAC GGCCTGAAAG AGCTTTGGGA CAAGCTCACG GGGTGGGTGA 660CCGGACTGTT CTCTCGAGGG TGGTCGAACC TGGAGTCCTT CTTTGCGGGC GTCCCCGGCT 720TGACCGGCGC GACCAGCGGC TTGTCGCAAG TGACTGGCTT GTTCGGTGCG GCCGGTCTGT 780CCGCATCGTC GGGCTTGGCT CACGCGGATA GCCTGGCGAG CTCAGCCAGC TTGCCCGCCC 840TGGCCGGCAT TGGGGGCGGG TCCGGTTTTG GGGGCTTGCC GAGCCTGGCT CAGGTCCATG 900CCGCCTCAAC TCGGCAGGCG CTACGGCCCC GAGCTGATGG CCCGGTCGGC GCCGCTGCCG 960AGCAGGTCGG CGGGCAGTCG CAGCTGGTCT CCGCGCAGGG TTCCCAAGGT ATGGGCGGAC 1020CCGTAGGCAT GGGCGGCATG CACCCCTCTT CGGGGGCGTC GAAAGGGACG ACGACGAAGA 1080AGTACTCGGA AGGCGCGGCG GCGGGCACTG AAGACGCCGA GCGCGCGCCA GTCGAAGCTG 1140ACGCGGGCGG TGGGCAAAAG GTGCTGGTAC GAAACGTCGT CTAACGGCAT GGCGAGCCAA 1200 392 amino acids amino acid single linear protein 138Met Ser Arg Ala Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly 1 5 10 15Leu Tyr Asp Leu Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu 20 25 30Tyr Ser Ser Leu Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala 35 40 45Ala Phe Pro Gly Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala 50 55 60Gly Lys Asn Arg Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu65 70 75 80Asp Arg Gln Leu Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln 85 90 95Thr Thr Arg Asp Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val 100 105 110Arg Pro Val Ala Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala 115 120 125Leu Ser Ala Ala Phe Gln Ala Pro Phe Cys Ala Gly Ala Met Ala Val 130 135 140Val Gly Gly Ala Leu Ala Tyr Leu Val Val Lys Thr Leu Ile Asn Ala145 150 155 160Thr Gln Leu Leu Lys Leu Leu Ala Lys Leu Ala Glu Leu Val Ala Ala 165 170 175Ala Ile Ala Asp Ile Ile Ser Asp Val Ala Asp Ile Ile Lys Gly Thr 180 185 190Leu Gly Glu Val Trp Glu Phe Ile Thr Asn Ala Leu Asn Gly Leu Lys 195 200 205Glu Leu Trp Asp Lys Leu Thr Gly Trp Val Thr Gly Leu Phe Ser Arg 210 215 220Gly Trp Ser Asn Leu Glu Ser Phe Phe Ala Gly Val Pro Gly Leu Thr225 230 235 240Gly Ala Thr Ser Gly Leu Ser Gln Val Thr Gly Leu Phe Gly Ala Ala 245 250 255Gly Leu Ser Ala Ser Ser Gly Leu Ala His Ala Asp Ser Leu Ala Ser 260 265 270Ser Ala Ser Leu Pro Ala Leu Ala Gly Ile Gly Gly Gly Ser Gly Phe 275 280 285Gly Gly Leu Pro Ser Leu Ala Gln Val His Ala Ala Ser Thr Arg Gln 290 295 300Ala Leu Arg Pro Arg Ala Asp Gly Pro Val Gly Ala Ala Ala Glu Gln305 310 315 320Val Gly Gly Gln Ser Gln Leu Val Ser Ala Gln Gly Ser Gln Gly Met 325 330 335Gly Gly Pro Val Gly Met Gly Gly Met His Pro Ser Ser Gly Ala Ser 340 345 350Lys Gly Thr Thr Thr Lys Lys Tyr Ser Glu Gly Ala Ala Ala Gly Thr 355 360 365Glu Asp Ala Glu Arg Ala Pro Val Glu Ala Asp Ala Gly Gly Gly Gln 370 375 380Lys Val Leu Val Arg Asn Val Val385 390 439 base pairs nucleic acid single linear cDNA 139ACGTTTACCC ATGCCGTCGG TGCAGAGCAA CGCCAGACAA CACAAAGTAG TCTAATTCCG 60TTATAAAGCA GACATTTCCG TGGTTATGTA GAAGATGTCG ACCGATCAGA TGAAGCGATC 120CGCGTCAGGT GGTATCCGAT GTCTTTTGTG ACCATCCAGC CGGTGGTCTT GGCAGCCGCG 180ACGGGGGACT TGCCGACGAT CGGTACCGCC GTGAGTGCTC GGAACACAGC CGTCTGTGCC 240CCGACGACGG GGGTGTTACC CCCTGCTGCC AATGACGTGT CGGTCCTGAC GGCGGCCCGG 300TTCACCGCGC ACACCAAGCA CTACCGAGTG GTGAGTAAGC CGGCCGCGCT GGTCCATGGC 360ATGTTCGTGG CCCTCCCGGC GGCCACCGCC GATGCGTATG CGACCACCGA GGCCGTCAAT 420GTGGTCGCGA CCGGTTAAG 439 1441 base pairs nucleic acid single linear cDNA 140GAGGTTGCTG GCAATGGATT TCGGGCTTTT ACCTCCGGAA GTGAATTCAA GCCGAATGTA 60TTCCGGTCCG GGGCCGGAGT CGATGCTAGC CGCCGCGGCC GCCTGGGACG GTGTGGCCGC 120GGAGTTGACT TCCGCCGCGG TCTCGTATGG ATCGGTGGTG TCGACGCTGA TCGTTGAGCC 180GTGGATGGGG CCGGCGGCGG CCGCGATGGC GGCCGCGGCA ACGCCGTATG TGGGGTGGCT 240GGCCGCCACG GCGGCGCTGG CGAAGGAGAC GGCCACACAG GCGAGGGCAG CGGCGGAAGC 300GTTTGGGACG GCGTTCGCGA TGACGGTGCC ACCATCCCTC GTCGCGGCCA ACCGCAGCCG 360GTTGATGTCG CTGGTCGCGG CGAACATTCT GGGGCAAAAC AGTGCGGCGA TCGCGGCTAC 420CCAGGCCGAG TATGCCGAAA TGTGGGCCCA AGACGCTGCC GTGATGTACA GCTATGAGGG 480GGCATCTGCG GCCGCGTCGG CGTTGCCGCC GTTCACTCCA CCCGTGCAAG GCACCGGCCC 540GGCCGGGCCC GCGGCCGCAG CCGCGGCGAC CCAAGCCGCC GGTGCGGGCG CCGTTGCGGA 600TGCACAGGCG ACACTGGCCC AGCTGCCCCC GGGGATCCTG AGCGACATTC TGTCCGCATT 660GGCCGCCAAC GCTGATCCGC TGACATCGGG ACTGTTGGGG ATCGCGTCGA CCCTCAACCC 720GCAAGTCGGA TCCGCTCAGC CGATAGTGAT CCCCACCCCG ATAGGGGAAT TGGACGTGAT 780CGCGCTCTAC ATTGCATCCA TCGCGACCGG CAGCATTGCG CTCGCGATCA CGAACACGGC 840CAGACCCTGG CACATCGGCC TATACGGGAA CGCCGGCGGG CTGGGACCGA CGCAGGGCCA 900TCCACTGAGT TCGGCGACCG ACGAGCCGGA GCCGCACTGG GGCCCCTTCG GGGGCGCGGC 960GCCGGTGTCC GCGGGCGTCG GCCACGCAGC ATTAGTCGGA GCGTTGTCGG TGCCGCACAG 1020CTGGACCACG GCCGCCCCGG AGATCCAGCT CGCCGTTCAG GCAACACCCA CCTTCAGCTC 1080CAGCGCCGGC GCCGACCCGA CGGCCCTAAA CGGGATGCCG GCAGGCCTGC TCAGCGGGAT 1140GGCTTTGGCG AGCCTGGCCG CACGCGGCAC GACGGGCGGT GGCGGCACCC GTAGCGGCAC 1200CAGCACTGAC GGCCAAGAGG ACGGCCGCAA ACCCCCGGTA GTTGTGATTA GAGAGCAGCC 1260GCCGCCCGGA AACCCCCCGC GGTAAAAGTC CGGCAACCGT TCGTCGCCGC GCGGAAAATG 1320CCTGGTGAGC GTGGCTATCC GACGGGCCGT TCACACCGCT TGTAGTAGCG TACGGCTATG 1380GACGACGGTG TCTGGATTCT CGGCGGCTAT CAGAGCGATT TTGCTCGCAA CCTCAGCAAA 1440G 1441 99 amino acids amino acid single linear protein 141Met Ser Phe Val Thr Ile Gln Pro Val Val Leu Ala Ala Ala Thr Gly 1 5 10 15Asp Leu Pro Thr Ile Gly Thr Ala Val Ser Ala Arg Asn Thr Ala Val 20 25 30Cys Ala Pro Thr Thr Gly Val Leu Pro Pro Ala Ala Asn Asp Val Ser 35 40 45Val Leu Thr Ala Ala Arg Phe Thr Ala His Thr Lys His Tyr Arg Val 50 55 60Val Ser Lys Pro Ala Ala Leu Val His Gly Met Phe Val Ala Leu Pro65 70 75 80Ala Ala Thr Ala Asp Ala Tyr Ala Thr Thr Glu Ala Val Asn Val Val 85 90 95Ala Thr Gly 423 amino acids amino acid single linear protein 142Met Asp Phe Gly Leu Leu Pro Pro Glu Val Asn Ser Ser Arg Met Tyr 1 5 10 15Ser Gly Pro Gly Pro Glu Ser Met Leu Ala Ala Ala Ala Ala Trp Asp 20 25 30Gly Val Ala Ala Glu Leu Thr Ser Ala Ala Val Ser Tyr Gly Ser Val 35 40 45Val Ser Thr Leu Ile Val Glu Pro Trp Met Gly Pro Ala Ala Ala Ala 50 55 60Met Ala Ala Ala Ala Thr Pro Tyr Val Gly Trp Leu Ala Ala Thr Ala65 70 75 80Ala Leu Ala Lys Glu Thr Ala Thr Gln Ala Arg Ala Ala Ala Glu Ala 85 90 95Phe Gly Thr Ala Phe Ala Met Thr Val Pro Pro Ser Leu Val Ala Ala 100 105 110Asn Arg Ser Arg Leu Met Ser Leu Val Ala Ala Asn Ile Leu Gly Gln 115 120 125Asn Ser Ala Ala Ile Ala Ala Thr Gln Ala Glu Tyr Ala Glu Met Trp 130 135 140Ala Gln Asp Ala Ala Val Met Tyr Ser Tyr Glu Gly Ala Ser Ala Ala145 150 155 160Ala Ser Ala Leu Pro Pro Phe Thr Pro Pro Val Gln Gly Thr Gly Pro 165 170 175Ala Gly Pro Ala Ala Ala Ala Ala Ala Thr Gln Ala Ala Gly Ala Gly 180 185 190Ala Val Ala Asp Ala Gln Ala Thr Leu Ala Gln Leu Pro Pro Gly Ile 195 200 205Leu Ser Asp Ile Leu Ser Ala Leu Ala Ala Asn Ala Asp Pro Leu Thr 210 215 220Ser Gly Leu Leu Gly Ile Ala Ser Thr Leu Asn Pro Gln Val Gly Ser225 230 235 240Ala Gln Pro Ile Val Ile Pro Thr Pro Ile Gly Glu Leu Asp Val Ile 245 250 255Ala Leu Tyr Ile Ala Ser Ile Ala Thr Gly Ser Ile Ala Leu Ala Ile 260 265 270Thr Asn Thr Ala Arg Pro Trp His Ile Gly Leu Tyr Gly Asn Ala Gly 275 280 285Gly Leu Gly Pro Thr Gln Gly His Pro Leu Ser Ser Ala Thr Asp Glu 290 295 300Pro Glu Pro His Trp Gly Pro Phe Gly Gly Ala Ala Pro Val Ser Ala305 310 315 320Gly Val Gly His Ala Ala Leu Val Gly Ala Leu Ser Val Pro His Ser 325 330 335Trp Thr Thr Ala Ala Pro Glu Ile Gln Leu Ala Val Gln Ala Thr Pro 340 345 350Thr Phe Ser Ser Ser Ala Gly Ala Asp Pro Thr Ala Leu Asn Gly Met 355 360 365Pro Ala Gly Leu Leu Ser Gly Met Ala Leu Ala Ser Leu Ala Ala Arg 370 375 380Gly Thr Thr Gly Gly Gly Gly Thr Arg Ser Gly Thr Ser Thr Asp Gly385 390 395 400Gln Glu Asp Gly Arg Lys Pro Pro Val Val Val Ile Arg Glu Gln Pro 405 410 415Pro Pro Gly Asn Pro Pro Arg 420 97 amino acids amino acid single linear protein 143Met Ser Leu Leu Asp Ala His Ile Pro Gln Leu Val Ala Ser Gln Ser 1 5 10 15Ala Phe Ala Ala Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala 20 25 30Glu Gln Ala Ala Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ser 35 40 45Ala Ala Phe Gln Ala Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys 50 55 60Val Asn Thr Leu Leu Asp Val Ala Gln Ala Asn Leu Gly Glu Ala Ala65 70 75 80Gly Thr Tyr Val Ala Ala Asp Ala Ala Ala Ala Ser Thr Tyr Thr Gly 85 90 95Phe 99 amino acids amino acid single linear protein 144Cys Arg Leu Cys Leu Asp Ser His Leu Arg Val Val Ala Leu Pro Ala 1 5 10 15Gly Gln Pro Gly Arg Leu Val Gln Ala Ile Gly Pro Ala Gln Glu Arg 20 25 30Asp Val Gly Gln Thr Arg Cys Thr Arg Thr Gly Leu Asp Xaa Val Ser 35 40 45Ala Leu Thr Ala Ala Gln Phe Ala Ala His Ala Gln Ile Tyr Gln Ala 50 55 60Val Ser Ala Gln Ala Ala Ala Ile His Glu Met Phe Val Asn Thr Leu65 70 75 80Gln Xaa Xaa Ser Gly Ser Tyr Ala Ala Thr Glu Ala Ala Asn Ala Ala 85 90 95Ala Ala Gly

Claims

1. An isolated polypeptide comprising an amino acid sequence as set forth in SEQ ID NO:109.

2. The polypeptide of claim 1, which is a soluble peptide.

3. The polypeptide of claim 1, which is produced by a recombinant DNA method.

4. The polypeptide of claim 1, which is produced by a chemical synthetic method.

5. The polypeptide of claim 1, which is purified from a cell source or culture supernatant.

6. The polypeptide of claim 1, which reacts with an antibody-containing biological sample.

7. The polypeptide of claim 1, which is a Mycobacterium tuberculosis antigen.

8. The polypeptide of claim 1, which is fused with a second heterologous polypeptide.

9. An isolated polypeptide from Mycobacterium tuberculosis encoded by a polynucleotide that hybridizes under moderately stringent conditions to a second polynucleotide that is complementary to the nucleotide sequence of SEQ ID NO:108,wherein said moderately stringent conditions comprise prewashing in a solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50-65° C. overnight in a solution comprising 5×SSC; and washing twice at 65° C. for 20 minutes with each of 2×, 0.5×, and 0.2×SSC containing 0.1% SDS.

10. The polypeptide of claim 9, encoded by a nucleic acid sequence comprising nucleotide sequence of SEQ ID NO:108.

11. The polypeptide of claim 9, comprising an amino acid sequence of SEQ ID NO:109.

12. A composition comprising:(a) an isolated polypeptide of claim 9, and (b) a physiological acceptable carrier.

13. The composition of claim 9, further comprising an adjuvant.