Meningococcal antigens

Abstract

The invention provides proteins from Neisseria meningitidis (strains A & B), including amino acid sequences, the corresponding nucleotide sequences, expression data, and serological data. The proteins are useful antigens for vaccines, immunogenic compositions, and/or diagnostics.

Description

This invention relates to antigens from the bacterium Neisseria meningitidis.

BACKGROUND

Neisseria meningitidis is a non-motile, gram negative diplococcus human pathogen. It colonises the pharynx, causing meningitis and, occasionally, septicaemia in the absence of meningitis. It is closely related to N.gonorrhoeae , although one feature that clearly differentiates meningococcus from gonococcus is the presence of a polysaccharide capsule that is present in all pathogenic meningococci.

N.meningitidis causes both endemic and epidemic disease. In the United States the attack rate is 0.6-1 per 100,000 persons per year, and it can be much greater during outbreaks (see Lieberman et al. (1996) Safety and Immunogenicity of a Serogroups A/C Neisseria meningitidis Oligosaccharide-Protein Conjugate Vaccine in Young Children. JAMA 275(19):1499-1503; Schuchat et al (1997) Bacterial Meningitis in the United States in 1995 . N Engl J Med 337(14):970-976). In developing countries, endemic disease rates are much higher and during epidemics incidence rates can reach 500 cases per 100,000 persons per year. Mortality is extremely high, at 10-20% in the United States, and much higher in developing countries. Following the introduction of the conjugate vaccine against Haemophilus influenzae, N. meningitidis is the major cause of bacterial meningitis at all ages in the United States (Schuchat et al (1997) supra).

Based on the organism's capsular polysaccharide, 12 serogroups of N.meningitidis have been identified. Group A is the pathogen most often implicated in epidemic disease in sub-Saharan Africa. Serogroups B and C are responsible for the vast majority of cases in the United States and in most developed countries. Serogroups W135 and Y are responsible for the rest of the cases in the United States and developed countries. The meningococcal vaccine currently in use is a tetravalent polysaccharide vaccine composed of serogroups A, C, Y and W135. Although efficacious in adolescents and adults, it induces a poor immune response and short duration of protection, and cannot be used in infants [eg. Morbidity and Mortality weekly report, Vol. 46, No. RR-5 (1997)]. This is because polysaccharides are T-cell independent antigens that induce a weak immune response that cannot be boosted by repeated immunization. Following the success of the vaccination against H.influenzae , conjugate vaccines against serogroups A and C have been developed and are at the final stage of clinical testing (Zollinger WD “New and Improved Vaccines Against Meningococcal Disease” in: New Generation Vaccines , supra, pp. 469-488; Lieberman et al (1996) supra; Costantino et al (1992) Development and phase I clinical testing of a conjugate vaccine against meningococcus A and C. Vaccine 10:691-698).

Meningococcus B remains a problem, however. This serotype currently is responsible for approximately 50% of total meningitis in the United States, Europe, and South America. The polysaccharide approach cannot be used because the menB capsular polysaccharide is a polymer of α(2-8)-linked N-acetyl neuraminic acid that is also present in mammalian tissue. This results in tolerance to the antigen; indeed, if an immune response were elicited, it would be anti-self, and therefore undesirable. In order to avoid induction of autoimmunity and to induce a protective immune response, the capsular polysaccharide has, for instance, been chemically modified substituting the N-acetyl groups with N-propionyl groups, leaving the specific antigenicity unaltered (Romero & Outschoorn (1994) Current status of Meningococcal group B vaccine candidates: capsular or non-capsular? Clin Microbiol Rev 7(4):559-575).

Alternative approaches to menB vaccines have used complex mixtures of outer membrane proteins (OMPs), containing either the OMPs alone, or OMPs enriched in porins, or deleted of the class 4 OMPs that are believed to induce antibodies that block bactericidal activity. This approach produces vaccines that are not well characterized. They are able to protect against the homologous strain, but are not effective at large where there are many antigenic variants of the outer membrane proteins. To overcome the antigenic variability, multivalent vaccines containing up to nine different porins have been constructed (eg. Poolman J T (1992) Development of a meningococcal vaccine. Infect. Agents Dis . 4:13-28). Additional proteins to be used in outer membrane vaccines have been the opa and opc proteins, but none of these approaches have been able to overcome the antigenic variability (eg. Ala'Aldeen & Borriello (1996) The meningococcal transferrin-binding proteins 1 and 2 are both surface exposed and generate bactericidal antibodies capable of killing homologous and heterologous strains. Vaccine 14(1):49-53).

A certain amount of sequence data is available for meningococcal and gonococcal genes and proteins (eg. EP-A-0467714, WO96/29412), but this is by no means complete. The provision of further sequences could provide an opportunity to identify secreted or surface-exposed proteins that are presumed targets for the immune system and which are not antigenically variable. For instance, some of the identified proteins could be components of efficacious vaccines against meningococcus B, some could be components of vaccines against all meningococcal serotypes, and others could be components of vaccines against all pathogenic Neisseriae.

THE INVENTION

The invention provides proteins comprising the N.meningitidis amino acid sequences disclosed in the examples.

It also provides proteins comprising sequences homologous (ie. having sequence identity) to the N.meningitidis amino acid sequences disclosed in the examples. Depending on the particular sequence, the degree of sequence identity is preferably greater than 50% (eg. 60%, 70%, 80%, 90%, 95%, 99% or more). These homologous proteins include mutants and allelic variants of the sequences disclosed in the examples. Typically, 50% identity or more between two proteins is considered to be an indication of functional equivalence. Identity between the proteins is preferably determined by the Smith-Waterman homology search algorithm as implemented in the MPSRCH program (Oxford Molecular), using an affine gap search with parameters gap open penalty=12 and gap extension penalty=1.

The invention further provides proteins comprising fragments of the N.meningitidis amino acid sequences disclosed in the examples. The fragments should comprise at least n consecutive amino acids from the sequences and, depending on the particular sequence, n is 7 or more (eg. 8, 10, 12, 14, 16, 18, 20 or more). Preferably the fragments comprise an epitope from the sequence.

The proteins of the invention can, of course, be prepared by various means (eg. recombinant expression, purification from cell culture, chemical synthesis etc.) and in various forms (eg. native, fusions etc.). They are preferably prepared in substantially pure form (ie. substantially free from other N.meningitidis or host cell proteins).

According to a further aspect, the invention provides antibodies which bind to these proteins. These may be polyclonal or monoclonal and may be produced by any suitable means.

According to a further aspect, the invention provides nucleic acid comprising the N.meningitidis nucleotide sequences disclosed in the examples. In addition, the invention provides nucleic acid comprising sequences homologous (ie. having sequence identity) to the N.meningitidis nucleotide sequences disclosed in the examples.

Furthermore, the invention provides nucleic acid which can hybridise to the N.meningitidis nucleic acid disclosed in the examples, preferably under “high stringency” conditions (eg. 65° C. in a at 0.1×SSC, 0.5% SDS solution).

Nucleic acid comprising fragments of these sequences are also provided. These should comprise at least n consecutive nucleotides from the N.meningitidis sequences and, depending on the particular sequence, n is 10 or more (eg 12, 14, 15, 18, 20, 25, 30, 35, 40 or more).

According to a further aspect, the invention provides nucleic acid encoding the proteins and protein fragments of the invention.

It should also be appreciated that the invention provides nucleic acid comprising sequences complementary to those described above (eg. for antisense or probing purposes).

Nucleic acid according to the invention can, of course, be prepared in many ways (eg. by chemical synthesis, from genomic or cDNA libraries, from the organism itself etc.) and can take various forms (eg. single stranded, double stranded, vectors, probes etc.).

In addition, the term “nucleic acid” includes DNA and RNA, and also their analogues, such as those containing modified backbones, and also peptide nucleic acids (PNA) etc.

According to a further aspect, the invention provides vectors comprising nucleotide sequences of the invention (eg. expression vectors) and host cells transformed with such vectors.

According to a further aspect, the invention provides compositions comprising protein, antibody, and/or nucleic acid according to the invention. These compositions may be suitable as vaccines, for instance, or as diagnostic reagents, or as immunogenic compositions.

The invention also provides nucleic acid, protein, or antibody according to the invention for use as medicaments (eg. as vaccines) or as diagnostic reagents. It also provides the use of nucleic acid, protein, or antibody according to the invention in the manufacture of: (i) a medicament for treating or preventing infection due to Neisserial bacteria; (ii) a diagnostic reagent for detecting the presence of Neisserial bacteria or of antibodies raised against Neisserial bacteria; and/or (iii) a reagent which can raise antibodies against Neisserial bacteria. Said Neisserial bacteria may be any species or strain (such as N.gonorrhoeae ) but are preferably N.meningitidis , especially strain A, strain B or strain C.

The invention also provides a method of treating a patient, comprising administering to the patient a therapeutically effective amount of nucleic acid, protein, and/or antibody according to the invention.

According to further aspects, the invention provides various processes.

A process for producing proteins of the invention is provided, comprising the step of culturing a host cell according to the invention under conditions which induce protein expression.

A process for producing protein or nucleic acid of the invention is provided, wherein the protein or nucleic acid is synthesised in part or in whole using chemical means.

A process for detecting polynucleotides of the invention is provided, comprising the steps of: (a) contacting a nucleic probe according to the invention with a biological sample under hybridizing conditions to form duplexes; and (b) detecting said duplexes.

A process for detecting proteins of the invention is provided, comprising the steps of: (a) contacting an antibody according to the invention with a biological sample under conditions suitable for the formation of an antibody-antigen complexes; and (b) detecting said complexes.

Unlike the sequences disclosed in PCT/IB98/01665, the sequences disclosed in the present application are believed not to have any significant homologs in N.gonorrhoeae . Accordingly, the sequences. of the present invention also find use in the preparation of reagents for distinguishing between N.meningitidis and N.gonorrhoeae

A summary of standard techniques and procedures which may be employed in order to perform the invention (eg. to utilise the disclosed sequences for vaccination or diagnostic purposes) follows. This summary is not a limitation on the invention but, rather, gives examples that may be used, but are not required.

General

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature eg. Sambrook Molecular Cloning; A Laboratory Manual, Second Edition (1989); DNA Cloning, Volumes I and ii (D. N Glover ed. 1985); Oligonucleotide Synthesis (M. J. Gait ed, 1984); Nucleic Acid Hybridization (B. D. Hames & S. J. Higgins eds. 1984); Transcription and Translation (B. D. Hames & S. J. Higgins eds. 1984); Animal Cell Culture (R. I. Freshney ed. 1986); Immobilized Cells and Enzymes (IRL Press, 1986); B. Perbal, A Practical Guide to Molecular Cloning (1984); the Methods in Enzymology series (Academic Press, Inc.), especially volumes 154 & 155 ; Gene Transfer Vectors for Mammalian Cells (J. H. Miller and M. P. Calos eds. 1987, Cold Spring Harbor Laboratory); Mayer and Walker, eds. (1987), Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); Scopes, (1987) Protein Purification: Principles and Practice , Second Edition (Springer-Verlag, N.Y.), and Handbook of Experimental Immunology, Volumes I-IV (D. M. Weir and C. C. Blackwell eds 1986).

Standard abbreviations for nucleotides and amino acids are used in this specification.

All publications, patents, and patent applications cited herein are incorporated in full by reference.

In particular, the contents of UK patent applications 9800760.2, 9819015.0 and 9822143.5 are incorporated herein.

Definitions

A composition containing X is “substantially free of” Y when at least 85% by weight of the total X+Y in the composition is X. Preferably, X comprises at least about 90% by weight of the total of X+Y in the composition, more preferably at least about 95% or even 99% by weight.

The term “comprising” means “including” as well as “consisting” eg. a composition “comprising” X may consist exclusively of X or may include something additional to X, such as X+Y.

A “conserved” Neisseria amino acid fragment or protein is one that is present in a particular Neisserial protein in at least x% of Neisseria. The value of x may be 50% or more, e.g., 66%, 75%, 80%, 90%, 95% or even 100% (i.e. the amino acid is found in the protein in question in all Neisseria). In order to determine whether an animo acid is “conserved” in a particular Neisserial protein, it is necessary to compare that amino acid residue in the sequences of the protein in question from a plurality of different Neisseria (a reference population). The reference population may include a number of different Neisseria species or may include a single species. The reference population may include a number of different serogroups of a particular species or a single serogroup. A preferred reference population consists of the 5 most common Neisseria.

The term “heterologous” refers to two biological components that are not found together in nature. The components may be host cells, genes, or regulatory regions, such as promoters. Although the heterologous components are not found together in nature, they can function together, as when a promoter heterologous to a gene is operably linked to the gene. Another example is where a Neisserial sequence is heterologous to a mouse host cell. A further examples would be two epitopes from the same or different proteins which have been assembled in a single protein in an arrangement not found in nature.

An “origin of replication” is a polynucleotide sequence that initiates and regulates replication of polynucleotides, such as an expression vector. The origin of replication behaves as an autonomous unit of polynucleotide replication within a cell, capable of replication under its own control. An origin of replication may be needed for a vector to replicate in a particular host cell. With certain origins of replication, an expression vector can be reproduced at a high copy number in the presence of the appropriate proteins within the cell. Examples of origins are the autonomously replicating sequences, which are effective in yeast; and the viral T-antigen, effective in COS-7 cells.

A “mutant” sequence is defined as DNA, RNA or amino acid sequence differing from but having sequence identity with the native or disclosed sequence. Depending on the particular sequence, the degree of sequence identity between the native or disclosed sequence and the mutant sequence is preferably greater than 50% (eg. 60%, 70%, 80%, 90%, 95%, 99% or more, calculated using the Smith-Waterman algorithm as described above). As used herein, an “allelic variant” of a nucleic acid molecule, or region, for which nucleic acid sequence is provided herein is a nucleic acid molecule, or region, that occurs essentially at the same locus in the genome of another or second isolate, and that, due to natural variation caused by, for example, mutation or recombination, has a similar but not identical nucleic acid sequence. A coding region allelic variant typically encodes a protein having similar activity to that of the protein encoded by the gene to which it is being compared. An allelic variant can also comprise an alteration in the 5′ or 3′ untranslated regions of the gene, such as in regulatory control regions (eg. see U.S. Pat. No. 5,753,235).

Expression Systems

The Neisserial nucleotide sequences can be expressed in a variety of different expression systems; for example those used with mammalian cells, baculoviruses, plants, bacteria, and yeast.

i. Mammalian Systems

Mammalian expression systems are known in the art. A mammalian promoter is any DNA sequence capable of binding mammalian RNA polymerase and initiating the downstream (3′) transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiating region, which is usually placed proximal to the 5′ end of the coding sequence, and a TATA box, usually located 25-30 base pairs (bp) upstream of the transcription initiation site. The TATA box is thought to direct RNA polymerase II to begin RNA synthesis at the correct site. A mammalian promoter will also contain an upstream promoter element, usually located within 100 to 200 bp upstream of the, TATA box. An upstream promoter element determines the rate at which transcription is initiated and can act in either orientation [Sambrook et al. (1989) “Expression of Cloned Genes in Mammalian Cells.” In Molecular Cloning. A Laboratory Manual , 2 nd ed.].

Mammalian viral genes are often highly expressed and have a broad host range; therefore sequences encoding mammalian viral genes provide particularly useful promoter sequences. Examples include the SV40 early promoter, mouse mammary tumor virus LTR promoter, adenovirus major late promoter (Ad MLP), and herpes simplex virus promoter. In addition, sequences derived from non-viral genes, such as the murine metallotheionein gene, also provide useful promoter sequences. Expression may be either constitutive or regulated (inducible), depending on the promoter can be induced with glucocorticoid in hormone-responsive cells.

The presence of an enhancer element (enhancer), combined with the promoter elements described above, will usually increase expression levels. An enhancer is a regulatory DNA sequence that can stimulate transcription up to 1000-fold when linked to homologous or heterologous promoters, with synthesis beginning at the normal RNA start site. Enhancers are also active when they are placed upstream or downstream from the transcription initiation site, in either normal or flipped orientation, or at a distance of more than 1000 nucleotides from the promoter [Maniatis et al. (1987) Science 236:1237; Alberts et al. (1989) Molecular Biology of the Cell , 2nd ed.]. Enhancer elements derived from viruses may be particularly useful, because they usually have a broader host range. Examples include the SV40 early gene enhancer [Dijkema et al (1985) EMBO J . 4:761] and the enhancer/promoters derived from the long terminal repeat (LTR) of the Rous Sarcoma Virus [Gormnan et al. (1982b) Proc. Natl. Acad. Sci . 79:6777] and from human cytomegalovirus [Boshart et al. (1985) Cell 41:521]. Additionally, some enhancers are regulatable and become active only in the presence of an inducer, such as a hormone or metal ion [Sassone-Corsi and Borelli (1986) Trends Genet . 2:215; Maniatis et al. (1987) Science 236:1237].

A DNA molecule may be expressed intracellularly in mammalian cells. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus of the recombinant protein will always be a methionine, which is encoded by the ATG start codon. If desired, the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide.

Alternatively, foreign proteins can also be secreted from the cell into the growth media by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provides for secretion of the foreign protein in mammalian cells. Preferably, there are processing sites encoded between the leader fragment and the foreign gene that can be cleaved either in vivo or in vitro. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell. The adenovirus triparite leader is an example of a leader sequence that provides for secretion of a foreign protein in mammalian cells.

Usually, transcription termination and polyadenylation sequences recognized by mammalian cells are regulatory regions located 3′ to the translation stop codon and thus, together with the promoter elements, flank the coding sequence. The 3′ terminus of the mature mRNA is formed by site-specific post-transcriptional cleavage and polyadenylation [Birnstiel et al. (1985) Cell 41:349; Proudfoot and Whitelaw (1988) “Ternmination and 3′ end processing of eukaryotic RNA. In Transcription and splicing (ed. B. D. Hames and D. M. Glover); Proudfoot (1989) Trends Biochem. Scip . 14:105]. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA. Examples of transcription terminater/polyadenylation signals include those derived from SV40 [Sambrook et al (1989) “Expression of cloned genes in cultured mammalian cells.” In Molecular Cloning: A Laboratory Manual].

Usually, the above described components, comprising a promoter, polyadenylation signal, and transcription termination sequence are put together into expression constructs. Enhancers, introns with fimctional splice donor and acceptor sites, and leader sequences may also be included in an expression construct, if desired. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as mammalian cells or bacteria. Mammalian replication systems include those derived from animal viruses, which require trans-acting factors to replicate. For example, plasmids containing the replication systems of papovaviruses, such as SV40 [Gluzman (1981) Cell 23:175] or polyomavirus, replicate to extremely high copy number in the presence of the appropriate viral T antigen. Additional examples of mammalian replicons include those derived from bovine papillomavirus and Epstein-Barr virus. Additionally, the replicon may have two replicaton systems, thus allowing it to be maintained, for example, in mammalian cells for expression and in a prokaryotic host for cloning and amplification. Examples of such mammalian-bacteria shuttle vectors include pMT2 [Kaufinan et al. (1989) Mol. Cell. Biol . 9:946] and pHEBO [Shimizu et al. (1986) Mol. Cell. Biol . 6:1074].

The transformation procedure used depends upon the host to be transformed. Methods for introduction of heterologous polynucleotides into mammalian cells are known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and direct microinjection of the DNA into nuclei.

Mammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC), including but not limited to, Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (eg. Hep G2), and a number of other cell lines.

ii. Baculovirus Systems

The polynucleotide encoding the protein can also be inserted into a suitable insect expression vector, and is operably linked to the control elements within that vector. Vector construction employs techniques which are known in the art. Generally, the components of the expression system include a transfer vector, usually a bacterial plasmid, which contains both a fragment of the baculovirus genome, and a convenient restriction site for insertion of the heterologous gene or genes to be expressed; a wild type baculovirus with a sequence homologous to the baculovirus-specific fragment in the transfer vector (this allows for the homologous recombination of the heterologous gene in to the baculovirus genome); and appropriate insect host cells and growth media.

After inserting the DNA sequence encoding the protein into the transfer vector, the vector and the wild type viral genome are transfected into an insect host cell where the vector and viral genome are allowed to recombine. The packaged recombinant virus is expressed and recombinant plaques are identified and purified. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, inter alia, Invitrogen, San Diego Calif. (“MaxBac” kit). These techniques are generally known to those skilled in the art and fully described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No . 1555 (1987) (hereinafter “Summers and Smith”).

Prior to inserting the DNA sequence encoding the protein into the baculovirus genome, the above described components, comprising a promoter, leader (if desired), coding sequence of interest, and transcription termination sequence, are usually assembled into an intermediate transplacement construct (transfer vector). This construct may contain a single gene and operably linked regulatory elements; multiple genes, each with its owned set of operably linked regulatory elements; or multiple genes, regulated by the same set of regulatory elements. Intermediate transplacement constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as a bacterium. The replicon will have a replication system, thus allowing it to be maintained in a suitable host for cloning and amplification.

Currently, the most commonly used transfer vector for introducing foreign genes into AcNPV is pAc373. Many other vectors, known to those of skill in the art, have also been designed. These include, for example, pVL985 (which alters the polyhedrin start codon from ATG to ATT, and which introduces a BamHI cloning site 32 basepairs downstream from the ATT; see Luckow and Summers, Virology (1989) 17:31.

The plasmid usually also contains the polyhedrin polyadenylation signal (Miller et al. (1988) Ann. Rev. Microbiol ., 42:177) and a prokaryotic ampicillin-resistance (amp) gene and origin of replication for selection and propagation in E. coli.

Baculovirus transfer vectors usually contain a baculovirus promoter. A baculovirus promoter is any DNA sequence capable of binding a baculovirus RNA polymerase and initiating the downstream (5′ to 3′) transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiation region which is usually placed proximal to the 5′ end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site and a transcription initiation site. A baculovirus transfer vector may also have a second domain called an enhancer, which, if present, is usually distal to the structural gene. Expression may be either regulated or constitutive.

Structural genes, abundantly transcribed at late times in a viral infection cycle, provide particularly useful promoter sequences. Examples include sequences derived from the gene encoding the viral polyhedron protein, Friesen et al., (1986) “The Regulation of Baculovirus Gene Expression,” in: The Molecular Biology of Baculoviruses (ed. Walter Doerfler);, EPO Publ. Nos. 127 839 and 155 476; and the gene encoding the p10 protein, Vlak et al., (1988), J. Gen. Virol . 69:765.

DNA encoding suitable signal sequences can be derived from genes for secreted insect or baculovirus proteins, such as the baculovirus polyhedrin gene (Carbonell et al. (1988) Gene , 73:409). Alternatively, since the signals for mammalian cell posttranslational modifications (such as signal peptide cleavage, proteolytic cleavage, and phosphorylation) appear to be recognized by insect cells, and the signals required for secretion and nuclear accumulation also appear to be conserved between the invertebrate cells and vertebrate cells, leaders of non-insect origin, such as those derived from genes encoding human cc-interferon, Maeda et al., (1985), Nature 315:592; human gastrin-releasing peptide, Lebacq-Verheyden et al., (1988), Molec. Cell. Biol . 8:3129; human IL-2, Smith et al., (1985) Proc. Nat'l Acad. Sci. USA , 82:8404; mouse IL-3, (Miyajima et al., (1987) Gene 58:273; and human glucocerebrosidase, Martin et al. (1988) DNA , 7:99, can also be used to provide for secretion in insects.

A recombinant polypeptide or polyprotein may be expressed intracellularly or, if it is expressed with the proper regulatory sequences, it can be secreted. Good intracellular expression of nonfused foreign proteins usually requires heterologous genes that ideally have a short leader sequence containing suitable translation initiation signals preceding an ATG start signal. If desired, methionine at the N-terminus may be cleaved from the mature protein by in vitro incubation with cyanogen bromide.

Alternatively, recombinant polyproteins or proteins which are not naturally secreted can be secreted from the insect cell by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provides for secretion of the foreign protein in insects. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the translocation of the protein into the endoplasmic reticulum.

After insertion of the DNA sequence and/or the gene encoding the expression product precursor of the protein, an insect cell host is co-transformed with the heterologous DNA of the transfer vector and the genomic DNA of wild type baculovirus—usually by co-transfection. The promoter and transcription termination sequence of the construct will usually comprise a 2-5 kb section of the baculovirus genome. Methods for introducing heterologous DNA into the desired site in the baculovirus virus are known in the art. (See Summers and Smith supra; Ju et al. (1987); Smith et al., Mol. Cell. Biol . (1983) 3:2156; and Luckow and Summers (1989)). For example, the insertion can be into a gene such as the polyhedrin gene, by homologous double crossover recombination; insertion can also be into a restriction enzyme site engineered into the desired baculovirus gene. Miller et al., (1989), Bioessays 4:91. The DNA sequence, when cloned in place of the polyhedrin gene in the expression vector, is flanked both 5′ and 3′ by polyhedrin-specific sequences and is positioned downstream of the polyhedrin promoter.

The newly formed baculovirus expression vector is subsequently packaged into an infectious recombinant baculovirus. Homologous recombination occurs at low frequency (between about 1% and about 5%); thus, the majority of the virus produced after cotransfection is still wild-type virus. Therefore, a method is necessary to identify recombinant viruses. An advantage of the expression system is a visual screen allowing recombinant viruses to be distinguished. The polyhedrin protein, which is produced by the native virus, is produced at very high levels in the nuclei of infected cells at late times after viral infection. Accumulated polyhedrin protein forms occlusion bodies that also contain embedded particles. These occlusion bodies, up to 15 μm in size, are highly refractile, giving them a bright shiny appearance that is readily visualized under the light microscope. Cells infected with recombinant viruses lack occlusion bodies. To distinguish recombinant virus from wild-type virus, the transfection supernatant is plaqued onto a monolayer of insect cells by techniques known to those skilled in the art. Namely, the plaques are screened under the light microscope for the presence (indicative of wild-type virus) or absence (indicative of recombinant virus) of occlusion bodies. “Current Protocols in Microbiology” Vol. 2 (Ausubel et al. eds) at 16.8 (Supp. 10, 1990); Summers and Smith, supra; Miller et al. (1989).

Recombinant baculovirus expression vectors have been developed for infection into several insect cells. For example, recombinant baculoviruses have been developed for, inter alia: Aedes aegypti, Autographa californica, Bombyx mori, Drosophila melanogaster, Spodoptera frugiperda , and Trichoplusia ni (WO 89/046699; Carbonell et al., (1985) J. Virol . 56:153; Wright (1986) Nature 321:718; Smith et al., (1983) Mol. Cell. Biol . 3:2156; and see generally, Fraser, et al. (1989) In Vitro Cell. Dev. Biol . 25:225).

Cells and cell culture media are commercially available for both direct and fusion expression of heterologous polypeptides in a baculovirus/expression system; cell culture technology is generally known to those skilled in the art. See, eg. Summers and Smith supra.

The modified insect cells may then be grown in an appropriate nutrient medium, which allows for stable maintenance of the plasmid(s) present in the modified insect host. Where the expression product gene is under inducible control, the host may be grown to high density, and expression induced. Alternatively, where expression is constitutive, the product will be continuously expressed into the medium and the nutrient medium must be continuously circulated, while removing the product of interest and augmenting depleted nutrients. The product may be purified by such techniques as chromatography, eg. HPLC, affinity chromatography, ion exchange chromatography, etc.; electrophoresis; density gradient centrifugation; solvent extraction, or the like. As appropriate, the product may be further purified, as required, so as to remove substantially any insect proteins which are also secreted in the medium or result from lysis of insect cells, so as to provide a product which is at least substantially free of host debris, eg. proteins, lipids and polysaccharides.

In order to obtain protein expression, recombinant host cells derived from the transforrnants are incubated under conditions which allow expression of the recombinant protein encoding sequence. These conditions will vary, dependent upon the host cell selected. However, the conditions are readily ascertainable to those of ordinary skill in the art, based upon what is known in the art.

iii. Plant Systems

There are many plant cell culture and whole plant genetic expression systems known in the art. Exemplary plant cellular genetic expression systems include those described in patents, such as: U.S. Pat. No. 5,693,506; U.S. Pat. No. 5,659,122; and U.S. Pat. No. 5,608,143. Additional examples of genetic expression in plant cell culture has been described by Zenk, Phytochemistry 30:3861-3863 (1991). Descriptions of plant protein signal peptides may be found in addition to the references described above in Vaulcombe et al., Mol. Gen. Genet . 209:33-40 (1987); Chandler et al., Plant Molecular Biology 3:407-418 (1984); Rogers, J. Biol. Chem . 260:3731-3738 (1985); Rothstein et al., Gene 55:353-356 (1987); Whittier et al., Nucleic Acids Research 15:2515-2535 (1987); Wirsel et al., Molecular Microbiology 3:3-14 (1989); Yu et al., Gene 122:247-253 (1992). A description of the regulation of plant gene expression by the phytohormone, gibberellic acid and secreted enzymes induced by gibberellic acid can be found in R. L. Jones and J. MacMillin, Gibberellins: in: Advanced Plant Physiology ,. Malcolm B. Wilkins, ed., 1984 Pitman Publishing Limited, London, pp. 21-52. References that describe other metabolically-regulated genes: Sheen, Plant Cell , 2:1027-1038(1990); Maas et al., EMBO J . 9:3447-3452 (1990); Benkel and Hickey, Proc. Natl. Acad. Sci . 84:1337-1339 (1987)

Typically, using techniques known in the art, a desired polynucleotide sequence is inserted into an expression cassette comprising genetic regulatory elements designed for operation in plants. The expression cassette is inserted into a desired expression vector with companion sequences upstream and downstream from the expression cassette suitable for expression in a plant host. The companion sequences will be of plasmid or viral origin and provide necessary characteristics to the vector to permit the vectors to move DNA from an original cloning host, such as bacteria, to the desired plant host. The basic bacterial/plant vector construct will preferably provide a broad host range prokaryote replication origin; a prokaryote selectable marker; and, for Agrobacterium transformations, T DNA sequences for Agrobacterium-mediated transfer to plant chromosomes. Where the heterologous gene is not readily amenable to detection, the construct will preferably also have a selectable marker gene suitable for determining if a plant cell has been transformed. A general review of suitable markers, for example for the members of the grass family, is found in Wilmink and Dons, 1993 , Plant Mol. Biol. Reptr , 11 (2):165-185.

Sequences suitable for permitting integration of the heterologous sequence into the plant genome are also recommended. These might include transposon sequences and the like for homologous recombination as well as Ti sequences which permit random insertion of a heterologous expression cassette into a plant genome. Suitable prokaryote selectable markers include resistance toward antibiotics such as ampicillin or tetracycline. Other DNA sequences encoding additional functions may also be present in the vector, as is known in the art.

The nucleic acid molecules of the subject invention may be included into an expression cassette for expression of the protein(s) of interest. Usually, there will be only one expression cassette, although two or more are feasible. The recombinant expression cassette will contain in addition to the heterologous protein encoding sequence the following elements, a promoter region, plant 5′ untranslated sequences, initiation codon depending upon whether or not the structural gene comes equipped with one, and a transcription and translation termination sequence. Unique restriction enzyme sites at the 5′ and 3′ ends of the cassette allow for easy insertion into a pre-existing vector.

A heterologous coding sequence may be for any protein relating to the present invention. The sequenceencoding the protein of interest will encode a signal peptide which allows processing and translocation of the protein, as appropriate, and will usually lack any sequence which might result in the binding of the desired protein of the invention to a membrane. Since, for the most part, the transcriptional initiation region will be for a gene which is expressed and translocated during germination, by employing the signal peptide which provides for translocation, one may also provide for translocation of the protein of interest. In this way, the protein(s) of interest will be translocated from the cells in which they are expressed and may be efficiently harvested. Typically secretion in seeds are across the aleurone or scutellar epithelium layer into the endosperm of the seed. While it is not required that the protein be secreted from the cells in which the protein is produced, this facilitates the isolation and purification of the recombinant protein.

Since the ultimate expression of the desired gene product will be in a eucaryotic cell it is desirable to determine whether any portion of the cloned gene contains sequences which will be processed out as introns by the host's splicosome machinery. If so, site-directed rnutagenesis of the “intron” region may be conducted to prevent losing a portion of the genetic message as a false intron code, Reed and Maniatis, Cell 41:95-105, 1985.

The vector can be microinjected directly into plant cells by use of micropipettes to mechanically transfer the recombinant DNA. Crossway, Mol. Gen. Genet , 202:179-185, 1985. The genetic material may also be transferred into the plant cell by using polyethylene glycol, Krens, et al., Nature , 296, 72-74, 1982. Another method of introduction of nucleic acid segments is high velocity ballistic penetration by small particles with the nucleic acid either within the matrix of small beads or particles, or on the surface, Klein, et al., Nature , 327, 70-73, 1987 and Knudsen and Muller, 1991 , Planta , 185:330-336 teaching particle bombardment of barley endosperm to create transgenic barley. Yet another method of introduction would be fusion of protoplasts with other entities, either minicells, cells, lysosomes or other fusible lipid-surfaced bodies, Fraley, et al., Proc. Natl. Acad. Sci. USA , 79, 1859-1863, 1982.

The vector may also be introduced into the plant cells by electroporation. (Fromm et al., Proc. Natl Acad. Sci. USA 82:5824, 1985). In this technique, plant protoplasts are electroporated in the presence of plasmids containing the gene construct. Electrical impulses of high field strength reversibly permeabilize biomembranes allowing the introduction of the plasmids. Electroporated plant protoplasts reform the cell wall, divide, and form plant callus.

All plants from which protoplasts can be isolated and cultured to give whole regenerated plants can be transformed by the present invention so that whole plants are recovered which contain the transferred gene. It is known that practically all plants can be regenerated from cultured cells or tissues, including but not limited to all major species of sugarcane, sugar beet, cotton, fruit and other trees, legumes and vegetables. Some suitable plants include, for example, species from the genera Fragaria, Lotus, Medicago, Onobrychis, Trifolium, Trigonella, Vigna, Citrus, Linum, Geranium, Manihot, Daucus, Arabidopsis, Brassica, Raphanus, Sinapis, Atropa, Capsicum, Datura, Hyoscyamus, Lycopersion, Nicotiana, Solanum, Petunia, Digitalis, Majorana, Cichorium, Helianthus, Lactuca, Bromus, Asparagus, Antirrhinum, Hererocallis, Nemesia, Pelargonium, Panicum, Pennisetum, Ranunculus, Senecio, Salpiglossis, Cucumis, Browaalia, Glycine, Lolium, Zea, Triticum, Sorghum, and Datura.

Means for regeneration vary from species to species of plants, but generally a suspension of transformed protoplasts containing copies of the heterologous gene is first provided. Callus tissue is formed and shoots may be induced from callus and subsequently rooted. Alternatively, embryo formation can be induced from the protoplast suspension. These embryos germinate as natural embryos to form plants. The culture media will generally contain various amino acids and hormones, such as auxin and cytokinins. It is also advantageous to add glutamic acid and proline to the medium, especially for such species as corn and alfalfa. Shoots and roots normally develop simultaneously. Efficient regeneration will depend on the medium, on the genotype, and on the history of the culture. If these three variables are controlled, then regeneration is fully reproducible and repeatable.

In some plant cell culture systems, the desired protein of the invention may be excreted or alternatively, the protein may be extracted from the whole plant. Where the desired protein of the invention is secreted into the medium, it may be collected. Alternatively, the embryos and embryoless-half seeds or other plant tissue may be mechanically disrupted to release any secreted protein between cells and tissues. The mixture may be suspended in a buffer solution to retrieve soluble proteins. Conventional protein isolation and purification methods will be then used to purify the recombinant protein. Parameters of time, temperature pH, oxygen, and volumes will be adjusted through routine methods to optimize expression and recovery of heterologous protein.

iv. Bacterial Systems

Bacterial expression techniques are known in the art. A bacterial promoter is any DNA sequence capable of binding bacterial RNA polymerase and initiating the downstream (3′) transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiation region which is usually placed proximal to the 5′ end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site and a transcription initiation site. A bacterial promoter may also have a second domain called an operator, that may overlap an adjacent RNA polymerase binding site at which RNA synthesis begins. The operator permits negative regulated (inducible) transcription, as a gene repressor protein may bind the operator and thereby inhibit transcription of a specific gene. Constitutive expression may occur in the absence of negative regulatory elements, such as the operator. In addition, positive regulation may be achieved by a gene activator protein binding sequence, which, if present is usually proximal (5′) to the RNA polymerase binding sequence. An example of a gene activator protein is the catabolite activator protein (CAP), which helps initiate transcription of the lac operon in Escherichia coli ( E. coli ) [Raibaud et al. (1984) Annu. Rev. Genet . 18:173]. Regulated expression may therefore be either positive or negative, thereby either enhancing or reducing transcription.

Sequences encoding metabolic pathway enzymes provide particularly useful promoter sequences. Examples include promoter sequences derived from sugar metabolizing enzymes, such as galactose, lactose (lac) [Chang et al. (1977) Nature 198:1056], and maltose. Additional examples include promoter sequences derived from biosynthetic enzymes such as tryptophan (trp) [Goeddel et al. (1980) Nuc. Acids Res . 8:4057; Yelverton et al. (1981) Nucl. Acids Res . 9:731; U.S. Pat. No. 4,738,921; EP-A-0036776 and EP-A-0121775]. The g-laotamase (bla) promoter system [Weissmann (1981) “The cloning of interferon and othermistakes.” In Interferon 3 (ed. I. Gresser)], bacteriophage lambda PL [Shimatake et al. (1981) Nature 292:128] and T5 [U.S. Pat. No. 4,689,406] promoter systems also provide useful promoter sequences.

In addition, synthetic promoters which do not occur in nature also function as bacterial promoters. For example, transcription activation sequences of one bacterial or bacteriophage promoter may be joined with the operon sequences of another bacterial or bacteriophage promoter, creating a synthetic hybrid promoter [U.S. Pat. No. 4,551,433]. For example, the tac promoter is a hybrid trp-lac promoter comprised of both trp promoter and lac operon sequences that is regulated by the lac repressor [Amann et al. (1983) Gene 25:167; de Boer et al. (1983) Proc. Natl. Acad. Sci . 80:21]. Furthermore, a bacterial promoter can include naturally occurring promoters of non-bacterial origin that have the ability to bind bacterial RNA polymerase and initiate transcription. A naturally occurring promoter of non-bacterial origin can also be coupled with a compatible RNA polymerase to produce high levels of expression of some genes in prokaryotes. The bacteriophage T7 RNA polymerase/promoter system is an example of a coupled promoter system [Studier et al. (1986) J. Mol. Biol . 189:113; Tabor et al. (1985) Proc. Natl. Acad. Sci . 82:1074]. In addition, a hybrid promoter can also be comprised of a bacteriophage promoter and an E. coli operator region (EPO-A-0 267 851).

In addition to a fumctioning promoter sequence, an efficient ribosome binding site is also useful for the expression of foreign genes in prokaryotes. In E. coli , the ribosome binding site is called the Shine-Dalgamo (SD) sequence and includes an initiation codon (ATG) and a sequence 3-9 nucleotides in length located 3-11 nucleotides upstream of the initiation codon [Shine et al. (1975) Nature 254:34]. The SD sequence is thought to promote binding of mRNA to the ribosome by the pairing of bases between the SD sequence and the 3′ and of E. coli 16S rRNA [Steitz et al. (1979) “Genetic signals and nucleotide sequences in messenger RNA.” In Biological Regulation and Development: Gene Expression (ed. R. F. Goldberger)]. To express eukaryotic genes and prokaryotic genes with weak ribosome-binding site [Sambrook et al. (1989) “Expression of cloned genes in Escherichia coli .” In Molecular Cloning: A Laboratory Manual].

A DNA molecule may be expressed intracellularly. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus will always be a methionine, which is encoded by the ATG start codon. If desired, methionine at the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide or by either in vivo on in vitro incubation with a bacterial methionine N-terminal peptidase (EPO-A-0 219 237).

Fusion proteins provide an alternative to direct expression. Usually, a DNA sequence encoding the N-terminal portion of an endogenous bacterial protein, or other stable protein, is fused to the 5′ end of heterologous coding sequences. Upon expression, this construct will provide a fusion of the two amino acid sequences. For example, the bacteriophage lambda cell gene can be linked at the 5′ terminus of a foreign gene and expressed in bacteria. The resulting fusion protein preferably retains a site for a processing enzyme (factor Xa) to cleave the bacteriophage protein from the foreign gene [Nagai et al. (1984) Nature 309:810]. Fusion proteins can also be made with sequences from the lacZ [Jia et al. (1987) Gene 60:197], trpE [Allen et al. (1987) J. Biotechnol. 5:93; Makoff et al. (1989) J. Gen. Microbiol . 135:11], and Chey [EP-A-0 324 647] genes. The DNA sequence at the junction of the two amino acid sequences may or may not encode a cleavable site. Another example is a ubiquitin fusion protein. Such a fusion protein is made with the ubiquitin region that preferably retains a site for a processing enzyme (eg. ubiquitin specific processing-protease) to cleave the ubiquitin from the foreign protein. Through this method, native foreign protein can be isolated [Miller et al. (1989) Bio/Technology 7:698].

Alternatively, foreign proteins can also be secreted from the cell by creating chimeric DNA molecules that encode a fusion protein comprised of a signal peptide sequence fragment that provides for secretion of the foreign protein in bacteria [U.S. Pat. No. 4,336,336]. The signal sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell. The protein is either secreted into the growth media (gram-positive bacteria) or into the periplasmic space, located between the inner and outer membrane of the cell (gram-negative bacteria). Preferably there are processing sites, which can be cleaved either in vivo or in vitro encoded between the signal peptide fragment and the foreign gene.

DNA encoding suitable signal sequences can be derived from genes for secreted bacterial proteins, such as the E. coli outer membrane protein gene (ompA) [Masui et al. (1983), in: Experimental Manipulation of Gene Expression ; Ghrayeb et al. (1984) EMBO J . 3:2437] and the E. coli alkaline phosphatase signal sequence (phoA) [Oka et al. (1985) Proc. Natl. Acad. Sci . 82:7212]. As an additional example, the signal sequence of the alpha-amylase gene from various Bacillus strains can be used to secrete heterologous proteins from B. subtilis [Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 244 042].

Usually, transcription termination sequences recognized by bacteria are regulatory regions located 3′ to the translation stop codon, and thus together with the promoter flank the coding sequence. These sequences direct the transcription of an mRNA which can be translated into the polypeptide encoded by the DNA. Transcription termination sequences frequently include DNA sequences of about 50 nucleotides capable of forming stem loop structures that aid in terminating transcription. Examples include transcription termination sequences derived from genes with strong promoters, such as the trp gene in E. coli as well as other biosynthetic genes.

Usually, the above described components, comprising a promoter, signal sequence (if desired), coding sequence of interest, and transcription termination sequence, are put together into expression constructs. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as bacteria. The replicon will have a replication system, thus allowing it to be maintained in a prokaryotic host either for expression or for cloning and amplification. In addition, a replicon may be either a high or low copy number plasmid. A high copy number plasmid will generally have a copy number ranging from about 5 to about 200, and usually about 10 to about 150. A host containing a high copy number plasmid will preferably contain at least about 10, and more preferably at least about 20 plasmids. Either a high or low copy number vector may be selected, depending upon the effect of the vector and the foreign protein on the host.

Alternatively, the expression constructs can be integrated into the bacterial genome with an integrating vector. Integrating vectors usually contain at least one sequence homologous to the bacterial chromosome that allows the vector to integrate. Integrations appear to result from recombinations between homologous DNA in the vector and the bacterial chromosome. For example, integrating vectors constructed with DNA from various Bacillus strains integrate into the Bacillus chromosome (EP-A-0 127 328). Integrating vectors may also be comprised of bacteriophage or transposon sequences.

Usually, extrachromosomal and integrating expression constructs may contain selectable markers to allow for the selection of bacterial strains that have been transformed. Selectable markers can be expressed in the bacterial host and may include genes which render bacteria resistant to drugs such as ampicillin, chloramphenicol, erythromycin, kanamycin (neomycin), and tetracycline [Davies et al. (1978) Annu. Rev. Microbiol . 32:469]. Selectable markers may also include biosynthetic genes, such as those in the histidine, tryptophan, and leucine biosynthetic pathways.

Alternatively, some of the above described components can be put together in transformation vectors. Transformation vectors are usually comprised of a selectable market that is either maintained in a replicon or developed into an integrating vector, as described above.

Expression and transformation vectors, either extra-chromosomal replicons or integrating vectors, have been developed for transformation into many bacteria. For example, expression vectors have been developed for, inter alia, the following bacteria: Bacillus subtilis [Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 036 259 and EP-A-0 063 953; WO 84/04541 ], Escherichia coli [Shimatake et al. (1981) Nature 292:128; Amaann et al. (1985) Gene 40:183; Studier et al. (1986) J. Mol. Biol . 189:113; EP-A-0 036 776, EP-A-0 136 829 and EP-A-0 136 907 ], Streptococcus cremoris [Powell et al. (1988) Appl. Environ. Microbiol . 54:655 ]; Streptococcus lividans [Powell et al. (1988) Appl. Environ. Microbiol . 54:655], Streptomyces lividans [U.S. Pat. No. 4,745,056].

Methods of introducing exogenous DNA into bacterial hosts are well-known in the art, and usually include either the transformation of bacteria treated with CaCl 2 or other agents, such as divalent cations and DMSO. DNA can also be introduced into bacterial cells by electroporation. Transformation procedures usually vary with the bacterial species to be transformed. See eg. [Masson et al. (1989) FEMS Microbiol. Lett . 60:273; Palva et al. (1982) Proc. Natl. Acad. Sci. USA 79:5582; EP-A-0 036 259 and EP-A-0 063 953; WO 84/04541, Bacillus], [Miller et al. (1988) Proc. Natl. Acad. Sci . 85:856; Wang et al. (1990) J. Bacteriol . 172:949, Campylobacter], [Cohen et al. (1973) Proc. Natl. Acad. Sci . 69:2110; Dower et al. (1988) Nucleic Acids Res . 16:6127; Kushner (1978) “An improved method for transformation of Escherichia coli with ColE1-derived plasmids. In Genetic Engineering: Proceedings of the International Symposium on Genetic Engineering (eds. H. W. Boyer and S. Nicosia); Mandel et al. (1970) J. Mol. Biol . 53:159; Taketo (1988) Biochim. Biophys. Acta 949:318; Escherichia], [Chassy et al. (1987) FEMS Microbiol. Lett . 44:173 Lactobacillus]; [Fiedler et al. (1988) Anal. Biochem 170:38, Pseudomonas]; [Augustin et al. (1990) FEMS Microbiol. Lett . 66:203, Staphylococcus], [Barany et al. (1980) J. Bacteriol . 144:698; Harlander (1987) “Transformation of Streptococcus lactis by electroporation, in: Streptococcal Genetics (ed. J. Ferretti and R. Curtiss III); Perry et al. (1981) Infect. Immun . 32:1295; Powell et al. (1988) Appl. Environ. Microbiol . 54:655; Somkuti et al. (1987) Proc. 4 th Evr. Cong. Biotechnology 1:412, Streptococcus].

v. Yeast Expression

Yeast expression systems are also known to one of ordinary skill in the art. A yeast promoter is any DNA sequence capable of binding yeast RNA polymerase and initiating the downstream (3′) transcription of a coding sequence (eg. structural gene) into mRNA. A promoter will have a transcription initiation region which is usually placed proximal to the 5′ end of the coding sequence. This transcription initiation region usually includes an RNA polymerase binding site (the “TATA Box”) and a transcription initiation site. A yeast promoter may also have a second domain called an upstream activator sequence (UAS), which, if present, is usually distal to the structural gene. The UAS permits regulated (inducible) expression. Constitutive expression occurs in the absence of a UAS. Regulated expression may be either positive or negative, thereby either enhancing or reducing transcription.

Yeast is a fermenting organism with an active metabolic pathway, therefore sequences encoding enzymes in the metabolic pathway provide particularly useful promoter sequences. Examples include alcohol dehydrogenase (ADH) (EP-A-0 284 044), enolase, glucokinase, glucose-6-phosphate isomerase, glyceraldehyde-3-phosphate-dehydrogenase (GAP or GAPDH), hexokinase, phosphofructokinase, 3-phosphoglycerate mutase, and pyruvate kinase (PyK) (EPO-A-0 329 203). The yeast PHO5 gene, encoding acid phosphatase, also provides useful promoter sequences [Myanohara et al. (1983) Proc. Natl. Acad. Sci. USA 80:1].

In addition, synthetic promoters which do not occur in nature also fuinction as yeast promoters. For example, UAS sequences of one yeast promoter may be joined with the transcription activation region of another yeast promoter, creating a synthetic hybrid promoter. Examples of such hybrid promoters include the ADH regulatory sequence linked to the GAP transcription activation region (U.S. Pat. Nos. 4,876,197 and 4,880,734). Other examples of hybrid promoters include promoters which consist of the regulatory sequences of either the ADH2, GAL4, GAL10, OR PHO5 genes, combined with the transcriptional activation region of a glycolytic enzyme gene such as GAP or PyK (EP-A-0 164 556). Furthermore, a yeast promoter can include naturally occurring promoters of non-yeast origin that have the ability to bind yeast RNA polymerase and initiate transcription. Examples of such promoters include, inter alia, [Cohen et al. (1980) Proc. Natl. Acad. Sci. USA 77:1078; Henikoff et al. (1981) Nature 283:835; Hollenberg et al. (1981) Curr. Topics Microbiol. Immunol . 96:119; Hollenberg et al. (1979) “The Expression of Bacterial Antibiotic Resistance Genes in the Yeast Saccharomyces cerevisiae ,” in: Plasmids of Medical, Environmental and Commercial Importance (eds. K. N. Timmis and A. Puhler); Mercerau-Puigalon et al. (1980) Gene 11:163; Panthier et al. (1980) Curr. Genet . 2:109;].

A DNA molecule may be expressed intracellularly in yeast. A promoter sequence may be directly linked with the DNA molecule, in which case the first amino acid at the N-terminus of the recombinant protein will always be a methionine, which is encoded by the ATG start codon. If desired, methionine at the N-terminus may be cleaved from the protein by in vitro incubation with cyanogen bromide.

Fusion proteins provide an alternative for yeast expression systems, as well as in mammalian, baculovirus, and bacterial expression systems. Usually, a DNA sequence encoding the N-terminal portion of an endogenous yeast protein, or other stable protein, is fused to the 5′ end of heterologous coding sequences. Upon expression, this construct will provide a fusion of the two amino acid sequences. For example, the yeast or human superoxide dismutase (SOD) gene, can be linked at the 5′ terminus of a foreign gene and expressed in yeast. The DNA sequence at the junction of the two amino acid sequences may or may not encode a cleavable site. See eg. EP-A-0 196 056. Another example is a ubiquitin fusion protein. Such a fusion protein is made with the ubiquitin region that preferably retains a site for a processing enzyme (eg. ubiquitin-specific processing protease) to cleave the ubiquitin from the foreign protein. Through this method, therefore, native foreign protein can be isolated (eg. WO88/024066).

Alternatively, foreign proteins can also be secreted from the cell into the growth media by creating chimeric DNA molecules that encode a fusion protein comprised of a leader sequence fragment that provide for secretion in yeast of the foreign protein. Preferably, there are processing sites encoded between the leader fragment and the foreign gene that can be cleaved either in vivo or in vitro. The leader sequence fragment usually encodes a signal peptide comprised of hydrophobic amino acids which direct the secretion of the protein from the cell.

DNA encoding suitable signal sequences can be derived from genes for secreted yeast proteins, such as the yeast invertase gene (EP-A-0 012 873; JPO. 62,096,086) and the A-factor gene (U.S. Pat. No. 4,588,684). Alternatively, leaders of non-yeast origin, such as an interferon leader, exist that also provide for secretion in yeast (EP-A-0 060 057).

A preferred class of secretion leaders are those that employ a fragment of the yeast alpha-factor gene, which contains both a “pre” signal sequence, and a “pro” region. The types of alpha-factor fragments that can be employed include the full-length pre-pro alpha factor leader (about 83 amino acid residues) as well as truncated alpha-factor leaders (usually about 25 to about 50 amino acid residues) (U.S. Pat. Nos. 4,546,083 and 4,870,008; EP-A-0 324 274). Additional leaders employing an alpha-factor leader fragment that provides for secretion include hybrid alpha-factor leaders made with a presequence of a first yeast, but a pro-region from a second yeast alphafactor. (eg. see WO 89/02463.)

Usually, transcription termination sequences recognized by yeast are regulatory regions located 3′ to the translation stop codon, and thus together with the promoter flank the coding sequence. These sequences direct the transcription of an niRNA which can be translated into the polypeptide encoded by the DNA. Examples of transcription terminator sequence and other yeast-recognized termination sequences, such as those coding for glycolytic enzymes.

Usually, the above described components, comprising a promoter, leader (if desired), coding sequence of interest, and transcription termination sequence, are put together into expression constructs. Expression constructs are often maintained in a replicon, such as an extrachromosomal element (eg. plasmids) capable of stable maintenance in a host, such as yeast or bacteria. The replicon may have two replication systems, thus allowing it to be maintained, for example, in yeast for expression and in a prokaryotic host for cloning and amplification. Examples of such yeast-bacteria shuttle vectors include YEp24 [Botstein et al. (1979) Gene 8:17-24], pC1/1 [Brake et al. (1984) Proc. Natl. Acad. Sci USA 81:4642-4646], and YRp17 [Stinchcomb et al. (1982) J. Mol. Biol . 158:157]. In addition, a replicon may be either a high or low copy number plasmid. A high copy number plasmid will generally have a copy number ranging from about 5 to about 200, and usually about 10 to about 150. A host containing a high copy number plasmid will preferably have at least about 10, and more preferably at least about 20. Enter a high or low copy number vector may be selected, depending upon the effect of the vector and the foreign protein on the host. See eg. Brake et al., supra.

Alternatively, the expression constructs can be integrated into the yeast genome with an integrating vector. Integrating vectors usually contain at least one sequence homologous to a yeast chromosome that allows the vector to integrate, and preferably contain two homologous sequences flanking the expression construct. Integrations appear to result from recombinations between homologous DNA in the vector and the yeast chromosome [Orr-Weaver et al. (1983) Methods in Enzymol . 101:228-245]. An integrating vector may be directed to a specific locus in yeast by selecting the appropriate homologous sequence for inclusion in the vector. See Orr-Weaver et al., supra. One or more expression construct may integrate, possibly affecting levels of recombinant protein produced [Rine et al. (1983) Proc. Natl. Acad. Sci. USA 80:6750]. The chromosomal sequences included in the vector can occur either as a single segment in the vector, which results in the integration of the entire vector, or two segments homologous to adjacent segments in the chromosome and flanking the expression construct in the vector, which can result in the stable integration of only the expression construct.

Usually, extrachromosomal and integrating expression constructs may contain selectable markers to allow for the selection of yeast strains that have been transformed. Selectable markers may include biosynthetic genes that can be expressed in the yeast host, such as ADE2, HIS4, LEU2, TRP1, and ALG7, and the G418 resistance gene, which confer resistance in yeast cells to tunicamycin and G418, respectively. In addition, a suitable selectable marker may also provide yeast with the ability to grow in the presence of toxic compounds, such as metal. For example, the presence of CUP1 allows yeast to grow in the presence of copper ions [Butt et al. (1987) Microbiol, Rev . 51:351].

Alternatively, some of the above described components can be put together into transformation vectors. Transformation vectors are usually comprised of a selectable marker that is either maintained in a replicon or developed into an integrating vector, as described above.

Expression and transformation vectors, either extrachromosomal replicons or integrating vectors, have been developed for transformation into many yeasts. For example, expression vectors have been developed for, inter alia, the following yeasts: Candida albicans [Kurtz, et al. (1986) Mol. Cell. Biol . 6:142 ], Candida maltosa [Kunze, et al. (1985) J. Basic Microbiol . 25:141 ]. Hansenula polymorpha [Gleeson, et al. (1986) J. Gen. Microbiol . 132:3459; Roggenkamp et al. (1986) Gen. Genet . 202:302 ], Kluyveromyces fragilis [Das, et al. (1984) J. Bacteriol . 158:1165 ], Kluyveromyces lactis [De Louvencourt et al. (1983) J. Bacteriol . 154:737; Van den Berg et al. (1990) Bio/Technology 8:135 ], Pichia guillerimondii [Kunze et al. (1985) J. Basic Microbiol . 25:141 ], Pichia pastoris [Cregg, et al. (1985) Mol. Cell. Biol . 5:3376; U.S. Pat. Nos. 4,837,148 and 4,929,555 ], Saccharomyces cerevisiae [Hinnen et al. (1978) Proc. Natl. Acad. Sci. USA 75:1929; Ito et al. (1983) J. Bacteriol . 153:163 ], Schizosaccharomyces pombe [Beach and Nurse (1981) Nature 300:706], and Yarrowia lipolytica [Davidow, et al. (1985) Curr. Genet . 10:380471 Gaillardin, et al. (1985) Curr. Genet . 10:49].

Methods of introducing exogenous DNA into yeast hosts are well-known in the art, and usually include either the transformation of spheroplasts or of intact yeast cells treated with alkali cations. Transformation procedures usually vary with the yeast species to be transformed. See eg. [Kurtz et al. (1986) Mol. Cell. Biol . 6:142; Kunze et al. (1985) J. Basic Microbiol . 25:141; Candida]; [Gleeson et al. (1986) J. Gen. Microbiol . 132:3459; Roggenkamp et al. (1986) Mol. Gen. Genet 202:302; Hansenula]; [Das et al. (1984) J. Bacteriol . 158:1165; De Louvencourt et al. (1983) J. Bacteriol . 154:1165; Van den Berg et al. (1990) Bio/Technology 8:135; Kluyveromyces]; [Cregg et al. (1985) Mol. Cell. Biol . 5:3376; Kunze et al. (1985) J. Basic Microbiol . 25:141; U.S. Pat. Nos. 4,837,148 and 4,929,555; Pichia]; [Hinnen et al. (1978) Proc. Natl. Acad. Sci. USA Ito et al. (1983) J. Bacteriol . 153:163 Saccharomyces]; [Beach and Nurse (1981) Nature 300:706; Schizosaccharomyces]; [Davidow et al. (1985) Curr. Genet . 10:39; Gaillardin et al. (1985) Curr. Genet . 10:49; Yarrowia].

Antibodies

As used herein, the term “antibody” refers to a polypeptide or group of polypeptides composed of at least one antibody combining site. An “antibody combining site” is the three-dimensional binding space with an internal surface shape and charge distribution complementary to the features of an epitope of an antigen, which allows a binding of the antibody with the antigen. “Antibody” includes, for example, vertebrate antibodies, hybrid antibodies, chimeric antibodies, humanised antibodies, altered antibodies, univalent antibodies, Fab proteins, and single domain antibodies. Antibodies against the proteins of the invention are useful for affinity chromatography, immunoassays, and distinguishing/identifying Neisserial proteins.

Antibodies to the proteins of the invention, both polyclonal and monoclonal, may be prepared by conventional methods. In general, the protein is first used to immunize a suitable animal, preferably a mouse, rat, rabbit or goat. Rabbits and goats are preferred for the preparation of polyclonal sera due to the volume of serum obtainable, and the availability of labeled anti-rabbit and anti-goat antibodies. Immunization is generally performed by mixing or emulsifying the protein in saline, preferably in an adjuvant such as Freund's complete adjuvant, and injecting the mixture or emulsion parenterally (generally subcutaneously or intramuscularly). A dose of 50-200 μg/injection is typically sufficient. Immunization is generally boosted 2-6 weeks later with one or more injections of the protein in saline, preferably using Freund's incomplete adjuvant. One may alternatively generate antibodies by in vitro immunization using methods known in the art, which for the purposes of this invention is considered equivalent to in vivo immunization. Polyclonal antisera is obtained by bleeding the immunized animal into a glass or plastic container, incubating the blood at 25° C. for one hour, followed by incubating at 4° C. for 2-18 hours. The serum is recovered by centrifugation (eg. 1,000 g for 10 minutes). About 20-50 ml per bleed may be obtained from rabbits.

Monoclonal antibodies are prepared using the standard method of Kohler & Milstein [ Nature (1975) 256:495-96], or a modification thereof Typically, a mouse or rat is immunized as described above. However, rather than bleeding the animal to extract serum, the spleen (and optionally several large lymph nodes) is removed and dissociated into single cells. If desired, the spleen cells may be screened (after removal of nonspecifically adherent cells) by applying a cell suspension to a plate or well coated with the protein antigen. B-cells expressing membrane-bound immunoglobulin specific for the antigen bind to the plate, and are not rinsed away with the rest of the suspension. Resulting B-cells, or all dissociated spleen cells, are then induced to fuse with myeloma cells to form hybridomas, and are cultured in a selective medium (eg. hypoxanthine, aminopterin, thymidine medium, “HAT”). The resulting hybridomas are plated by limiting dilution, and are assayed for the production of antibodies which bind specifically to the immunizing antigen (and which do not bind to unrelated antigens). The selected MAb-secreting hybridomas are then cultured either in vitro (eg. in tissue culture bottles or hollow fiber reactors), or in vivo (as ascites in mice).

If desired, the antibodies (whether polyclonal or monoclonal) may be labeled using conventional techniques. Suitable labels include fluorophores, chromophores, radioactive atoms (particularly 32 P and 125 I), electron-dense reagents, enzymes, and ligands having specific binding partners. Enzymes are typically detected by their activity. For example, horseradish peroxidase is usually detected by its ability to convert 3,3′,5,5′-tetramethylbenzidine (TMB) to a blue pigment, quantifiable with a spectrophotometer. “Specific binding partner” refers to a protein capable of binding a ligand molecule with high specificity, as for example in the case of an antigen and a monoclonal antibody specific therefor. Other specific binding partners include biotin and avidin or streptavidin, IgG and protein A, and the numerous receptor-ligand couples known in the art. It should be understood that the above description is not meant to categorize the various labels into distinct classes, as the same label may serve in several different modes. For example, 125 I may serve as a radioactive label or as an electron-dense reagent. HRP may serve as enzyme or as antigen for a MAb. Further, one may combine various labels for desired effect. For example, MAbs and avidin also require labels in the practice of this invention: thus, one might label a MAb with biotin, and detect its presence with avidin labeled with 125 I, or with an anti-biotin MAb labeled with HRP. Other permutations and possibilities will be readily apparent to those of ordinary skill in the art, and are considered as equivalents within the scope of the instant invention.

Pharmaceutical Compositions

Pharmaceutical compositions can comprise either polypeptides, antibodies, or nucleic acid of the invention. The pharmaceutical compositions will comprise a therapeutically effective amount of either polypeptides, antibodies, or polynucleotides of the claimed invention.

The term “therapeutically effective amount” as used herein refers to an amount of a therapeutic agent to treat, ameliorate, or prevent a desired disease or condition, or to exhibit a detectable therapeutic or preventative effect. The effect can be detected by, for example, chemical markers or antigen levels. Therapeutic effects also include reduction in physical symptoms, such as decreased body temperature. The precise effective amount for a subject will depend upon the subject's size and health, the nature and extent of the condition, and the therapeutics or combination of therapeutics selected for administration. Thus, it is not useful to specify an exact effective amount in advance. However, the effective amount for a given situation can be determined by routine experimentation and is within the judgement of the clinician.

For purposes of the present invention, an effective dose will be from about 0.01 mg/kg to 50 mg/kg or 0.05 mg/kg to about 10 mg/kg of the DNA constructs in the individual to which it is administered.

A pharmaceutical composition can also contain a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” refers to a carrier for administration of a therapeutic agent, such as antibodies or a polypeptide, genes, and other therapeutic agents. The term refers to any pharmaceutical carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity. Suitable carriers may be large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, and inactive virus particles. Such carriers are well known to those of ordinary skill in the art.

Pharmaceutically acceptable salts can be used therein, for example, mineral acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, and the like; and the salts of organic acids such as acetates, propionates, malonates, benzoates, and the like. A thorough discussion of pharmaceutically acceptable excipients is available in Remington's Pharmaceutical Sciences (Mack Pub. Co., N.J. 1991).

Pharmaceutically acceptable carriers in therapeutic compositions may contain liquids such as water, saline, glycerol and ethanol. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles. Typically, the therapeutic compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. Liposomes are included within the definition of a pharmaceutically acceptable carrier.

Delivery Methods

Once formulated, the compositions of the invention can be administered directly to the subject. The subjects to be treated can be animals; in particular, human subjects can be treated.

Direct delivery of the compositions will generally be accomplished by injection, either subcutaneously, intraperitoneally, intravenously or intramuscularly or delivered to the interstitial space of a tissue. The compositions can also be administered into a lesion. Other modes of administration include oral and pulmonary administration, suppositories, and transdermal or transcutaneous applications (eg. see WO98/20734), needles, and gene guns or hyposprays. Dosage treatment may be a single dose schedule or a multiple dose schedule.

Vaccines

Vaccines according to the invention may either be prophylactic (ie. to prevent infection) or therapeutic (ie. to treat disease after infection).

Such vaccines comprise immunising antigen(s), immunogen(s), polypeptide(s), protein(s) or nucleic acid, usually in combination with “pharmaceutically acceptable carriers,” which include any carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition. Suitable carriers are typically large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, lipid aggregates (such as oil droplets or liposomes), and inactive virus particles. Such carriers are well known to those of ordinary skill in the art. Additionally, these carriers may function as immunostimulating agents (“adjuvants”). Furthermore, the antigen or immunogen may be conjugated to a bacterial toxoid, such as a toxoid from diphtheria, tetanus, cholera, H. pylori , etc. pathogens.

Preferred adjuvants to enhance effectiveness of the composition include, but are not limited to: (1) aluminum salts (alum), such as aluminum hydroxide, aluminum phosphate, aluminum sulfate, etc; (2) oil-in-water emulsion formulations (with or without other specific immunostimulating agents such as muramyl peptides (see below) or bacterial cell wall components), such as for example (a) MF59™ (WO 90/14837; Chapter 10 in Vaccine design: the subunit and adjuvant approach , eds. Powell & Newman, Plenum Press 1995), containing 5% Squalene, 0.5% Tween 80, and 0.5% Span 85 (optionally containingvarious amounts of MTP-PE (see below), although not required) formulated into submicron particles using a microfluidizer such as Model 110Y microfluidizer (Microfluidics, Newton, Mass.), (b) SAF, containing 10% Squalane, 0.4% Tween 80, 5% pluronic-blocked polymer L121, and thr-MDP (see below) either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion, and (c) Ribi™ adjuvant system (RAS), (Ribi Immunochem, Hamilton, Mont.) containing 2% Squalene, 0.2% Tween 80, and one or more bacterial cell wall components from the group consisting of monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL+CWS (Detoxam); (3) saponin adjuvants, such as Stimulon™ (Cambridge Bioscience, Worcester, Mass.) may be used or particles generated therefrom such as ISCOMs (immunostimulating complexes); (4) Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA); (5) cytokines, such as interleukins (eg. IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12, etc.), interferons (eg. gamma interferon), macrophage colony stimulating factor (M-CSF), tumor necrosis factor (TNF), etc; and (6) other substances that act as immunostimulating agents to enhance the effectiveness of the composition. Alum and MF59™ are preferred.

As mentioned above, muramyl peptides include, but are not limited to, N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-normnuramyl- L -alanyl- D -isoglutamine (nor-MDP), N-acetylmuramyl- L -alanyl- D -isoglutaminyl- L -alanine-2-(1′-2′-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamine (MTP-PE), etc.

The immunogenic compositions (eg. the immunising antigen/immunogen/polypeptide/protein/ nucleic acid, pharmaceutically acceptable carrier, and adjuvant) typically will contain diluents, such as water, saline, glycerol, ethanol, etc. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles.

Typically, the immunogenic compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. The preparation also may be emulsified or encapsulated in liposomes for enhanced adjuvant effect, as discussed above under pharmaceutically acceptable carriers.

Immunogenic compositions used as vaccines comprise an immunologically effecfive amount of the antigenic or immunogenic polypeptides, as well as any other of the above-mentioned components, as needed. By “immunologically effective amount”, it is meant that the administration of that amount to an individual, either in a single dose or as part of a series, is effective for treatment or prevention. This amount varies depending upon the health and physical condition of the individual to be treated, the taxonomic group of individual to be treated (eg. nonhuman primate, primate, etc.), the capacity of the individual's immune system to synthesize antibodies, the degree of protection desired, the formulation of the vaccine, the treating doctor's assessment of the medical situation, and other relevant factors. It is expected that the amount will fall in a relatively broad range that can be determined through routine trials.

The immunogenic compositions are conventionally administered parenterally, eg. by injection, either subcutaneously, intramuscularly, or transdermally/transcutaneously (eg. WO98/20734). Additional formulations suitable for other modes of administration include oral and pulmonary formulations, suppositories, and transdermal applications. Dosage treatment may be a single dose schedule or a multiple dose schedule. The vaccine may be administered in conjunction with other immunoregulatory agents.

As an alternative to protein-based vaccines, DNA vaccination may be employed [eg. Robinson & Torres (1997) Seminars in Immunology 9:271-283; Donnelly et al. (1997) Annu Rev Immunol 15:617-648; see later herein].

Gene Delivery Vehicles

Gene therapy vehicles for delivery of constructs including a coding sequence of a therapeutic of the invention, to be delivered to the mammal for expression in the mammal, can be administered either locally or systemically. These constructs can utilize viral or non-viral vector approaches in in vivo or ex vivo modality. Expression of such coding sequence can be induced using endogenous mammalian or heterologous promoters. Expression of the coding sequence in vivo can be either constitutive or regulated.

The invention includes gene delivery vehicles capable of expressing the contemplated nucleic acid sequences. The gene delivery vehicle is preferably a viral vector and, more preferably, a retroviral, adenoviral, adeno-associated viral (AAV), herpes viral, or alphavirus vector. The viral vector can also be an astrovirus, coronavirus, orthomyxovirus, papovavirus, paramyxovirus, parvovirus, picomavirus, poxvirus, or togavirus viral vector. See generally, Jolly (1994) Cancer Gene Therapy 1:51-64; Kimura (1994) Human Gene Therapy 5:845-852; Connelly (1995) Human Gene Therapy 6:185-193; and Kaplitt (1994) Nature Genetics 6:148-153.

Retroviral vectors are well known in the art and we contemplate that any retroviral gene therapy vector is employable in the invention, including B, C and D type retroviruses, xenotropic retroviruses (for example, NZB-X1, NZB-X2 and NZB9-1 (see O'Neill (1985) J. Virol . 53:160) polytropic retroviruses eg, MCF and MCF-MLV (see Kelly (1983) J. Virol . 45:291), spumaviruses and lentiviruses. See RNA Tumor Viruses, Second Edition, Cold Spring Harbor Laboratory, 1985.

Portions of the retroviral gene therapy vector may be derived from different retroviruses. For example, retrovector LTRs may be derived from a Murine Sarcoma Virus, a tRNA binding site from a Rous Sarcoma Virus, a packaging signal from a Murine Leukemia Virus, and an origin of second strand synthesis from an Avian Leukosis Virus.

These recombinant retroviral vectors may be used to generate transduction competent retroviral vector particles by introducing them into appropriate packaging cell lines (see U.S. Pat. No. 5,591,624). Retrovirus vectors can be constructed for site-specific integration into host cell DNA by incorporation of a chimeric integrase enzyme into the retroviral particle (see WO96/37626). It is preferable that the recombinant viral vector is a replication defective recombinant virus.

Packaging cell lines suitable for use with the above-described retrovirus vectors are well known in the art, are readily prepared (see WO95/30763 and WO92/05266), and can be used to create producer cell lines (also termed vector cell lines or “VCLs”) for the production of recombinant vector particles. Preferably, the packaging cell lines are made from human parent cells (eg. HT1080 cells) or mink parent cell lines, which eliminates inactivation in human serum.

Preferred retroviruses for the construction of retroviral gene therapy vectors include Avian Leukosis Virus, Bovine Leukemia, Virus, Murine Leukemia Virus, Mink-Cell Focus-Inducing Virus, Murine Sarcoma Virus, Reticuloendotheliosis Virus and Rous Sarcoma Virus. Particularly preferred Murine Leukemia Viruses include 4070A and 1504A (Hartley and Rowe (1976) J Virol 19:19-25), Abelson (ATCC No. VR-999), Friend (ATCC No. VR-245), Graffi, Gross (ATCC Nol VR-590), Kirsten, Harvey Sarcoma Virus and Rauscher (ATCC No. VR-998) and Moloney Murine Leukemia Virus (ATCC No. VR-190). Such retroviruses may be obtained from depositories or collections such as the American Type Culture Collection (“ATCC”) in Rockville, Maryland or isolated from known sources using commnonly available techniques.

Exemplary known retroviral gene therapy vectors employable in this invention include those described in patent applications GB2200651, EP0415731, EP0345242, EP0334301, WO89/02468; WO89/05349, WO89/09271, WO90/02806, WO90/07936, WO94/03622, WO93/25698, WO93/25234, WO93/11230, WO93/10218, WO91/02805, WO91/02825, WO95/07994, U.S. Pat. No. 5,219,740, U.S. Pat. No. 4,405,712, U.S. Pat. No. 4,861,719, U.S. Pat. No. 4,980,289, U.S. Pat. No. 4,777,127, U.S. Pat. No. 5,591,624. See (1993) Cancer Res 53:3860-3864; Vile (1993) Cancer Res 53:962-967; Ram (1993) Cancer Res 53 (1993) 83-88; Takamiya (1992) J Neurosci Res 33:493-503; Baba (1993) J Neurosurg 79:729-735; Mann (1983) Cell 33:153; Cane (1984) Proc Natl Acad Sci 81:6349; and Miller (1990) Human Gene Therapy 1.

Human adenoviral gene therapy vectors are also known in the art and employable in this invention. See, for example, Berkner (1988) Biotechniques 6:616 and Rosenfeld (1991) Science 252:431, and WO93/07283, WO93/06223, and WO93/07282. Exemplary known adenoviral gene therapy vectors employable in this invention include those described in the above referenced documents and in WO94/12649, WO93/03769, WO93/19191, WO94/28938, WO95/11984, WO95/00655, WO95/27071, WO95/29993, WO95/34671, WO96/05320, WO94/08026, WO94/11506, WO93/06223, WO94/24299, WO95/14102, WO95/24297, WO95/02697, WO94/28152, WO94/24299, WO95/09241, WO95/25807, WO95/05835, WO94/18922 and WO95/09654. Alternatively, administration of DNA linked to killed adenovirus as described in Curiel (1992) Hum. Gene Ther . 3:147-154 may be employed. The gene delivery vehicles of the invention also include adenovirus associated virus (AAV) vectors. Leading and preferred examples of such vectors for use in this invention are the AAV-2 based vectors disclosed in Srivastava, WO93/09239. Most preferred AAV vectors comprise the two AAV inverted terminal repeats in which the native D-sequences are modified by substitution of nucleotides, such that at least 5 native nucleofides and up to 18 native nucleotides, preferably at least 10 native nucleotides up to 18 native nucleotides, most preferably 10 native nucleotides are retained and the remaining nucleotides of the D-sequence are deleted or replaced with non-native nucleotides. The native D-sequences of the AAV inverted terminal repeats are sequences of 20 consecutive nucleotides in each AAV inverted terminal repeat (ie. there is one sequence at each end) which are not involved in HP formation. The non-native replacement nucleotide may be any nucleotide other than the nucleotide found in the native D-sequence in the same position. Other employable exemplary AAV vectors are pWP-19, pWN-1, both of which are disclosed in Nahreini (1993) Gene 124:257-262. Another example of such an AAV vector is psub20l (see Samulski (1987) J. Virol . 61:3096). Another exemplary AAV vector is the Double-D ITR vector. Construction of the Double-D ITR vector is disclosed in U.S. Pat. No. 5,478,745. Still other vectors are those disclosed in Carter U.S. Pat. No. 4,797,368 and Muzyczka U.S. Pat. No. 5,139,941, Chartejee U.S. Pat. No. 5,474,935, and Kotin WO94/288157. Yet a further example of an AAV vector employable in this invention is SSV9AFABTKneo, which contains the AFP enhancer and albumin promoter and directs expression predominantly in the liver. Its structure and construction are disclosed in Su (1996) Human Gene Therapy 7:463-470. Additional AAV gene therapy vectors are described in U.S. Pat. No. 5,354,678, U.S. Pat. No. 5,173,414, U.S. Pat. No. 5,139,941, and U.S. Pat. No. 5,252,479.

The gene therapy vectors of the invention also include herpes vectors. Leading and preferred examples are herpes simplex virus vectors containing a sequence encoding a thymidine kinase polypeptide such as those disclosed in U.S. Pat. No. 5,288,641 and EP0176170 (Roizman). Additional exemplary herpes simplex virus vectors include HFEM/ICP6-LacZ disclosed in WO95/04139 (Wistar Institute), pHSVlac described in Geller (1988) Science 241:1667-1669 and in WO90/09441 and WO92/07945, HSV Us3::pgC-lacZ described in Fink (1992) Human Gene Therapy 3:11-19 and HSV 7134, 2 RH 105 and GAL4 described in EP 0453242 (Breakefield), and those deposited with the ATCC as accession numbers ATCC VR-977 and ATCC VR-260.

Also contemplated are alpha virus gene therapy vectors that can be employed in this invention. Preferred alpha virus vectors are Sindbis viruses vectors. Togaviruses, Semliki Forest virus (ATCC VR-67; ATCC VR-1247), Middleberg virus (ATCC VR-370), Ross River virus (ATCC VR-373; ATCC VR-1246), Venezuelan equine encephalitis virus (ATCC VR923; ATCC VR-1250; ATCC VR-1249; ATCC VR-532), and those described in U.S. Pat. Nos. 5,091,309, 5,217,879, and WO92/10578. More particularly, those alpha virus vectors described in U.S. Ser. No. 08/405,627, filed Mar. 15, 1995, WO94/21792, WO92/10578, WO95/07994, U.S. Pat. No. 5,091,309 and U.S. Pat. No. 5,217,879 are employable. Such alpha viruses may be obtained from depositories or collections such as the ATCC in Rockville, Md. or isolated from known sources using commonly available techniques. Preferably, alphavirus vectors with reduced cytotoxicity are used (see U.S. Ser. No. 08/679640).

DNA vector systems such as eukaryotic layered expression systems are also useful for expressing the nucleic acids of the invention. See WO95/07994 for a detailed description of eukaryotic layered expression systems. Preferably, the eukaryotic layered expression systems of the invention are derived from alphavirus vectors and most preferably from Sindbis viral vectors.

Other viral vectors suitable for use in the present invention include those derived from poliovirus, for example ATCC VR-58 and those described in Evans, Nature 339 (1989) 385 and Sabin (1973) J. Biol. Standardization 1:115; rhinovirus, for example ATCC VR-1110 and those described in Arnold (1990) J Cell Biochem L401; pox viruses such as canary pox virus or vaccinia virus, for example ATCC VR-111 and ATCC VR-2010 and those described in Fisher-Hoch (1989) Proc Natl Acad Sci 86:317; Flexner (1989) Ann NY Acad Sci 569:86, Flexner (1990) Vaccine 8:17; in U.S. Pat. No. 4,603,112 and U.S. Pat. No. 4,769,330 and WO89/01973; SV40 virus, for example ATCC VR-305 and those described in Mulligan (1979) Nature 277:108 and Madzak (1992) J Gen Virol 73:1533; influenza virus, for example ATCC VR-797 and recombinant influenza viruses made employing reverse genetics techniques as described in U.S. Pat. No. 5,166,057 and in Enami (1990) Proc Natl Acad Sci 87:3802-3805; Enami & Palese (1991) J Virol 65:2711-2713 and Luytjes (1989) Cell 59:110, (see also McMichael (1983) NEJ Med 309:13, and Yap (1978) Nature 273:238 and Nature (1979) 277:108); human immunodeficiency virus as described in EP-0386882 and in Buchschacher (1992) J. Virol . 66:2731; measles virus, for example ATCC VR-67 and VR-1247 and those described in EP-0440219; Aura virus, for example ATCC VR-368; Bebaru virus, for example ATCC VR-600 and ATCC VR-1240; Cabassou virus, for example ATCC VR-922; Chikungunya virus, for example ATCC VR-64 and ATCC VR-1241; Fort Morgan Virus, for example ATCC VR-924; Getah virus, for example ATCC VR-369 and ATCC VR-1243; Kyzylagach virus, for example ATCC VR-927; Mayaro virus, for example ATCC VR-66; Mucambo virus, for example ATCC VR-580 and ATCC VR-1244; Ndumu virus, for example ATCC VR-371; Pixuna virus, for example ATCC VR-372 and ATCC VR-1245; Tonate virus, for example ATCC VR-925; Triniti virus, for example ATCC VR-469; Una virus, for example ATCC VR-374; Whataroa virus, for example ATCC VR-926; Y-62-33 virus, for example ATCC VR-375; O'Nyong virus, Eastern encephalitis virus, for example ATCC VR-65 and ATCC VR-1242; Western encephalitis virus, for example ATCC VR-70, ATCC VR-1251, ATCC VR-622 and ATCC VR-1252; and coronavirus, for example ATCC VR-740 and those described in Hamre (1966) Proc Soc Exp Biol Med 121:190.

Delivery of the compositions of this invention into cells is not limited to the above mentioned viral vectors. Other delivery methods and media may be employed such as, for example, nucleic acid expression vectors, polycationic condensed DNA linked or unlinked to killed adenovirus alone, for example see U.S. Ser. No. 08/366,787, filed Dec. 30, 1994 and Curiel (1992) Hum Gene Ther 3:147-154 ligand linked DNA, for example see Wu (1989) J Biol Chem 264:16985-16987, eucaryotic cell delivery vehicles cells, for example see U.S. Ser. No. 08/240,030, filed May 9, 1994, and U.S. Ser. No. 08/404,796, deposition of photopolymerized hydrogel materials, hand-held gene transfer particle gun, as described in U.S. Pat. No. 5,149,655, ionizing radiation as described in U.S. Pat. No. 5,206,152 and in WO92/11033, nucleic charge neutralization or filsion with cell membranes. Additional approaches are described in Philip (1994) Mol Cell Biol 14:2411-2418 and in Woffendin (1994) Proc Natl Acad Sci 91:1581-1585.

Particle mediated gene transfer may be employed, for example see U.S. Ser. No. 60/023,867. Briefly, the sequence can be inserted into conventional vectors that contain conventional control sequences for high level expression, and then incubated with synthetic gene transfer molecules such as polymeric DNA-binding cations like polylysine, protamine, and albumin, linked to cell targeting ligands such as asialoorosomucoid, as described in Wu & Wu (1987) J. Biol. Chem . 262:4429-4432, insulin as described in Hucked (1990) Biochem Pharmacol 40:253-263, galactose as described in Plank (1992) Bioconjugate Chem 3:533-539, lactose or transferrin.

Naked DNA may also be employed. Exemplary naked DNA introduction methods are described in WO 90/11092 and U.S. Pat. No. 5,580,859. Uptake efficiency rnay be improved using biodegradable latex beads. DNA coated latex beads are efficiently transported into cells after endocytosis initiation by the beads. The method may be improved further by treatment of the beads to increase hydrophobicity and thereby facilitate disruption of the endosome and release of the DNA into the cytoplasm.

Liposomes that can act as gene delivery vehicles are described in U.S. Pat. No. 5,422,120, WO95/13796, WO94/23697, WO91/14445 and EP-524,968. As described in U.S. Ser. No. 60/023,867, on non-viral delivery, the nucleic acid sequences encoding a polypeptide can be inserted into conventional vectors that contain conventional control sequences for; high level expression, and then be incubated with synthetic gene transfer molecules such as polymeric DNA-binding cations like polylysine, protamine, and albumin, linked to cell targeting ligands such as asialoorosomucoid, insulin, galactose, lactose, or transferrin. Other delivery systems include the use of liposomes to encapsulate DNA comprising the gene under the control of a variety of tissue-specific or ubiquitously-active promoters. Further non-viral delivery suitable for use includes mechanical delivery systems such as the approach described in Woffendin et al (1994) Proc. Natl. Acad. Sci. USA 91(24):11581-11585. Moreover, the coding sequence and the product of expression of such can be delivered through deposition of photopolymerized hydrogel materials. Other conventional methods for gene delivery that can be used for delivery of the coding sequence include, for example, use of hand-held gene transfer particle gun, as described in U.S. Pat. No. 5,149,655; use of ionizing radiation for activating transferred gene, as described in U.S. Pat. No. 5,206,152 and WO92/11033

Exemplary liposome and polycationic gene delivery vehicles are those described in U.S. Pat. Nos. 5,422,120 and 4,762,915; in WO 95/13796; WO94/23697; and WO91/14445; in EP-0524968; and in Stryer, Biochemistry, pages 236-240 (1975) W. H. Freeman, San Francisco; Szoka (1980) Biochem Biophys Acta 600: 1; Bayer (1979) Biochem Biophys Acta 550:464; Rivnay (1987) Meth Enzymol 20 149:119; Wang (1987) Proc Natl Acad Sci 84:7851; Plant (1989) Anal Biochem 176:420.

A polynucleotide composition can comprises therapeutically effective amount of a gene therapy vehicle, as the term is defined above. For purposes of the present invention, an effective dose will be from about 0.01 mg/kg to 50 mg/kg or 0.05 mg/kg to about 10 mg/kg of the DNA constructs in the individual to which it is administered.

Delivery Methods

Once formulated, the polynucleotide compositions of the invention can be administered (1) directly to the subject; (2) delivered ex vivo, to cells derived from the subject; or (3) in vitro for expression of recombinant proteins. The subjects to be treated can be mammals or birds. Also, human subjects can be treated.

Direct delivery of the compositions will generally be accomplished by injection, either subcutaneously, intraperitoneally, intravenously or intramuscularly or delivered to the interstitial space of a tissue. The compositions can also be administered into a lesion. Other modes of administration include oral and pulmonary administration, suppositories, and transdermal or transcutaneous applications (eg. see WO98/20734), needles, and gene guns or hyposprays. Dosage treatment may be a single dose schedule or a multiple dose schedule.

Methods for the ex vivo delivery and reimplantation of transformed cells into a subject are known in the art and described in eg. WO93/14778. Examples of cells useful in ex vivo applications include, for example, stem cells, particularly hematopoetic, lymph cells, macrophages, dendritic cells, or tumor cells.

Generally, delivery of nucleic acids for both ex vivo and in vitro applications can be accomplished by the following procedures, for example, dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and direct microinjection of the DNA into nuclei, all well known in the art.

Polynucleotide and Polypeptide Pharmaceutical Compositions

In addition to the pharmaceutically acceptable carriers and salts described above, the following additional agents can be used with polynucleotide and/or polypeptide compositions.

A.Polypeptides

One example are polypeptides which include, without limitation: asioloorosomucoid (ASOR); transferrin; asialoglycoproteins; antibodies; antibody fragments; ferritin; interleukins; interferons, granulocyte, macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), stem cell factor and erythropoietin. Viral antigens, such as envelope proteins, can also be used. Also, proteins from other invasive organisms, such as the 17 amino acid peptide from the circumsporozoite protein of plasmodium falciparum known as RII.

B.Hormones, Vitamins, etc.

Other groups that can be included are, for example: hormones, steroids, androgens, estrogens, thyroid hormone, or vitamins, folic acid.

C.Polyalkylenes, Polysaccharides, etc.

Also, polyalkylene glycol can be included with the desired polynucleotides/polypeptides. In a preferred embodiment, the polyalkylene glycol is polyethlylene glycol. In addition, mono-, di-, or polysaccharides can be included. In a preferred embodiment of this aspect, the polysaccharide is dextran or DEAE-dextran. Also, chitosan and poly(lactide-co-glycolide)

D.Lipids, and Liposomes

The desired polynucleotide/polypeptide can also be encapsulated in lipids or packaged in liposomes prior to delivery to the subject or to cells derived therefrom.

Lipid encapsulation is generally accomplished using liposomes which are able to stably bind or entrap and retain nucleic acid. The ratio of condensed polynucleotide to lipid preparation can vary but will generally be around 1:1 (mg DNA:micromoles lipid), or more of lipid. For a review of the use of liposomes as carriers for delivery of nucleic acids, see, Hug and Sleight (1991) Biochim. Biophys. Acta . 1097:1-17; Straubinger (1983) Meth. Enzymol . 101:512-527.

Liposomal preparations for use in the present invention include cationic (positively charged), anionic (negatively charged) and neutral preparations. Cationic liposomes have been shown to mediate intracellular delivery of plasmid DNA (Felgner (1987) Proc. Natl. Acad. Sci. USA 20 84:7413-7416); mRNA (Malone (1989) Proc. Natl. Acad. Sci. USA 86:6077-6081); and purified transcription factors (Debs (1990) J. Biol. Chem . 265:10189-10192), in functional form.

Cationic liposomes are readily available. For example, N[1-2,3-dioleyloxy)propyl]-N,N,N-triethylammonium (DOTMA) liposomes are available under the trademark Lipofectin, from GIBCO BRL, Grand Island, N.Y. (See, also, Feigner supra). Other commercially available liposomes include transfectace (DDAB/DOPE) and DOTAP/DOPE (Boerhinger). Other cationic liposomes can be prepared from readily available materials using techniques well known in the art. See, eg. Szoka (1978) Proc. Natl. Acad. Sci. USA 75:4194-4198; WO90/11092 for a description of the synthesis of DOTAP (1,2-bis(oleoyloxy)-3-(trimethylammonio)propane) liposomes.

Similarly, anionic and neutral liposomes are readily available, such as from Avanti Polar Lipids (Birmingham, Ala.), or can be easily prepared using readily available materials. Such materials include phosphatidyl choline, cholesterol, phosphatidyl ethanolamine, dioleoylphosphatidyl choline (DOPC), dioleoylphosphatidyl glycerol (DOPG), dioleoylphoshatidyl ethanolamine (DOPE), among others. These materials can also be mixed with the DOTMA and DOTAP starting materials in appropriate ratios. Methods for making liposomes using these materials are well known in the art.

The liposomes can comprise multilammelar vesicles (MLVs), small unilamellar vesicles (SUVs), or large unilamellar vesicles (LUVs). The various liposome-nucleic acid complexes are prepared using methods known in the art. See eg. Straubinger (1983) Meth. Immunol . 101:512-527; Szoka (1978) Proc. Natl. Acad. Sci. USA 75:4194-4198; Papahadjopoulos (1975) Biochim. Biophys. Acta 394:483; Wilson (1979) Cell 17:77); Deamer & Bangham (1976) Biochim. Biophys. Acta 443:629; Ostro (1977) Biochem. Biophys. Res. Commun . 76:836; Fraley (1979) Proc. Natl. Acad. Sci. USA 76:3348); Enoch & Strittmatter (1979) Proc. Natl. Acad. Sci. USA 76:145; Fraley (1980) J. Biol. Chem . (1980) 255:10431; Szoka & Papahadjopoulos (1978) Proc. Natl. Acad. Sci. USA 75:145; and Schaefer-Ridder (1982) Science 215:166.

E.Lipoproteins

In addition, lipoproteins can be included with the polynucleotide/polypeptide to be delivered. Examples of lipoproteins to be utilized include: chylomicrons, HDL, IDL, LDL, and VLDL. Mutants, fragments, or fusions of these proteins can also be used. Also, modifications of naturally occurring lipoproteins can be used, such as acetylated LDL. These lipoproteins can target the delivery of polynucleotides to cells expressing lipoprotein receptors. Preferably, if lipoproteins are including with the polynucleotide to be delivered, no other targeting ligand is included in the composition.

Naturally occurring lipoproteins comprise a lipid and a protein portion. The protein portion are known as apoproteins. At the present, apoproteins A, B, C, D, and E have been isolated and identified. At least two of these contain several proteins, designated by Roman numerals, AI, AII, AIV; CI, CII, CIII.

A lipoprotein can comprise more than one apoprotein. For example, naturally occurring chylomicrons comprises of A, B, C, and E, over time these lipoproteins lose A and acquire C and E apoproteins. VLDL comprises A, B, C, and E apoproteins, LDL comprises apoprotein B; and HDL comprises apoproteins A, C, and E.

The amino acid of these apoproteins are known and are described in, for example, Breslow (1985) Annu Rev. Biochem 54:699; Law (1986) Adv. Exp Med. Biol. 151:162; Chen (1986) J Biol Chem 261:12918; Kane (1980) Proc Natl Acad Sci USA 77:2465; and Utermann (1984) Hum Genet 65:232.

Lipoproteins contain a variety of lipids including, triglycerides, cholesterol (free and esters), and phospholipids. The composition of the lipids varies in naturally occurring lipoproteins. For example, chylomicrons comprise mainly triglycerides. A more detailed description of the lipid content of naturally occurring lipoproteins can be found, for example, in Meth. Enzymol . 128 (1986). The composition of the lipids are chosen to aid in conformation of the apoprotein for receptor binding activity. The composition of lipids can also be chosen to facilitate hydrophobic interaction and association with the polynucleotide binding molecule.

Naturally occurring lipoproteins can be isolated from serum by ultracentrifugation, for instance. Such methods are described in Meth. Enzymol . (supra); Pitas (1980) J. Biochem . 255:5454-5460 and Mahey (1979) J Clin. Invest 64:743-750. Lipoproteins can also be produced by in vitro or recombinant methods by expression of the apoprotein genes in a desired host cell. See, for example, Atkinson (1986) Annu Rev Biophys Chem 15:403 and Radding (1958) Biochim Biophys Acta 30:443. Lipoproteins can also be purchased from commercial suppliers, such as Biomedical Techniologies, Inc., Stoughton, Mass., USA. Further description of lipoproteins can be found in Zuckermann et al. PCT/US97/14465.

F.Polycationic Agents

Polycationic agents can be included, with or without lipoprotein, in a composition with the desired polynucleotide/polypeptide to be delivered.

Polycationic agents, typically, exhibit a net positive charge at physiological relevant pH and are capable of neutralizing the electrical charge of nucleic acids to facilitate delivery to a desired location. These agents have both in vitro, ex vivo, and in vivo applications. Polycationic agents can be used to deliver nucleic acids to a living subject either intramuscularly, subcutaneously, etc.

The following are examples of useful polypeptides as polycationic agents: polylysine, polyarginine, polyornithine, and protamine. Other examples include histones, protamines, human serum albumin, DNA binding proteins, non-histone chromosomal proteins, coat proteins from DNA viruses, such as (X174, transcriptional factors also contain domains that bind DNA and therefore may be useful as nucleic aid condensing agents. Briefly, transcriptional factors such as C/CEBP, c-jun, c-fos, AP-1, AP-2, AP-3, CPF, Prot-1, Sp-1, Oct-1, Oct-2, CREP, and TFIID contain basic domains that bind DNA sequences.

Organic polycationic agents include: spermine, spermidine, and purtrescine.

The dimensions and of the physical properties of a polycationic agent can be extrapolated from the list above, to construct other polypeptide polycationic agents or to produce synthetic polycationic agents.

Synthetic polycationic agents which are usefuil include, for example, DEAE-dextran, polybrene. Lipofectin™, and lipofectAMINE™ are monomers that form polycationic complexes when combined with polynucleotides/polypeptides.

Immunodiagnostic Assays

Neisserial antigens of the invention can be used in immunoassays to detect antibody levels (or, conversely, anti-Neisserial antibodies can be used to detect antigen levels). Immunoassays based on well defined, recombinant antigens can be developed to replace invasive diagnostics methods. Antibodies to Neisserial proteins within biological samples, including for example, blood or serum samples, can be detected. Design of the immunoassays is subject to a great deal of variation, and a variety of these are known in the art. Protocols for the immunoassay may be based, for example, upon competition, or direct reaction, or sandwich type assays. Protocols may also, for example, use solid supports, or may be by immunoprecipitation. Most assays involve the use of labeled antibody or polypeptide; the labels may be, for example, fluorescent, chemiluminescent, radioactive, or dye molecules. Assays which amplify the signals from the probe are also known; examples of which are assays which utilize biotin and avidin, and enzyme-labeled and mediated immunoassays, such as ELISA assays.

Kits suitable for immunodiagnosis and containing the appropriate labeled reagents are constructed by packaging the appropriate materials, including the compositions of the invention, in suitable containers, along with the remaining reagents and materials (for example, suitable buffers, salt solutions, etc.) required for the conduct of the assay, as well as suitable set of assay instructions.

Nucleic Acid Hybridisation

“Hybridization” refers to the association of two nucleic acid sequences to one another by hydrogen bonding. Typically, one sequence will be fixed to a solid support and the other will be free in solution. Then, the two sequences will be placed in contact with one another under conditions that favor hydrogen bonding. Factors that affect this bonding include: the type and volume of solvent; reaction temperature; time of hybridization; agitation; agents to block the non-specific attachment of the liquid phase sequence to the solid support (Denhardt's reagent or BLOTTO); concentration of the sequences; use of compounds to increase the rate of association of sequences (dextran sulfate or polyethylene glycol); and the stringency of the washing conditions following hybridization. See Sambrook et al. [supra] Volume 2, chapter 9, pages 9.47 to 9.57.

“Stringency” refers to conditions in a hybridization reaction that favor association of very similar sequences over sequences that differ. For example, the combination of temperature and salt concentration should be chosen that is approximately 120 to 200° C. below the calculated Tm of the hybrid under study. The temperature and salt conditions can often be determined empirically in preliminary experiments in which samples of genomic DNA immobilized on filters are hybridized to the sequence of interest and then washed under conditions of different stringencies. See Sambrook et al. at page 9.50.

Variables to consider when performing, for example, a Southern blot are (1) the complexity of the DNA being blotted and (2) the homology between the probe and the sequences being detected. The total amount of the fragment(s) to be studied can vary a magnitude of 10, from 0.1 to 1 μg for a plasmid or phage digest to 10 −9 to 10 −8 g for a single copy gene in a highly complex eukaryotic genome. For lower complexity polynucleotides, substantially shorter blotting, hybridization, and exposure times, a smaller amount of starting polynucleotides, and lower specific activity of probes can be used. For example, a single-copy yeast gene can be detected with an exposure time of only 1 hour starting with 1 μg of yeast DNA, blotting for two hours, and hybridizing for 4-8 hours with a probe of 10 8 cpm/μg. For a single-copy mammalian gene a conservative approach would start with 10 μg of DNA, blot overnight, and hybridize overnight in the presence of 10% dextran sulfate using a probe of greater than 10 8 cpm/μg, resulting in an exposure time of ˜24 hours.

Several factors can affect the melting temperature (Tm) of a DNA-DNA hybrid between the probe and the fragment of interest, and consequently, the appropriate conditions for hybridization and washing. In many cases the probe is not 100% homologous to the fragment. Other commonly encountered variables include the length and total G+C content of the hybridizing sequences and the ionic strength and formamide content of the hybridization buffer. The effects of all of these factors can be approximated by a single equation:

Tm =81+16.6(log 10 Ci)+0.4[%( G+C )]−0.6(%formamide)−600 /n −1.5(%mismatch).

where Ci is the salt concentration (monovalent ions) and n is the length of the hybrid in base pairs (slightly modified from Meinkoth & Wahl (1984) Anal. Biochem . 138: 267-284).

In designing a hybridization experiment, some factors affecting nucleic acid hybridization can be conveniently altered. The temperature of the hybridization and washes and the salt concentration during the washes are the simplest to adjust. As the temperature of the hybridization increases (ie. stringency), it becomes less likely for hybridization to occur between strands that are nonhomologous, and as a result, background decreases. If the radiolabeled probe is not completely homologous with the immobilized fragment (as is frequently the case in gene family and interspecies hybridization experiments), the hybridization temperature must be reduced, and background will increase. The temperature of the washes affects the intensity of the hybridizing band and the degree of background in a similar manner. The stringency of the washes is also increased with decreasing salt concentrations.

In general, convenient hybridization temperatures in the presence of 50% formamide are 42° C. for a probe with is 95% to 100% homologous to the target fragment, 37° C. for 90% to 95% homology, and 32° C. for 85% to 90% homology. For lower homologies, formamide content should be lowered and temperature adjusted accordingly, using the equation above. If the homology between the probe and the target fragment are not known, the simplest approach is to start with both hybridization and wash conditions which are nonstringent. If non-specific bands or high background are observed after autoradiography, the filter can be washed at high stringency and reexposed. If the time required for exposure makes this approach impractical, several hybridization and/or washing stringencies should be tested in parallel.

Nucleic Acid Probe Assays

Methods such as PCR, branched DNA probe assays, or blotting techniques utilizing nucleic acid probes according to the invention can determine the presence of cDNA or mRNA. A probe is said to “hybridize” with a sequence of the invention if it can form a duplex or double stranded complex, which is stable enough to be detected.

The nucleic acid probes will hybridize to the Neisserial nucleotide sequences of the invention (including both sense and antisense strands). Though many different nucleotide sequences will encode the amino acid sequence, the native Neisserial sequence is preferred because it is the actual sequence present in cells. mRNA represents a coding sequence and so a probe should be complementary to the coding sequence; single-stranded cDNA is complementary to mRNA, and so a cDNA probe should be complementary to the non-coding sequence.

The probe sequence need not be identical to the Neisserial sequence (or its complement)—some variation in the sequence and length can lead to increased assay sensitivity if the nucleic acid probe can form a duplex with target nucleotides, which can be detected. Also, the nucleic acid probe can include additional nucleotides to stabilize the formed duplex. Additional Neisserial sequence may also be helpful as a label to detect the formed duplex. For example, a non-complementary nucleotide sequence may be attached to the 5′ end of the probe, with the remainder of the probe sequence being complementary to a Neisserial sequence. Alternatively, non-complementary bases or longer sequences can be interspersed into the probe, provided that the probe sequence has sufficient complementarity with the a Neisserial sequence in order to hybridize therewith and thereby form a duplex which can be detected.

The exact length and sequence of the probe will depend on the hybridization conditions, such as temperature, salt condition and the like. For example, for diagnostic applications, depending on the complexity of the analyte sequence, the nucleic acid probe typically contains at least 10-20 nucleotides, preferably 15-25, and more preferably at least 30 nucleotides, although it may be shorter than this. Short primers generally require cooler temperatures to form sufficiently stable hybrid complexes with the template.

Probes may be produced by synthetic procedures, such as the triester method of Matteucci et al. [ J. Am. Chem. Soc . (1981) 103:3185], or according to Urdea et al. [ Proc. Natl. Acad. Sci. USA (1983) 80: 7461], or using commercially available automated oligonucleotide synthesizers.

The chemical nature of the probe can be selected according to preference. For certain applications, DNA or RNA are appropriate. For other applications, modifications may be incorporated eg. backbone modifications, such as phosphorothioates or methylphosphonates, can be used to increase in vivo half-life, alter RNA affinity, increase nuclease resistance etc. [eg. see Agrawal & Iyer (1995) Curr Opin Biotechnol 6:12-19; Agrawal (1996) TIBTECH 14:376-387]; analogues such as peptide nucleic acids may also be used [eg. see Corey (1997) TIBTECH 15:224-229; Buchardt et al. (1993) TIBTECH 11:384-386].

Alternatively, the polymerase chain reaction (PCR) is another well-known means for detecting small amounts of target nucleic acids. The assay is described in: Mullis et al. [ Meth. Enzytmnol . (1987) 155: 335-350]; U.S. Pat. Nos. 4,683,195 and 4,683,202. Two “primer” nucleotides hybridize with the target nucleic acids and are used to prime the reaction. The primers can comprise sequence that does not hybridize to the sequence of the amplification target (or its complement) to aid with duplex stability or, for example, to incorporate a convenient restriction site. Typically, such sequence will flank the desired Neisserial sequence.

A thermostable polymerase creates copies of target nucleic acids from the primers using the original target nucleic acids as a template. After a threshold amount of target nucleic acids are generated by the polymerase, they can be detected by more traditional methods, such as Southern blots. When using the Southern blot method, the labelled probe will hybridize to the Neisserial sequence (or its complement).

Also, mRNA or cDNA can be detected by traditional blotting techniques described in Sambrook et al [supra]. mRNA, or cDNA generated from mRNA using a polymerase enzyme, can be purified and separated using gel electrophoresis. The nucleic acids on the gel are then blotted onto a solid support, such as nitrocellulose. The solid support is exposed to a labelled probe and then washed to remove any unhybridized probe. Next, the duplexes containing the labeled probe are detected. Typically, the probe is labelled with a radioactive moiety.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-E show biochemical data and sequence analysis pertaining to ORF 40. FIG. 1A shows the results of affinity purification and FIG. 1B shows the results of expression of the GST-fusion in E. coli , where M2 and M1 are molecular weight markers, and Arrows indicate the position of the main recombinant product. FIG. 1C shows the results of FACS analysis of the sera of mice that were immunized with the purified protein. FIG. 1D shows the results of bactericidal assay where a diamond (♦) shows preimmune data; a triangle (▴) shows GST control data; and a circle (&Circlesolid;) shows data with recombinant N.meningitidis protein. FIG. 1E shows computer analysis showing a hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower). The AMPHI program has been used to predict T-cell epitopes (Gao et al. (1989) J. Immunol . 143:3007; Roberts et al. (1996) AIDS Res Hum Retrovir 12:593; Quakyi et al. (1992) Scad J. Immunol suppl . 11:9) and is available in the Protean Package of DNASTAR, Inc. (1228 South Park Street, Madison, Wis. 53715 USA).

FIGS. 2A-E show biochemical data and sequence analysis pertaining to ORF 38-1. FIG. 2A shows the results of affinity purification and FIG. 2B shows the results of expression of the GST-fusion in E. coli , where M2 and M1 are molecular weight markers, and Arrows indicate the position of the main recombinant product. FIG. 2C shows the results of FACS analysis of the sera of mice that were immunized with the purified protein. FIG. 2D shows the results of bactericidal assay where a diamond (♦) shows preimmune data; a triangle (▴) shows GST control data; and a circle (&Circlesolid;) shows data with recombinant N.meningitidis protein. FIG. 2E shows plots of hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower).

FIGS. 3A-D show biochemical data and sequence analysis pertaining to ORF 44-1. FIG. 3A shows the results of affinity purification and FIG. 3B shows the results of expression of the GST-fusion in E. coli , where M2 and M1 are molecular weight markers, and Arrows indicate the position of the main recombinant product. FIG. 3C shows the results of bactericidal assay where a diamond (+) shows preimmune data; a triangle (▴) shows GST control data; and a circle (&Circlesolid;) shows data with recombinant N.meningitidis protein. FIG. 3D shows plots of hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower).

FIG. 4A shows the results of affinity purification of ORF 52, where M1 is a molecular weight marker, and the arrow indicates the position of the main recombinant product. FIG. 4B shows a hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower).

FIG. 5 shows a hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower) for ORF 114.

FIG. 6 shows a hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower) for ORF 41.

FIG. 7 shows a hydrophilicity plot (upper), an antigenic index plot (middle), and an AMPHI analysis (lower) for ORF 124.

FIG. 8 shows an alignment comparison of amino acid sequences for ORF 40 for several strains of Neisseria (zn07 — 1, SEQ ID NO:96; zn20 — 1, SEQ ID NO:104; zn21 — 1, SEQ ID NO:105; zn06 — 1, SEQ ID NO:95; zn19 — 1, SEQ ID NO:103; zn03 — 1, SEQ ID NO:93; zn 1 8 — 1, SEQ ID NO:102; zn11_ass, SEQ ID NO:99; zn02 — 1, SEQ ID NO:92; zn04 — 1, SEQ ID NO:94; zn16 — 1, SEQ ID NO:101; zn14 — 1, SEQ ID NO:100; z2491, SEQ ID NO:91; zn10 — 1, SEQ ID NO:98; zn22 — 1, SEQ ID NO:106; zn23 — 1, SEQ ID NO: 107; zn28_ass, SEQ ID NO: 110; zn24 — 1, SEQ ID NO: 108; zn25_ass, SEQ ID NO:109; zn08 — 1, SEQ ID NO:97; zn29_ass, SEQ ID NO:111). Dark shading indicates regions of homology, and gray shading indicates the conservation of amino acids with similar characteristics. The Figure demonstrates a high degree of conservation among the various strains, further confirming its utility as an antigen for both vaccines and diagnostics.

EXAMPLES

The examples describe nucleic acid sequences which have been identified in N.meningitidis , along with their putative translation products. Not all of the nucleic acid sequences are complete ie. they encode less than the full-length wild-type protein. It is believed at present that none of the DNA sequences described herein have significant homologs in N.gonorrhoeae.

The examples are generally in the following format:

a nucleotide sequence which has been identified in N.meningitidis (strain B)

the putative translation product of this sequence

a computer analysis of the translation product based on database comparisons

a corresponding gene and protein sequence identified in N.meningitidis (strain A)

a description of the characteristics of the proteins which indicates that they might be suitably antigenic

results of biochemical analysis (expression, purification, ELISA, FACS etc.)

The examples typically include details of sequence homology between species and strains. Proteins that are similar in sequence are generally similar in both structure and function, and the homology often indicates a common evolutionary origin. Comparison with sequences of proteins of known function is widely used as a guide for the assignment of putative protein function to a new sequence and has proved particularly useful in whole-genome analyses.

Sequence comparisons were performed at NCBI (http://www.ncbi.nlm.nih.gov) using the algorithms BLAST, BLAST2, BLASTn, BLASTp, tBLASTn, BLASTx, & tBLASTx [eg. see also Altschul et al. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25:2289-3402]. Searches were performed against the following databases: non-redundant GenBank+EMBL+DDBJ+PDB sequences and non-redundant GenBank CDS translations+PDB+SwissProt+SPupdate+PIR sequences.

Dots within nucleotide sequences (eg. position 288 in Example 12) represent nucleotides which have been arbitrarily introduced in order to maintain a reading frame. In the same way, double-underlined nucleotides were removed. Lower case letters (eg. position 589 in Example 12) represent ambiguities which arose during alignment of independent sequencing reactions (some of the nucleotide sequences in the examples are derived from combining the results of two or more experiments).

Nucleotide sequences were scanned in all six reading frames to predict the presence of hydrophobic domains using an algorithm based on the statistical studies of Esposti et al. [Critical evaluation of the hydropathy of membrane proteins (1990) Eur J Biochem 190:207-219]. These domains represent potential transmembrane regions or hydrophobic leader sequences.

Open reading frames were predicted from fragmented nucleotide sequences using the program ORFFINDER (NCBI).

Underlined amino acid sequences indicate possible transmembrane domains or leader sequences in the ORFs, as predicted by the PSORT algorithm (http://www.psort.nibb.ac.jp). Functional domains were also predicted using the MOTIFS program (GCG Wisconsin & PROSITE).

Various tests can be used to assess the in vivo immunogenicity of the proteins identified in the examples. For example, the proteins can be expressed recombinantly and used to screen patient sera by immunoblot. A positive reaction between the protein and patient serum indicates that the patient has previously mounted an immune response to the protein in question ie. the protein is an immunogen. This method can also be used to identify immunodominant proteins.

The recombinant protein can also be conveniently used to prepare antibodies eg. in a mouse. These can be used for direct confirmation that a protein is located on the cell-surface. Labelled antibody (eg. fluorescent labelling for FACS) can be incubated with intact bacteria and the presence of label on the bacterial surface confirms the location of the protein.

In particular, the following methods (A) to (S) were used to express, purify and biochemically characterise the proteins of the invention:

A) Chromosomal DNA Preparation

N.meningitidis strain 2996 was grown to exponential phase in 100 ml of GC medium, harvested by centrifugation, and resuspended in 5 ml buffer (20% Sucrose, 50 mM Tris-HCl, 50 mM EDTA, pH8). After 10 minutes incubation on ice, the bacteria were lysed by adding 10 ml lysis solution (50 mM NaCl, 1% Na-Sarkosyl, 50 μg/ml Proteinase K), and the suspension was incubated at 37° C. for 2 hours. Two phenol extractions (equilibrated to pH 8) and one ChCl 3 /isoamylalcohol (24:1) extraction were performed. DNA was precipitated by addition of 0.3M sodium acetate and 2 volumes ethanol, and was collected by centrifugation. The pellet was washed once with 70% ethanol and redissolved in 4 ml buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8). The DNA concentration was measured by reading the OD at 260 nm.

B) Oligonucleotide Design

Synthetic oligonucleotide primers were designed on the basis of the coding sequence of each ORF, using (a) the meningococcus B sequence when available, or (b) the gonococcus/meningococcus A sequence, adapted to the codon preference usage of meningococcus as necessary. Any predicted signal peptides were omitted, by deducing the 5′-end amplification primer sequence immediately downstream from the predicted leader sequence.

The 5′ primers included two restriction enzyme recognition sites (BamHI-NdeI, BamHI-NheI, or EcoRI-NheI, depending on the gene's own restriction pattern); the 3′ primers included a XhoI restriction site. This procedure was established in order to direct the cloning of each amplification product (corresponding to each ORF) into two different expression systems: pGEX-KG (using either BamHI-XhoI or EcoRI-XhoI), and pET21b+ (using either NdeI-XhoI or NheI-XhoI).

5′-end primer tail:	CGC GGATCCCATATG	(BamHI-NdeI)	(SEQ ID NO:134)
	CGC GGATCCGCTAGC	(BamHI-NheI)	(SEQ ID NO:135)
	CCG GAATTC TA GCTAGC	(EcoRI-NheI)	(SEQ ID NO:136)
3′-end primer tail:	CCCG CTCGAG	(XhoI)	(SEQ ID NO:137)

As well as containing the restriction enzyme recognition sequences, the primers included nucleotides which hybridised to the sequence to be amplified. The number of hybridizing nucleotides depended on the melting temperature of the whole primer, and was determined for each primer using the formulae:

T                                                                        m                                                                     = 4 (G+C) + 2 (A+T) (tail excluded)
T                                                                        m                                                                     = 64.9 + 0.41 (% GC) − 600/N (whole primer)

The average melting temperature of the selected oligos were 65-70° C. for the whole oligo and 50-55° C. for the hybridising region alone.

Table I shows the forward and reverse primers used for each amplification. Oligos were synthesized by a Perkin Elmer 394 DNA/RNA Synthesizer, eluted from the columns in 2 ml NH 40 H, and deprotected by 5 hours incubation at 56° C. The oligos were precipitated by addition of 0.3M Na-Acetate and 2 volumes ethanol. The samples were then centrifuged and the pellets resuspended in either 100 μl or 1 ml of water. OD 260 was determined using a Perkin Elmer Lambda Bio spectrophotometer and the concentration was determined and adjusted to 2-10 μmol/μl.

TABLE I
PCR primers
ORF	Primer	Sequence	Restriction sites
ORF 38	Forward	CGC GGATCCCATATG -TCGCCGCAAAATTCCGA <SEQ ID 112>	BamHI-NdeI
	Reverse	CCCG CTCGAG -TTTTGCCGCGTTAAAAGC <SEQ ID 113>	XhoI
ORF 40	Forward	CGC GGATCCCATATG -ACCGTGAAGACCGCC <SEQ ID 114>	BamHI-NdeI
	Reverse	CCCG CTCGAG -CCACTGATAACCGACAGA <SEQ ID 115>	XhoI
ORF 41	Forward	CGC GGATCCCATATG -TATTTGAAACAGCTCCAAG <SEQ ID 116>	BamHI-NdeI
	Reverse	CCCG CTCGAG -TTCTGGGTGAATGTTA <SEQ ID 117>	XhoI
ORF 44	Forward	GC GGATCCCATATG -GGCACGGACAACCCC <SEQ ID 118>	BamHI-NdeI
	Reverse	CCCG CTCGAG -ACGTGGGGAACAGTCT <SEQ ID 119>	XhoI
ORF 51	Forward	GC GGATCCCATATG -AAAAATATTCAAGTAGTTGC <SEQ ID 120>	BamHI-NdeI
	Reverse	CCCG CTCGAG -AAGTTTGATTAAACCCG <SEQ ID 121>	XhoI
ORF 52	Forward	CGC GGATCCCATATG -TGCCAACCGCAATCCG <SEQ ID 122>	BamHI-NdeI
	Reverse	CCCG CTCGAG -TTTTTCCAGCTCCGGCA <SEQ ID 123>	XhoI
ORF 56	Forward	GC GGATCCCATATG -GTTATCGGAATATTACTCG <SEQ ID 124>	BamHI-NdeI
	Reverse	CCCG CTCGAG -GGCTCCAGAAGCTGG <SEQ ID 125>	XhoI
ORF 69	Forward	CGC GGATCCCATATG -CGGACGTGGTTGGTTTT <SEQ ID 126>	BamHI-NdeI
	Reverse	CCCG CTCGAG -ATATCTTCCGTTTTTTTCAC <SEQ ID 127>	XhoI
ORF 82	Forward	CGC GGATCCGCTAGC -GTAAATTTATTATTTTTAGAA <SEQ ID 128>	BamHI-NheI
	Reverse	CCCG CTCGAG -TTCCAACTCATTGAAGTA <SEQ ID 129>	XhoI
ORF 114	Forward	CGC GGATCCCATATG -AATAAAGGTTTACATCGCAT <SEQ ID 130>	BamHI-NheI
	Reverse	CCCG CTCGAG -AATCGCTGCACCGGCT <SEQ ID 131>	XhoI
ORF 124	Forward	CGC GGATCCCATATG -ACTGCCTTTTCGACA <SEQ ID 132>	BamHI-NheI
	Reverse	CCCG CTCGAG -GCGTGAAGCGTCAGGA <SEQ ID 133>	XhoI

C) Amplification

The standard PCR protocol was as follows: 50-200 ng of genomic DNA were used as a template in the presence of 20-40 μM of each oligo, 400-800 μM dNTPs solution, 1×PCR buffer (including 1.5 mM MgCl 2 ), 2.5 units TaqI DNA polymerase (using Perkin-Elmer AmpliTaQ, GIBCO Platinum, Pwo DNA polymerase, or Tahara Shuzo Taq polymerase).

In some cases, PCR was optimised by the addition of 10 μl DMSO or 50 μl 2M betaine.

After a hot start (adding the polymerase during a preliminary 3 minute incubation of the whole mix at 95° C.), each sample underwent a double-step amplification: the first 5 cycles were performed using as the hybridization temperature the one of the oligos excluding the restriction enzymes tail, ollowed by 30 cycles performed according to the hybridization temperature of the whole length oligos. The cycles were followed by a final 10 minute extension step at 72° C.

The standard cycles were as follows:

	Denaturation	Hybridisation	Elongation
	First 5 cycles	30 seconds	30 seconds	30-60 seconds
		95° C.	50-55° C.	72° C.
	Last 30 cycles	30 seconds	30 seconds	30-60 seconds
		95° C.	65-70° C.	72° C.

The elongation time varied according to the length of the ORF to be amplified.

The amplifications were performed using either a 9600 or a 2400 Perkin Elmer GeneAmp PCR System. To check the results, 1/10 of the amplification volume was loaded onto a 1-1.5% agarose gel and the size of each amplified fragment compared with a DNA molecular weight marker.

The amplified DNA was either loaded directly on a 1% agarose gel or first precipitated with ethanol and resuspended in a suitable volume to be loaded on a 1% agarose gel. The DNA fragment corresponding to the right size band was then eluted and purified from gel, using the Qiagen Gel Extraction Kit, following the instructions of the manufacturer. The final volume of the DNA fragment was 30 μl or 50 μl of either water or 10 mM Tris, pH 8.5.

D) Digestion of PCR Fragments

The purified DNA corresponding to the amplified fragment was split into 2 aliquots and double-digested with:

NdeI/XhoI or NheI/XhoI for cloning into pET-21b+ and further expression of the protein as a C-terminus His-tag fusion

BamHI/XhoI or EcoRI/XhoI for cloning into pGEX-KG and further expression of the protein as N-terminus GST fusion.

EcoRI/PstI, EcoRI/SalI, SalI/PstI for cloning into pGex-His and further expression of the protein as N-terminus His-tag fusion

Each purified DNA fragment was incubated (37° C. for 3 hours to overnight) with 20 units of each restriction enzyme (New England Biolabs ) in a either 30 or 40 μl final volume in the presence of the appropriate buffer. The digestion product was then purified using the QIAquick PCR purification kit, following the manufacturer's instructions, and eluted in a final volume of 30 or 50 μl of either water or 10 mM Tris-HCl, pH 8.5. The final DNA concentration was determined by 1% agarose gel electrophoresis in the presence of titrated molecular weight marker.

E) Digestion of the Cloning Vectors (pET22B, pGEX-KG, pTRC-His A, and pGex-His)

10 μg plasmid was double-digested with 50 units of each restriction enzyme in 200 μl reaction volume in the presence of appropriate buffer by overnight incubation at 37° C. After loading the whole digestion on a 1% agarose gel, the band corresponding to the digested vector was purified from the gel using the Qiagen QIAquick Gel Extraction Kit and the DNA was eluted in 50 μl of 10 mM Tris-HCl, pH 8.5. The DNA concentration was evaluated by measuring OD 260 of the sample, and adjusted to 50 μg/μl. 1 μl of plasmid was used for each cloning procedure.

The vector pGEX-His is a modified pGEX-2T vector carrying a region encoding six histidine residues upstream to the thrombin cleavage site and containing the multiple cloning site of the vector pTRC99 (Pharmacia).

F) Cloning

The fragments corresponding to each ORF, previously digested and purified, were ligated in both pET22b and pGEX-KG. In a final volume of 20 μl, a molar ratio of 3:1 fragrnent/vector was ligated using 0.5 μl of NEB T4 DNA ligase (400 units/μl), in the presence of the buffer supplied by the manufacturer. The reaction was incubated at room temperature for 3 hours. In some experiments, ligation was performed using the Boehringer “Rapid Ligation Kit”, following the manufacturer's instructions.

In order to introduce the recombinant plasmid in a suitable strain, 100 μl E. coli DH5 competent cells were incubated with the ligase reaction solution for 40 minutes on ice, then at 37° C. for 3 minutes, then, after adding 800 μl LB broth, again at 37° C. for 20 minutes. The cells were then centrifuged at maximum speed in an Eppendorf microfuge and resuspended in approximately 200 μl of the supernatant. The suspension was then plated on LB ampicillin (100 mg /ml).

The screening of the recombinant clones was performed by growing 5 randomly-chosen colonies overnight at 37° C. in either 2 ml (PGEX or pTC clones) or 5 ml (pET clones) LB broth+100 μg/ml ampicillin. The cells were then pelletted and the DNA extracted using the Qiagen QIAprep Spin Miniprep Kit, following the manufacturer's instructions, to a final volume of 30 μl. 5 μl of each individual miniprep (approximately 1 g ) were digested with either NdeI/XhoI or BamHI/XhoI and the whole digestion loaded onto a 1-1.5% agarose gel (depending on the expected insert size), in parallel with the molecular weight marker (1 Kb DNA Ladder, GIBCO). The screening of the positive clones was made on the base of the correct insert size.

G) Expression

Each ORF cloned into the expression vector was transformed into the strain suitable for expression of the recombinant protein product. 1 μl of each construct was used to transform 30 μl of E.coli BL21 (PGEX vector), E.coli TOP 10 (PTRC vector) or E.coli BL21-DE3 (pET vector), as described above. In the case of the pGEX-His vector, the same E.coli strain (W3110) was used for initial cloning and expression. Single recombinant colonies were inoculated into 2 ml LB+Amp (100 μg/ml), incubated at 37° C. overnight, then diluted 1:30 in 20 ml of LB+Amp (100 μg/ml) in 100 ml flasks, making sure that the OD 600 ranged between 0.1 and 0.15. The flasks were incubated at 30° C. into gyratory water bath shakers until OD indicated exponential growth suitable for induction of expression (0.4-0.8 OD for pET and pTRC vectors; 0.8-1 OD for pGEX and pGEX-His vectors). For the pET, pTRC and pGEX-His vectors, the protein expression was induced by addition of 1 mM IPTG, whereas in the case of pGEX system the final concentration of IPTG was 0.2 mM. After 3 hours incubation at 30° C., the final concentration of the sample was checked by OD. In order to check expression, 1 ml of each sample was removed, centrifuged in a microfuge, the pellet resuspended in PBS, and analysed by 12% SDS-PAGE with Coomassie Blue staining. The whole sample was centrifuged at 6000g and the pellet resuspended in PBS for further use.

H) GST-fusion Proteins Large-scale Purification

A single colony was grown overnight at 37° C. on LB+Amp agar plate. The bacteria were inoculated into 20 ml of LB+Amp liquid culture in a water bath shaker and grown overnight. Bacteria were diluted 1:30 into 600 ml of fresh medium and allowed to grow at the optimal temperature (20-37° C.) to OD 550 0.8-1. Protein expression was induced with 0.2 mM IPTG followed by three hours incubation. The culture was centrifuged at 8000 rpm at 4° C. The supernatant was discarded and the bacterial pellet was resuspended in 7.5 ml cold PBS. The cells were disrupted by sonication on ice for 30 sec at 40W using a Branson sonifier B-15, frozen and thawed twice and centrifuged again. The supernatant was collected and mixed with 150 μl Glutatione-Sepharose 4B resin (Pharmacia) (previously washed with PBS) and incubated at room temperature for 30 minutes. The sample was centrifuged at 700 g for 5 minutes at 4° C. The resin was washed twice with 10 ml cold PBS for 10 minutes, resuspended in 1 ml cold PBS, and loaded on a disposable column. The resin was washed twice with 2 ml cold PBS until the flow-through reached OD 280 of 0.02-0.06. The GST-fusion protein was eluted by addition of 700 μl cold Glutathione elution buffer (10 mM reduced glutathione, 50 mM Tris-HCl) and fractions collected until the OD 280 was 0.1. 21 μl of each fraction were loaded on a 12% SDS gel using either Biorad SDS-PAGE Molecular weight standard broad range (M1) (200, 116.25, 97.4, 66.2, 45, 31, 21.5, 14.4, 6.5 kDa) or Amersham Rainbow Marker (M2) (220, 66, 46, 30, 21.5, 14.3 kDa) as standards. As the MW of GST is 26 kDa, this value must be added to the MW of each GST-fusion protein.

I) His-fusion Solubility Analysis

To analyse the solubility of the His-fusion expression products, pellets of 3 ml cultures were resuspended in buffer M1 [500 μl PBS pH 7.2]. 25 μl lysozyme (10 mg/ml) was added and the bacteria were incubated for 15 min at 4° C. The pellets were sonicated for 30 sec at 40W using a Branson sonifier B-15, frozen and thawed twice and then separated again into pellet and supernatant by a centrifugation step. The supernatant was collected and the pellet was resuspended in buffer M2 [8M urea, 0.5M NaCl, 20 mM imidazole and 0.1M NaH 2 PO 4 ] and incubated for 4 hours at 4° C. After centrifugation, the supernatant was collected and the pellet was resuspended in buffer M3 [6M guanidinium-HCl, 0.5M NaCl, 20 mM imidazole and 0.1M NaH 2 PO 4 ] overnight at 4° C. The supernatants from all steps were analysed by SDS-PAGE.

J) His-fusion Large-scale Purification

A single colony was grown overnight at 37° C. on a LB+Amp agar plate. The bacteria were inoculated into 20 ml of LB+Amp liquid culture and incubated overnight in a water bath shaker. Bacteria were diluted 1:30 into 600 ml fresh medium and allowed to grow at the optimal temperature (20-37° C.) to OD 550 0.6-0.8. Protein expression was induced by addition of 1 mM IPTG and the culture further incubated for three hours. The culture was centrifuged at 8000 rpm at 4° C., the supernatant was discarded and the bacterial pellet was resuspended in 7.5 ml of either (i) cold buffer A (300 mM NaCl, 50 mM phosphate buffer, 10 mM imidazole, pH 8) for soluble proteins or (ii) buffer B (urea 8M, 10 mM Tris-HCl, 100 mM phosphate buffer, pH 8.8) for insoluble proteins.

The cells were disrupted by sonication on ice for 30 sec at 40W using a Branson sonifier B-15, frozen and thawed two times and centrifuged again.

For insoluble proteins, the supernatant was stored at −20° C., while the pellets were resuspended in 2 ml buffer C (6M guanidine hydrochloride, 100 mM phosphate buffer, 10 mM Tris-HCl, pH 7.5) and treated in a homogenizer for 10 cycles. The product was centrifuged at 13000 rpm for 40 minutes.

Supernatants were collected and mixed with 150 μl Ni 2+ -resin (Pharmacia) (previously washed with either buffer A or buffer B, as appropriate) and incubated at room temperature with gentle agitation for 30 minutes. The sample was centrifuged at 700 g for 5 minutes at 4° C. The resin was washed twice with 10 ml buffer A or B for 10 minutes, resuspended in 1 ml buffer A or B and loaded on a disposable column. The resin was washed at either (i) 4° C. with 2 ml cold buffer A or (ii) room temperature with 2 ml buffer B, until the flow-through reached OD 280 of 0.02-0.06.

The resin was washed with either (i) 2 ml cold 20 mM imidazole buffer (300 mM NaCl, 50 mM phosphate buffer, 20 mM imidazole, pH 8) or (ii) buffer D (urea 8M, 10 mM Tris-HCl, 100 mM phosphate buffer, pH 6.3) until the flow-through reached the O.D 280 of 0.02-0.06. The His-fusion protein was eluted by addition of 700 μl of either (i) cold elution buffer A (300 mM NaCl, 50 mM phosphate buffer, 250 mM imidazole, pH 8) or (ii) elution buffer B (urea 8M, 10 mM Tris-HCl, 100 mM phosphate buffer, pH 4.5) and fractions collected until the OD 280 was 0.1. 21 μl of each fraction were loaded on a 12% SDS gel.

K) His-fusion Proteins Renaturation

10% glycerol was added to the denatured proteins. The proteins were then diluted to 20 μg/ml using dialysis buffer I (10% glycerol, 0.5M arginine, 50 mM phosphate buffer, 5 mM reduced glutathione, 0.5 mM oxidised glutathione, 2M urea, pH 8.8) and dialysed against the same buffer at 4° C. for 12-14 hours. The protein was further dialysed against dialysis buffer II (10% glycerol, 0.5M arginine, 50 mM phosphate buffer, 5 mM reduced glutathione, 0.5 mM oxidised glutathione, pH 8.8) for 12-14 hours at 4° C. Protein concentration was evaluated using the formula:

Protein (mg/ml)=(1.55 ×OD 280 )−(0.76 ×OD 260 )

L) His-fusion Large-scale Purification

500 ml of bacterial cultures were induced and the fusion proteins were obtained soluble in buffer M1, M2 or M3 using the procedure described above. The crude extract of the bacteria was loaded onto a Ni-NTA superflow column (Qiagen) equilibrated with buffer M1, M2 or M3 depending on the solubilization buffer of the fusion proteins. Unbound material was eluted by washing the column with the same buffer. The specific protein was eluted with the corresponding buffer containing 500 mM imidazole and dialysed against the corresponding buffer without imidazole. After each run the columns were sanitized by washing with at least two column volumes of 0.5 M sodium hydroxide and reequilibrated before the next use.

M) Mice Immunisations

20 μg of each purified protein were used to immunise mice intraperitoneally. In the case of ORF 44, CD1 mice were imnmunised with Al(OH) 3 as adjuvant on days 1, 21 and 42, and immune response was monitored in samples taken on day 56. For ORF 40, CD1 mice were immunised using Freund's adjuvant, rather than Al(OH) 3 , and the same immunisation protocol was used, except that the immune response was measured on day 42, rather than 56. Similarly, for ORF 38, CD1 mice were immunised with Freund's adjuvant, but the immune response was measured on day 49.

N) ELISA Assay (Sera Analysis)

The acapsulated MenB M7 strain was plated on chocolate agar plates and incubated overnight at 37° C. Bacterial colonies were collected from the agar plates using a sterile dracon swab and inoculated into 7 ml of Mueller-Hinton Broth (Difco) containing 0.25% Glucose. Bacterial growth was monitored every 30 minutes by following OD 620 . The bacteria were let to grow until the OD reached the value of 0.3-0.4. The culture was centrifuged for 10 minutes at 10000 rpm. The supernatant was discarded and bacteria were washed once with PBS, resuspended in PBS containing 0.025% formaldehyde, and incubated for 2 hours at room temperature and then overnight at 4° C. with stirring. 100 μl bacterial cells were added to each well of a 96 well Greiner plate and incubated overnight at 4° C. The wells were then washed three times with PBT washing buffer (0.1% Tween-20 in PBS). 200 μl of saturation buffer (2.7% Polyvinylpyrrolidone 10 in water) was added to each well and the plates incubated for 2 hours at 37° C. Wells were washed three times with PBT. 200 μl of diluted sera (Dilution buffer: 1% BSA, 0.1% Tween-20, 0.1% NaN 3 in PBS) were added to each well and the plates incubated for 90 minutes at 37° C. Wells were washed three times with PBT. 100 μl of HRP-conjugated rabbit anti-mouse (Dako) serum diluted 1:2000 in dilution buffer were added to each well and the plates were incubated for 90 minutes at 37° C. Wells were washed three times with PBT buffer. 100 μl of substrate buffer for HRP (25 ml of citrate buffer pH5, 10 mg of O-phenildiamine and 10 μl of H 2 O) were added to each well and the plates were left at room temperature for 20 minutes. 100 μl H 2 SO 4 was added to each well and OD 490 was followed. The ELISA was considered positive when OD 490 was 2.5 times the respective pre-immune sera.

O) FACScan Bacteria Binding Assay Procedure

The acapsulated MenB M7 strain was plated on chocolate agar plates and incubated overnight at 37° C. Bacterial colonies were collected from the agar plates using a sterile dracon swab and inoculated into 4 tubes containing 8 ml each Mueller-Hinton Broth (Difco) containing 0.25% glucose. Bacterial growth was monitored every 30 minutes by following OD 620 . The bacteria were let to grow until the OD reached the value of 0.35-0.5. The culture was centrifuged for 10 minutes at 4000 rpm. The supernatant was discarded and the pellet was resuspended in blocking buffer (1% BSA, 0.4% NaN 3 ) and centrifuged for 5 minutes at 4000 rpm. Cells were resuspended in blocking buffer to reach OD 620 of 0.07. 100 μl bacterial cells were added to each well of a Costar 96 well plate. 100 μl of diluted (1:200) sera (in blocking buffer) were added to each well and plates incubated for 2 hours at 4° C. Cells were centrifuged for 5 minutes at 4000 rpm, the supernatant aspirated and cells washed by addition of 200 μl/well of blocking buffer in each well. 100 μl of R-Phicoerytrin conjugated F(ab) 2 goat anti-mouse, diluted 1:100, was added to each well and plates incubated for 1 hour at 4° C. Cells were spun down by centrifuigation at 4000 rpm for 5 minutes and washed by addition of 200 μl/well of blocking buffer. The supernatant was aspirated and cells resuspended in 200 μl/well of PBS, 0.25% formaldehyde. Samples were transferred to FACScan tubes and read. The condition for FACScan setting were: FL1 on, FL2 and FL3 off; FSC-H threshold:92; FSC PMT Voltage: E 02; SSC PMT: 474; Amp. Gains 7.1; FL-2 PMT: 539; compensation values: 0.

P) OMV Preparations

Bacteria were grown overnight on 5 GC plates, harvested with a loop and resuspended in 10 ml 20 mM Tris-HCl. Heat inactivation was performed at 56° C. for 30 minutes and the bacteria disrupted by sonication for 10 minutes on ice (50% duty cycle, 50% output). Unbroken cells were removed by centrifugation at 5000 g for 10 minutes and the total cell envelope fraction recovered by centrifugation at 50000 g at 4° C. for 75 minutes. To extract cytoplasmic membrane proteins from the crude outer membranes, the whole fraction was resuspended in 2% sarkosyl (Sigma) and incubated at room temperature for 20 minutes. The suspension was centrifuged at 1000 g for 10 minutes to remove aggregates, and the supernatant further ultracentrifuged at 50000 g for 75 minutes to pellet the outer membranes. The outer membranes were resuspended in 10 mM Tris-HCl, pH8 and the protein concentration measured by the Bio-Rad Protein assay, using BSA as a standard.

Q) Whole Extracts Preparation

Bacteria were grown overnight on a GC plate, harvested with a loop and resuspended in 1 ml of 20 mM Tris-HCl. Heat inactivation was performed at 56° C. for 30 minutes.

R) Western Blotting

Purified proteins (500 ng/lane), outer membrane vesicles (5 kg) and total cell extracts (25 μg) derived from MenB strain 2996 were loaded on 15% SDS-PAGE and transferred to a nitrocellulose membrane. The transfer was performed for 2 hours at 150 mA at 4° C., in transferring buffer (0.3% Tris base, 1.44% glycine, 20% methanol). The membrane was saturated by overnight incubation at 4° C. in saturation buffer (10% skimmed milk, 0.1% Triton X100 in PBS). The membrane was washed twice with washing buffer (3% skimmed milk, 0.1% Triton X100 in PBS) and incubated for 2 hours at 37° C. with mice sera diluted 1:200 in washing buffer. The membrane was washed twice and incubated for 90 minutes with a 1:2000 dilution of horseradish peroxidase labelled anti-mouse Ig. The membrane was washed twice with 0.1% Triton X100 in PBS and developed with the Opti-4CN Substrate Kit (Bio-Rad). The reaction was stopped by adding water.

S) Bactericidal Assay

MC58 strain was grown overnight at 37° C. on chocolate agar plates. 5-7 colonies were collected and used to inoculate 7 ml Mueller-Hinton broth. The suspension was incubated at 37° C. on a nutator and let to grow until OD 620 was 0.5-0.8. The culture was aliquoted into sterile 1.5 ml Eppendorf tubes and centrifuged for 20 minutes at maximum speed in a microfuge. The pellet was washed once in Gey's buffer (Gibco) and resuspended in the same buffer to an OD 620 of 0.5, diluted 1:20000 in Gey's buffer and stored at 25° C.

50 μl of Gey's buffer/1% BSA was added to each well of a 96-well tissue culture plate. 25 μl of diluted mice sera (1:100 in Gey's buffer/0.2% BSA) were added to each well and the plate incubated at 4° C. 25 μl of the previously described bacterial suspension were added to each well. 25 μl of either heat-inactivated (56° C. waterbath for 30 minutes) or normal baby rabbit complement were added to each well. Immediately after the addition of the baby rabbit complement, 22 μl of each sample/well were plated on Mueller-Hinton agar plates (time 0). The 96-well plate was incubated for 1 hour at 37° C. with rotation and then 22 μl of each sample/well were plated on Mueller-Hinton agar plates (time 1) After overnight incubation the colonies corresponding to time 0 and time 1 hour were counted.

Table II gives a summary of the cloning, expression and purification results.

	TABLE II
	Cloning, expression and purification
		PCR/	His-fusion	GST-fusion
	ORF	cloning	expression	expression	Purification
	orf 38	+	+	+	His-fusion
	orf 40	+	+	+	His-fusion
	orf 41	+	n.d.	n.d.
	orf 44	+	+	+	His-fusion
	orf 51	+	n.d.	n.d.
	orf 52	+	n.d.	+	GST-fusion
	orf 56	+	n.d.	n.d.
	orf 69	+	n.d.	n.d.
	orf 82	+	n.d.	n.d.
	orf 114	+	n.d.	+	GST-fusion
	orf 124	+	n.d.	n.d.

Example 1

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 1>:

1   ..ACACTGTTGT TTGCAACGGT TCAGGCAAGT GCTAACCAA                                                                        T                                                                     GAAGAGCAAG
51  AAGAAGATTT ATATTTAGAC CCCGTACAAC GCACTGTTGC CGTGTTGATA
101 GTCAATTCCG ATAAAGAAGG CACGGGAGAA AAAGAAAAAG TAGAAGAAAA
151 TTCAGATTGG GCAGTATATT TCAACGAGAA AGGAGTACTA ACAGCCAGAG
201 AAATCACCyT CAAAGCCGGC GACAACCTGA AAATCAAACA AAACGGCACA
251 AACTTCACCT ACTCGCTGAA AAAAGACCTC AcAGATCTGA CCAGTGTTGG
301 AACTGAAAAA TTATCGTTTA GCGCAAACGG CAATAAAGTC AACATcACAA
351 GCGACACCAA AGGCTTGAAT TTTGCGAAAG AAACGGCTGG sACGAACGgC
401 GACACCACGG TTCATCTGAA CGGTATTGGT TCGACTTTGA CCGATACGCT
451 GCTGAATACC GGAGCGACCA CAAACGTAAC CAACGACAAC GTTACCGATG
501 ACGAGAAAAA ACGTGCGGCA AGCGTTAAAG ACGTATTAAA CGCTGGCTGG
551 AACATTAAAG GCGTTAAACC CGGTACAACA GCTTCCGATA ACGTTGATTT
601 CGTCCGCACT TACGACACAG TCGAGTTCTT GAGCGCAGAT ACGAAAACAA
651 CGACTGTTAA TGTGGAAAGC AAAGACAACG GCAAGAAAAC CGAAGTTAAA
701 ATCGGTGCGA AGACTTCTGT TATTAAAGAA AAAGAC...

This corresponds to the amino acid sequence <SEQ ID 2; ORF40>:

1   ..TLLFATVQAS ANQEEQEEDL YLDPVQRTVA VLIVNSDKEG TGEKEKVEEN
51  SDWAVYFNEK GVLTAREITX KAGDNLKIKQ NGTNFTYSLK KDLTDLTSVG
101 TEKLSFSANG NKVNITSDTK GLNFAKETAG TNGDTTVHLN GIGSTLTDTL
151 LNTGATTNVT NDNVTDDEKK RAASVKDVLN AGWNIKGVKP GTTASDNVDF
201 VRTYDTVEFL SADTKTTTVN VESKDNGKKT EVKIGAKTSV IKEKD...

Further work revealed the complete DNA sequence <SEQ ID 3>:

1    ATGAACAAAA TATACCGCAT CATTTGGAAT AGTGCCCTCA ATGCCTGGGT
51   CGTCGTATCC GAGCTCACAC GCAACCACAC CAAACGCGCC TCCGCAACCG
101  TGAAGACCGC CGTATTGGCG ACACTGTTGT TTGCAACGGT TCAGGCAAGT
151  GCTAACAATG AAGAGCAAGA AGAAGATTTA TATTTAGACC CCGTACAACG
201  CACTGTTGCC GTGTTGATAG TCAATTCCGA TAAAGAAGGC ACGGGAGAAA
251  AAGAAAAAGT AGAAGAAAAT TCAGATTGGG CAGTATATTT CAACGAGAAA
301  GGAGTACTAA CAGCCAGAGA AATCACCCTC AAAGCCGGCG ACAACCTGAA
351  AATCAAACAA AACGGCACAA ACTTCACCTA CTCGCTGAAA AAAGACCTCA
401  CAGATCTGAC CAGTGTTGGA ACTGAAAAAT TATCGTTTAG CGCAAACGGC
451  AATAAAGTCA ACATCACAAG CGACACCAAA GGCTTGAATT TTGCGAAAGA
501  AACGGCTGGG ACGAACGGCG ACACCACGGT TCATCTGAAC GGTATTGGTT
551  CGACTTTGAC CGATACGCTG CTGAATACCG GAGCGACCAC AAACGTAACC
601  AACGACAACG TTACCGATGA CGAGAAAAAA CGTGCGGCAA GCGTTAAAGA
651  CGTATTAAAC GCTGGCTGGA ACATTAAAGG CGTTAAACCC GGTACAACAG
701  CTTCCGATAA CGTTGATTTC GTCCGCACTT ACGACACAGT CGAGTTCTTG
751  AGCGCAGATA CGAAAACAAC GACTGTTAAT GTGGAAAGCA AAGACAACGG
801  CAAGAAAACC GAAGTTAAAA TCGGTGCGAA GACTTCTGTT ATTAAAGAAA
851  AAGACGGTAA GTTGGTTACT GGTAAAGACA AAGGCGAGAA TGGTTCTTCT
901  ACAGACGAAG GCGAAGGCTT AGTGACTGCA AAAGAAGTGA TTGATGCAGT
951  AAACAAGGCT GGTTGGAGAA TGAAAACAAC AACCGCTAAT GGTCAAACAG
1001 GTCAAGCTGA CAAGTTTGAA ACCGTTACAT CAGGCACAAA TGTAACCTTT
1051 GCTAGTGGTA AAGGTACAAC TGCGACTGTA AGTAAAGATG ATCAAGGCAA
1101 CATCACTGTT ATGTATGATG TAAATGTCGG CGATGCCCTA AACGTCAATC
1151 AGCTGCAAAA CAGCGGTTGG AATTTGGATT CCAAAGCGGT TGCAGGTTCT
1201 TCGGGCAAAG TCATCAGCGG CAATGTTTCG CCGAGCAAGG GAAAGATGGA
1251 TGAAACCGTC AACATTAATG CCGGCAACAA CATCGAGATT ACCCGCAACG
1301 GTAAAAATAT CGACATCGCC ACTTCGATGA CCCCGCAGTT TTCCAGCGTT
1351 TCGCTCGGCG CGGGGGCGGA TGCGCCCACT TTGAGCGTGG ATGGGGACGC
1401 ATTGAATGTC GGCAGCAAGA AGGACAACAA ACCCGTCCGC ATTACCAATG
1451 TCGCCCCGGG CGTTAAAGAG GGGGATGTTA CAAACGTCGC ACAACTTAAA
1501 GGCGTGGCGC AAAACTTGAA CAACCGCATC GACAATGTGG ACGGCAACGC
1551 GCGTGCGGGC ATCGCCCAAG CGATTGCAAC CGCAGGTCTG GTTCAGGCGT
1601 ATTTGCCCGG CAAGAGTATG ATGGCGATCG GCGGCGGCAC TTATCGCGGC
1651 GAAGCCGGTT ACGCCATCGG CTACTCCAGT ATTTCCGACG GCGGAAATTG
1701 GATTATCAAA GGCACGGCTT CCGGCAATTC GCGCGGCCAT TTCGGTGCTT
1751 CCGCATCTGT CGGTTATCAG TGGTAA

This corresponds to the amino acid sequence <SEQ ID 4; ORF40-1>:

1   MNKIYRIIWN SALNAWVVVS ELTRNHTKRA SATVKTAVLA TLLFATVQAS
51  ANNEEQEEDL YLDPVQRTVA VLIVNSDKEG TGEKEKVEEN SDWAVYFNEK
101 GVLTAREITL KAGDNLKIKQ NGTNFTYSLK KDLTDLTSVG TEKLSFSANG
151 NKVNITSDTK GLNFAKETAG TNGDTTVHLN GIGSTLTDTL LNTGATTNVT
201 NDNVTDDEKK RAASVKDVLN AGWNIKGVKP GTTASDNVDF VRTYDTVEFL
251 SADTKTTTVN VESKDNGKKT EVKIGAKTSV IKEKDGKLVT GKDKGENGSS
301 TDEGEGLVTA KEVIDAVNKA GWRMKTTTAN GQTGQADKFE TVTSGTNVTF
351 ASGKGTTATV SKDDQGNITV MYDVNVGDAL NVNQLQNSGW NLDSKAVAGS
401 SGKVISGNVS PSKGKMDETV NINAGNNIEI TRNGKNIDIA TSMTPQFSSV
451 SLGAGADAPT LSVDGDALNV GSKKDNKPVR ITNVAPGVKE GDVTNVAQLK
501 GVAQNLNNRI DNVDGNARAG IAQAIATAGL VQAYLPGKSM MAIGGGTYRG
551 EAGYAIGYSS ISDGGNWIIK GTASGNSRGH FGASASVGYQ W*

Further work identified the corresponding gene in strain A of N.meningilidis <SEQ ID 5 >:

1    ATGAACAAAA TATACCGCAT CATTTGGAAT AGTGCCCTCA ATGCCTGNGT
51   CGCCGTATCC GAGCTCACAC GCAACCACAC CAAACGCGCC TCCGCAACCG
101  TGAAGACCGC CGTATTGGCG ACACTGTTGT TTGCAACGGT TCAGGCGAAT
151  GCTACCGATG AAGATGAAGA AGAAGAGTTA GAATCCGTAC AACGCTCTGT
201  CGTAGGGAGC ATTCAAGCCA GTATGGAAGG CAGCGGCGAA TTGGAAACGA
251  TATCATTATC AATGACTAAC GACAGCAAGG AATTTGTAGA CCCATACATA
301  GTAGTTACCC TCAAAGCCGG CGACAACCTG AAAATCAAAC AAAACACCAA
351  TGAAAACACC AATGCCAGTA GCTTCACCTA CTCGCTGAAA AAAGACCTCA
401  CAGGCCTGAT CAATGTTGAN ACTGAAAAAT TATCGTTTGG CGCAAACGGC
451  AAGAAAGTCA ACATCATAAG CGACACCAAA GGCTTGAATT TCGCGAAAGA
501  AACGGCTGGG ACGAACGGCG ACACCACGGT TCATCTGAAC GGTATCGGTT
551  CGACTTTGAC CGATACGCTT GCGGGTTCTT CTGCTTCTCA CGTTGATGCG
601  GGTAACCNAA GTACACATTA CACTCGTGCA GCAAGTATTA AGGATGTGTT
651  GAATGCGGGT TGGAATATTA AGGGTGTTAA ANNNGGCTCA ACAACTGGTC
701  AATCAGAAAA TGTCGATTTC GTCCGCACTT ACGACACAGT CGAGTTCTTG
751  AGCGCAGATA CGNAAACAAC GACNGTTAAT GTGGAAAGCA AAGACAACGG
801  CAAGAGAACC GAAGTTAAAA TCGGTGCGAA GACTTCTGTT ATTAAAGAAA
851  AAGACGGTAA GTTGGTTACT GGTAAAGGCA AAGGCGAGAA TGGTTCTTCT
901  ACAGACGAAG GCGAAGGCTT AGTGACTGCA AAAGAAGTGA TTGATGCAGT
951  AAACAAGGCT GGTTGGAGAA TGAAAACAAC AACCGCTAAT GGTCAAACAG
1001 GTCAAGCTGA CAAGTTTGAA ACCGTTACAT CAGGCACAAA TGTAACCTTT
1051 GCTAGTGGTA AAGGTACAAC TGCGACTGTA AGTAAAGATG ATCAAGGCAA
1101 CATCACTGTT ATGTATGATG TAAATGTCGG CGATGCCCTA AACGTCAATC
1151 AGCTGCAAAA CAGCGGTTGG AATTTGGATT CCAAAGCGGT TGCAGGTTCT
1201 TCGGGCAAAG TCATCAGCGG CAATGTTTCG CCGAGCAAGG GAAAGATGGA
1251 TGAAACCGTC AACATTAATG CCGGCAACAA CATCGAGATT AGCCGCAACG
1301 GTAAAAATAT CGACATCGCC ACTTCGATGG CGCCGCAGTT TTCCAGCGTT
1351 TCGCTCGGCG CGGGGGCAGA TGCGCCCACT TTAAGCGTGG ATGACGAGGG
1401 CGCGTTGAAT GTCGGCAGCA AGGATGCCAA CAAACCCGTC CGCATTACCA
1451 ATGTCGCCCC GGGCGTTAAA GANGGGGATG TTACAAACGT CNCACAACTT
1501 AAAGGCGTGG CGCAAAACTT GAACAACCGC ATCGACAATG TGGACGGCAA
1551 CGCGCGTGCN GGCATCGCCC AAGCGATTGC AACCGCAGGT CTGGTTCAGG
1601 CGTATCTGCC CGGCAAGAGT ATGATGGCGA TCGGCGGCGG CACTTATCGC
1651 GGCGAAGCCG GTTACGCCAT CGGCTACTCC AGTATTTCCG ACGGCGGAAA
1701 TTGGATTATC AAAGGCACGG CTTCCGGCAA TTCGCGCGGC CATTTCGGTG
1751 CTTCCGCATC TGTCGGTTAT CAGTGGTAA

This encodes a protein having amino acid sequence <SEQ ID 6; ORF40a>:

1   MNKIYRIIWN SALNAXVAVS ELTRNHTKRA SATVKTAVLA TLLFATVQAN
51  ATDEDEEEEL ESVQRSVVGS IQASMEGSGE LETISLSMTN DSKEFVDPYI
101 VVTLKAGDNL KIKQNTNENT NASSFTYSLK KDLTGLINVX TEKLSFGANG
151 KKVNIISDTK GLNFAKETAG TNGDTTVHLN GIGSTLTDTL AGSSASHVDA
201 GNXSTHYTRA ASIKDVLNAG WNIKGVKXGS TTGQSENVDF VRTYDTVEFL
251 SADTXTTTVN VESKDNGKRT EVKIGAKTSV IKEKDGKLVT GKGKGENGSS
301 TDEGEGLVTA KEVIDAVNKA GWRMKTTTAN GQTGQADKFE TVTSGTNVTF
351 ASGKGTTATV SKDDQGNITV MYDVNVGDAL NVNQLQNSGW NLDSKAVAGS
401 SGKVISGNVS PSKGKMDETV NINAGNNIEI SRNGKNIDIA TSMAPQFSSV
451 SLGAGADAPT LSVDDEGALN VGSKDANKPV RITNVAFGVK XGDVTNVXQL
501 KGVAQNLNNR IDNVDGNARA GIAQAIATAG LVQAYLPGKS MMAIGGGTYR
551 GEAGYAIGYS SISDGGNWII KGTASGNSRG HFGASASVGY QW*

The originally-identified partial strain B sequence (ORF40 (SEQ ID NO:2)) shows 65.7% identity over a 254aa overlap with ORF40a (SEQ ID NO:138):

          10        20        30
orf40.pep TLLFATVQASANQEEQEEDLYLDPVQRTVA
          |||||||||:|::|::||:|  : |||:|
orf40a    SALNAXVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL--ESVQRSV-
          20        30        40        50        60
          40        50         60        70        80
orf40.pep VLIVNSDKEGTGEKEKVEEN-SDWAVYFNEKGVLTAREITXKAGDNLKIKQN------GT
          |  :::: ||:|| | :  : :: :  | :  ::    :| |||||||||||      ::
orf40a    VGSIQASMEGSGELETISLSMTNDSKEFVDPYIV----VTLKAGDNLKIKQNTNENTNAS
          70        80        90       100           110       120
          90       100       110       120       130       140
orf40.pep NFTYSLKKDLTDLTSVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNGDTTVHLNGIG
          :|||||||||| | :| ||||||:|||:|||| |||||||||||||||||||||||||||
orf40a    SFTYSLKKDLTGLINVXTEKLSFGANGKKVNIISDTKGLNFAKETAGTNGDTTVHLNGIG
          130       140       150       160       170       180
          150       160       170       180       190         200
orf40.pep STLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTA--SDNVDFV
          ||||||| :::|: :|   | :  :  ||||:|||||||||||||| |:|:  |:|||||
orf40a    STLTDTLAGSSAS-HVDAGNXST-HYTRAASIKDVLNAGWNIKGVKXGSTTGQSENVDFV
          190        200        210       220       230       240
          210       220       230       240
orf40.pep RTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKD
          ||||||||||||| |||||||||||||:||||||||||||||||
orf40a    RTYDTVEFLSADTXTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSST
          250       260       270       280       290       300

The complete strain B sequence (ORF40-1 (SEQ ID NO:4)) and ORF40a (SEQ ID NO:6) show 83.7% identity in 601 aa overlap:

            10        20        30        40        50        60
orf40-1.pep MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVKTAVLATLLFATVQASANNEEQEEDL
            ||||||||||||||| |:|||||||||||||||||||||||||||||||:|::|::||:|
orf40a      MNKIYRIIWNSALNAXVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL
            10        20        30        40        50        60
            70        80        90        100       110      119
orf40-1.pep YLDPVQRTVAVLIVNSDKEGTGEKEKVEEN-SDWAVYFNEKGVLTAREITLKAGDNLKIK
            : |||:| |  :::: ||:|| | :  : :: :  | :  ::    :|||||||||||
orf4Oa      --ESVQRSV-VGSIQASMEGSGELETISLSMTNDSKEFVDPYIV----VTLKAGDNLKIK
            70        80        90       100           110
            120             130       140       150       160       170
orf40-1.pep QN------GTNFTYSLKKDLTDLTSVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNG
            ||      :::|||||||||| | :| ||||||:|||:|||| |||||||||||||||||
orf40a      QNTNENTNASSFTYSLKKDLTGLINVXTEKLSFGANGKKVNIISDTKGLNFAKETAGTNG
            120       130       140       150       160       170
            180       190       200       210       220       230
orf40-1.pep DTTVHLNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTT
            ||||||||||||||||| :::|: :|   | :  :  ||||:|||||||||||||| |:|
orf40a      DTTVHLNGIGSTLTDTLAGSSAS-HVDAGNXST-HYTRAASIKDVLNAGWNIKGVKXGST
            180       190        200        210       220       230
            240       250       260       270       280       290
orf40-1.pep A--SDNVDFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTG
            :  |:|||||||||||||||||| |||||||||||||:||||||||||||||||||||||
orf40a      TGQSENVDFVRTYDTVEFLSADTXTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTG
            240       250       260       270       280       290
            300       310       320       330       340       350
orf40-1.pep KDKGENGSSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFA
            | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf40a      KGKGENGSSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFA
            300       310       320       330       340       350
            360       370       380       390       400       410
orf40-1.pep SGKGTTATVSKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSP
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf40a      SGKGTTATVSKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSP
            360       370       380       390       400       410
            420       430       440       450       460        470
orf40-1.pep SKGKMDETVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDGD-ALNV
            |||||||||||||||||||:||||||||||||:|||||||||||||||||||| : ||||
orf40a      SKGKMDETVNINAGNNIEISRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNV
            420       430       440       450       460       470
            480       490       500       510       520       530
orf40-1.pep GSKKDNKPVRITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGL
            ||| ||||||||||||||| |||||| |||||||||||||||||||||||||||||||||
orf40a      GSKDANKPVRITNVAPGVKXGDVTNVXQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGL
            480       490       500       510       520       530
            540       550       560       570       580       590
orf40-1.pep VQAYLPGKSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQ
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf40a      VQAYLPGKSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQ
            540       550       560       570       580       590
orf40-1.pep WX
            ||
orf40a      WX

Computer analysis of these two amino acid sequences gave the following results:

Homology With Hsf Protein Encoded by the Type b Surface Fibrils Locus of H.influenzae (Accession Number U41852)

ORF40 (SEQ ID NO:2) and Hsf protein (SEQ ID NO:139) show 54% aa identity (SEQ ID NO:140) in 251 aa overlap:

Orf40	1	TLLFATVQASANQEEQEEDLYLDPVQRTVAVLIVNSDXXXXXXXXXXXXNSDWAVYFNEK	60
		TLLFATVQA+A  E++E    LDPV RT  VL  +SD            NS+W +YF+ K
Hsf	41	TLLFATVQANATDEDEE----LDPVVRTAPVLSFHSDKEGTGEKEVTE-NSNWGIYFDNK	95
Orf40	61	GVLTAREITXKAGDNLKIKQN------GTNFTYSLKKDLTDLTSVGTEKLSFSANGNKVN	114
		GVL A  IT KAGDNLKIKQN       ++FTYSLKKDLTDLTSV TEKLSF ANG+KV+
Hsf	96	GVLKAGAITLKAGDNLKIKQNTDESTNASSFTYSLKKDLTDLTSVATEKLSFGANGDKVD	155
Orf40	115	ITSDTKGLNFAKETAGTNGDTTVHLNGIGSTLTDTLLNTGAXXXXXXXXXXXXEKKRAAS	174
		ITSD  GL  AK      G+  VHLNG+ STL D + NTG             EK RAA+
Hsf	156	ITSDANGLKLAK-----TGNGNVHLNGLDSTLPDAVTNTGVLSSSSFTPNDV-EKTRAAT	209
Orf40	175	VKDVLNAGWNIKGVKPGTTASDNVDFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKI	234
		VKDVLNAGWNIKG K      ++VD V  Y+ VEF++ D  T  V + +K+NGK TEVK
Hsf	210	VKDVLNAGWNIKGAKTAGGNVESVDLVSAYNNVEFITGDKNTLDVVLTAKENGKTTEVKF	269
Orf40	235	GAKTSVIKEKD	245
		KTSVIKEKD
Hsf	270	TPKTSVIKEKD	280

ORF40a also shows homology to Hsf:

g1|1666683 (U41852) hsf gene product [ Haemophilus influenzae ] Length = 2353
Score = 153 (67.7 bits), Expect = 1.5e-116, Sum P(11) = 1.5e-116
Identities = 33/36 (91%), Positives = 34/36 (94%)
Query:	16	VAVSELTRNHTKRASATVKTAVLATLLFATVQANAT	51	(SEQ ID NO:141)
		V VSELTR HTKRASATV+TAVLATLLFATVQANAT		(SEQ ID NO:142)
Sbjct:	17	VVVSELTRTHTKRASATVETAVLATLLFATVQANAT	52	(SEQ ID NO:143)
Score = 161 (71.2 bits), Expect = 1.5e-116, Sum P(11) = 1.5e-116
Identities = 32/38 (84%), Positives = 36/38 (94%)
Query:	101	VTLKAGDNLKIKQNTNENTNASSFTYSLKKDLTGLINV	138	(SEQ ID NO:144)
		+TLKAGDNLKIKQNT+E+TNASSFTYSLKKDLT L +V		(SEQ ID NO:145)
Sbjct:	103	ITLKAGDNLKIKQNTDESTNASSFTYSLKKDLTDLTSV	140	(SEQ ID NO:146)
Score = 110 (48.7 bits), Expect = 1.5e-116, Sum P(11) = 1.5e-116
Identities = 21/29 (72%), Positives = 25/29 (86%)
Query:	138	VTEKLSFGANGKKVNIISDTKGLNFAKET	166	(SEQ ID NO:147)
		V++KLS G NG KVNI SDTKGLNFAK++		(SEQ ID NO:148)
Sbjct:	1439	VSDKLSLGTNGNKVNITSDTKGLNFAKDS	1467	(SEQ ID NO:149)
Score = 85 (37.6 bits), Expect = 1.5e-116, Sum P(11) = 1.5e-116
Identities = 18/32 (56%), Positives = 20/32 (62%)
Query:	169	TNGDTTVHLNGIGSTLTDTLAGSSASHVDAGN	200	(SEQ ID NO:150)
		T  D  +HLNGI STLTDTL  S A+    GN		(SEQ ID NO:151)
Sbjct:	1469	TGDDANIHLNGIASTLTDTLLNSGATTNLGGN	1500	(SEQ ID NO:152)
Score = 92 (40.7 bits), Expect = 1.5e-116, Sum P(11) = 1.5e-116
Identities = 16/19 (84%), Positives = 19/19 (100%)
Query:	206	RAASIKDVLNAGWNIKGVK	224	(SEQ ID NO:153)
		RAAS+KDVLNAGWN++GVK		(SEQ ID NO:154)
Sbjct:	1509	RAASVKDVLNAGWNVRGVK	1527	(SEQ ID NO:155)
Score = 90 (39.8 bits), Expect = 1.5e-116, Sum P(11) = 1.5e-116
Identities = 17/28 (60%), Positives = 20/28 (71%)
Query:	226	STTGQSENVDFVRTYDTVEFLSADTTTT	253	(SEQ ID NO:156)
		S   Q EN+DFV TYDTV+F+S D  TT		(SEQ ID NO:157)
Sbjct:	1530	SANNQVENIDFVATYDTVDFVSGDKDTT	1557	(SEQ ID NO:158)

Based on homology with Hsf, it was predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

ORF40-1 (61 kDa) was cloned in pET and pGex vectors and expressed in E.coli , as described above. The products of protein expression and purification were analyzed by SDS-PAGE. FIG. 1A shows the results of affinity purification of the His-fuision protein, and FIG. 1B shows the results of expression of the GST-fusion in E.coli . Purified His-fusion protein was used to immunise mice, whose sera were used for FACS analysis (FIG. 1 C), a bactericidal assay (FIG. 1 D), and ELISA (positive result). These experiments confirm that ORF40-1 is a surface-exposed protein, and that it is a useful immunogen.

FIG. 1E shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF40-1.

Example 2

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 7>

1   ATGTTACGTt TGACTGCtTT AGCCGTATGC ACCGCCCTCG CTTTGGGCGC
51  GTGTTCGCCG CAAAATTCCG ACTCTGCCCC ACAAGCCAAA GaACAGGCGG
101 TTTCCGCCGC ACAAACCGAA GgCGCGTCCG TTACCGTCAA AACCGCGCGC
151 GGCGACGTTC AAATACCGCA AAACCCCGAA CGCATCGCCG TTTACGATTT
201 GGGTATGCTC GACACCTTGA GCAAACTGGG CGTGAAAACC GGTTTGTCCG
251 TCGATAAAAA CCGCCTGCCG TATTTAGAGG AATATTTCAA AACGACAAAA
301 CCTGCCGGCA CTTTGTTCGA GCCGGATTAC GAAACGCTCA ACGCTTACAA
351 ACCGCAGCTC ATCATCATCG GCAGCCGCGC CgCCAAGGCG TTTGACAAAT
401 TGAAcGAAAT CGCGCCGACC ATCGrmwTGA CCGCCGATAC CGCCAACCTC
451 AAAGAAAGTG CCAArGAGGC ATCGACGCTG GCGCAAATCT TC..

This corresponds to the amino acid sequence <SEQ ID 8; ORF38>:

1   MLRLTAL                                                                        AVC TALALGAC                                                                    SP QNSDSAPQAK EQAVSAAQTE GASVTVKTAR
51  GDVQIPQNPE RIAVYDLGML DTLSKLGVKT GLSVDKNRLF YLEEYFKTTK
101 PAGTLFEPDY ETLNAYKPQL IIIGSRAAKA FDKLNEIAPT IXXTADTANL
151 KESAKEASTL AQIF..

Further work revealed the complete nucleotide sequence <SEQ ID 9>:

1   ATGTTACGTT TGACTGCTTT AGCCGTATGC ACCGCCCTCG CTTTGGGCGC
51  GTGTTCGCCG CAAAATTCCG ACTCTGCCCC ACAAGCCAAA GAACAGGCGG
101 TTTCCGCCGC ACAAACCGAA GGCGCGTCCG TTACCGTCAA AACCGCGCGC
151 GGCGACGTTC AAATACCGCA AAACCCCGAA CGCATCGCCG TTTACGATTT
201 GGGTATGCTC GACACCTTGA GCAAACTGGG CGTGAAAACC GGTTTGTCCG
251 TCGATAAAAA CCGCCTGCCG TATTTAGAGG AATATTTCAA AACGACAAAA
301 CCTGCCGGCA CTTTGTTCGA GCCGGATTAC GAAACGCTCA ACGCTTACAA
351 ACCGCAGCTC ATCATCATCG GCAGCCGCGC CGCCAAGGCG TTTGACAAAT
401 TGAACGAAAT CGCGCCGACC ATCGAAATGA CCGCCGATAC CGCCAACCTC
451 AAAGAAAGTG CCAAAGAGCG CATCGACGCG CTGGCGCAAA TCTTCGGCAA
501 ACAGGCGGAA GCCGACAAGC TGAAGGCGGA AATCGACGCG TCTTTTGAAG
551 CCGCGAAAAC TGCCGCACAA GGTAAGGGCA AAGGTTTGGT GATTTTGGTC
601 AACGGCGGCA AGATGTCGGC TTTCGGCCCG TCTTCACGCT TGGGCGGCTG
651 GCTGCACAAA GACATCGGCG TTCCCGCTGT CGATGAATCA ATTAAAGAAG
701 GCAGCCACGG TCAGCCTATC AGCTTTGAAT ACCTGAAAGA GAAAAATCCC
751 GACTGGCTGT TTGTCCTTGA CCGAAGCGCG GCCATCGGCG AAGAGGGTCA
801 GGCGGCGAAA GACGTGTTGG ATAATCCGCT GGTTGCCGAA ACAACCGCTT
851 GGAAAAAAGG ACAGGTCGTG TACCTCGTTC CTGAAACTTA TTTGGCAGCC
901 GGTGGCGCGC AAGAGCTGCT GAATGCAAGC AAACAGGTTG CCGACGCTTT
951 TAACGCGGCA AAATAA

This corresponds to the amino acid sequence <SEQ ID 10; ORF38-1>:

1   MLRLTALAVC TALALGA                                                                    CSP QNSDSAPQAK EQAVSAAQTE GASVTVKTAR
51  GDVQIPQNPE RIAVYDLGML DTLSKLGVKT GLSVDKNRLP YLEEYFKTTK
101 PAGTLFEPDY ETLNAYKPQL IIIGSRAAKA FDKLNEIAPT IEMTADTANL
151 KESAKERIDA LAQIFGKQAE ADKLKAEIDA SFEAAKTAAQ GKGKGLVILV
201 NGGKMSAFGP SSRLGGWLHK DIGVPAVDES IKEGSHGQPI SFEYLKEKNP
251 DWLFVLDRSA AIGEEGQAAK DVLDNPLVAE TTAWKKGQVV YLVPETYLAA
301 GGAQELLNAS KQVADAFNAA K*

Computer analysis of this amino acid sequence reveals a putative prokaryotic membrane lipoprotein lipid attachment site (underlined).

Further work identified the corresponding gene in strain A of N.meningitidis <SEQ ID 11>:

1   ATGTTACGTT TGACTGCTTT AGCCGTATGC ACCGCCCTCG CTTTGGGCGC
51  GTGTTCGCCG CAAAATTCCG ACTCTGCCCC ACAAGCCAAA GAACAGGCGG
101 TTTCCGCCGC ACAATCCGAA GGCGTGTCCG TTACCGTCAA AACGGCGCGC
151 GGCGATGTTC AAATACCGCA AAACCCCGAA CGTATCGCCG TTTACGATTT
201 GGGTATGCTC GACACCTTGA GCAAACTGGG CGTGAAAACC GGTTTGTCCG
251 TCGATAAAAA CCGCCTGCCG TATTTAGAGG AATATTTCAA AACGACAAAA
301 CCTGCCGGAA CTTTGTTCGA GCCGGATTAC GAAACGCTCA ACGCTTACAA
351 ACCGCAGCTC ATCATCATCG GCAGCCGCGC AGCCAAAGCG TTTGACAAAT
401 TGAACGAAAT CGCGCCGACC ATCGAAATGA CCGCCGATAC CGCCAACCTC
451 AAAGAAAGTG CCAAAGAGCG TATCGACGCG CTGGCGCAAA TCTTCGGCAA
501 AAAGGCGGAA GCCGACAAGC TGAAGGCGGA AATCGACGCG TCTTTTGAAG
551 CCGCGAAAAC TGCCGCGCAA GGCAAAGGCA AGGGTTTGGT GATTTTGGTC
601 AACGGCGGCA AGATGTCCGC CTTCGGCCCG TCTTCACGAC TGGGCGGCTG
651 GCTGCACAAA GACATCGGCG TTCCCGCTGT TGACGAAGCC ATCAAAGAAG
701 GCAGCCACGG TCAGCCTATC AGCTTTGAAT ACCTGAAAGA GAAAAATCCC
751 GACTGGCTGT TTGTCCTTGA CCGCAGCGCG GCCATCGGCG AAGAGGGTCA
801 GGCGGCGAAA GACGTGTTGA ACAATCCGCT GGTTGCCGAA ACAACCGCTT
851 GGAAAAAAGG ACAAGTCGTT TACCTTGTTC CTGAAACTTA TTTGGCAGCC
901 GGTGGCGCGC AAGAGCTACT GAATGCAAGC AAACAGGTTG CCGACGCTTT
951 TAACGCGGCA AAATAA

This encodes a protein having amino acid sequence <SEQ ID 12; ORF38a>:

1   MLRLTALAVC TALALGA                                                                    CSF QNSDSAFQAK EQAVSAAQSE GVSVTVKTAR
51  GDVQIPQNPE RIAVYDLGML DTLSKLGVKT GLSVDKNRLP YLEEYFKTTK
101 FAGTLFEPDY ETLNAYKFQL IIIGSRAAKA FDKLNEIAPT IEMTADTANL
151 KESAKERIDA LAQIFGKKAE ADKLKAEIDA SFEAAKTAAQ GKGKGLVILV
201 NGGKMSAFGP SSRLGGWLHK DIGVPAVDEA IKEGSHGQPI SFEYLKEKNP
251 DWLFVLDRSA AIGEEGQAAK DVLNNPLVAE TTAWKKGQVV YLVFETYLAA
301 GGAQELLNAS KQVADAFNAA K*

The originally-identified partial strain B sequence (ORF38 (SEQ ID NO:8)) shows 95.2% identity over a 165aa overlap with ORF38a (SEQ ID NO:159):

          10        20        30        40        50        60
orf38.pep MLRLTALAVCTALALGA                                                                    CSPQNSDSAPQAKEQAVSAAQTEGASVTVKTARGDVQIPQNPE
          ||||||||||||||||||||||||||||||||||||||:||:||||||||||||||||||
orf38a    MLRLTALAVCTALALGA                                                                    CSPQNSDSAPQAKEQAVSAAQSEGVSVTVKTARGDVQXPQNPE
          10        20        30        40        50        60
          70        80        90       100       110       120
orf38.pep RIAVYDLGMLDTLSKLGVKTGLSVDKNRLPYLEEYFKTTKPAGTLFEPDYETLNAYKPQL
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ort38a    RIAVYDLGMLDTLSKLGVKTGLSVDKNRLPYLEEYFKTTKPAGTLFEPDYETLNAYKPQL
          70        80        90       100       110       120
          130       140       150        160
orf38.pep IIIGSRAAKAFDKLNEIAPTIXXTADTANLKESAKE-ASTLAQIF
          |||||||||||||||||||||  |||||||||||||  ::|||||
orf38a    IIIGSRAAKAFDKLNEIAPTIEMTADTANLKESAKERIDALAQIFGKKAEADKLKAEIDA
          130       140       150        160      170       180
orf38a    SFEAAKTAAQGKGKGLVILVNGGKMSAFGPSSRLGGWLHKDIGVPAVDEAIKEGSHGQPI
          190       200       210       220       230       240

The complete strain B sequence (ORF38-1 (SEQ ID NO: 10)) and OR38a (SEQ ID NO: 12) show 98.4% identity in 321 aa overlap:

orf38a.pep MLRLTALAVCTALALGACSPQNSDSAPQAKEQAVSAAQSEGVSVTVKTARGDVQIPQNPE
           ||||||||||||||||||||||||||||||||||||||:||:||||||||||||||||||
orf38-1    MLRLTALAVCTALALGACSPQNSDSAFQAKEQAVSAAQTEGASVTVKTARGDVQIPQNFE
orf38a.pep RIAVYDLGMLDTLSKLGVKTGLSVDKNRLPYLEEYFKTTKPAGTLFEFDYETLNAYKPQL
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf38-1    RIAVYDLGNLDTLSKLGVKTGLSVDKNRLPYLEEYFKTTKPAGTLFEPDYETLNAYKPQL
orf38a.pep IIIGSFAAKAFDKLNEIAFTIEMTADTANLKESAKERIDALAQIFGKKAEADKLKAEIDA
           |||||||||||||||||||||||||||||||||||||||||||||||:||||||||||||
orf38-l    IIIGSRAAKAFDKLNEIAPTIEMTADTANLKESAKERIDALAQIFGKQAEADKLKAEIDA
orf38a.pep SFEAAKTAAQGKGKGLVILVNGGKMSAFGPSSRLGGWLHKDIGVFAVDEAIKEGSHGQPI
           |||||||||||||||||||||||||||||||||||||||||||||||||:||||||||||
orf38-1    SFEAAKTAAQGKGKGLVILVNGGKMSAFGPSSRLGGWLHKDIGVPAVDESIKEGSHGQPI
orf38a.pep SFEYLKEKNFDWLFVLDRSAAIGEEGQAAKDVLNNFLVAETTAWKKGQVVYLVPETYLAA
           |||||||||||||||||||||||||||||||||:||||||||||||||||||||||||||
orf38-1    SFEYLKEKNPDWLFVLDRSAAIGEEGQAAKDVLDNPLVAETTAWKKGQVVYLVPETYLAA
orf38a.pep GGAQELLNASKQVADAFNAAK
           |||||||||||||||||||||
orf38-1    GGAQELLNASKQVADAFNAAK

Computer analysis of these sequences revealed the following:

Homology With a Lipoprotein (Lipo) of C.jejuni (Accession Number X82427)

ORF38 (SEQ ID NO:160) and lipo (SEQ ID NO:162) show 38% aa identity (SEQ ID NO:161) in 96 aa overlap:

Orf38: 40	EGASVTVKTARGDVQIPQNPERIAVYDLGMLDTLSKLGVKTGLS-VDKNRLPYLEEYFKT	98
	EG S  VK + G+ + P+NP ++ + DLG+LDT   L +   ++ V    LP   + FK
Lipo: 51	EGDSFLVKDSLGENKTPKNPSKVVILDLGILDTFDALKLNDKVAGVFAKNLPKYLQQFKN	110
Orf38: 99	TKPAGTLFEPDYETLNAYKPQLIIIGSRAAKAFDKL	134
	G + + D+E +NA KP LIII  R +K +DKL
Lipo: 111	KPSVGGVQQVDFEAINALKPDLIIISGRQSKFYDKL	146

Based on this analysis, it was predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

ORF38-1 (32 kDa) was cloned in pET and pGex vectors and expressed in E.coli , as described above. The products of protein expression and purification were analyzed by SDS-PAGE. FIG. 2A shows the results of affinity purification of the His-fusion protein, and FIG. 2B shows the results of expression of the GST-fusion in E.coli . Purified His-fusion protein was used to immunise mice, whose sera were used for Western blot analysis ( FIG. 2C ) and FACS analysis (FIG. 2 D). These experiments confirm that ORF38-1 is a surface-exposed protein, and that it is a useful imrnunogen.

FIG. 2E shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF38-1.

Example 3

The following N.meningitidis DNA sequence was identified <SEQ ID 13>:

1   ATGAAACTTC TGACCACCGC AATCCTGTCT TCCGCAATCG CGCTCAGCAG
51  TATGGCTGCC GCCGCTGGCA CGGACAACCC CACTGTTGCA AAAAAAACCG
101 TCAGCTACGT CTGCCAGCAA GGTAAAAAAG TCAAAGTAAC CTACGGCTTC
151 AACAAACAGG GTCTGACCAC ATACGCTTCC GCCGTCATCA ACGGCAAACG
201 CGTGCAAATG CCTGTCAATT TGGACAAATC CGACAATGTG GAAACATTCT
251 ACGGCAAAGA AGGCGGTTAT GTTTTGGGTA CCGGCGTGAT GGATGGCAAA
301 TCCTACCGCA AACAGCCCAT TATGATTACC GCACCTGACA ACCAAATCGT
351 CTTCAAAGAC TGTTCCCCAC GTTAA

This corresponds to the amino acid sequence <SEQ ID 14; ORF44>:

1   MKLLTTAILS SAIALSSMAA AA                                                                    GTDNPTVA KKTVSYVCQQ GKKVKVTYGF
51  NKQGLTTYAS AVINGKRVQM PVNLDKSDNV ETFYGKEGGY VLGTGVMDGK
101 SYRKQPIMIT AFDNQIVFKD CSPR*

Computer analysis of this amino acid sequence predicted the leader peptide shown underlined.

Further work identified the corresponding gene in strain A of N.meningitidis <SEQ ID 15>:

1   ATGAAACTTC TGACCACCGC AATCCTGTCT TCCGCAATCG CGCTCAGCAG
51  TATGGCTGCT GCTGCCGGCA CGAACAACCC CACCGTTGCC AAAAAAACCG
101 TCAGCTACGT CTGCCAGCAA GGTAAAAAAG TCAAAGTAAC CTACGGCTTT
151 AACAAACAGG GCCTGACCAC ATACGCTTCC GCCGTCATCA ACGGCAAACG
201 TGTGCAAATG CCTGTCAATT TGGACAAATC CGACAATGTG GAAACATTCT
251 ACGGCAAAGA AGGCGGTTAT GTTTTGGGTA CCGGCGTGAT GGATGGCAAA
301 TCCTATCGCA AACAGCCTAT TATGATTACC GCACCTGACA ACCAAATCGT
351 CTTCAAAGAC TGTTCCCCAC GTTAA

This encodes a protein having amino acid sequence <SEQ ID 16; ORF44a>:

1   MKLLTTAILS SAIALSSMA                                                                    A AAGTNNPTVA KKTVSYVCQQ GKKVKVTYGF
51  NKQGLTTYAS AVINGKRVQM PVNLDKSDNV ETFYGKEGGY VLGTGVMDGK
101 SYRKQPIMIT APDNQIVFKD CSPR*

The strain B sequence (ORF44 (SEQ ID NO: 14)) shows 99.2% identity over a 124aa overlap with ORF44a (SEQ ID NO:16):

          10        20        30        40        50        60
orf44.pep MKLLTTAILSSAIALSSMA                                                                    AAAGTDNPTVAKKTVSYVCQQGKKVKVTYGFNKQGLTTYAS
          ||||||||||||||||||||||||:|||||||||||||||||||||||||||||||||||
orf44a    MKLLTTAILSSAIALSSMA                                                                    AAAGTNNPTVAKKTVSYVCQQGKKVKVTYGFNKQGLTTYAS
          10        20        30        40        50        60
          70        80        90       100       110       120
orf44.pep AVINGKRVQMPVNLDKSDNVETFYGKEGGYVLGTGVMDGKSYRKQPIMITAPDNQIVFKD
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf44a    AVINGKRVQMPVNLDKSDNVETFYGKEGGYVLGTGVMDGKSYRKQPIMITAPDNQIVFKD
          70        80        90       100       110       120
ort44.pep CSPRX
          |||||
ort44a    CSPRX

Computer analysis gave the following results:

Homology With the LecA Adhesin of Eikenella corrodens (Accession Number D78153)

ORF44 (SEQ ID NO:163) and LecA (SEQ ID NO:165) protein show 45% aa identity (SEQ ID NO: 164) in 91 aa overlap:

Orf44 33  TVSYVCQQGKKVKVTYGFNKQGLTTYASAVINGKRVQMPVNLDKSDNVETFYGKEGGYVL  92
      +V+YVCQQG+++ V Y FN  G+ T A   +N + +++P NL  SDNV+T +    GY L
LecA  135 SVAYVCQQGRRLNVNYRFNSAGVPTSAELRVNNRNLRLPYNLSASDNVDTVF-SANGYRL 193
Orf44 93  GTGVMDGKSYRKQPIMITAPDNQIVFKDCSP 123
      T  MD  +YR Q I+++AP+ Q+++KDCSP
LecA  194 TTNAMDSANYRSQDIIVSAPNGQMLYKDCSP 224

Based on homology with the adhesin, it was predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

ORF44-4 (11.2 kDa) was cloned in pET and pGex vectors and expressed in E.coli , as described above. The products of protein expression and purification were analyzed by SDS-PAGE. FIG. 3A shows the results of affinity purification of the His-fusion protein, and FIG. 3B shows the results of expression of the GST-fusion in E.coli . Purified His-fusion protein was used to immunise mice, whose sere were used for ELISA, which gave positive results, and for a bactericidal assay (FIG. 3 C). These experiments confirm that ORF44-1 is a surface-exposed protein, and that it is a usefuel immunogen.

FIG. 3D showa plots of hydrophilicity, antigenic index, and AMPHI regions for ORF44-1.

Example 4

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 17>

1   ..GGCACCGAAT TCAAAACCAC CCTTTCCGGA GCCGACATAC AGGCAGGGGT
51  GGGTGAAAAA GCCCGAGCCG ATGCGAAAAT TATCCTAAAA GGCATCGTTA
101 ACCGCATCCA AACCGAAGAA AAGCTGGAAT CCAACTCGAC CGTATGGCAA
151 AAGCAGGCCG GAAGCGGCAG CACGGTTGAA ACGCTGAAGC TACCGAGCTT
201 TGAAGGGCCG GCACTGCCTA AGCTGACCGC TCCCGGCGGC TATATCGCCG
251 ACATCCCCAA AGGCAACCTC AAAACCGAAA TCGAAAAGCT GGCCAAACAG
301 CCCGAATATG CCTATCTGAA ACAGCTTCAG ACGGTCAAGG ACGTGAACTG
351 GAACCAAGTA CAGCTCGCTT ACGACAAATG GGACTATAAA CAGGAAGGCC
401 TAACCGGAGC CGGAGCCGCA ATTANCGCAC TGGCCGTTAC CGTGGTCACC
451 TCAGGCGCAG GAACCGGAGC CGTATTGGGA TTAANACGNG TGGCCGCCGC
501 CGCAACCGAT GCAGCATTT...

This corresponds to the amino acid sequence <SEQ ID 18; ORF49>:

1   ..GTEFKTTLSG ADIQAGVGEK ARADAKIILK GIVNRIQTEE KLESNSTVWQ
51  KQAGSGSTVE TLKLPSFEGP ALPKLTAPGG YIADIPKGNL KTEIEKLAKQ
101 PEYAYLKQLQ TVKDVNWNQV QLAYDKWDYK QE                                                                        GLTGAGAA IXALAVTVV                                                                    T
151 SGAGTGAVLG LXRVAAAATD AAF..

Further work revealed the complete nucleotide sequence <SEQ ID 19>:

1    ATGCAACTGC TGGCAGCCGA AGGCATTCAC CAACACCAAT TGAATGTTCA
51   GAAAAGTACC CGTTTCATCG GCATCAAAGT GGGTAAAAGC AATTACAGCA
101  AAAACGAGCT GAACGAAACC AAACTGCCCG TACGCGTTAT CGCCCAAACA
151  GCCAAAACCC GTTCCGGCTG GGATACCGTA CTCGAAGGCA CCGAATTCAA
201  AACCACCCTT TCCGGAGCCG ACATACAGGC AGGGGTGGGT GAAAAAGCCC
251  GAGCCGATGC GAAAATTATC CTAAAAGGCA TCGTTAACCG CATCCAAACC
301  GAAGAAAAGC TGGAATCCAA CTCGACCGTA TGGCAAAAGC AGGCCGGAAG
351  CGGCAGCACG GTTGAAACGC TGAAGCTACC GAGCTTTGAA GGGCCGGCAC
401  TGCCTAAGCT GACCGCTCCC GGCGGCTATA TCGCCGACAT CCCCAAAGGC
451  AACCTCAAAA CCGAAATCGA AAAGCTGGCC AAACAGCCCG AATATGCCTA
501  TCTGAAACAG CTTCAGACGG TCAAGGACGT GAACTGGAAC CAAGTACAGC
551  TCGCTTACGA CAAATGGGAC TATAAACAGG AAGGCCTAAC CGGAGCCGGA
601  GCCGCAATTA TCGCACTGGC CGTTACCGTG GTCACCTCAG GCGCAGGAAC
651  CGGAGCCGTA TTGGGATTAA ACGGTGCGGC CGCCGCCGCA ACCGATGCAG
701  CATTTGCCTC TTTGGCCAGC CAGGCTTCCG TATCGTTCAT CAACAACAAA
751  GGCAATATCG GTAACACCCT GAAAGAGCTG GGCAGAAGCA GCACGGTGAA
801  AAATCTGATG GTTGCCGTCG CTACCGCAGG CGTAGCCGAC AAAATCGGTG
851  CTTCGGCACT GAACAATGTC AGCGATAAGC AGTGGATCAA CAACCTGACC
901  GTCAACCTGG CCAATGCGGG CAGTGCCGCA CTGATTAATA CCGCTGTCAA
951  CGGCGGCAGC CTGAAAGACA ATCTGGAAGC GAATATCCTT GCGGCTTTGG
1001 TGAATACTGC GCATGGAGAG GCAGCAAGTA AAATCAAACA GTTGGATCAG
1051 CACTACATTG CCCATAAGAT TGCCCATGCC ATAGCGGGCT GTGCGGCAGC
1101 GGCGGCGAAT AAGGGCAAGT GTCAAGATGG TGCGATCGGT GCGGCGGTCG
1151 GTGAAATCCT TGGCGAAACC CTACTGGACG GCAGAGACCC TGGCAGCCTG
1201 AATGTGAAGG ACAGGGCAAA AATCATTGCT AAGGCGAAGC TGGCAGCAGG
1251 GGCGGTTGCG GCGTTGAGTA AGGGGGATGT GAGTACGGCG GCGAATGCGG
1301 CTGCTGTGGC GGTAGAGAAT AATTCTTTAA ATGATATACA GGATCGTTTG
1351 TTGAGTGGAA ATTATGCTTT ATGTATGAGT GCAGGAGGAG CAGAAAGCTT
1401 TTGTGAGTCT TATCGACCAC TGGGCTTGCC ACACTTTGTA AGTGTTTCAG
1451 GAGAAATGAA ATTACCTAAT AAATTCGGGA ATCGTATGGT TAATGGAAAA
1501 TTAATTATTA ACACTAGAAA TGGCAATGTA TATTTCTCTG TAGGTAAAAT
1551 ATGGAGTACT GTAAAATCAA CAAAATCAAA TATAAGTGGG GTATCTGTCG
1601 GTTGGGTTTT AAATGTTTCC CCTAATGATT ATTTAAAAGA AGCATCTATG
1651 AATGATTTCA GAAATAGTAA TCAAAATAAA GCCTATGCAG AAATGATTTC
1701 CCAGACTTTG GTAGGTGAGA GTGTTGGTGG TAGTCTTTGT CTGACAAGAG
1751 CCTGCTTTTC GGTAAGTTCA ACAATATCTA AATCTAAATC TCCTTTTAAA
1801 GATTCAAAAA TTATTGGGGA AATCGGTTTG GGAAGTGGTG TTGCTGCAGG
1851 AGTAGAAAAA ACAATATACA TAGGTAACAT AAAAGATATT GATAAATTTA
1901 TTAGTGCAAA CATAAAAAAA TAG

This corresponds to the amino acid sequence <SEQ ID 20; ORF49-1>:

1   MQLLAAEGIH QHQLNVQKST RFIGIKVGKS NYSKNELNET KLPVRVIAQT
51  AKTRSGWDTV LEGTEFKTTL SGADIQAGVG EKARADAKII LKGIVNRIQT
101 EEKLESNSTV WQKQAGSGST VETLKLPSFE GPALPKLTAP GGYIADIPKG
151 NLKTEIEKLA KQPEYAYLKQ LQTVKDVNWN QVQLAYDKWD YKQEG                                                                        LTGAG
201 AAIIALAVTV VT                                                                    SGAGTGAV LGLNGAAAAA TDAAFASLAS QASVSFINNK
251 GNIGNTLKEL GRSSTVKNLM VAVATAGVAD KIGASALNNV SDKQWINNLT
301 VNLANAGSAA LINTAVNGGS LKDNLEANIL AALVNTAHGE AASKIKQLDQ
351 HYIAHKIAHA IAGCAAAAAN KGKCQDGAIG AAVGEILGET LLDGRDPGSL
401 NVKDRAKIIA KAKLAAGAVA ALSKGDVSTA ANAAAVAVEN NSLNDIQDRL
451 LSGNYALCMS AGGAESFCES YRPLGLPHFV SVSGEMKLPN KFGNRMVNGK
501 LIINTRNGNV YFSVGKIWST VKSTKSNISG VSVGWVLNVS PNDYLKEASM
551 NDFRNSNQNK AYAEMISQTL VGESVGGSLC LTRACFSVSS TISKSKSPFK
601 DSKIIGEIGL GSGVAAGVEK TIYIGNIKDI DKFISANIKK *

Computer analysis predicts a transmembrane domain and also indicates that ORF49 has no significant amino acid homology with known proteins. A corresponding ORF from N.meningitidis strain A was, however, identified:

ORF49 (SEQ ID NO:18) shows 86.1% identity over a 173aa overlap with an ORF (ORF49a (SEQ ID NO:166)) from strain A of N.meningitidis :

          10        20        30
orf49.pep GTEFKTTLSGADIQAGVGEKARADAKIILK
          ||||||||:|||||||| ||||:|||||||
orf49a    SKNELNETKLPVRVVAQXAATRSGWDTVLEGTEFKTTLAGADIQAGVXEKARVDAKIILK
          40        50        60        70        80        90
          40        50        60        70        80        90
orf49.pep GIVNRIQTEEKLESNSTVWQKQAGSGSTVETLKLPSFEGPALPKLTAPGGYIADIPKGNL
          |||||||:|||||:|||||||||| |||:|||||||||:|: |||:||||||:|||||||
orf49a    GIVNRIQSEEKLETNSTVWQKQAGRGSTIETLKLPSFESPTPPKLSAPGGYIVDIPKGNL
          100       110       120       130       140       150
          100       110       120       130       140       150
orf49.pep KTEIEKLAKQPEYAYLKQLQTVKDVNWNQVQLAYDKWDYKQEG                                                                        LTGAGAAIXALAVTVVT
          |||||||:||||||||||||::|::||||||||||:||||||||| ||||| ||||||||
orf49a    KTEIEKLSKQPEYAYLKQLQVAKNINWNQVQLAYDRWDYKQEG                                                                        LTEAGAAIIALAVTVVT
          160       170       180       190       200       210
          160       170
orf49.pep SGAGTGAVLGLXRVAAAATDAAF
          |||||||||||  : ||||||||
orf49a    SGAGTGAVLGLNGAXAAATDAAFASLASQASVSFINNKGDVGKTLKELGRSSTVKNLVVA
          220       230       240       250       260       270

ORF49-1 (SEQ ID NO:168) and ORF49a (SEQ ID NO:167) show 83.2% identity in 457 aa overlap:

orf49.pep XQLLAEEGIHKHELDVQKSRRFIGIKVGXSNYSKNELNETKLPVRVVAQXAATRSGWDTV
          |||| ||||:|:|:|||| |||||||| |||||||||||||||||:||:| ||||||||
orf49-1   MQLLAAEGIHQHQLNVQKSTRFIGIKVGKSNYSKNELNETKLPVRVIAQTAKTRSGWDTV
orf49.pep LEGTEFKTTLAGADIQAGVXEKARVDAKIILKGIVNRIQSEEKLETNSTVWQKQAGRGST
          ||||||||||:|||||||| ||||:||||||||||||||:|||||:|||||||||| |||
orf49-1   LEGTEFKTTLSGADIQAGVGEKARADAKIILKGIVNRIQTEEKLESNSVWTQKQAGSGST
orf49.pep IETLKLPSFESPTPPKLSAPGGYIVDIPKGNLKTEIEKLSKQPEYAYLKQLQVAKNINWN
          :|||||||||:|: |||:||||||:||||||||||||||:||||||||||||::|::|||
orf49-1   VETLKLPSFEGPALPKLTAPGGYIADIPKGNLKTEIEKLAKQPEYAYLKQLQTVKDVNWN
orf49.pep QVQLAYDRWDYKQEGLTEAGAAIIALAVTVVTSGAGTGAVLGLNGAXAAATDAAFASLAS
          |||||||:||||||||| |||||||||||||||||||||||||||| |||||||||||||
orf49-1   QVQLAYDKWDYKQEGLTGAGAAIIALAVTVVTSGAGTGAVLGLNGAAAAATDAAFASLAS
orf49.pep QASVSFINNKGDVGKTLKELGRSSTVKNLVVAAATAGVADKIGASALXNVSDKQWINNLT
          |||||||||||::|:||||||||||||||:||:|||||||||||||| ||||||||||||
orf49-1   QASVSFINNKGNIGNTLKELGRSSTVKNLMVAVATAGVADKIGASALNNVSDKQWINNLT
orf49.pep VNLANAGSAALINTAVNGGSLKDXLEANILAALVNTAHGEAASKIKQLDQHYIVHKIAHA
          ||||||||||||||||||||||| |||||||||||||||||||||||||||||:||||||
orf49-1   VNLANAGSAALINTAVNGGSLKDNLEANILAALVNTAHGEAASKIKQLDQHYIAHKIAHA
orf49.pep IAGCAAAAANKGKCQDGAIGAAVGEIVGEALTNGKNPDTLTAKEREQILAYSKLVAGTVS
          ||||||||||||||||||||||||||:||:| :|::| :|::|:| :|:| :||:||:|:
orf49-1   IAGCAAAAANKGKCQDGAIGAAVGEILGETLLDGRDPGSLNVKDRAKIIAKAKLAAGAVA
orf49.pep GVVGGDVNAAANAAEVAVKNNQLSDXEGREFDNEMTACAKQNXPQLCRKNTVKKYQNVAD
          ::  |||::||||| |||:||:|:| : | :::::: |
orf49-1   ALSKGDVSTAANAAAVAVENNSLNDIQDRLLSGNYALCMSAGGAESFCESYRPLGLPHFV
orf49.pep KRLAASIAICTDISRSTECRTIRKQHLIDSRSLHSSWEAGLIGKDDEWYKLFSKSYTQAD
orf49-1   SVSGEMKLPNKFGNRMVNGKLIINTRNGNVYFSVGKIWSTVKSTKSNISGVSVGWVLNVS

The complete length ORF49a nucleotide sequence <SEQ ID 21> is:

1    NTGCAACTGC TGGCAGAAGA AGGCATCCAC AAGCACGAGT TGGATGTCCA
51   AAAAAGCCGC CGCTTTATCG GCATCAAGGT AGGTNAGAGC AATTACAGTA
101  AAAACGAACT GAACGAAACC AAATTGCCTG TCCGCGTCGT CGCCCAAANT
151  GCAGCCACCC GTTCAGGCTG GGATACCGTG CTCGAAGGTA CCGAATTCAA
201  AACCACGCTG GCCGGTGCCG ACATTCAGGC AGGTGTANGC GAAAAAGCCC
251  GTGTCGATGC GAAAATTATC CTCAAAGGCA TTGTGAACCG TATCCAGTCG
301  GAAGAAAAAT TAGAAACCAA CTCAACCGTA TGGCAGAAAC AGGCCGGACG
351  CGGCAGCACT ATCGAAACGC TAAAACTGCC CAGCTTCGAA AGCCCTACTC
401  CGCCCAAATT GTCCGCACCC GGCGGNTATA TCGTCGACAT TCCGAAAGGC
451  AATCTGAAAA CCGAAATCGA AAAGCTGTCC AAACAGCCCG AGTATGCCTA
501  TCTGAAACAG CTCCAAGTAG CGAAAAACAT CAACTGGAAT CAGGTGCAGC
551  TTGCTTACGA CAGATGGGAC TACAAACAGG AGGGCTTAAC CGAAGCAGGT
601  GCGGCGATTA TCGCACTGGC CGTTACCGTG GTCACCTCAG GCGCAGGAAC
651  CGGAGCCGTA TTGGGATTAA ACGGTGCGNC CGCCGCCGCA ACCGATGCAG
701  CATTCGCCTC TTTGGCCAGC CAGGCTTCCG TATCGTTCAT CAACAACAAA
751  GGCGATGTCG GCAAAACCCT GAAAGAGCTG GGCAGAAGCA GCACGGTGAA
801  AAATCTGGTG GTTGCCGCCG CTACCGCAGG CGTAGCCGAC AAAATCGGCG
851  CTTCGGCACT GANCAATGTC AGCGATAAGC AGTGGATCAA CAACCTGACC
901  GTCAACCTAG CCAATGCGGG CAGTGCCGCA CTGATTAATA CCGCTGTCAA
951  CGGCGGCAGC CTGAAAGACA NTCTGGAAGC GAATATCCTT GCGGCTTTGG
1001 TCAATACCGC GCATGGAGAA GCAGCCAGTA AAATCAAACA GTTGGATCAG
1051 CACTACATAG TCCACAAGAT TGCCCATGCC ATAGCGGGCT GTGCGGCAGC
1101 GGCGGCGAAT AAGGGCAAGT GTCAGGATGG TGCGATAGGT GCGGCTGTGG
1151 GCGAGATAGT CGGGGAGGCT TTGACAAACG GCAAAAATCC TGACACTTTG
1201 ACAGCTAAAG AACGCGAACA GATTTTGGCA TACAGCAAAC TGGTTGCCGG
1251 TACGGTAAGC GGTGTGGTCG GCGGCGATGT AAATGCGGCG GCGAATGCGG
1301 CTGAGGTAGC GGTGAAAAAT AATCAGCTTA GCGACNAAGA GGGTAGAGAA
1351 TTTGATAACG AAATGACTGC ATGCGCCAAA CAGAATANTC CTCAACTGTG
1401 CAGAAAAAAT ACTGTAAAAA AGTATCAAAA TGTTGCTGAT AAAAGACTTG
1451 CTGCTTCGAT TGCAATATGT ACGGATATAT CCCGTAGTAC TGAATGTAGA
1501 ACAATCAGAA AACAACATTT GATCGATAGT AGAAGCCTTC ATTCATCTTG
1551 GGAAGCAGGT CTAATTGGTA AAGATGATGA ATGGTATAAA TTATTCAGCA
1601 AATCTTACAC CCAAGCAGAT TTGGCTTTAC AGTCTTATCA TTTGAATACT
1651 GCTGCTAAAT CTTGGCTTCA ATCGGGCAAT ACAAAGCCTT TATCCGAATG
1701 GATGTCCGAC CAAGGTTATA CACTTATTTC AGGAGTTAAT CCTAGATTCA
1751 TTCCAATACC AAGAGGGTTT GTAAAACAAA ATACACCTAT TACTAATGTC
1801 AAATACCCGG AAGGCATCAG TTTCGATACA AACCTANAAA GACATCTGGC
1851 AAATGCTGAT GGTTTTAGTC AAGAACAGGG CATTAAAGGA GCCCATAACC
1901 GCACCAATNT TATGGCAGAA CTAAATTCAC GAGGAGGANG NGTAAAATCT
1951 GAAACCCANA CTGATATTGA AGGCATTACC CGAATTAAAT ATGAGATTCC
2001 TACACTAGAC AGGACAGGTA AACCTGATGG TGGATTTAAG GAAATTTCAA
2051 GTATAAAAAC TGTTTATAAT CCTAAAAANT TTTNNGATGA TAAAATACTT
2101 CAAATGGCTC AANATGCTGN TTCACAAGGA TATTCAAAAG CCTCTAAAAT
2151 TGCTCAAAAT GAAAGAACTA AATCAATATC GGAAAGAAAA AATGTCATTC
2201 AATTCTCAGA AACCTTTGAC GGAATCAAAT TTAGANNNTA TNTNGATGTA
2251 AATACAGGAA GAATTACAAA CATTCACCCA GAATAATTTA A

This encodes a protein having amino acid sequence <SEQ ID 22>:

1   XQLLAEEGIH KHELDVQKSR RFIGIKVGXS NYSKNELNET KLPVRVVAQX
51  AATRSGWDTV LEGTEFKTTL AGADIQAGVX EKARVDAKII LKGIVNRIQS
101 EEKLETNSTV WQKQAGRGST IETLKLPSFE SPTPPKLSAP GGYIVDIPKG
151 NLKTEIEKLS KQPEYAYLKQ LQVAKNINWN QVQLAYDRWD YKQEG                                                                        LTEAG
201 AAIIALAVTV VT                                                                    SGAGTGAV LGLNGAXAAA TDAAFASLAS QASVSFINNK
251 GDVGKTLKEL GRSSTVKNLV VAAATAGVAD KIGASALXNV SDKQWINNLT
301 VNLANAGSAA LINTAVNGGS LKDXLEANIL AALVNTAHGE AASKIKQLDQ
351 HYIVHKIAHA IAGCAAAAAN KGKCQDGAIG AAVGEIVGEA LTNGKNPDTL
401 TAKEREQILA YSKLVAGTVS GVVGGDVNAA ANAAEVAVKN NQLSDXEGRE
451 FDNEMTACAK QNXPQLCRKN TVKKYQNVAD KRLAASIAIC TDISRSTECR
501 TIRKQHLIDS RSLHSSWEAG LIGKDDEWYK LFSKSYTQAD LALQSYHLNT
551 AAKSWLQSGN TKPLSEWMSD QGYTLISGVN PRFIPIPRGF VKQNTPITNV
601 KYPEGISFDT NLXRHLANAD GFSQEQGIKG AHNRTNXMAE LNSRGGXVKS
651 ETXTDIEGIT RIKYEIPTLD RTGKPDGGFK EISSIKTVYN PKXFXDDKIL
701 QMAQXAXSQG YSKASKIAQN ERTKSISERK NVIQFSETFD GIKFRXYXDV
751 NTGRITNIHP E*

Based on the presence of a putative transmembrane domain, it is predicted that these proteins from N.meningitidis , and their epitopes, could be useful antigens for vaccines or diagnostics.

Example 5

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 23>

1   CGGATCGTTG TAGGTTTGCG GATTTCTTGC GCCGTAGTCA CCGTAGTCCC
51  AAGTATAACC CAAGGCTTTG TCTTCGCCTT TCATTCCGAT AAGGGATATG
101 ACGCTTTGGT CGGTATAGCC GTCTTGGGAA CCTTTGTCCA CCCAACGCAT
151 ATCTGCCTGC GGATTCTCAT TGCCGCTTCT TGGCTGCTGA TTTTTCTGCC
201 TTCGCGTTTT TCAACTTCGC GCTTGAGGGC TTCGGCATAT TTGTCGGCCA
251 ACGCCATTTC TTTCGGATGC AGCTGCCTAT TGTTCCAATC TACATTCGCA
301 CCCACCACAG CACCACCACT ACCACCAGTT GCATAG

This corresponds to the amino acid sequence <SEQ ID 24; ORF50>:

1   R                                                                        IVVGLRISC AVVTVVPS                                                                    IT QGFVFAFHSD KGYDALVGIA VLGTFVHPTH
51  ICLRILIAAS WLLIFLP                                                                    SRF STSRLRASAY LSANAISFGC SCLLFQSTFA
101 PTTAPPLPPV A*

Computer analysis predicts two transmembrane domains and also indicates that ORF50 has no significant amino acid homology with known proteins.

Based on the presence of a putative transmembrane domain, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 6

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 25>

1    AAGTTTGACT TTACCTGGTT TATTCCGGCG GTAATCAAAT ACCGCCGGTT
51   GTTTTTTGAA GTATTGGTGG TGTCGGTGGT GTTGCAGCTG TTTGCGCTGA
101  TTACGCCTCT GTTTTTCCAA GTGGTGATGG ACAAGGTGCT GGTACATCGG
151  GGATTCTCTA CTTTGGATGT GGTGTCGGTG GCTTTGTTGG TGGTGTCGCT
201  GTTTGAGATT GTGTTGGGCG GTTTGCGGAC GTATCTGTTT GCACATACGA
251  CTTCACGTAT TGATGTGGAA TTGGGCGCGC GTTTGTTCCG GCATCTGCTT
301  TCCCTGCCTT TATCCTATTT CGAGCACAGA CGAGTGGGTG ATACGGTGGC
351  TCGGGTGCGG GAATTGGAGC AGATTCGCAA TTTCTTGACC GGTCAGGCCC
401  TGACTTCGGT GTTGGATTTG GCGTTTTCGT TTATCTTTCT GGCGGTGATG
451  TGGTATTACA GCTCCACTCT GACTTGGGTG GTATTGGCTT CGTTG.....
1451 .......... .......... .......... .......... ..........
1501 .......... .......... .......... .......... ..ATTTGCGC
1551 CAACCGGACG GTGCTGATTA TCGCCCACCG TCTGTCCACT GTTAAAACGG
1601 CACACCGGAT CATTGCCATG GATAAAGGCA GGATTGTGGA AGCGGGAACA
1651 CAGCAGGAAT TGCTGGCGAA CG..AACGGA TATTACCGCT ATCTGTATGA
1701 TTTACAGAAC GGGTAG

This corresponds to the amino acid sequence <SEQ ID 26; ORF39>:

1   KFDFTWFIPA VIKYRRLFFE VLVVSVVLQL FALITPLFFQ VVMDKVLVHR
51  GFSTLDVVSV ALLVVSLFEI VLGGLRTYLF AHTTSRIDVE LGARLFRHLL
101 SLPLSYFEHR RVGDTVARVR ELEQIRNFLT GQALTSVLDL AFSFIFLAVM
151 WYYSSTLTWV VLASL..... .......... .......... ..........
501 .......... ....ICANRT VLIIAHRLST VKTAHRIIAM DKGRIVEAGT
551 QQELLANXNG YYRYLYDLQN G*

Further work revealed the complete nucleotide sequence <SEQ ID 27>:

1    ATGTCTATCG TATCCGCACC GCTCCCCGCC CTTTCCCCCC TCATCATCCT
51   CGCCCATTAC CACGGCATTG CCGCCAATCC TGCCGATATA CAGCATGAAT
101  TTTGTACTTC CGCACAGAGC GATTTAAATG AAACGCAATG GCTGTTAGCC
151  GCCAAATCTT TGGGATTGAA GGCAAAGGTA GTCCGCCAGC CTATTAAACC
201  TTTGGCTATG GCGACTTTAC CCGCATTGGT ATGGTGTGAT CACGGCAACC
251  ATTTCATTTT GGCCAAAACA GACGGTGAGG GTGAGCATGC CCAATTTTTG
301  ATACAGGATT TGGTTACGAA TAAGTCTGCG GTATTGTCTT TTGCCGAATT
351  TTCTAACAGA TATTCGGGCA AACTGATATT GGTTGCTTCC CGCGCTTCGG
401  TATTGGGCAG TTTGGCAAAG TTTGACTTTA CCTGGTTTAT TCCGGCGGTA
451  ATCAAATACC GCCGGTTGTT TTTTGAAGTA TTGGTGGTGT CGGTGGTGTT
501  GCAGCTGTTT GCGCTGATTA CGCCTCTGTT TTTCCAAGTG GTGATGGACA
551  AGGTGCTGGT ACATCGGGGA TTCTCTACTT TGGATGTGGT GTCGGTGGCT
601  TTGTTGGTGG TGTCGCTGTT TGAGATTGTG TTGGGCGGTT TGCGGACGTA
651  TCTGTTTGCA CATACGACTT CACGTATTGA TGTGGAATTG GGCGCGCGTT
701  TGTTCCGGCA TCTGCTTTCC CTGCCTTTAT CCTATTTCGA GCACAGACGA
751  GTGGGTGATA CGGTGGCTCG GGTGCGGGAA TTGGAGCAGA TTCGCAATTT
801  CTTGACCGGT CAGGCGCTGA CTTCGGTGTT GGATTTGGCG TTTTCGTTTA
851  TCTTTCTGGC GGTGATGTGG TATTACAGCT CCACTCTGAC TTGGGTGGTA
901  TTGGCTTCGT TGCCTGCCTA TGCGTTTTGG TCGGCATTTA TCAGTCCGAT
951  ACTGCGGACG CGTCTGAACG ATAAGTTCGC GCGCAATGCA GACAACCAGT
1001 CGTTTTTAGT AGAAAGCATC ACTGCGGTGG GTACGGTAAA GGCGATGGCG
1051 GTGGAGCCGC AGATGACGCA GCGTTGGGAC AATCAGTTGG CGGCTTATGT
1101 GGCTTCGGGA TTTCGGGTAA CGAAGTTGGC GGTGGTCGGC CAGCAGGGGG
1151 TGCAGCTGAT TCAGAAGCTG GTGACGGTGG CGACGTTGTG GATTGGCGCA
1201 CGGCTGGTAA TTGAGAGCAA GCTGACGGTG GGGCAGCTGA TTGCGTTTAA
1251 TATGCTCTCG GGACAGGTGG CGGCGCCTGT TATCCGTTTG GCGCAGTTGT
1301 GGCAGGATTT CCAGCAGGTG GGGATTTCGG TGGCGCGTTT GGGGGATATT
1351 CTGAATGCGC CGACCGAGAA TGCGTCTTCG CATTTGGCTT TGCCCGATAT
1401 CCGGGGGGAG ATTACGTTCG AACATGTCGA TTTCCGCTAT AAGGCGGACG
1451 GCAGGCTGAT TTTGCAGGAT TTGAACCTGC GGATTCGGGC GGGGGAAGTG
1501 CTGGGGATTG TGGGACGTTC GGGGTCGGGC AAATCCACAC TCACCAAATT
1551 GGTGCAGCGT CTGTATGTAC CGGAGCAGGG ACGGGTGTTG GTGGACGGCA
1601 ACGATTTGGC TTTGGCCGCT CCTGCCTGGC TGCGGCGGCA GGTCGGCGTG
1651 GTCTTGCAGG AGAATGTGCT GCTCAACCGC AGCATACGCG ACAATATCGC
1701 GCTGACGGAT ACGGGTATGC CGCTGGAACG CATTATCGAA GCAGCCAAAC
1751 TGGCGGGCGC ACACGAGTTT ATTATGGAGC TGCCGGAAGG CTACGGCACC
1801 GTGGTGGGCG AACAAGGGGC CGGCTTGTCG GGCGGACAGC GGCAGCGTAT
1851 TGCGATTGCC CGCGCGTTAA TCACCAATCC GCGCATTCTG ATTTTTGATG
1901 AAGCCACCAG CGCGCTGGAT TATGAAAGTG AACGAGCGAT TATGCAGAAC
1951 ATGCAGGCCA TTTGCGCCAA CCGGACGGTG CTGATTATCG CCCACCGTCT
2001 GTCCACTGTT AAAACGGCAC ACCGGATCAT TGCCATGGAT AAAGGCAGGA
2051 TTGTGGAAGC GGGAACACAG CAGGAATTGC TGGCGAAGCC GAACGGATAT
2101 TACCGCTATC TGTATGATTT ACAGAACGGG TAG

This corresponds to the amino acid sequence <SEQ ID 28; ORF39-1>:

1   MSIVSAPLPA LSALIILAHY HGIAANPADI QHEFCTSAQS                                                                     DLNETQWLLA
51  AKSLGLKAKV VRQPIKRLAM ATLPALVWCD DGNHFILAKT DGEGERAQFL
101 IQDLVTNKSA VLSFAEFSNR YSGKLILVAS RASVLGSLAK FDFTWFIPAV
151 IKYRRL                                                                        FFEV LVVSVVLQLF ALIT                                                                    PLFFQV VMDKVLVHRG FSTLD                                                                        VVSVA
201 LLVVSLFEIV LG                                                                    GLRTYLFA HTTSRIDVEL GARLFRHLLS LPLSYFEHRR
251 VGDTVARVRE LEQIRNFLTG QA                                                                        LTSVLDLA FSFIFLAVM                                                                    W YYSSTLTW                                                                        VV
301 LASLPAYAFW SAFIS                                                                    PILRT RLNDKFARNA DNQSFLVESI TAVGTVKAMA
351 VEPQMTQRWD NQLAAYVASG FRVTKLAVVG QQGVQLIQKL VTVATLWIGA
401 RLVIFSKLTV GQLIAFNMLS GQVAAPVIRL AQLWQDFQQV GISVARLGDI
451 LNAPTENASS HLALPDIRGE ITFEHVDFRY KADGRLILQD LNLRIRAGEV
501 LGIVGRSGSG KSTLTKLVQR LYVFEQGRVL VDGNDLALAA PAWLRRQVGV
551 VLQENVLLNR SIRDNIALTD TGMPLERIIE AAKLAGAHEF IMELPEGYGT
601 VVGEQGAGLS GGQRQRIAIA RALITNPPIL IFDEATSALD YESERAIMQN
651 MQAICANRTV LIIAHRLSTV KTAHRIIAMD KGRIVEAGTQ QELLAKFNGY
701 YRYLYDLQNG *

Computer analysis of this amino acid sequence gave the following results:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF39 (SEQ ID NO:169) shows 100% identity over a 165aa overlap with an ORF (ORF39a (SEQ ID NO:170)) from strain A of N.meningitidis :

orf39.pep KFDFTWFIPAVIKYRRLFFEVINVSVVLQL
          ||||||||||||||||||||||||||||||
orf39a    AVLSFAEFSNRYSGKLILVASRASVLGSLAKFDFTWFIPAVIKYRRLFFEVLVVSVVLQL
          110       120       130       140       150       160
          40        50        60        70        80        90
orf39.pep FALI                                                                    TPLFFQVVMDKVLVHRGFSTL                                                                        DVVSVALLVVSLFEIVLG                                                                    GLRTYLFAHTTSRIDVE
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a    FALI                                                                    TPLFFQVVMDKVLVHRGFSTL                                                                        DVVSVALLVVSLFEIVLG                                                                    GLRTYLFAHTTSRIDVE
          170       180       190       200       210       220
          100       110       120       130       140       150
orf39.pep LGARLFRHLLSLPLSYFEHRRVGDTVARVRELEQIRNFLTGQA                                                                        LTSVLDLAFSFIFLAVM
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a    LGARLFRHLLSLFLSYFEHRRVGDTVARVRELEQIRNFLTGQA                                                                        LTSVLDLAFSFIFLAVM
          230       240       250       260       270       280
          160       170       180       190       200       210
orf39.pep WYYSSTLTW                                                                        VVLASL                                                                    XXXXXXXXXXXXXXXXXXXXXXXXXXXXICANRTVLIIAHRLSTV
          |||||||||||||||
orf39a    WYYSSTLTW                                                                        VVLASLFAYAFWSAFI                                                                    SPILRTRLNDKFARNADNQSFLVESITAVGTVKAM
          290       300       310       320       330       340

ORF39-1 (SEQ ID NO:28) and ORF39a (SEQ ID NO:30) show 99.4% identity in 710 aa overlap:

orf39-1.pep MSIVSAFLFALSALIILAHYHGIAANFADIQHEFCTSAQSDLNETQWLLAAKSLGLKAKV
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      MSIVSAPLPALSALIILAHYHGIAANPADIQHEFCTSAQSDLNETQWLLAAKSLGLKAKV
orf39-1.pep VRQPIKRLAMATLFALVWCDDGNHFILAKTDGEGEHAQFLIQDLVTNKSAVLSFAEFSNR
            |||||||||||||||||||||||||||||||| |||||:|||||:|||||||||||||||
orf39a      VRQPIKRLAMATLPALVWCDDGNRFILAKTDGGGEHAQYLIQDLTTNKSAVLSFAEFSNR
orf39-1.pep YSGKLILVASRASVLGSLAKFDFTWFIPAVIKYRRLFFEVLVVSVVLQLFALITPLFFQV
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      YSGKLILVASRASVLGSLAKFDFTWFIFAVIKYRRLFFEVLVVSVVLQLFALITPLFFQV
orf39-1.pep VMDKVLVHRGFSTLDVVSVALLVVSLFEIVLGGLRTYLFAHTTSRIDVELGARLFRHLLS
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      VMDKVLVHRGFSTLDVVSVALLVVSLFEIVLGGLRTYLFAHTTSRIDVELGARLFRHLLS
orf39-1.pep LPLSYFERRRVGDTVARVRELEQIRNFLTGQALTSVLDLAFSFIFLAVMWYYSSTLTWVV
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      LFLSYFEHRRVGDTVARVRELEQIRNFLTGQALTSVLDLAFSFIFLAVMWYYSSTLTWVV
orf39-1.pep LASLPAYAPWSAFISPILRTRLNDKFARNADNQSFLVESITAVGTVKAMAVEPQMTQRWD
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      LASLPAYAFWSAFISFILRTRLNDKFARNADNQSFLVESITAVGTVKAMAVEFQMTQRWD
orf39-1.pep NQLAAYVASGFRVTKLAVVGQQGVQLIQKLVTVATLWIGARLVIESKLTVGQLIAFNMLS
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      NQLAAYVASGFRVTKLAVVGQQGVQLIQKLVTVATLWIGARLVIESKLTVGQLIAFNMLS
orf39-1-pep GQVAAFVIRLAQLWQDFQQVGISVARLCDILNAPTENASSHLALPDIRGEITFEHVDFRY
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      GQVAAFVIRLAQLWQDFQQVGISVARLGDILNAPTENASSHLALFDIRGEITFEHVDFRY
orf39-1.pep KADGRLILQDLNLRIRAGEVLGIVGRSGSGKSTLTKLVQRLYVPEQGRVLVDGNDLALAA
            |||||||||||||||||||||||||||||||||||||||||||| |||||||||||||||
orf39a      KADGRLILQDLNLRIRAGEVLGIVGRSGSGKSTLTKLVQRLYVPAQGRVLVDGNDLALAA
orf39-1.pep PAWLRRQVGVVLQENVLLNRSIRDNIALTDTGMFLERIIEAAKLAGAHEFIMELFEGYGT
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      PAWLRRQVGVVLOENVLLNRSIRDNIALTDTGMFLERIIEAAKLAGAHEFIMELPEGYGT
orf39-1.pep VVGEQGAGLSGGQRQRIAIARALITNPRILIFDEATSALDYESERAIMQNMQAICANRTV
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      VVGEQGAGLSGGQRQRIAIARALITNPRILIFDEATSALDYESERAIMQNMQAICANRTV
orf39-1.pep LIIAHRLSTVKTAHRIIAMDKGRIVEAGTQQELLAKPNGYYRYLYDLQNGX
            |||||||||||||||||||||||||||||||||||||||||||||||||||
orf39a      LIIAHRLSTVKTAHRIIAMDKGRIVEAGTQQELLAKFNGYYRYLYDLQNGX

The complete length ORF39a nucleotide sequence <SEQ ID 29>is:

1    ATGTCTATCG TATCCGCACC GCTCCCCGCC CTTTCCGCCC TCATCATCCT
51   CGCCCATTAC CACGGCATTG CCGCCAATCC TGCCGATATA CAGCATGAAT
101  TTTGTACTTC CGCACAGAGC GATTTAAATG AAACGCAATG GCTGTTAGCC
151  GCCAAATCTT TGGGATTGAA GGCAAAGGTA GTCCGCCAGC CTATTAAACG
201  TTTGGCTATG GCGACTTTAC CCGCATTGGT ATGGTGTGAT GACGGCAACC
251  ATTTTATTTT GGCTAAAACA GACGGTGGGG GTGAGCATGC CCAATATCTA
301  ATACAGGATT TAACTACGAA TAAGTCTGCG GTATTGTCTT TTGCCGAATT
351  TTCTAACAGA TATTCGGGCA AACTGATATT GGTTGCTTCC CGCGCTTCGG
401  TATTGGGCAG TTTGGCAAAG TTTGACTTTA CCTGGTTTAT TCCGGCGGTA
451  ATCAAATACC GCCGGTTGTT TTTTGAAGTA TTGGTGGTGT CGGTGGTGTT
501  GCAGCTGTTT GCGCTGATTA CGCCTCTGTT TTTCCAAGTG GTGATGGACA
551  AGGTGCTGGT ACATCGGGGA TTCTCTACTT TGGATGTGGT GTCGGTGGCT
601  TTGTTGGTGG TGTCGCTGTT TGAGATTGTG TTGGGCGGTT TGCGGACGTA
651  TCTGTTTGCA CATACGACTT CACGTATTGA TGTGGAATTG GGCGCGCGTT
701  TGTTCCGGCA TCTGCTTTCC CTGCCTTTAT CCTATTTCGA GCACAGACGA
751  GTGGGTGATA CGGTGGCTCG GGTGCGGGAA TTGGAGCAGA TTCGCAATTT
801  CTTGACCGGT CAGGCGCTGA CTTCGGTGTT GGATTTGGCG TTTTCGTTTA
851  TCTTTCTGGC GGTGATGTGG TATTACAGCT CCACTCTGAC TTGGGTGGTA
901  TTGGCTTCGT TGCCTGCCTA TGCGTTTTGG TCGGCATTTA TCAGTCCGAT
951  ACTGCGGACG CGTCTGAACG ATAAGTTCGC GCGCAATGCA GACAACCAGT
1001 CGTTTTTAGT AGAAAGCATC ACTGCGGTGG GTACGGTAAA GGCGATGGCG
1051 GTGGAGCCGC AGATGACGCA GCGTTGGGAC AATCAGTTGG CGGCTTATGT
1101 GGCTTCGGGA TTTCGGGTAA CGAAGTTGGC GGTGGTCGGC CAGCAGGGGG
1151 TGCAGCTGAT TCAGAAGCTG GTGACGGTGG CGACGTTGTG GATTGGCGCA
1201 CGGCTGGTAA TTGAGAGCAA GCTGACGGTG GGGCAGCTGA TTGCGTTTAA
1251 TATGCTCTCG GGACAGGTGG CGGCGCCTGT TATCCGTTTG GCGCAGTTGT
1301 GGCAGGATTT CCAGCAGGTG GGGATTTCGG TGGCGCGTTT GGGGGATATT
1351 CTGAATGCGC CGACCGAGAA TGCGTCTTCG CATTTGGCTT TGCCCGATAT
1401 CCGGGGGGAG ATTACGTTCG AACATGTCGA TTTCCGCTAT AAGGCGGACG
1451 GCAGGCTGAT TTTGCAGGAT TTGAACCTGC GGATTCGGGC GGGGGAAGTG
1501 CTGGGGATTG TGGGACGTTC GGGGTCGGGC AAATCCACAC TCACCAAATT
1551 GGTGCAGCGT CTGTATGTAC CGGCGCAGGG ACGGGTGTTG GTGGACGGCA
1601 ACGATTTGGC TTTGGCCGCT CCTGCTTGGC TGCGGCGGCA GGTCGGCGTG
1651 GTCTTGCAGG AGAATGTGCT GCTCAACCGC AGCATACGCG ACAATATCGC
1701 GCTGACGGAT ACGGGTATGC CGCTGGAACG CATTATCGAA GCAGCCAAAC
1751 TGGCGGGCGC ACACGAGTTT ATTATGGAGC TGCCGGAAGG CTACGGCACC
1801 GTGGTGGGCG AACAAGGGGC CGGCTTGTCG GGCGGACAGC GGCAGCGTAT
1851 TGCGATTGCC CGCGCGTTAA TCACCAATCC GCGCATTCTG ATTTTTGATG
1901 AAGCCACCAG CGCGCTGGAT TATGAAAGTG AACGAGCGAT TATGCAGAAC
1951 ATGCAGGCCA TTTGCGCCAA CCGGACGGTG CTGATTATCG CCCACCGTCT
2001 GTCCACTGTT AAAACGGCAC ACCGGATCAT TGCCATGGAT AAAGGCAGGA
2051 TTGTGGAAGC GGGAACACAG CAGGAATTGC TGGCGAAGCC GAACGGATAT
2101 TACCGCTATC TGTATGATTT ACAGAACGGG TAG

This encodes a protein having amino acid sequence <SEQ ID 30>:

1   MSIVSAPLPA LSALIILAHY HGIAANFADI QHEFCTSAQS                                                                     DLNETQWLLA
51  AKSLGLKAKV VRQPIKRLAM ATLFALVWCD DGNHFILAKT DGGGEHAQYL
101 IQDLTTNKSA VLSFAEFSNR YSGKLILVAS RASVLGSLAK FDFTWFIPAV
151 IKYRRL                                                                        FFEV LVVSVVLQLF ALI                                                                    TPLFFQV VMDKVLVHRG FSTLD                                                                        VVSVA
201 LLVVSLFEIV LG                                                                    GLRTYLFA HTTSRIDVEL GARLFRHLLS LFLSYFEHRR
251 VGDTVARVRE LEQIRNFLTG QA                                                                        LTSVLDLA FSFIFLAVM                                                                    W YYSSTLTW                                                                        VV                                                                    n
301 LASLPAYAFW SAFI                                                                    SPILRT RLNDKFARNA DNQSFLVESI TAVGTVKAMA
351 VEPQMTQRWD NQLAAYVASG FRVTKLAVVG QQGVQLIQKL VTVATLWIGA
401 RLVIESKLTV GQLIAFNMLS GQVAAPVIRL AQLWQDFQQV GISVARLGDI
451 LNAPTENASS HLALFDIRGE ITFEHVDFRY KADGRLILQD LNLRIRAGEV
501 LGIVGRSGSG KSTLTKLVQR LYVPAQGRVL VDGNDLALAA FAWLRRQVGV
551 VLQENVLLNR SIRDNIALTD TGMPLERIIE AAKLAGAHEF IMELPEGYGT
601 VVGEQGAGLS GGQRQRIAIA RALITNFRIL IFDEATSALD YESERAIMQN
651 MQAICANRTV LIIAHRLSTV KTAMRIIAMD KGRIVEAGTQ QELLAKPNGY
701 YRYLYDLQNG *

ORF39a is homologous to a cytolysin from A.pleuropneumoniae :

sp|P26760|RT1B_ACTPL RTX-I TOXIN DETERMINANT B (TOXIN RTX-I SECRETION ATP-
BINDING PROTEIN) (APX-IB) (HLY-IB) (CYTOLYSIN IB) (CLY-IB)
>gi|97137|pir||D43599 cytolysin IB - Actinobacillus pleuropneuxnoriae (serotype 9)
>gi|38944 (X61112) ClyI-B protein Actinabacillus pleuropneumoniae Length = 707
Score = 931 bits (2379), Expect = 0.0
Identities = 472/690 (68%), Positives = 540/690 (77%) Gaps =3/690 (0%)
Query: 20	YHGIAANPADIQMEFCTSAQSDLNETQWXXXXXXXXXXXXVVRQFIKRLAMATLPALVWC	79
	YH/IA/NP/+++H+F    +  L+ T W             V++ I RIA   LPALVW
Sbjct: 20	YHNIAVNPEELKHKFDLEGKG-LDLTAWLLAAKSLELKAKQVKKAIDRLAFIALFALVWR	78
Query: 80	DDGNHFILAKTDGGGEHAQYLIQDLTTNKSAVLSFAEFSNRYSGKLILVASRASVLGSLA	139
	+DG HFIL K D   E  +YLI DL T+   +L  AEF + Y GKLILVASRAS++G LA
Sbjct: 79	EDGKHFILTKIDN--EAKKYLIFDLETHNPRILEQAEFESLYQGKLILVASRASIVGKLA	136
Query: 140	KFDFTWFIPAVIKYRRXXXXXXXXXXXXXXXXXITPLFFQVVMDKVLVHRGFXXXXXXXX	199
	KFDFTWFIPAVIKYR+                 ITFLFFQVVMDKVLVHRGF
Sbjct: 137	KFDFTWFIPAVIKYRKIFIETLIVSIFLQIFALITFLFFQVVMDKVLVHRGFSTLNVITV	196
Query: 200	XXXXXXXFEIVLGGLRTYLFAHTTSRIDVELGARLFRHLLSLPLSYFEHRRVGDTVARVR	259
	FEIVL GLRTY+FAH+TSRTDVELGARLFRHLL+LP+SYFE+RRVGDTVARVR
Sbjct: 197	ALAIVVLFEIVLNGLRTYIFAHSTSRIDVELGARLFRHLLALFISYFENRRVGDTVARVR	256
Query: 260	ELEQIRNFLTGQALTSVLDLAFSFIFLAVMWYYSSTLTWVVLASLFAYAFWSAFISFILR	319
	EL+QIRNFLTGQALTSVLDL FSFIF AVMWYYS  LT V+L SLP Y  WS FISPILR
Sbjct: 257	ELDQIRNFLTGQALTSVLDLMFSFIFFAVMWYYSFKLTLVILGSLPFYMGWSIFISPILR	316
Query: 320	TRLNDKFARNADNQSFLVESITAVGTVKANAVEFQMTQRWDNQLAAYVASGFRVTKLAVV	379
	RL++KFAR ADNQSFLVES+TA+ T+KA+AV PQMT  WD QLA YV++GFRVT LA +
Sbjct: 317	RRLDEKFARGADNQSFLVESVTAINTIKALAVTPQMTNTWDKQLASYVSAGFRVTTLATI	376
Query: 380	GQQGVQLIQKLVTVATLWIGARLVIESKLTVGQLIAFNMLSGQVAAPVIRLAQLWQDFQQ	439
	GQQGVQ IQK+V V TLW+GA LVI   L++GQLIAFNMLSGQV AFVIRLAQLWQDFQQ
Sbjct: 377	GQQGVQFIQKVVMVITLWLGAHLVISGDLSIGQLIAFNMLSGQVIAFVIRLAQLWQDFQQ	436
Query: 440	VGISVARLGDILNAFTENASSHLALPDIRGEITFEHVDFRYKADGRLILQDLNLRIRAGE	499
	VGISV RLGD+LN+FTE+    LALP+I+G+ITF ++ FRYK D  +IL D+NL I+ GE
Sbjct: 437	VGISVTRLGDVLNSPTESYQGKLALPEIKGDITFRNIRFRYKFDAFVILNDVNLSIQQGE	496
Query: 500	VLGIVGRSGSGKSTLTKLVQRLYVFAQGRVLVDGNDLALAAPAWLRRQVGVVLQENVLLN	559
	V+GIVGRSGSGKSTLTKL+QR Y+P  G+VL+DG+DLALA P WLRRQVGVVLQ+NVLLN
Sbjct: 497	VIGIVGRSGSGKSTLTKLIQRFYIFENGQVLIDGHDLALADPNWLRRQVGVVLQDNVLLN	556
Query: 560	RSIRDNIALTDTGMPLERIIEAAKLAGAHEFIMELPEGYGTVVGEQGAGLSGGQRQRIAI	619
	RSIRDNIAL D GMF+E+I+ AAKLAGAHEFI EL EGY T+VGEQGAGLSGGQRQRIAI
Sbjct: 557	RSIRDNIALADPGMPMEKIVHAAKLAGAHEFISELREGYNTIVGEQGAGLSGGQRQRIAI	616
Query: 620	ARALITNFRILIFDEATSALDYESERAIMQNMQAICANRTVLIIAHRLSTVKTAHRITAM	679
	ARAL+ NP+ILIFDEATSALDYESE  IM+NM  IC  RTV+IIAHRLSTVK A RII M
Sbjct: 617	ARALVNNPKILIFDEATSALDYESEHIIMRNMHQICKGRTVIIIAHRLSTVKNADRIIVM	676
Query: 680	DKGRIVEAGTQQELLAKPNGYYRYLYDLQN	709	(SEQ ID NO:171)
	+KG+IVE G  +ELLA PNG Y YL+ LQ+		(SEQ ID NO:172)
Sbjct: 677	EKGQIVEQGKHKELLADPNGLYHYLHQLQS	706	(SEQ ID NO:173)

Homology With the HlyB Leucotoxin Secretion ATP-binding Protein of Haemophilus actinomycetemcomitans (Accession Number X53955)

ORF39 (SEQ ID NO:174) and HlyB (SEQ ID NO:176) protein show 71% and 69% amino acid identity (SEQ ID NO:175) in 167 and 55 overlap at the N- and C-terminal regions, respectively:

Orf39	1	KFDFTWFIPAVIKYRRXXXXXXXXXXXXXXXXXITPLFFQVVMDKVLVHRGFXXXXXXXX	60
		KFDFTWFIPAVIKYR+                 ITPLFFQVVMDKVLVHRGF
HlyB	137	KFDFTWFIPAVIKYRKIFIETLIVSIFLQIFALITPLFFQVVMDKVLVMRGFSTLNVITV	196
Orf39	61	XXXXXXXFEIVLGGLRTYLFAHTTSRIDVELGARLFRHLLSLPLSYFEHRRVGDTVARVR	120
		FEI+LGGLRTY+FAH+TSRIDVELGARLFRHLL+LP+SYFE RRVGDTVARVR
HlyB	197	ALAIVVLFEIILGGLRTYVFAHSTSRIDVELGARLFRHLLALPISYFEARRVGDTVARVR	256
Orf39	121	ELEQIRNFLTGQALTSVLDLAFSFIFLAVMWYYSSTLTWVVLASLIC	167
		EL+QIRNFLTGQALTS+LDL FSFIF AVMWYYS  LT VVL SL C
HlyB	257	ELDQIRNFLTGQALTSILDLLFSFIFFAVMWYYSFKLTLVVLGSLPC	303
Orf39	166	ICANRTVLIIAHRLSTVKTAHRIIAMDKGRIVEAGTQQELLANXNGYYRYLYDLQ	220
		IC NRTVLIIAHRLSTVK A RII MDKG I+E G  QELL +  G Y YL+ LQ
HlyB	651	ICQNRTVLIIAHRLSTVKNADRIIVHDKGEIIEQGKHQELLKDEKGLYSYLHQLQ	705

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 7

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 31>

1   ATGAAATACT TGATCCGCAC CGCCTTACTC GCAGTCGCAG CCGCCGGCAT
51  CTACGCCTGC CAACCGCAAT CCGAAGCCGC AGTGCAAGTC AAGGCTGAAA
101 ACAGCCTGAC CGCTATGCGC TTAGCCGTCG CCGACAAACA GGCAGAGATT
151 GACGGGTTGA ACGCCCAAAk sGACGCCGAA ATCAGA ...

This corresponds to the amino acid sequence <SEQ ID 32; ORF52>:

1  MKYL                                                                        IRTALL AVAAAGIYAC QPQSE                                                                    AAVQV KAENSLTAMR LAVADKQAEI
51 DGLNAQXDAE IR..

Further work revealed the complete nucleotide sequence <SEQ ID 33>:

1   ATGAAATACT TGATCCGCAC CGCCTTACTC GCAGTCGCAG CCGCCGGCAT
51  CTACGCCTGC CAACCGCAAT CCGAAGCCGC AGTGCAAGTC AAGGCTGAAA
101 ACAGCCTGAC CGCTATGCGC TTAGCCGTCG CCGACAAACA GGCAGAGATT
151 GACGGGTTGA ACGCCCAAAT CGACGCCGAA ATCAGACAAC GCGAAGCCGA
201 AGAATTGAAA GACTACCGAT GGATACACGG CGACGCGGAA GTGCCGGAGC
251 TGGAAAAATG A

This corresponds to the amino acid sequence <SEQ ID 34; ORF52-1>:

1  MKYLIRTALL AVAAAGIYA                                                                    C QFQSEAAVQV KAENSLTAMR LAVADKQAEI
51 DGLNAQIDAE IRQREAEELK DYRWIHGDAE VPELEK*

Computer analysis of this amino acid sequence predicts a prokaryotic membrane lipoprotein lipid attachment site (underlined).

ORF52-1 (7 kDa) was cloned in the pGex vectors and expressed in E.coli , as described above. The products of protein expression and purification were analyzed by SDS-PAGE. FIG. 4A shows the results of affinity purification of the GST-fusion. FIG. 4B shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF52-1.

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 8

The following DNA sequence was identified in N.meningitidis <SEQ ID 35>

1   ATGGTTATCG GAATATTACT CGCATCAAGC AAGCATGCTC TTGTCATTAC
51  TCTATTGTTA AATCCCGTCT TCCATGCATC CAGTTGCGTA TCGCGTTsGG
101 CAATACGGAA TAAAAtCTGC TGTTCTGCTT TGGCTAAATT TGCCAAATTG
151 TTTATTGTTT CTTTAGGaGC AGCTTGCTTA GCCGCCTTCG CTTTCGACAA
201 CGCCCCCACA GGCGCTTCCC AAGCgTTGCC TACCGTTACC GCACCCGTGG
251 CGATTCCCGC GCCCGCTTCG GCAGCCTGA

This corresponds to the amino acid sequence <SEQ ID 36; ORF56>:

1  MVIGILLASS KHALVITLLL NPVFHASSCV SRXAIRNKIC CSALAKFAKL
51 FIVSLG                                                                    AACL AAFAFDNAPT GASQALPTVT APVAIPAPAS AA*

Further work revealed the complete nucleotide sequence <SEQ ID 37>:

1   ATGGCTTGTA CAGGTTTGAT GGTTTTTCCG TTAATGGTTA TCGGAATATT
51  ACTTGCATCA AGCAAGCCTG CTCCTTTCCT TACTCTATTG TTAAATCCCG
101 TCTTCCATGC ATCCAGTTGC GTATCGCGTT GGGCAATACG GAATAAAATC
151 TGCTGTTCTG CTTTGGCTAA ATTTGCCAAA TTGTTTATTG TTTCTTTAGG
201 AGCAGCTTGC TTAGCCGCCT TCGCTTTCGA CAACGCCCCC ACAGGCGCTT
251 CCCAAGCGTT GCCTACCGTT ACCGCACCCG TGGCGATTCC CGCGCCCGCT
301 TCGGCAGCCT GA

This corresponds to the amino acid sequence <SEQ ID 38; ORF56-1>:

1   MACTGLMVFP LMVIGILLAS SKPAPFLTLL LNFVFHASSC VSRWAIPNKL
51  CCSALAKFAK LFIVSLGAAC LAAFAFDNAP TGASQALPTV TAPVAIPAPA
101 SAA*

Computer analysis of this amino acid sequence predicts a leader peptide (underlined) and suggests that ORF56 might be a membrane or periplasmic protein.

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 9

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 39>

1   ATGTTCAGTA TTTTAAATGT CTTTCTTCAT TGTATTCTGG CTTGTGTAGT
51  CTCTGGTGAG ACGCCTACTA TATTTGGTAT CCTTGCTCTT TTTTACTTAT
101 TGTATCTTTC TTATCTTGCT GTTTTTAAGA TTTTCTTTTC TTTTTTCTTA
151 GACAGAGTTT CACTCCGGTC TCCCAGGCTG GAGTGCAAAT GGCATGACCC
201 TTTGGCTCAC TGGCTCACGG CCACTTCTGC TATTCTGCCG CCTCAGCCTC
251 CAGGG ...

This corresponds to the amino acid sequence <SEQ ID 40; ORF63>:

1  M                                                                        FSILNVFLH CILACVVS                                                                    GE                                                                         TPTIFGILAL FYLLYLSYLA                                                                     VFKIFFSFFL
51 DRVSLRSPRL ECKWHDPLAH WLTATSAILP PQPPG...

Computer analysis of this amino acid sequence predicts a transmembrane region.

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 10

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 41>

1   ..GTGCGGACGT GGTTGGTTTT TTGGTTGCAG CGTTTGAAAT ACCCGTTGTT
51  GCTTTGGATT GCGGATATGT TGCTGTACCG GTTGTTGGGC GGCGCGGAAA
101 TCGAATGCGG CCGTTGCCCT GTGCCGCCGA TGACGGATTG GCAGCATTTT
151 TTGCCGGCGA TGGGAACGGT GTCGGCTTGG GTGGCGGTGA TTTGGGCATA
201 CCTGATGATT GAAAGTGAAA AAAACGGAAG ATATTGA

This corresponds to the amino acid sequence <SEQ ID 42; ORF69>:

1  ..VRTWLVFWLQ RLKYPLLLWI ADMLLYRLLG GAEIECGRCP VPPMTDWQHF
51 LPA                                                                        MGTVSAW VAVIWAYLMI                                                                     ESEKNGRY*

Computer analysis of this amino acid sequence predicts a transmembrane region.

A corresponding ORF from strain A of N.meningitidis was also identified:

Homology With a Predicted ORF from N.meningitidis (Strain A)

ORF69 (SEQ ID NO:42) shows 96.2% identity over a 78aa overlap with an ORF (ORF69a (SEQ ID NO:44)) from strain A of N.meningitidis :

          10        20        30        40        50        60
orf69.pep VRTWLVFWLQRLKYP                                                                        LLLWIADMLLYRLLGGA                                                                    EIECGRCPVPPMTDWQHFLPA                                                                        MGTVSAW
          |||||||||||||||||| |||||||||||||||||||||||||||||||:|:||||:||
orf69a    VRTWLVFWLQRLKYP                                                                        LLLCIADMLLYRLLGGA                                                                    EIECGRCPVPPMTDWQHFLPT                                                                        MGTVAAW
          10        20        30        40        50        60
          70       79
orf69.pep VAVIWAYLMI                                                                    ESEKNGRYX
          |||||||||||||||||||
orf69a    VAVIWAYLMI                                                                    ESEKNGRYX
          70

The ORF69a nucleotide sequence <SEQ ID 43>is:

1   GTGCGGACGT GGTTGGTTTT TTGGTTGCAG CGTTTGAAAT ACCCGTTGTT
51  GCTTTGTATT GCGGATATGC TGCTGTACCG GTTGTTGGGC GGCGCGGAAA
101 TCGAATGCGG CCGTTGCCCT GTACCGCCGA TGACGGATTG GCAGCATTTT
151 TTGCCGACGA TGGGAACGGT GGCGGCTTGG GTGGCGGTGA TTTGGGCATA
201 CCTGATGATT GAAAGTGAAA AAAACGGAAG ATATTGA

This encodes a protein having amino acid sequence <SEQ ID 44>:

1  VRTWLVFWLQ RLKYP                                                                        LLLCI ADMLLYRLLG GA                                                                    EIECGRCP VPPMTDWCHF
51 LPT                                                                        MGTVAAW VAVIWAYLMI                                                                    ESEKNGRY*

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 11

The following DNA sequence was identified in N.meningitidis <SEQ ID 45>

1   ATGTTTCAAA ATTTTGATTT GGGCGTGTTC CTGCTTGCCG TCCTCCCCGT
51  GCTGCCCTCC ATTACCGTCT CGCACGTGGC GCGCGGCTAT ACGGCGCGCT
101 ACTGGGGAGA CAACACTGCC GAACAATACG GCAGGCTGAC ACTGAACCCC
151 CTGCCCCATA TCGATTTGGT CGGCACAATC ATCgTACCGC TGCTTACTTT
201 GATGTTCACG CCCTTCCTGT TCGGCTGGGC GCGTCCGATT CCTATCGATT
251 CGCGCAACTT CCGCAACCCG cGCCTTGCCT GGCGTTGCGT TGCCGCGTCC
301 GGCCCGCTGT CGAATCTAGC GATGGCTGTw CTGTGGGGCG TGGTTTTGGT
351 GCTGACTCCG TATGTCGGCG GGGCGTATCA GATGCCGTTG GCTCAAATGG
401 CAAACTACGG TATTCTGATC AATGCGATTC TGTTCGCGCT CAACATCATC
451 CCCATCCTGC CTTGGGACGG CGGCATTTTC ATCGACACCT TCCTGTCGGC
501 GAAATATTCG CAAGCGTTCC GCAAAATCGA ACCTTATGGG ACGTGGATTA
551 TCCTACTGCT GATGCTGACC sGGGTTTTGG GTGCGTTTAT wGCACCGATT
601 sTGCGGmTGc GTGATTGCrT TTGTGCAGAT GTwCGTCTGA CTGGCTTTCA
651 GACGGCATAA

This corresponds to the amino acid sequence <SEQ ID 46; ORF77>:

1   MFQNFDLGVF LLAVLPVLPS ITVSHVA                                                                    RGY TARYWGDNTA EQYGRLTLNP
51  LP                                                                        HIDLVGTI IVPLLTLMF                                                                    T PFLFGWARPI PIDSRNFRNP RLAWRCVAAS
101 GP                                                                        LSNLAMAV LWGVVLVLT                                                                    P YVGGAYQMPL AQ                                                                        MANYGILI NAILFALNII
151 PILPWDGGIF IDTFLSAKYS QAFRKIEPY                                                                        G TWIILLLMLT XVLGAF                                                                    IAPI
201 XRXRDCXCAD VRLTGFQTA*

Further work revealed the complete nucleotide sequence <SEQ ID 47>:

1   ATGTTTCAAA ATTTTGATTT GGGCGTGTTT CTGCTTGCCG TCCTGCCCGT
51  GCTGCTCTCC ATTACCGTCA GGGAGGTGGC GCGCGGCTAT ACGGCGCGCT
101 ACTGGGGAGA CAACACTGCC GAACAATACG GCAGGCTGAC ACTGAACCCC
151 CTGCCCCATA TCGATTTGGT CGGCACAATC ATCGTACCGC TGCTTACTTT
201 GATGTTCACG CCCTTCCTGT TCGGCTGGGC GCGTCCGATT CCTATCGAPT
251 CGCGCAACTT CCGCAACCCG CGCCTTGCCT GGCGTTGCGT TGCCGCGTCC
301 GGCCCGCTGT CGAATCTAGC GATGGCTGTT CTGTGGGGCG TGGTTTTGGT
351 GCTGACTCCG TATGTCGGCG GGGCGTATCA GATGCCGTTG GCTCAAATGG
401 CAAACTACGG TATTCTGATC AATGCGATTC TGTTCGCGCT CAACATCATC
451 CCCATCCTGC CTTGGGACGG CGGCATTTTC ATCGACACCT TCCTGTCGGC
501 GAAATATTCG CAAGCGTTCC GCAAAATCGA ACCTTATGGG ACGTGGATTA
551 TCCTACTGCT GATGCTGACC GGGGTTTTGG GTGCGTTTAT TGCACCGATT
601 GTGCGGCTGG TGATTGCGTT TGTGCAGATG TTCGTCTGA

This corresponds to the amino acid sequence <SEQ ID 48; ORF77-1>:

1   MFQNFDLGVF LLAVLPVLLS ITVREVA                                                                    RGY TARYWGDNTA EQYGRLTLNP
51  LP                                                                        HIDLVGTI IVPLLTLMF                                                                    T PFLFGWARPI PIDSRNFRNP RLAWRCVAAS
101 GP                                                                        LSNLAMAV LWGVVLVLT                                                                    P YVGGAYQMPL AQ                                                                        MANYGILI NAILFALNII
151 PIL                                                                    PWDGGIF IDTFLSAKYS QAFRKIFPY                                                                        G TWIILLLMLT GVLGAF                                                                    IAPI
201 VRLVIAFVQM FV*

Computer analysis of this amino acid sequence reveals a putative leader sequence and several transmembrane domains.

A corresponding ORF from strain A of N.meningitidis was also identified:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF77 (SEQ ID NO:46) shows 96.5% identity over a 173aa overlap with an ORF (ORF77a (SEQ ID NO:50)) from strain A of N.meningitidis :

          10        20        30        40        50        60
orf77.pep MFQNFDLGVFLLAVLPVLPSITVSHVARGYTARYWGDNTAEQYGR                                                                        LTLNPLPHIDLVGTI
          |||||||||||||||||||||||||||||||||
orf77a    RGYTARYWGDNTAEQYGR                                                                        LTLNPLPHIDLVGTI
          10        20        30
          70        80        90       100       110       120
orf77.pep IV                                                                    PLLTLMFTPFLFGWARPIPIDSRNFRNPRLAWRCVAASGPLSN                                                                        LAMAVLWGVVLVLTP
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf77a    IV                                                                    PLLTLMFTPFLFGWARPIPIDSRNFRNPRLAWRCVAASGPLSN                                                                        LAMAVLWGVVLVLTP
          40        50        60        70        80        90
          130       140       150       160       170       180
orf77.pep YV                                                                    GGAYQMPLAQMANY                                                                        GILINAILFALNIIPIL                                                                    PWDGGIFIDTFLSAKYSQAFRKIEPY                                                                        G
          |||||||||||||||| ||||||| ||||||||||||||||||||||| |||||||||||
orf77a    YV                                                                    GGAYQMPLAQMANY                                                                        XILINAILXALNIIPIL                                                                    PWDGGIFIDTFLSAKXSQAFRKIEFY                                                                        G
          100       110       120       130       140       150
          190       200       210       220
orf77.pep TWIILLLMLTXVLGAF                                                                    IAPIXRXRDCXCADVRLTGFQTAX
          |||| ||||| |||| ||||
orf77a    TWIIXLLMLTGVLGAX                                                                    IAPIVQLVIAFVQMFVX
          160       170       180

ORF77-1 (SEQ ID NO:48) and ORF77a (SEQ ID NO:50) show 96.8% identity in 185 aa overlap:

            10        20        30        40        50        60
orf77-1.pep MFQNFDLGVFLLAVLPVLLSITVREVARGYTARYWGDNTAEQYGRLTLNPLPHIDLVGTI
            |||||||||||||||||||||||||||||||||
orf77a      RGYTARYWGDNTAEQYGRLTLNPLPHIDLVGTI
            10        20        30
            70        80        90       100       110       120
orf77-1.pep IVPLLTLMFTPFLFGWARPIPIDSRNFRNPRLAWRCVAASGPLSNLAMAVLWGVVLVLTP
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf77a      IVPLLTLMFTPFLFGWARPIPIDSRNFRNFRLAWRCVAASGPLSNLAMAVLWGVVLVLTP
            40        50        60        70        80        90
            130       140       150       160       170       180
orf77-1.pep YVGGAYQMPLAQMANYGILINAILFALNIIPILPWDGGIFIDTFLSAKYSQAFRKIEPYG
            |||||||||||||||| ||||||| ||||||||||||||||||||||| |||||||||||
orf77a      YVGGAYQMPLAQMANYXILINAILXALNIIPILPWDGGIFIDTFLSAKXSQAFRKIEPYG
            100       110       120       130       140       150
            190       200       210
orf77-1.pep TWIILLLMLTGVLGAFIAPIVRLVIAFVQMFVX
            |||| |||||||||| |||||:|||||||||||
orf77a      TWIIXLLMLTGVLGAXIAPIVQLVIAFVQMFVX
            160       170       180

A partial ORF77a nucleotide sequence <SEQ ID 49>was identified:

1   CGCGGCTATA CAGCGCGCTA CTGGGGTGAC AACACTGCCG AACAATACGG
51  CAGGCTGACA CTGAACCCCC TGCCCCATAT CGATTTGGTC GGCACAATCA
101 TCGTACCGCT GCTTACTTTG ATGTTTACGC CCTTCCTGTT CGGCTGGGCG
151 CGTCCGATTC CTATCGATTC GCGCAACTTC CGCAACCCGC GCCTTGCCTG
201 GCGTTGCGTT GCCGCGTCCG GCCCGCTGTC GAATCTGGCG ATGGCTGTTC
251 TGTGGGGCGT GGTTTTGGTG CTGACTCCGT ATGTCGGTGG GGCGTATCAG
301 ATGCCGTTGG CNCAAATGGC AAACTACNNN ATTCTGATCA ATGCGATTCT
351 GTNCGCGCTC AACATCATCC CCATCCTGCC TTGGGACGGC GGCATTTTCA
401 TCGACACCTT CCTGTCGGCN AAATANTCGC AAGCGTTCCG CAAAATCGAA
451 CCTTATGGGA CGTGGATTAT CCNGCTGCTT ATGCTGACCG GGGTTTTGGG
501 TGCGTNTATT GCACCGATTG TGCAGCTGGT GATTGCGTTT GTGCAGATGT
551 TCGTCTGA

This encodes a protein having amino acid sequence <SEQ ID 50>:

1   RGYTARYWGD NTAEQYGR                                                                        LT LNPLFHIDLV GTIIV                                                                    PLLTL MFTPFLFGWA
51  RPIPIDSRNF RNPRLAWRCV AASGPLSN                                                                        LA MAVLWGVVLV LTPYV                                                                    GGAYQ
101 MPLAQNANY                                                                        X ILINAILXAL NIIPIL                                                                    PWDG GIFIDTFLSA KXSQAFRKIE
151 PY                                                                        GTWIIXLL MLTGVLGAX                                                                    I APIVQLVIAF VQMFV*

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 12

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 51>

1   ATGAACCTGA TTTCACGTTA CATCATCCGT CAAATGGCGG TTATGGCGGT
51  TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT
101 ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGCTG
151 GGCTACACCG CCCTCAAAAT GCCCGCCCGC GCCTACGAAC TGATTCCCCT
201 CGCCGTCCTT ATCGGCGGAC TGGTCTCCCT CAGCCAGCTT GCCGCCGGCA
251 GCGAACTGAC CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG
301 TTGATTCTGT CGCAGTTCGG TTTTATTTTT GCTATTGCCA CCGTCGCGCT
351 CGGCGAATGG GTTGCGCCCA CACTGAGCCA AAAAGCCGAA AACATCAAAG
401 CCGCCGCCAT CAACGGCAAA ATCAGCACCG GCAATACCGG CCTTTGGCTG
451 AAAGAAAAAA ACAGCGTGAT CAATGTGCGC GAAATGTTGC CCGACCAT..

This corresponds to the amino acid sequence <SEQ ID 52; ORF112>:

1   MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEML
51  GYTALKMPAR AYE                                                                        LIPLAVL IGGLVSLSQL                                                                     AAGSELTVIK ASGMSTKK                                                                        LL
101 LILSQFGFIF AIATV                                                                    ALGEW VAPTLSQKAE NIKAAAINGK ISTGNTGLWL
151 KEKNSVINVR EMLPDH...

Further work revealed further partial nucleotide sequence <SEQ ID 53>:

1   ATGAACCTGA TTTCACGTTA CATCATCCGT CAAATGGCGG TTATGGCGGT
51  TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT
101 ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGCTG
151 gGCTACACCG CCCTCAAAAT GCCCGCCCGC GCCTACGAAC TGATTCCCCT
201 CGCCGTCCTT ATCGGCGGAC TGGTCTCCCT CAGCCAGCTT GCCGCCGGCA
251 GCGAACTGAC CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG
301 TTGATTCTGT CGCAGTTCGG TTTTATTTTT GCTATTGCCA CCGTCGCGCT
351 CGGCGAATGG GTTGCGCCCA CACTGAGCCA AAAAGCCGAA AACATCAAAG
401 CCGCCGCCAT CAACGGCAAA ATCAGCACCG GCAATACCGG CCTTTGGCTG
451 AAAGAAAAAA ACAGCrTkAT CAATGTGCGC GAAATGTTGC CCGACCATAC
501 GCTTTTGGGC ATCAAAATTT GGGCGCGCAA CGATAAAAAC GAATTGGCAG
551 AGGCAGTGGA AGCCGATTCC GCCGTTTTGA ACAGCGACGG CAGTTGGCAG
601 TTGAAAAACA TCCGCCGCAG CACGCTTGGC GAAGACAAAG TCGAGGTCTC
651 TATTGCGGCT GAAGAAAACT GGCCGATTTC CGTCAAACGC AACCTGATGG
701 ACGTATTGCT CGTCAAACCC GACCAAATGT CCGTCGGCGA ACTGACCACC
751 TACATCCGCC ACCTCCAAAA CAACAGCCAA AACACCCGAA TCTACGCCAT
801 CGCATGGTGG CGCAAATTGG TTTACCCCGC CGCAGCCTGG GTGATGGCGC
851 TCGTCGCCTT TGCCTTTACC CCGCAAACCA CCCGCCACGG CAATATGGGC
901 TTAAAACTCT TCGGCGGCAT CTGTsTCGGA TTGCTGTTCC ACCTTGCCGG
951 ACGGCTCTTT GGGTTTACCA GCCAACTCGG...

This corresponds to the amino acid sequence <SEQ ID 54; ORF112-1>:

1   MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEML
51  GYTALKMPAR A                                                                    YE                                                                        LIPLAVL IGGLVSLSQL                                                                     AAGSELTVIK ASGMSTKKLL
101 LILSQFGFIF AIATV                                                                    ALGEW VAPTLSQKAE NIKAAAINGK ISTGNTGLWL
151 KEKNSXTNVR EMLPDHTLLG IKIWARNDKN ELAEAVEADS AVLNSDGSWQ
201 LKNIRRSTLG EDKVEVSIAA EENWPISVKR NLMDVLLVKP DQMSVGELTT
251 YIRHLQNNSQ NTRIYAIAWW RK                                                                        LVYPAAAW VMALVAFAF                                                                    T PQTTRHGN                                                                        MG
301 LKLFGGICXG LLFHL                                                                    AGRLF GFTSQL...

Computer analysis of this amino acid sequence predicts two transmembrane domains.

A corresponding ORF from strain A of N.meningitidis was also identified:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF112 (SEQ ID NO:52) shows 96.4% identity over a 166aa overlap with an ORF (ORF112a (SEQ ID NO: 177)) from strain A of N.meningitidis :

           10        20        30        40        50        60
orf112.pep MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR
           ||||||||||||||||||||||||||||||||||||||||||||||||| ||||||| ||
orf112a    MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMXGYTALKMXAR
           10        20        30        40        50        60
           70        80        90       100       110       120
orf112.pep AYELIPLAVLIGGLVSLSQLAAGSELTVIKASGMSTKKLLLILSQFGFIFAIATVALGEW
           ||||:||||||||||| |||||||||:|||||||||||||||||||||||||||||||||
orf112a    AYELMPLAVLIGGLVSXSQLAAGSELXVIKASGMSTKKLLLILSQFGFIFAIATVALGEW
           70        80        90       100       110       120
           130       140       150       160
orf112.pep VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSVINVREMLPDH
           |||||||||||||||||||||||||||||||||||:||||||||||
orf112a    VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSIINVREMLPDHTLLGIKIWARNDKN
           130       140       150       160       170       180
orf112a    ELAEAVEADSAVLNSDGSWQLKNIRRSTLGEDKVEVSIAAEEXWPISVKRNLMDVLLVKP
           190       200       210       220       230       240

A partial ORF112a nucleotide sequence <SEQ ID 55> was identified:

1    ATGAACCTGA TTTCACGTTA CATCATCCGT CAAATGGCGG TTATGGCGGT
51   TTACGCGCTC CTTGCCTTCC TCGCTTTGTA CAGCTTTTTT GAAATCCTGT
101  ACGAAACCGG CAACCTCGGC AAAGGCAGTT ACGGCATATG GGAAATGNTG
151  GGNTACACCG CCCTCAAAAT GNCCGCCCGC GCCTACGAAC TGATGCCCCT
201  CGCCGTCCTT ATCGGCGGAC TGGTCTCTNT CAGCCAGCTT GCCGCCGGCA
251  GCGAACTGAN CGTCATCAAA GCCAGCGGCA TGAGCACCAA AAAGCTGCTG
301  TTGATTCTGT CGCAGTTCGG TTTTATTTTT GCTATTGCCA CCGTCGCGOT
351  CGGCGAATGG GTTGCGCCCA CACTGAGCCA AAAAGCCGAA AACATCAAAG
401  CCGCGGCCAT CAACGGCAAA ATCAGTACCG GCAATACCGG CCTTTGGCTG
451  AAAGAAAAAA ACAGCATTAT CAATGTGCGC GAAATGTTGC CCGACCATAC
501  CCTGCTGGGC ATTAAAATCT GGGCCCGCAA CGATAAAAAC GAACTGGCAG
551  AGGCAGTGGA AGCCGATTCC GCCGTTTTGA ACAGCGACGG CAGTTGGCAG
601  TTGAAAAACA TCCGCCGCAG CACGCTTGGC GAAGACAAAG TCGAGGTCTC
651  TATTGCGGCT GAAGAAAANT GGCCGATTTC CGTCAAACGC AACCTGATGG
701  ACGTATTGCT CGTCAAACCC GACCAAATGT CCGTCGGCGA ACTGACCACC
751  TACATCCGCC ACCTCCAAAN NNACAGCCAA AACACCCGAA TCTACGCCAT
801  CGCATGGTGG CGCAAATTGG TTTACCCCGC CGCAGCCTGG GTGATGGCGC
851  TCGTCGCCTT TGCCTTTACC CCGCAAACCA CCCGCCACGG CAATATGGGC
901  TTAAAANTCT TCGGCGGCAT CTGTCTCGGA TTGCTGTTCC ACCTTGCCGG
951  NCGGCTCTTC NGGTTTACCA GCCAACTCTA CGGCATCCCG CCCTTCCTCG
1001 NCGGCGCACT ACCTACCATA GCCTTCGCCT TGCTCGCCGT TTGGCTGATA
1051 CGCAAACAGG AAAAACGCTA A

This encodes a protein having amino acid sequence <SEQ ID 56>:

1   MNLISRYIIR QMAVMAVYAL LAFLALYSFF EILYETGNLG KGSYGIWEMX
51  GYTALKMXAR A                                                                    YE                                                                        LMPLAVL IGGLVSXSQ                                                                    L AAGSELXVIK ASGMSTKK                                                                        LL
101 LILSQFGFIF AIATV                                                                    ALGEW VAPTLSQKAE NTKAAAINGK ISTGNTGLWL
151 KEKNSIINVR EMLFDHTLLG IKIWARNDKN ELAEAVEADS AVLNSDGSWQ
201 LKNIRRSTLG EDKVEVSIAA EEXWPISVKR NLMDVLLVKP DQMSVGELTT
251 YIRHLQXXSQ NTRIYAIAWW RK                                                                        LVYPAAAW VMALVAFAF                                                                    T PQTTRHGN                                                                        MG
301 LKXFGGICLG LLFHL                                                                    AGRLF XFTSQLYGIP PFLXGALPTI AFALLAVWLI
351 RKQEKR*

ORF112a (SEQ ID NO:56) and ORF112-1 (SEQ ID NO:54) show 96.3% identity in 326 aa overlap:

orf112a.pep MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMXGYTALKMXAR
            ||||||||||||||||||||||||||||||||||||||||||||||||| ||||||| ||
orf112-1    MNLISRYIIRQMAVMAVYALLAFLALYSFFEILYETGNLGKGSYGIWEMLGYTALKMPAR
orf112a.pep AYELMPLAVLIGGLVSXSQLAAGSELXVIKASGMSTKKLLLILSQFGFIFAIATVALGEW
            ||||:||||||||||| |||||||||:|||||||||||||||||||||||||||||||||
orf112-1    AYELIPLAVLIGGLVSLSQLAAGSELTVIKASGMSTKKLLLILSQFGFIFAIATVALGEW
orf112a.pep VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSIINVREMLPDRTLLGIKIWARNDKN
            ||||||||||||||||||||||||||||||||||| ||||||||||||||||||||||||
orf112-1    VAPTLSQKAENIKAAAINGKISTGNTGLWLKEKNSXINVREMLPDHTLLGIKIWARNDKN
orf112a.pep ELAEAVEADSAVLNSDGSWQLKNIRRSTLGEDKVEVSIAAEEXWPISVKRNLMDVLLVKP
            |||||||||||||||||||||||||||||||||||||||||| |||||||||||||||||
orf112-1    ELAEAVEADSAVLNSDGSWQLKNIRRSTLGEDKVEVSIAAEENWPISVKRNLMDVLLVKP
orf112a.pep DQMSVGELTTYIRHLQXXSQNTRIYAIAWWRKLVYPAAAWVMALVAFAFTPQTTRHGNNG
            ||||||||||||||||  ||||||||||||||||||||||||||||||||||||||||||
orf1l2-1    DQMSVGELTTYIRHLQNNSQNTRIYAIAWWRKLVYPAAAWVMALVAFAFTPQTTRHGNMG
orf112a.pep LKXFGGICLGLLFHLAGRLFXFTSQLYGIPPFLXGALPTIAFALLAVWLIRKQEKRX
            || ||||| ||||||||||| |||||
orf112-1    LKLFGGICXGLLFHLAGRLFGFTSQL

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 13

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 57>

1   GCAGTAGCCG AAACTGCCAA CAGCCAGGGC AAAGGTAAAC AGGCAGGCAG
51  TTCGGTTTCT GTTTCACTGA AAACTTCAGG CGACCTTTGC GGCAAACTCA
101 AAACCACCCT TAAAACTTTG GTCTGCTCTT TGGTTTCCCT GAGTATGGTA
151 TTGCCTGCCC ATGCCCAAAT TACCACCGAC AAATCAGCAC CTAAAAACCA
201 GCAGGTCGTT ATCCTTAAAA CCAACACTGG TGCCCCCTTG GTGAATATCC
251 AAACTCCGAA TGGACGCGGA TTGAGCCACA ACCGCTA.TA CGCATTTGAT
301 GTTGACAACA AAGGGGCAGT GTTAAACAAC GACCGTAACA ATAATCCGTT
351 TGTGGTCAAA GGCAGTGCGC AATTGATTTT GAACGAGGTA CGCGGTACGG
401 CTAGCAAACT CAACGGCATC GTTACCGTAG GCGGTCAAAA GGCCGACGTG
451 ATTATTGCCA ACCCCAACGG CATTACCGTT AATGGCGGCG GCTTTAAAAA
501 TGTCGGTCGG GGCATCTTAA CTACCGGTGC GCCCCAAATC GGCAAAGACG
551 GTGCACTGAC AGGATTTGAT GTG                                                                        C                                                                    GTCAAG GCACATTGgA CCGTAGrAGC
601 AGCAGGTTGG AATGATAAAG GCGGAGCmrm yTACACCGGG GTACTTGCTC
651 GTGCAGTTGC TTTGCAGGGG AAATTwmmGG GTAAA.AACT GGCGGTTTCT
701 ACCGGTCCTC AGAAAGTAGA TTACGCCAGC GGCGAAATCA GTGCAGGTAC
751 GGCAGCGGGT ACGAAACCGA CTATTGCCCT TGATACTGCC GCACTGGGCG
801 GTATGTACGC CGACAGCATC ACACTGATTG CCAATGAAAA AGGCGTAGGC
851 GTCTAA

This corresponds to the amino acid sequence <SEQ ID 58; ORF114>:

1   AVAETANSQG KGKQAGSSVS VSLKTSGDLC GKLKTT                                                                        LKTL VCSLVSLSNV
51  LPA                                                                    HAQITTD KSAPKNQQVV ILKTNTGAPL VNIQTPNGRG LSHNRXYAFD
101 VDNKGAVLNN DRNNNPFVVK GSAQLILNEV RGTASKLNGI VTVGGQKADV
151 IIANPNGITV NGGGFKNVGR GILTTGAPQI GKDGALTGFD VVKAHWTVXA
201 AGWNDKGGAX YTGVLARAVA LQGKXXGKXL AVSTGPQKVD YASGEISAGT
251 AAGTKPTIAL DTAALGGMYA DSITLIANEK GVGV*

Further work revealed the complete nucleotide sequence <SEQ ID 59>:

1    ATGAATAAAG GTTTACATCG CATTATCTTT AGTAAAAAGC ACAGCACCAT
51   GGTTGCAGTA GCCGAAACTG CCAACAGCCA GGGCAAAGGT AAACAGGCAG
101  GCAGTTCGGT TTCTGTTTCA CTGAAAACTT CAGGCGACCT TTGCGGCAAA
151  CTCAAAACCA CCCTTAAAAC TTTGGTCTGC TCTTTGGTTT CCCTGAGTAT
201  GGTATTGCCT GCCCATGCCC AAATTACCAC CGACAAATCA GCACCTAAAA
251  ACCAGCAGGT CGTTATCCTT AAAACCAACA CTGGTGCCCC CTTGGTGAAT
301  ATCCAAACTC CGAATGGACG CGGATTGAGC CACAACCGCT ATACGCAGTT
351  TGATGTTGAC AACAAAGGGG CAGTGTTAAA CAACGACCGT AACAATAATC
401  CGTTTGTGGT CAAAGGCAGT GCGCAATTGA TTTTGAACGA GGTACGCGGT
451  ACGGCTAGCA AACTCAACGG CATCGTTACC GTAGGCGGTC AAAAGGCCGA
501  CGTGATTATT GCCAACCCCA ACGGCATTAC CGTTAATGGC GGCGGCTTTA
551  AAAATGTCGG TCGGGGCATC TTAACTACCG GTGCGCCCCA AATCGGCAAA
601  GACGGTGCAC TGACAGGATT TGATGTGCGT CAAGGCACAT TGACCGTAGG
651  AGCAGCAGGT TGGAATGATA AAGGCGGAGC CGACTACACC GGGGTACTTG
701  CTCGTGCAGT TGCTTTGCAG GGGAAATTAC AGGGTAAAAA CCTGGCGGTT
751  TCTACCGGTC CTCAGAAAGT AGATTACGCC AGCGGCGAAA TCAGTGCAGG
801  TACGGCAGCG GGTACGAAAC CGACTATTGC CCTTGATACT GCCGCACTGG
851  GCGGTATGTA CGCCGACAGC ATCACACTGA TTGCCAATGA AAAAGGCGTA
901  GGCGTCAAAA ATGCCGGCAC ACTCGAAGCG GCCAAGCAAT TGATTGTGAC
951  TTCGTCAGGC CGCATTGAAA ACAGCGGCCG CATCGCCACC ACTGCCGACG
1001 GCACCGAAGC TTCACCGACT TATCTCTCCA TCGAAACCAC CGAAAAAGGA
1051 GCGGCAGGCA CATTTATCTC CAATGGTGGT CGGATCGAGA GCAAAGGCTT
1101 ATTGGTTATT GAGACGGGAG AAGATATCAG CTTGCGTAAC GGAGCCGTGG
1151 TGCAGAATAA CGGCAGTCGC CCAGCTACCA CGGTATTAAA TGCTGGTCAT
1201 AATTTGGTGA TTGAGAGCAA AACTAATGTG AACAATGCCA AAGGCCCGGC
1251 TACTCTGTCG GCCGACGGCC GTACCGTCAT CAAGGAGGCC AGTATTCAGA
1301 CTGGCACTAC CGTATACAGT TCCAGCAAAG GCAACGCCGA ATTAGGCAAT
1351 AACACACGCA TTACCGGGGC AGATGTTACC GTATTATCCA ACGGCACCAT
1401 CAGCAGTTCC GCCGTAATAG ATGCCAAAGA CACCGCACAC ATCGAAGCAG
1451 GCAAACCGCT TTCTTTGGAA GCTTCAACAG TTACCTCCGA TATCCGCTTA
1501 AACGGAGGCA GTATCAAGGG CGGCAAGCAG CTTGCTTTAC TGGCAGACGA
1551 TAACATTACT GCCAAAACTA CCAATCTGAA TACTCCCGGC AATCTGTATG
1601 TTCATACAGG TAAAGATCTG AATTTGAATG TTGATAAAGA TTTGTCTGCC
1651 GCCAGCATCC ATTTGAAATC GGATAACGCT GCCCATATTA CCGGCACCAG
1701 TAAAACCCTC ACTGCCTCAA AAGACATGGG TGTGGAGGCA GGCTCGCTGA
1751 ATGTTACCAA TACCAATCTG CGTACCAACT CGGGTAATCT GCACATTCAG
1801 GCAGCCAAAG GCAATATTCA GCTTCGCAAT ACCAAGCTGA ACGCAGCCAA
1851 GGCTCTCGAA ACCACCGCAT TGCAGGGCAA TATCGTTTCA GACGGCCTTC
1901 ATGCTGTTTC TGCAGACGGT CATGTATCCT TATTGGCCAA CGGTAATGCC
1951 GACTTTACCG GTCACAATAC CCTGACAGCC AAGGCCGATG TCAATGCAGG
2001 ATCGGTTGGT AAAGGCCGTC TGAAAGCAGA CAATACCAAT ATCACTTCAC
2051 CTTCAGGAGA TATTACGTTG GTTGCCGGCA ACGGTATTCA GCTTGGTGAC
2101 GGAAAACAAC GCAATTCAAT CAACGGAAAA CACATCAGCA TCAAAAACAA
2151 CGGTGGTAAT GCCGACTTAA AAAACCTTAA CGTCCATGCC AAAAGCGGGG
2201 CATTGAACAT TCATTCCGAC CGGGCATTGA GCATAGAAAA TACCAAGCTG
2251 GAGTCTACCC ATAATACGCA TCTTAATGCA CAACACGAGC GGGTAACGCT
2301 CAACCAAGTA GATGCCTACG CACACCGTCA TCTAAGCATT ACCGGCAGCC
2351 AGATTTGGCA AAACGACAAA CTGCCTTCTG CCAACAAGCT GGTGGCTAAC
2401 GGTGTATTGG CACTCAATGC GCGCTATTCC CAAATTGCCG ACAACACCAC
2451 GCTGAGAGCG GGTGCAATCA ACCTTACTGC CGGTACCGCC CTAGTCAAGC
2501 GCGGCAACAT CAATTGGAGT ACCGTTTCGA CCAAAACTTT GGAAGATAAT
2551 GCCGAATTAA AACCATTGGC CGGACGGCTG AATATTGAAG CAGGTAGCGG
2601 CACATTAACC ATCGAACCTG CCAACCGCAT CAGTGCGCAT ACCGACCTGA
2651 GCATCAAAAC AGGCGGAAAA TTGCTGTTGT CTGCAAAAGG AGGAAATGCA
2701 GGTGCGCCTA GTGCTCAAGT TTCCTCATTG GAAGCAAAAG GCAATATCCG
2751 TCTGGTTACA GGAGAAACAG ATTTAAGAGG TTCTAAAATT ACAGCCGGTA
2801 AAAACTTGGT TGTCGCCACC ACCAAAGGCA AGTTGAATAT CGAAGCCGTA
2851 AACAACTCAT TCAGCAATTA TTTTCCTACA CAAAAAGCGG CTGAACTCAA
2901 CCAAAAATCC AAAGAATTGG AACAGCAGAT TGCGCAGTTG AAAAAAAGCT
2951 CGCCTAAAAG CAAGCTGATT CCAACCCTGC AAGAAGAACG CGACCGTCTC
3001 GCTTTCTATA TTCAAGCCAT CAACAAGGAA GTTAAAGGTA AAAAACCCAA
3051 AGGCAAAGAA TACCTGCAAG CCAAGCTTTC TGCACAAAAT ATTGACTTGA
3101 TTTCCGCACA AGGCATCGAA ATCAGCGGTT CCGATATTAC CGCTTCCAAA
3151 AAACTGAACC TTCACGCCGC AGGCGTATTG CCAAAGGCAG CAGATTCAGA
3201 GGCGGCTGCT ATTCTGATTG ACGGCATAAC CGACCAATAT GAAATTGGCA
3251 AGCCCACCTA CAAGAGTCAC TACGACAAAG CTGCTCTGAA CAAGCCTTCA
3301 CGTTTGACCG GACGTACAGG GGTAAGTATT CATGCAGCTG CGGCACTCGA
3351 TGATGCACGT ATTATTATCG GTGCATCCGA AATCAAAGCT CCCTCAGGCA
3401 GCATAGACAT CAAAGCCCAT AGTGATATTG TACTGGAGGC TGGACAAAAC
3451 GATGCCTATA CCTTCTTAAA AACCAAAGGT AAAAGCGGCA AAATCATCAG
3501 AAAAACCAAG TTTACCAGCA CCCGCGACCA CCTGATTATG CCAGCCCCCG
3551 TCGAGCTGAC CGCCAACGGC ATAACGCTTC AGGCAGGCGG CAACATCGAA
3601 GCTAATACCA CCCGCTTCAA TGCCCCTGCA GGTAAAGTTA CCCTGGTTGC
3651 GGGTGAAGAG CTGCAACTGC TGGCAGAAGA AGGCATCCAC AAGCACGAGT
3701 TGGATGTCCA AAAAAGCCGC CGCTTTATCG GCATCAAGGT AGGCAAGAGC
3751 AATTACAGTA AAAACGAACT GAACGAAACC AAATTGCCTG TCCGCGTCGT
3801 CGCCCAAACT GCAGCCACCC GTTCAGGCTG GGATACCGTG CTCGAAGGTA
3851 CCGAATTCAA AACCACGCTG GCCGGTGCGG ACATTCAGGC AGGTGTAGGC
3901 GAAAAAGCCC GTGCCGATGC GAAAATTATC CTCAAAGGCA TTGTGAACCG
3951 TATCCAGTCG GAAGAAAAAT TAGAAACCAA CTCAACCGTA TGGCAGAAAC
4001 AGGCCGGACG CGGCAGCACT ATCGAAACGC TGAAACTGCC CAGCTTCGAA
4051 AGCCCTACTC CGCCCAAACT GACCGCCCCC GGTGGCTATA TCGTCGACAT
4101 TCCGAAAGGC AATTTGAAAA CCGAAATCGA AAAGCTGGCC AAACAGCCCG
4151 AGTATGCCTA TCTGAAACAG CTCCAAGTAG CGAAAAACGT CAACTGGAAC
4201 CAGGTGCAAC TGGCTTACGA TAAATGGGAC TATAAGCAGG AAGGCTTAAC
4251 CAGAGCCGGT GCAGCGATTG TTACCATAAT CGTAACCGCA CTGACTTATG
4301 GATACGGCGC AACCGCAGCG GGCGGTGTAG CCGCTTCAGG AAGTAGTACA
4351 GCCGCAGCTG CCGGAACAGC CGCCACAACG ACAGCAGCAG CTACTACCGT
4401 TTCTACAGCG ACTGCCATGC AAACCGCTGC TTTAGCCTCC TTGTATAGCC
4451 AAGCAGCTGT ATCCATCATC AATAATAAAG GTGATGTCGG CAAAGCGTTG
4501 AAAGATCTCG GCACCAGTGA TACGGTCAAG CAGATTGTCA CTTCTGCCCT
4551 GACGGCGGGT GCATTAAATC AGATGGGCGC AGATATTGCC CAATTGAACA
4601 GCAAGGTAAG AACCGAACTG TTCAGCAGTA CGGGCAATCA AACTATTGCC
4651 AACCTTGGAG GCAGACTGGC TACCAATCTC AGTAATGCAG GTATCTCAGC
4701 TGGTATCAAT ACCGCCGTCA ACGGCGGCAG CCTGAAAGAC AACTTAGGCA
4751 ATGCCGCATT AGGAGCATTG GTTAATAGCT TCCAAGGAGA AGCCGCCAGC
4801 AAAATCAAAA CAACCTTCAG CGACGATTAT GTTGCCAAAC AGTTCGCCCA
4851 CGCTTTGGCT GGGTGTGTTA GCGGATTGGT ACAAGGAAAA TGTAAAGACG
4901 GGGCAATTGG CGCAGCAGTT GGGGAAATCG TAGCCGACTC CATGCTTGGC
4951 GGCAGAAACC CTGCTACACT CAGCGATGCG GAAAAGCATA AGGTTATCAG
5001 TTACTCGAAG ATTATTGCCG GCAGCGTGGC GGCACTCAAC GGCGGCGATG
5051 TGAATACTGC GGCGAATGCG GCTGAGGTGG CGGTAGTGAA TAATGCTTTG
5101 AATTTTGACA GTACCCCTAC CAATGCGAAA AAGCATCAAC QGCAGAAGCC
5151 CGACAAAACC GCACTGGAAA AAATTATCCA AGGTATTATG CCTGCACATG
5201 CAGCAGGTGC GATGACTAAT CCGCAGGATA AGGATGCTGC CATTTGGATA
5251 AGCAATATCC GTAATGGCAT CACAGGCCCG ATTGTGATTA CCAGCTATGG
5301 GGTTTATGCT GCAGGTTGGA CAGCTCCGCT GATCGGTACA GCGGGTAAAT
5351 TAGCTATCAG CACCTGCATG GCTAATCCTT CTGGTTGTAC TGTCATGGTC
5401 ACTCAGGCTG CCGAAGCGGG CGCGGGAATC GCCACGGGTG CGGTAACGGT
5451 AGGCAACGCT TGGGAAGCGC CTGTGGGGGC GTTGTCGAAA GCGAAGGCGG
5501 CCAAGCAGGC TATACCAACC CAGACAGTTA AAGAACTTGA TGGCTTACTA
5551 CAAGAATCAA AAAATATAGG TGCTGTAAAT ACACGAATTA ATATAGCGAA
5601 TAGTACTACT CGATATACAC CAATGAGACA AACGGGACAA CCGGTATCTG
5651 CTGGCTTTGA GCATGTTCTT GAGGGGCACT TCCATAGGCC TATTGCGAAT
5701 AACCGTTCAG TTTTTACCAT CTCCCCAAAT GAATTGAAGG TTATACTTCA
5751 AAGTAATAAA GTAGTTTCTT CTCCCGTATC GATGACTCCT GATGGCCAAT
5801 ATATGCGGAC TGTCGATGTA GGAAAAGTTA TTGGTACTAC TTCTATTAAA
5851 GAAGGTGGAC AACCCACAAC TACAATTAAA GTATTTACAG ATAAGTCAGG
5901 AAATTTGATT ACTACATACC CAGTAAAAGG AAACTAA

This corresponds to the amino acid sequence <SEQ ID 60; ORF114-1>:

1    MNKGLHRIIF SKKHSTMVAV AETANSQGKG KQAGSSVSVS LKTSGDLCGK
51   LKTTLKTLVC SLVSLSMVLP AHAQITTDKS APKNQQVVIL KTNTGAPLVN
101  IQTPNGRGLS HYNRYTQFDVD NKGAVLNNDR NNNPFVVKGS AQLILNEVRG
151  TASKLNGIVT VGGQKADVII ANPNGITVNG GGFKNVGRGI LTTGAPQIGK
201  DGALTGFDVR QGTLTVGAAG WNDKGGADYT GVLARAVALQ GKLQGKNLAV
251  STGPQKVDYA SGEISAGTAA GTKPTIALDT AALGGMYADS ITLIANEKGV
301  GVKNAGTLEA AKQLIVTSSG RIENSGRIAT TADGTEASPT YLSIETTEKG
351  AAGTFISNGG RIESKGLLVI ETGEDISLRN GAVVQNNGSR FATTVLNAGH
401  NLVIESKTNV NNAKGPATLS ADGRTVIKEA SIQTGTTVYS SSKGNAELGN
451  NTRITGADVT VLSNGTISSS AVIDAKDTAH IEAGKPLSLE ASTVTSDIRL
501  NGGSIKGGKQ LALLADDNIT AKTTNLNTPG NLYVHTGKDL NLNVDKDLSA
551  ASIHLKSDNA AHITGTSKTL TASKDMGVEA GSLNVTNTNL RTNSGNLHIQ
601  AAKGNIQLRN TKLNAAKALE TTALQGNIVS DGLHAVSADG HVSLLANGNA
651  DFTGHNTLTA KADVNAGSVG KGRLKADNTN ITSSSGDITL VAGNGIQLGD
701  GKQRNSINGK HISIKNNGGN ADLKNLNVHA KSGALNIHSD RALSIENTKL
751  ESTHNTHLNA QRERVTLNQV DAYAHRHLSI TGSQIWQNDK LPSANKLVAN
801  GVLALNARYS QIADNTTLRA GAINLTAGTA LVKRGNINWS TVSTKTLEDN
851  AELKPLAGRL NIEAGSGTLT IEPANRISAH TDLSIKTGGK LLLSAKGGNA
901  GAPSAQVSSL EAKGNIRLVT GETDLRGSKI TAGKNLVVAT TKGKLNIEAV
951  NNSFSNYFPT QKAAELNQKS KELEQQIAQL KKSSPKSKLI PTLQEERDRL
l00l AFYIQAINKE VKGKKPKGKE YLQAKLSAQN IDLISAQGIE ISGSDITASK
1051 KLNLHAAGVL PKAADSEAAA ILIDGITDQY EIGKPTYKSH YDKAALNKPS
1101 RLTGRTGVSI HAAAALDDAR IIIGASEIKA PSGSIDIKAH SDIVLEAGQN
1151 DAYTFLKTKG KSGKIIRKTK FTSTRDHLIM PAPVELTANG ITLQAGGNIE
1201 ANTTRFNAPA GKVTLVAGEE LQLLAEEGIH KHELDVQKSR RFIGIKVGKS
1251 NYSKNELNET KLPVRVVAQT AATRSGWDTV LEGTEFKTTL AGADIQAGVG
1301 EKARADAKII LKGIVNRIQS EEKLETNSTV WQKQAGRGST IETLKLPSFE
1351 SPTPPKLTAP GGYIVDIPKG NLKTEIEKLA KQP                                                                        EYAYLKQ LQVAKNVNWN
1401 QVQLAYDKWD YKQEGLTRAG AAIVTIIVTA LTYGYGATAA GGVAASGSST
1451 AAAAGTAATT TAAATTVSTA TAMQTAALAS LYSQAAVSII NNKGDVGKAL
1501 KDLGTSDTVK QIVTSALTAG ALNQMGADIA QLNSKVRTEL FSSTGNQTIA
1551 NLGGRLATNL SNAGISAGIN TAVNGGSLKD NLGNAALGAL VNSFQGEAAS
1601 KIKTTFSDDY VAKQFAHALA GCVSGLVQGK CKDGAIGAAV GEIVADSMLG
1651 GRNPATLSDA EKHKVISYSK IIAGSVAALN GGDVNTAANA AEVAVVNNAL
1701 NFDSTPTNAK KHQPQKPDKT ALEKIIQGIM PAHAAGAMTN PQDKDAAIWI
1751 SNIRNGITGP IVITSYGVYA AGWTAPLIGT AGKLAISTCM ANPSGCTVMV
1801 TQAAEAGAGI ATGAVTVGNA WEAPVGALSK AKAAKQAIPT QTVKELDGLL
1851 QESKNIGAVN TRINIANSTT RYTPMRQTGQ PVSAGFEHVL EGHFHRPIAN
1901 NRSVFTISPN ELKVILQSNK VVSSPVSMTP DGQYMRTVDV GKVIGTTSIK
1951 EGGQPTTTIK VFTDKSGNLI TTYPVKGN*

Computer analysis of this amino acid sequence predicts a transmembrane region and also gives the following results:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF114 (SEQ ID NO:58) shows 91.9% identity over a 284aa overlap with an ORF (ORF114a (SEQ ID NO:178)) from strain A of N.meningitidis :

           10        20        30        40
orf114.pep AVAETANSQGKGKQAGSSVSVSLKTSGDLCGKLKTTLKTLVC
           ||||||||||||||||||||||||||||||||||||||||||
orf114a    MNKGLHRIIFSKKHSTMVAVAETANSQGKGKQAGSSVSVSLKTSGDLCGKLKTTLKTLVC
           10        20        30        40        50        60
           50        60        70        80        90       100
orf114.pep SLVSLSMVLPAHAQITTDKSAPKNQQVVILKTNTGAPLVNIQTPNGRGLSHNRXYAFDVD
           |||||||      ||||||||||| ||||||||||||||||||||||||||||   ||||
orf114a    SLVSLSMXXXXXXQITTDKSAPKNXQVVILKTNTGAPLVNIQTPNGRGLSHNRYTQFDVD
           70        80        90       100       110       120
           110       120       130       140       150       160
orf114.pep NKGAVLNNDRNNNPFVVKGSAQLILNEVRGTASKLNGIVTVGGQKADVIIANPNGITVNG
           |||||||||||||||:||||||||||||||||||||||||||||||||||||||||||||
orf114a    NKGAVLNNDRNNNPFLVKGSAQLILNEVRGTASKLNGIVTVGGQKADVIIANPNGITVNG
           130       140       150       160       170       180
           170       180       190       200       210       220
orf114.pep GGFKNVGRGILTTGAPQIGKDGALTGFDVVKAHWTVXAAGWNDKGGAXYTGVLARAVALQ
           |||||||||||| |||||||||||||||| ::  || |||||||||| ||||||||||||
orf114a    GGFKNVGRGILTIGAPQIGKDGALTGFDVRQGTLTVGAAGWNDKGGADYTGVLARAVALQ
           190       200       210       220       230       240
           230       240       250       260       270       280
orf114.pep GKXXGKXLAVSTGPQKVDYASGEISAGTAAGTKPTIALDTAALGGMYADSITLIANEKGV
           ||  || |||||||||||||||||||||||||||||||||||||||||||||||| ||||
orf114a    GKLQGKNLAVSTGPQKVDYASGEISAGTAAGTKPTIALDTAALGGMYADSITLIAXEKGV
           250       260       270       280       290       300
orf114.pep GVX
           ||
orf114a    GVKNAGTLEAAKQLIVTSSGRIENSGRIATTADGTEASPTYLXIETTEKGAXGTFISNGG
           310       320       330       340       350       360

The complete length ORF114a nucleotide sequence <SEQ ID 61> is:

1    ATGAATAAAG GTTTACATCG CATTATCTTT AGTAAAAAGC ACAGCACCAT
51   GGTTGCAGTA GCCGAAACTG CCAACAGCCA GGGCAAAGGT AAACAGGCAG
101  GCAGTTCGGT TTCTGTTTCA CTGAAAACTT CAGGCGACCT TTGCGGCAAA
151  CTCAAAACCA CCCTTAAAAC CTTGGTCTGC TCTTTGGTTT CCCTGAGTAT
201  GGNATTNCNN NNCNNTNCCC AAATTACCAC CGACAAATCA GCACCTAAAA
251  ACCANCAGGT CGTTATCCTT AAAACCAACA CTGGTGCCCC CTTGGTGAAT
301  ATCCAAACTC CGAATGGACG CGGATTGAGC CACAACCGCT ATACGCAGTT
351  TGATGTTGAC AACAAAGGGG CAGTGTTAAA CAACGACCGT AACAATAATC
401  CGTTTCTGGT CAAAGGCAGT GCGCAATTGA TTTTGAACGA GGTACGCGGT
451  ACGGCTAGCA AACTCAACGG CATCGTTACC GTAGGCGGTC AAAAGGCCGA
501  CGTGATTATT GCCAACCCCA ACGGCATTAC CGTTAATGGC GGCGGCTTTA
551  AAAATGTCGG TCGGGGCATC TTAACTATCG GTGCGCCCCA AATCGGCAAA
601  GACGGTGCAC TGACAGGATT TGATGTGCCT CAAGGCACAT TGACCGTAGG
651  AGCAGCAGGT TGGAATGATA AAGGCGGAGC CGACTACACC GGGGTACTTG
701  CTCGTGCAGT TGCTTTGCAG GGGAAATTAC AGGGTAAAAA CCTGGCGGTT
751  TCTACCGGTC CTCAGAAAGT AGATTACGCC AGCGGCGAAA TCAGTGCAGG
801  TACGGCAGCG GGTACGAAAC CGACTATTGC CCTTGATACT GCCGCACTGG
851  GCGGTATGTA CGCCGACAGC ATCACACTGA TTGCCANTGA AAAAGGCGTA
901  GGCGTCAAAA ATGCCGGCAC ACTCGAAGCG GCCAAGCAAT TGATTGTGAC
951  TTCGTCAGGC CGCATTGAAA ACAGCGGCCG CATCGCCACC ACTGCCGACG
1001 GCACCGAAGC TTCACCGACT TATCTNNCNA TCGAAACCAC CGAAAAAGGA
1051 GCNNCAGGCA CATTTATCTC CAATGGTGGT CGGATCGAGA GCAAAGGCTT
1101 ATTGGTTATT GAGACGGGAG AAGATATCAN CTTGCGTAAC GGAGCCGTGG
1151 TGCAGAATAA CGGCAGTCGC CCAGCTACCA CGGTATTAAA TGCTGGTCAT
1201 AATTTGGTGA TTGAGAGTAA AACTAATGTG AACAATGCCA AAGGCTCGNC
1251 TAATCTGTCG GCCGGCGGTC GTACTACGAT CAATGATGCT ACTATTCAAG
1301 CGGGCAGTTC CGTGTACAGC TCCACCAAAG GCGATACTGA NTTGGGTGAA
1351 AATACCCGTA TTATTGCTGA AAACGTAACC GTATTATCTA ACGGTAGTAT
1401 TGGCAGTGCT GCTGTAATTG AGGCTAAAGA CACTGCACAC ATTGAATCGG
1451 GCAAACCGCT TTCTTTAGAA ACCTCGACCG TTGCCTCCAA CATCCGTTTG
1501 AACAACGGTA ACATTAAAGG CGGAAAGCAG CTTGCTTTAC TGGCAGACGA
1551 TAACATTACT GCCAAAACTA CCAATCTGAA TACTCCCGGC AATCTGTATG
1601 TTCATACAGG TAAAGATCTG AATTTGAATG TTGATAAAGA TTTGTCTGCC
1651 GCCAGCATCC ATTTGAAATC GGATAACGCT GCCCATATTA CCGGCACCAG
1701 TAAAACCCTC ACTGCCTCAA AAGACATGGG TGTGGAGGCA GGCTTGCTGA
1751 ATGTTACCAA TACCAATCTG CGTACCAACT CGGGTAATCT GCACATTCAG
1801 GCAGCCAAAG GCAATATTCA GCTTCGCAAT ACCAAGCTGA ACGCAGCCAA
1851 GGCTCTCGAA ACCACCGCAT TGCAGGGCAA TATCGTTTCA GACGGCCTTC
1901 ATGCTGTTTC TGCAGACGGT CATGTATCCT TATTGGCCAA CGGTAATGCC
1951 GACTTTACCG GTCACAATAC CCTGACAGCC AAGGCCGATG TCNATGCAGG
2001 ATCGGTTGGT AAAGGCCGTC TGAAAGCAGA CAATACCAAT ATCACTTCAT
2051 CTTCAGGAGA TATTACGTTG GTTGCCGNNN NCGGTATTCA GCTTGGTGAC
2101 GGAAAACAAC GCAATTCAAT CAACGGAAAA CACATCAGCA TCAAAAACAA
2151 CGGTGGTAAT GCCGACTTAA AAAACCTTAA CGTCCATGCC AAAAGCGGGG
2201 CATTGAACAT TCATTCCGAC CGGGCATTGA GCATAGAAAA TACNAAGCTG
2251 GAGTCTACCC ATAATACGCA TCTTAATGCA CAACACGAGC GGGTAACGCT
2301 CAACCAAGTA GATGCCTACG CACACCGTCA TCTAAGCATT ANCGGCAGCC
2351 AGATTTGGCA AAACGACAAA CTGCCTTCTG CCAACAAGCT GGTGGCTAAC
2401 GGTGTATTGG CANTCAATGC GCGCTATTCC CAAATTGCCG ACAACACCAC
2451 GCTGAGAGCG GGTGCAATCA ACCTTACTGC CGGTACCGCC CTAGTCAAGC
2501 GCGGCAACAT CAATTGGAGT ACCGTTTCGA CCAAGACTTT GGAAGATAAT
2551 GCCGAATTAA AACCATTGGC CGGACGGCTG AATATTGAAG CAGGTAGCGG
2601 CACATTAACC ATCGAACCTG CCAACCGCAT CAGTGCGCAT ACCGACCTGA
2651 GCATCAAAAC AGGCGGAAAA TTGCTGTTGT CTGCAAAAGG AGGAAATGCA
2701 GGTGCGCNTA GTGCTCAAGT TTCCTCATTG GAAGCAAAAG GCAATATCCG
2751 TCTGGTTACA GGAGNAACAG ATTTAAGAGG TTCTAAAATT ACAGCCGGTA
2801 AAAACTTGGT TGTCGCCACC ACCAAAGGCA AGTTGAATAT CGAAGCCGTA
2851 AACAACTCAT TCAGCAATTA TTTTCNTACA CAAAAAGNGN NNGNNCTCAA
2901 CCAAAAATCC AAAGAATTGG AACAGCAGAT TGCGCAGTTG AAAAAAAGCT
2951 CGCNTAAAAG CAAGCTGATT CCAACCCTGC AAGAAGAACG CGACCGTCTC
3001 GCTTTCTATA TTCAAGCCAT CAACAAGGAA GTTAAAGGTA AAAAACCCAA
3051 AGGCAAAGAA TACCTGCAAG CCAAGCTTTC TGCACAAAAT ATTGACTTGA
3101 TTTCCGCACA AGGCATCGAA ATCAGCGGTT CCGATATTAC CGCTTCCAAA
3151 AAACTGAACC TTCACGCCGC AGGCGTATTG CCAAAGGCAG CAGATTCAGA
3201 GGCGGCTGCT ATTCTGATTG ACGGCATAAC CGACCAATAT GAAATTGGCA
3251 AGCCCACCTA CAAGAGTCAC TACGACAAAG CTGCTCTGAA CAAGCCTTCA
3301 CGTTTGACCG GACGTACGGG GGTAAGTATT CATGCAGCTG CGGCACTCGA
3351 TGATGCACGT ATTATTATCG GTGCATCCGA AATCAAAGCT CCCTCAGGCA
3401 GCATAGACAT CAAAGCCCAT AGTGATATTG TACTGGAGGC TGGACAAAAC
3451 GATGCCTATA CCTTCTTANA AACCAAAGGT AAAAGCGGCA NAATNATCAG
3501 AAAAACNAAG TTTACCAGCA CCNGCGANCA CCTGATTATG CCAGCCCCNG
3551 TCGAGCTGAC CGCCAACGGT ATCACGCTTC AGGCAGGCGG CAACATCGAA
3601 GCTAATACCA CCCGCTTCAA TGCCCCTGCA GGTAAAGTTA CCCTGGTTGC
3651 GGGTGAANAG NTGCAACTGC TGGCAGAAGA AGGCATCCAC AAGCACGAGT
3701 TGGATGTCCA AAAAAGCCGC CGCTTTATCG GCATCAAGGT AGGTNAGAGC
3751 AATTACAGTA AAAACGAACT GAACGAAACC AAATTGCCTG TCCGCGTCGT
3801 CGCCCAAANT GCAGCCACCC GTTCAGGCTG GGATACCGTG CTCGAAGGTA
3851 CCGAATTCAA AACCACGCTG GCCGGTGCCG ACATTCAGGC AGGTGTANGC
3901 GAAAAAGCCC GTGTCGATGC GAAAATTATC CTCAAAGGCA TTGTGAACCG
3951 TATCCAGTCG GAAGAAAAAT TAGAAACCAA CTCAACCGTA TGGCAGAAAC
4001 AGGCCGGACG CGGCAGCACT ATCGAAACGC TAAAACTGCC CAGCTTCGAA
4051 AGCCCTACTC CGCCCAAATT GTCCGCACCC GGCGGNTATA TCGTCGACAT
4101 TCCGAAAGGC AATCTGAAAA CCGAAATCGA AAAGCTGTCC AAACAGCCCG
4151 AGTATGCCTA TCTGAAACAC CTCCAAGTAG CGAAAAACAT CAACTGGAAT
4201 CAGGTGCAGC TTGCTTACGA CAGATGGGAC TACAAACAGG AGGGCTTAAC
4251 CGAAGCAGGT GCGGCGATTA TCGCACTGGC CGTTACCGTG GTCACCTCAG
4301 GCGCAGGAAC CGGAGCCGTA TTGGGATTAA ACGGTGCGNC CGCCGCCGCA
4351 ACCGATGCAG CATTCGCCTC TTTGGCCAGC CAGGCTTCCG TATCGTTCAT
4401 CAACAACAAA GGCGATGTCG GCAAAACCCT GAAAGAGCTG GGCAGAAGCA
4451 GCACGGTGAA AAATCTGGTG GTTGCCGCCG CTACCGCAGG CGTAGCCGAC
4501 AAAATCGGCG CTTCGGCACT GANCAATGTC AGCGATAAGC AGTGGATCAA
4551 CAACCTGACC GTCAACCTAG CCAATGNCGG GCAGTGCCGC ACTGAttaa

This encodes a protein having amino acid sequence <SEQ ID 62>:

1    MNKGLHRIIF SKKHSTMVAV AETANSQGKG KQAGSSVSVS LKTSGDLCGK
51   LKTTLKTLVC SLVSLSMXXX XXXQITTDKS APKNXQVVIL KTNTGAPLVN
101  IQTPNGRGLS HNRYTQFDVD NKGAVLNNDR NNNPFLVKGS AQLILNEVRG
151  TASKLNGIVT VGGQKADVII ANPNGITVNG GGFKNVGRGI LTIGAPQIGK
201  DGALTGFDVR QGTLTVGAAG WNDKGGADYT GVLARAVALQ GKLQGKNLAV
251  STGPQKVDYA SGEISAGTAA GTKPTIALDT AALGGMYADS ITLIAXEKGV
301  GVKNAGTLEA AKQLIVTSSG RIENSGRIAT TADGTEASPT YLXIETTEKG
351  AXGTFISNGG RIESKGLLVI ETGEDIXLRN GAVVQNNGSR PATTVLNAGH
401  NLVIESKTNV NNAKGSXNLS AGGRTTINDA TIQAGSSVYS STKGDTXLGE
451  NTRIIAENVT VLSNGSIGSA AVIEAKDTAH IESGKPLSLE TSTVASNIRL
501  NNGNIKGGKQ LALLADDNIT AKTTNLNTPG NLYVHTGKDL NLNVDKDLSA
551  ASIHLKSDNA AHITGTSKTL TASKDMGVEA GLLNVTNTNL RTNSGNLHIQ
601  AAKGNIQLRN TKLNAAKALE TTALQGNIVS DGLHAVSADG HVSLLANGNA
651  DFTGHNTLTA KADVXAGSVG KGRLKADNTN ITSSSGDITL VAXXGIQLGD
701  GKQRNSINGK HISIKNNGGN ADLKNLNVHA KSGALNIHSD RALSIENTKL
751  ESTHNTHLNA QHERVTLNQV DAYAHRHLSI XGSQIWQNDK LPSANKLVAN
801  GVLAXNARYS QIADNTTLRA GAINLTAGTA LVKRGNINWS TVSTKTLEDN
851  AELKPLAGRL NIEAGSGTLT IEPANRISAH TDLSIKTGGK LLLSAKGGNA
901  GAXSAQVSSL EAKGNIRLVT GXTDLRGSKI TAGKNLVVAT TKGKLNIEAV
951  NNSFSNYFXT QKXXXLNQKS KELEQQIAQL KKSSXKSKLI PTLQEERDRL
1001 AFYIQAINKE VKGKKPKGKE YLQAKLSAQN IDLISAQGIE ISGSDITASK
1051 KLNLHAAGVL PKAADSEAAA ILIDGITDQY EIGKPTYKSH YDKAALNKPS
1101 RLTGRTGVSI HAAAALDDAR IIIGASEIKA PSGSIDIKAH SDIVLEAGQN
1151 DAYTFLXTKG KSGXXIRKTK FTSTXXHLIM PAPVELTANG ITLQAGGNIE
1201 ANTTRFNAPA GKVTLVAGEX XQLLAEEGIH KHELDVQKSR RFIGIKVGXS
1251 NYSKNELNET KLPVRVVAQX AATRSGWDTV LEGTEFKTTL AGADIQAGVX
1301 EKARVDAKII LKGIVNRIQS EEKLETNSTV WQKQAGRGST IETLKLPSFE
1351 SPTPPKLSAP GGYIVDIPKG NLKTEIEKLS KQP                                                                        EYAYLKQ LQVAKNINWN
1401 QVQLAYDRWD YKQEGLTEAG AAIIALAVTV VTSGAGTGAV LGLNGAXAAA
1451 TDAAFASLAS QASVSFINNK GDVGKTLKEL GRSSTVKNLV VAAATAGVAD
1501 KIGASALXNV SDKQWINNLT VNLANXGQCR TD*

ORF114-1 (SEQ ID NO: 179) and ORF 114a (SEQ ID NO:62) show 89.8% identity in 1564 aa overlap

orf114a.pep	MNKGLHRIIFSKKHSTMVAVAETANSQGKGKQAGSSVSVSLKTSGDLCGKLKTTLKTLVC
	||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	MNKGLHRIIFSKKHSTMVAVAETANSQGKGKQAGSSVSVSLKTSGDLCGKLKTTLKTLVC
orf114a.pep	SLVSLSMXXXXXXQITTDKSAPKNXQVVILKTNTGAPLVNIQTPNGRGLSHNRYTQFDVD
	|||||||      ||||||||||| |||||||||||||||||||||||||||||||||||
orf114-1	SLVSLSMVLPAHAQITTDKSAPKNQQVVILKTNTGAPLVNIQTPNGRGLSHNRYTQFDVD
orf114a.pep	NKGAVLNNDRNNNPFLVKGSAQLILNEVRGTASKLNGIVTVGGQKADVIIANPNGITVNG
	|||||||||||||||:||||||||||||||||||||||||||||||||||||||||||||
orf114-1	NKGAVLNNDRNNNPFVVKGSAQLILNEVRGTASKLNGIVTVGGQKADVIIANPNGITVNG
orf114a.pep	GGFKNVGRGILTIGAPQIGKDGALTGFDVRQGTLTVGAAGWNDKGGADYTGVLARAVALQ
	|||||||||||| |||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	GGFKNVGRGILTTGAPQIGKDGALTGFDVRQGTLTVGAAGWNDKGGADYTGVLARAVALQ
orf114a.pep	GKLQGKNLAVSTGPQKVDYASGEISAGTAAGTKPTIALDTAALGGMYADSITLIAXEKGV
	||||||||||||||||||||||||||||||||||||||||||||||||||||||| ||||
orf114-1	GKLQGKNLAVSTGPQKVDYASGEISAGTAAGTKPTIALDTAALGGMYADSITLIANEKGV
orf114a.pep	GVKNAGTLEAAKQLIVTSSGRIENSGRIATTADGTEASPTYLXIETTEKGAXGTFISNGG
	|||||||||||||||||||||||||||||||||||||||||| |||||||| ||||||||
orf114-1	GVKNAGTLEAAKQLIVTSSGRIENSGRIATTADGTEASPTYLSIETTEKGAAGTFISNGG
orf114a.pep	RIESKGLLVIETGEDIXLRNGAVVQNNGSRPATTVLNAGHNLVIESKTNVNNAKGSXNLS
	|||||||||||||||| ||||||||||||||||||||||||||||||||||||||  :||
orf114-1	RIESKGLLVIETGEDISLRNGAVVQNNGSRPATTVLNAGHNLVIESKTNVNNAKGPATLS
orf114a.pep	AGGRTTINDATIQAGSSVYSSTKGDTXLGENTRIIAENVTVLSNGSIGSAAVIEAKDTAH
	| |||:|::|:||:|::||||:||:: ||:|||| : :|||||||:|:|:|||:||||||
orf114-1	ADGRTVIKEASIQTGTTVYSSSKGNAELGNNTRITGADVTVLSNGTISSSAVIDAKDTAH
orf114a.pep	IESGKPLSLETSTVASNIRLNNGNIKGGKQLALLADDNITAKTTNLNTPGNLYVHTGKDL
	||:|||||||:|||:|:||||:|:||||||||||||||||||||||||||||||||||||
orf114-1	IEAGKPLSLEASTVTSDIRLNGGSIKGGKQLALLADDNITAKTTNLNTPGNLYVHTGKDL
orf114a.pep	NLNVDKDLSAASIHLKSDNAAHITGTSKTLTASKDMGVEAGLLNVTNTNLRTNSGNLHIQ
	||||||||||||||||||||||||||||||||||||||||| ||||||||||||||||||
orf114-1	NLNVDKDLSAASIHLKSDNAAHITGTSKTLTASKDMGVEAGSLNVTNTNLRTNSGNLHIQ
orf114a.pep	AAKGNIQLRNTKLNAAKALETTALQGNIVSDGLHAVSADGHVSLLANGNADFTGHNTLTA
	||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	AAKGNIQLRNTKLNAAKALETTALQGNIVSDGLHAVSADGHVSLLANGNADFTGHNTLTA
orf114a.pep	KADVXAGSVGKGRLKADNTNITSSSGDITLVAXXGIQLGDGKQRNSINGKHISIKNNGGN
	|||| |||||||||||||||||||||||||||  ||||||||||||||||||||||||||
orf114-1	KADVNAGSVGKGRLKADNTNITSSSGDITLVAGNGIQLGDGKQRNSINGKHISIKNNGGN
orf114a.pep	ADLKNLNVHAKSGALNIHSDRALSIENTKLESTHNTHLNAQHERVTLNQVDAYAHRHLSI
	||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	ADLKNLNVHAKSGALNIHSDRALSIENTKLESTHNTHLNAQHERVTLNQVDAYAHRHLSI
orf114a.pep	XGSQIWQNDKLPSANKLVANGVLAXNARYSQIADNTTLRAGAINLTAGTALVKRGNINWS
	:||||||||||||||||||||||| |||||||||||||||||||||||||||||||||||
orf114-1	TGSQIWQNDKLPSANKLVANGVLALNARYSQIADNTTLRAGAINLTAGTALVKRGNINWS
orf114a.pep	TVSTKTLEDNAELKPLAGRLNIEAGSGTLTIEPANRISAHTDLSIKTGGKLLLSAKGGNA
	||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	TVSTKTLEDNAELKPLAGRLNIEAGSGTLTIEPANRISAHTDLSIKTGGKLLLSAKGGNA
orf114a.pep	GAXSAQVSSLEAKGNIRLVTGXTDLRGSKITAGKNLVVATTKGKLNIEAVNNSFSNYFXT
	|| |||||||||||||||||| |||||||||||||||||||||||||||||||||||| |
orf114-1	GAPSAQVSSLEAKGNIRLVTGETDLRGSKITAGKNLVVATTKGKLNIEAVNNSFSNYFPT
orf114a.pep	QKXXXLNQKSKELEQQIAQLKKSSXKSKLIPTLQEERDRLAFYIQAINKEVKGKKPKGKE
	||   ||||||||||||||||||| |||||||||||||||||||||||||||||||||||
orf114-1	QKAAELNQKSKELEQQIAQLKKSSPKSKLIPTLQEERDRLAFYIQAINKEVKGKKPKGKE
orf114a.pep	YLQAKLSAQNIDLISAQGIEISGSDITASKKLNLHAAGVLPKAADSEAAAILIDGITDQY
	||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	YLQAKLSAQNIDLISAQGIEISGSDITASKKLNLHAAGVLPKAADSEAAAILIDGITDQY
orf114a.pep	EIGKPTYKSHYDKAALNKPSRLTGRTGVSIHAAAALDDARIIIGASEIKAPSGSIDIKAH
	||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf114-1	EIGKPTYKSHYDKAALNKPSRLTGRTGVSIHAAAALDDARIIIGASEIKAPSGSIDIKAH
orf114a.pep	SDIVLEAGQNDAYTFLXTKGKSGXXIRKTKFTSTXXHLIMPAPVELTANGITLQAGGNIE
	|||||||||||||||| ||||||  |||||||||  ||||||||||||||||||||||||
orf114-1	SDIVLEAGQNDAYTFLKTKGKSGKIIRKTKFTSTRDHLIMPAPVELTANGITLQAGGNIE
orf114a.pep	ANTTRFNAPAGKVTLVAGEXXQLLAEEGIHKHELDVQKSRRFIGIKVGXSNYSKNELNET
	|||||||||||||||||||  ||||||||||||||||||||||||||| |||||||||||
orf114-1	ANTTRFNAPAGKVTLVAGEELQLLAEEGIHKHELDVQKSRRFIGIKVGKSNYSKNELNET
orf114a.pep	KLPVRVVAQXAATRSGWDTVLEGTEFKTTLAGADIQAGVXEKARVDAKIILKGIVNRIQS
	|||||||||:||||||||||||||||||||||||||||| ||||:|||||||||||||||
orf114-1	KLPVRVVAQTAATRSGWDTVLEGTEFKTTLAGADIQAGVGEKARADAKIILKGIVNRIQS
orf114a.pep	EEKLETNSTVWQKQAGRGSTIETLKLPSFESPTPPKLSAPGGYIVDIPKGNLKTEIEKLS
	|||||||||||||||||||||||||||||||||||||:|||||||||||||||||||||:
orf114-1	EEKLETNSTVWQKQAGRGSTIETLKLFSFESFTFFKLTAPGGYIVDIPKGNLKTEIEKLA
orf114a.pep	KQPEYAYLKQLQVAKNINWNQVQLAYDRWDYKQEGLTEAGAAIIALAVTVVTSGAGTGAV
	||||||||||||||||:||||||||||:|||||||||:|||||::: | ::| | |: |:
orf114-1	KQPEYAYLKQLQVAKNVNWNQVQLAYDKWDYKQEGLTRAGAAIVTIIVTALTYGYGATAA
orf114a.pep	LGLNGA--------------XAAATD---------AAFASLASQASVSFINNKGDVGKTL	1477
	|: ::              :||||          ||:||| |||:||:|||||||||:|
orf114-1	GGVAASGSSTAAAAGTAATTTAAATTVSTATAMQTAALASLYSQAAVSIINNKGDVGKAL	1500
orf114a.pep	KELGRSSTVKNLVVAAATAGVADKIGA----------SALXNVSDKQWINNL----TVNL	1523
	|:|| |:|||::|::| |||: :::||          : | : : :| | ||    ::||
orf114-1	KDLGTSDTVKQIVTSALTAGALNQMGADIAQLNSKVRTELFSSTGNQTIANLGGRLATNL	1560
orf114a.pep	ANXGQCRTDX
	:| |
orf114-1	SNAGISAGINTAVN...

Homology with pspA Putative Secreted Protein of N.meningitidis (Accession Number AF030941)

ORF114 (SEQ ID NO:180) and pspA (SEQ ID NO:182) protein show 36% aa identity (SEQ ID NO:181) in 302aa overlap:

Orf114:	1	AVAETANSQGKGKQAGSSVSVSL----KTSGDXXXXXXXXXXXXXXXXXXXXXXXPAHAQ	56
		AVAE  +  GK  Q   + SV +      S                         PA A
pspA:	19	AVAENVHRDGKSMQDSEAASVRVTGAASVSSARAAFGFRMAAFSVMLALGVAAFSPAPAS	78
Orf114:	57	-ITTDKSAFKNQQVVILKTNTGAPLVNIQTFNGRGLSHNRXYAFDVDNKGAVLNNDRNN-	114
		I  DKSAPKNQQ VIL+T  G P VNIQTP+ +G+S NR   FDVD KG +LNN R+N
pspA:	79	GIIADKSAFKNQQAVILQTANGLPQVNIQTPSSQGVSVNRFKQFDVDEKGVILNNSRSNT	138
Orf114:	115	----------NPFVVKGSAQLILNEV-RGTASKLNGIVTVGGQKADVIIANPNGITVNGG	163
		NP + +G A++I+N++     S LNG + VGG++A+V++ANP+GI VNGG
pspA:	139	QTQLGGWIQGNPHLARGEARVIVNQIDSSNFSLLNGYIEVGGKRAEVVVANPSGIRVNGG	198
Orf114:	164	GFKNVGRGILTTGAPQIGKDGALTGFDVVKAHWTVXAAGWNDKGGAXYTGVLARAVALQG	223
		G  N     LT+G P +  +G LTGFDV      +   G  D   A YT +L+RA  +
pspA:	199	GLINAASVTLTSGVPVL-NNGNLTGFDVSSGKVVIGGKGL-DTSDADYTRILSKAAEINA	256
Orf114:	224	KXXGKXLAVSTGPQKVDYASGEISAGTAAGTK----PTIALDTAALGGMYADSITLIANE	279
		GK + V +G  K+D+        +A  +     PT+A+DTA LGGMYAD ITLI+ +
pspA:	257	GVWGKDVKVVSGKNKLDFDGSLAKTASAPSSSDSVTPTVAIDTATLGGMYADKITLISTD	316
Orf114:	280	KG	281
		G
p5pA:	317	NG	318

ORF114a is also homologous to pspA:

gi|2623258 (AF030941), putative secreted protein [Neisseria meningitidis] Length
= 2273
Score = 261 bits (659), Expect = 3e-68
Identities = 203/663 (30%), Positives = 314/663 (46%), Gaps = 76/663 (11%)
Query: 1	MNKGLHRIIFSKKHSTMVAVAETANSQGKGKQAGSSVSVSLK-----TSGDXXXXXXXXX	55
	MNK  +++IF+KK S M+AVAE  +  GK  Q  + SV +      +S
Sbjct: 1	MNKRCYKVIFNKKRSCMMAVAENVHRDGKSMQDSEAASVRVTGAASVSSARAAFGFRMAA	60
Query: 56	XXXXXXXXXXXXXXXXXXQITTDKSAPKNXQVVILKTNTGAPLVNIQTPNGRGLSHNRYT	115
	I  DKSAPKN Q VIL+T  G P VNIQTP+ +G+S NR+
Sbjct: 61	FSVMLALGVAAFSPAPASGIIADKSAPKNQQAVILQTANGLPQVNIQTPSSQGVSVNRFK	120
Query: 116	QFDVDNKGAVLNNDRNN-----------NPFLVKGSAQLILNEV-RGTASKLNGIVTVGG	163
	QFDVD KG +LNN R+N           NP L +G A++I+N++     S LNG + VGG
Sbjct: 121	QFDVDEKGVILNNSRSNTQTQLGGWIQGNPHLARGEARVIVNQIDSSNPSLLNGYIEVGG	180
Query: 164	QKADVIIANPNGITVNGGGFKNVGRGILTIGAPQIGKDGALTGFDVRQGTLTVGAAGWND	223
	++A+V++ANP+GI VNGGG  N     LT G P +  +G LTGFDV  G + +G  G  D
Sbjct: 181	KRAEVVVANPSGIRVNGGGLINAASVTLTSGVPVL-NNGNLTGFDVSSGKVVIGGKGL-D	238
Query: 224	KGGADYTGVLARAVALQGKLQGKNLAVSTGPQKVDYASGEISAGTAAGTK----PTIALD	279
	ADYT +L+RA  +   + GK++ V +G  K+D+        +A  +     PT+A+D
Sbjct: 239	TSDADYTRILSRAAEINAGVWGKDVKVVSGKNKLDFDGSLAKTASAFSSSDSVTPTVAID	298
Query: 280	TAALGGMYADSITLIAXEKGVGVKNAGTLEAAK-QLIVTSSGRIENSGRIATTADGTEAS	338
	TA LGGMYAD ITLI+ + G  ++N G + AA   + +++ G++ NSG I       +A+
Sbjct: 299	TATLGGMYADKITLISTDNGAVIRNKGRIFAATGGVTLSADGKLSNSGSI-------DAA	351
Query: 339	PTYLXTETTEKGAXGTFTSNGGRIESKGLLVIETGEDIXLRNGAVVQNNGSRPATTVLNA	398
	+  +T +        +  G I S    V++  + I  +  G +    GS     + +
Sbjct: 352	EITISAQTVD--------NRQGFIRSGKGSVLKVSDGINNQAGLI----GSAGLLDIRDT	399
Query: 399	GHNLVIESKTNVNNAKGS----XNLSAGGRTTINDATIQAGSSVYSSTKGDTXLGENTRI	454
	G     +S  ++NN  G+     ++S   ++  ND  + A   V S +  D   G+
Sbjct: 400	G-----KSSLHTNNTDGTIIAGKDVSLQAKSLDNDGILTAARDV-SVSLHDDFAGKRDIE	453
Query: 455	IAENVTVLSNGSTGSAAVIEAKDTAHIESGKPLSLETSTVASNTRLNNGNIKGGKQLALL	514
	+T  + G + +  +I+A DT  + + +  +  +  + S  R       G     L+
Sbjct: 454	AGRTLTFSTQGRLKNTRIIQAGDTVSLTAAQIDNTVSGKIQSGNRTGLNGKNGITNRGLI	513
Query: 515	ADDNIT-----AKTTNLNTPGNLYVMTGKDLNLNVDKDLSAASIHLKSDNAANTTGTSKT	569
	+ IT     AK+ N  T G +Y   G  + +  D  L+          AA
Sbjct: 514	NSNGITLLQTEAKSDNAGT-GRIY---GSRVAVKADTLLNREETVNGETKAA-------V	562
Query: 570	LTASKDMGVEAGXXXXXXXXXXXXSGNLHIQAA---KGNIQLRNTKL-NAAKALETTALQ	625
	+ A + + + A             SG+LHI +A      +Q  NT L N + A+E++
Sbjct: 563	IAARERLDTGAREIENREAALLSSSGDLHIGSALNGSRQVQGANTSLHNRSAAIESS---	619
Query: 626	GNI	628	(SEQ ID NO:183)
	GNI		(SEQ ID NO:184)
Sbjct: 620	GNI	622	(SEQ ID NO:185)
Score = 37.5 bits (85), Expect = 0.53
Identities = 87/432 (20%), Positives = 159/432 (36%), Gaps = 62/432 (14%)
Query: 239	LQGKLQGKNLAVSTGPQKVDYASGEISAGTAAGTKPTIALDTAALGGMYADSITLIAXEK	298
	LQG LQGKN+  + G    +  G I A  A   K        A   + + S T     +
Sbjct: 1023	LQGDLQGKNIFAAAGSDITN--TGSIGAENALLLK--------ASNNIESRSETRSNQNE	1072
Query: 299	GVGVKNAGTLEAAKQLIVTSSGRI--ENSGRIATTADGTEASPTYLXIETTEKGAXG-TF	355
	V+N G + A   L    +G +  +    I  TA            E T +   G T
Sbjct: 1073	QGSVRNIGRV-AGIYLTGRQNGSVLLDAGNNIVLTAS-----------ELTNQSEDGQTV	1120
Query: 356	ISNGGRIESKGLLVIETGEDIXLRNGAVVQNNGSRPATTVLNAGHNLVIESK-------T	408
	++ GG I S    +      I   +  V++   +   +T+   G NL + +K
Sbjct: 1121	LNAGGDIRSDTTGISRNQNTIFDSDNYVIRKEQNEVGSTIRTRG-NLSLNAKGDIRIRAA	1179
Query: 409	NVNNAKGSXNLSAGGRTTINDATIQAGSS--------VYSSTKGDTXLGENTRIIAENVT	460
	V + +G   L+AG      D  ++AG +        Y+   G     + TR +
Sbjct: 1180	EVGSEQGRLKLAAG-----RDIKVEAGKAHTETEDALKYTGRSGGGIKQKMTRHLKNQNG	1234
Query: 461	VLSNGSIGSAAVIEAKDTAHIESGKPLSLETSTVASNIRLNNGNIKGGKQLALLADDNIT	520
	+G++    +I         +G  +  +  T+ S    NN  +K  +  +  A+ N
Sbjct: 1235	QAVSGTLDGKEIILVSGRDITVTGSNIIADNHTILS--AKNNIVLKAAETRSRSAEMNKK	1292
Query: 521	AKTTNLNTPG-NLYVRTGKDLNLNVDKDLSAASIHLKSDN-------AAHITGTSKTLTA	572
	K+  + + G      + KD   N  + +S     + S N         H T T  T+++
Sbjct: 1293	EKSGLMGSGGIGFTAGSKKDTQTNRSETVSHTESVVGSLNGNTLISAGKHYTQTGSTISS	1352
Query: 573	SK-DMGVEAGXXXXXXXXXXXXSGNLHIQAAKG-----NIQLRNTKLNAAAALETTALQG	626
	+ D+G+ +G              +  +   KG     ++ + NT + A  A++     G
Sbjct: 1353	PQGDVGISSGKISIDAAQNRYSQESKQVYEQKGVTVAISVPVVNTVMGAVDAVKAVQTVG	1412
Query: 627	NIVSDGLHAVSA	638	(SEQ ID NO:186)
	+  ++A++A		(SEQ ID NO:187)
Sbjct: 1413	KSKNSRVNAMAA	1424	(SEQ ID NO:188)

Amino acids 1-1423 of ORF114-1 were cloned in the pGex vector and expressed in E.coli , as described above. ST-fusion expression was visible using SDS-PAGE, and FIG. 5 shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF114-1.

Based on these results, including the homology with the putative secreted protein of N.meningitidis and on the presence of a transmembrane domain, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 14

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 63>

1    CGCTTCATTC ATGATGAAGC AGTCGGCAGC AACATCGGCG GCGGCAAAAT
51   GATTGTTGCA GCCGGGCAGG ATATCAATGT ACGCGGCAnA ACCCTTATTT
101  CTGATAAGGG CATTGTTTTA AAAGCAGGAC ACGACATCGA TATTTCTACT
151  GCCCATAATC CCTATACCGC CAATCAATAC CACCACACCA wAAAwTCAGG
201  CGTCATGGGT ACTGGCGGAT TGGGCTTTAC TATCGGTAAC CGGAAAACTA
251  CCGATGACAC TGATCGTACC AATATTGTsC ATACAGGCAG CATTATAGGC
301  AGCCTGAaTG GAGACACCGT TACAGTTGCA GGAAACCGCT ACCGACAAAC
351  CGGCAGTACC GTCTCCAGCC CCGAGGGGCC CAATACCGTC ACAGCCAAAw
401  GCATAGATGT AGAGTTCGCA AACAACCGGT ATGCCACTGA CTACGcCCAT
451  ACCCA                                                                                                                                  g                                                                                                                          GGAA CAAAAAGGCC TTACCGTCGC CCTCAATGTC CCGGTTGTCC
501  AAGCTGCACA AAACTTCATA CAAGCAGCCC AAAATGTGGG CAAAAGTAAA
551  AATAAACGCG TTAATGCCAT GGCTGCAGCC AATGCTGCAT GGCAGAGTTA
601  TCAAGCAACC CAACAAATGC AACAATTTGC TCCAAGCAGC AGTGCGGGAC
651  AAGGTCAAAA CTACAATCAA AGCCCCAGTA TCAGTGTGTC CATTAC.TAC
701  GGCGAACAGA AAAGTCGTAA CGAGCAAAAA AGACATTACA CCGAAgCGGC
751  AgCAAGTCAA ATTATCGGCA AAGGGCAAAC CACACTTGCG GCAACAGGAA
801  GTGGGGAGCA GTCCAATATC AATATTACAG GTTCCGATGT CATCGGCCAT
851  GCAGGTACTC C.CTCATTGC CGACAACCAT ATCAGACTCC AATCTGCCAA
901  ACAGGACGGC AGCGAGCAAA GCAAAAACAA AAGCAGTGGT TGGAATGCAG
951  GCGTACGTnn CAAAATAGGC AACGGCATCA GGTTTGGAAT TACCGCCGGA
1001 GGAAATATCG GTAAAGGTAA AGAGCAAGGG GGAAGTACTA CCCACCGCCA
1051 CACCCATGTC GGCAGCACAA CCGGCAAAAC TACCATCCGA AGCGGCGGG                                                                                                                                  G
1101 GATACCACCC TCAAAGGTGT GCAGCTCATC GGCAAAGGCA TACAGGCAGA
1151 TACGCGCAAC CTGCATATAG AAAGTGTTCA AGATACTGAA ACCTATCAGA
1201 GCAAACAGCA AAACGGCAAT GTCCAAGTT                                                                                                                                  t                                                                                                                           ACTGTCGGTT ACGGATTCAG
1251 TGCAAGCGGC AGTTACCGCC AAAGCAAAGT CAAAGCAGAC CATGCCTCCG
1301 TAACCGGGCA AAgCGGTATT TATGCCGGAG AAGACGGCTA TCAAATyAAA
1351 GTyAGAGACA ACACAGACCT yAAGGGCGGT ATCATCACGT CTAGCCAAAG
1401 CGCAGAAGAT AAGGGCAAAA ACCTTTTTCA GACGGCCACC CTTACTGCCA
1451 GCGACATTCA AAACCACAGC CGCTACGAAG GCAGAAGCTT CGGCATAGGC
1501 GGCAGTTTCG ACCTGAACGG CGGCTGGGAC GGCACGGTTA CCGACAAACA
1551 AGGCAGGCCT ACCGACAGGA TAAGCCCGGC AGCCGGCTAC GGCAGCGACG
1601 GAGACAGCAA AAACAGCACC ACCCGCAGCG GCGTCAACAC CCACAACATA
1651 CACATCACCG ACGAAGCGGG ACAACTTGCC CGAACAGGCA GGACTGCAAA
1701 AGAAACCGAA GCGCGTATCT ACACCGGCAT CGACACCGAA ACTGCGGATC
1751 AACACTCAGG CCATCTGAAA AACAGCTTCG AC...

This corresponds to the amino acid sequence <SEQ ID 64; ORF116>:

1   RFIHDEAVGS NIGGGKMIVA AGQDINVRGX SLISDKGIVL KAGHDIDIST
51  AHNRYTGNEY HESXXSGVMG TGGLGFTIGN RKTTDDTDRT NIVHTGSIIG
101 SLNGDTVTVA GNRYRQTGST VSSFEGRNTV TAKXIDVEFA NNRYATDYAH
151 TQEQKGLTVA LNVPVVQAAQ NFIQAAQNVG KSKNKRVNAM AAANAAWQSY
201 QATQQMQQFA PSSSAGQGQN YNQSPSISVS IXYGEQKSRN EQKRHYTEAA
251 ASQIIGKGQT TLAATGSGEQ SNINITGSDV IGHAGTXLIA DNHIRLQSAK
301 QDGSEQSKNK SSGWNAGVRX KIGNGIRFGI TAGGNIGKGK EQGGSTTHRH
351 THVGSTTGKT TIRSGGDTTL KGVQLIGKGI QADTRNLHIE SVQDTETYQS
401 KQQNGNVQVT VGYGFSASGS YRQSKVKADH ASVTGQSGIY AGEDGYQIKV
451 RDNTDLKGGI ITSSQSAEDK GKNLFQTATL TASDIQNHSR YEGRSFGIGG
501 SFDLNGGWDG TVTDKQGRPT DRISPAAGYG SDGDSKNSTT RSGVNTHNIH
551 ITDEAGQLAR TGRTAKETEA RIYTGIDTET ADQHSGHLKN SFD...

Computer analysis of this amino acid sequence gave the following results:

Homology With pspA Putative Secreted Protein of N.meningitidis (Accession Number AF030941)

ORF116 (SEQ ID NO:189) and pspA (SEQ ID NO:191) protein show 38% aa identity (SEQ ID NO:190) in 502aa overlap:

Orf116: 6	EAVGSNIGGGKMTVAAGQDINVRGXSLISDKGIVLKAGHDIDISTAHNRYTGNEYHESXX	65
	+AV   + G ++I+ +G+DI V G ++I+D   +L A ++I +  A  R    E ++
PspA: 1235	QAVSGTLDGKEIILVSGRDITVTGSNIIADNHTILSAKNNIVLKAAETRSRSAEMNKKEK	1294
Orf116: 66	XXXXXXXXXXXXXXNRKXXXXXXRTNIVHTGSIIGSLNGDTVTVAGNRYRQTGSTVSSPE	125
	++K         + HT S++GSLNG+T+  AG  Y QTGST+SSP+
PspA: 1295	SGLMGSGGIGFTAGSKKDTQTNRSETVSHTESVVGSLNGNTLISAGKHYTQTGSTISSPQ	1354
Orf116: 126	GRNTVTAKXIDVEFANNRYATDYAHTQEQKGLTVALNVPXXXX---XXXXXXXXXXXGKS	182
	G   +++  I ++ A NRY+ +     EQKG+TVA++VP                  GKS
PspA: 1355	GDVGISSGK:SIDAAQNRYSQESKQVYEQKGVTVAISVPVVNTVMGAVDAVKAVQTVGKS	1414
Orf116: 183	KNKRVXXXXXXXXXWQSYQATQQMQQFA--PSSSAGQGQNYNQSPSISVSIXYGEQKSRN	240
	KN RV          +   +   +   A  P  +AGQG        ISVS+YGEQK+  +
PspA: 1415	KNSRVNAMAANALNKGVDSGVALYNAARNPKKAAGQG---------ISVSVTYGEQKNTS	1466
Orf116: 241	EQKRHYTEAAASQIIGKGQTTLAATGSGEQSNINITGSDVIGHAGTXLIADNHIRLQSAK	300
	E +   T+    +I G G+ +L A+G+G+ S I ITGSDV G  GT L A+N +++++A+
PspA: 1467	ESRIKGTQVQEGKITGGGKVSLTASGAGKDSRITITGSDVYGGKGTRLKAENAVQIEAAR	1526
Orf116: 301	QDGSEQSKNKSSGWNAGVRXKIGNGIRFGITAXXXXXXXXXXXXSTTHRHTHVGSTTGKT	360
	Q   E+S+NKS+G+NAGV   I  GI FG TA             T +R++H+GS   +T
PspA: 1527	QTHQERSENKSAGFNAGVAIAINKGISFGFTAGANYGKGYGNGDETAYRNSHIGSKDSQT	1586
Orf116: 361	TIRSGGDTTLKGVQLIGKGIQADTRNLHIESVQDTETYQSKQQNGNVQVTVGYGFSASGS	420
	I SGGDT +KG QL GKG+     +LHIES+QDT  ++ KQ+N + QVTVGYGFS  GS
PspA: 1587	AIESGGDTVIKGGQLKGKGVGVTAESLHIESLQDTAVFKGKQENVSAQVTVGYGFSVGGS	1646
Orf116: 421	YRQSKVKADRASVTGQSGIYAGEDGYQIKVRDNTDLKGGITTSSQSAEDKGKNLFQTATL	480
	Y +SK  +D+ASV  QSGI+AG DGY+I+V   T L G  + S     DK KNL +T+ +
PspA: 1647	YNRSKSSSDYASVNEQSGIFAGGDGYRIRVNGKTGLVGAAVVSD---ADKSKNLLKTSEI	1703
Orf116: 481	TASDIQNHSRYEGRSFGIGGSF	502
	DIQNH+     + G+ G F
PspA: 1704	WHKDIQNHASAAASALGLSGGF	1725

Based on homology with pspA, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 15

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 65>

1   ACGACCGGCA GCCTCGGCGG CATACTGGCC GGCGGCGGCA CTTCCCTTGC
51  CGCACCGTAT TTGGACAAAG CGGCGGAAAA CCTCGGTCCG GCGGGCAAAG
101 CGGCGGTCAA CGCACTGGGC GGTGCGGCCA TCGGCTATGC AACTGGTGGT
151 AGTGGTGGTG CTGTGGTGGG TGCGAATGTA GATTGGAACA ATAGGCAGCT
201 GCATCCGAAA GAAATGGCGT TGGCCGACAA ATATGCCGAA GCCCTCAAGC
251 GCGAAGTTGA AAAACGCGAA GGCAGAAAAA TCAGCAGCCA AGAAGCGGCA
301 ATGAGAATCC GCAGGCAGAT ATGCGTTGGG TGGACAAAGG TTCCCAAGAC
351 GGCTATACCG ACCAAAGCGT CATATCCCTT ATCGGAATGA

This corresponds to the amino acid sequence <SEQ ID 66; ORF 118>:

1   T                                                                        TGSLGGILA GGGTSLAAPY L                                                                    DKAAENLGP AGKAAVN                                                                        ALG GAAIGYATGG
51  SGGAVVGA                                                                    NV DWNNRQLHPK EMALADKYAE ALKREVEKRE GRKISSQEAA
101 MRIRRQICVG WTKVPKTAIP TKASYPLSE*

Computer analysis of this amino acid sequence reveals two putative transmembrane domains.

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 16

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 67>

1   CAATGCCGTC TGAAAAGCTC ACAATTTTAC AGACGGCATT TGTTATGCAA
51  GTACATATAC AGATTCCCTA TATACTGCCC AGrkGCGTGC GTgGCTGAAG
101 ACACCCCCTA CGCTTGCTAT TTGrAACAGC TCCAAGTCAC CAAAGACGTC
151 AACTGGAACC AGGTACwACT GGCGTACGAC AAATGGGACT ATAAACAGGA
201 AGGCTTAACC GGAGCCGGAG CAGCGATTAT TGCGCTGGCT GTTACCGTGG
251 TTACTGCGGG CGCGGGAgCC GGAGCCGCAC TGGGCTTAAA CGGCGCGGCc
301 GCAGCGGCAA CCGATGCCGC ATTCGCCTCG CTGGCCAGCC AGGCTTCCGT
351 ATCGCTCATC AaCAACAAAG GCAATATCGG TAaCACCCTG AAAGAGCTGG
401 GCAGAAGCAG CACGGTGAAA AATCTGATGG TTGCCGTCGc tACCGCAgGC
451 GTagCcgaCA AAATCGGTGC TTCGGCACTG AACAATGTCA GCGATAAGCA
501 GTGGATCAAC AACCTGACCG TCAACCTGGC CAATGCGGGC AGTGCCGCAC
551 TGATTAATAC CGCTGTCAAC GGCGGCAGCc tgAAAGACAA TCTGGAAGCG
601 AATATCCTTG CGGCTTTGGT GAATACTGCG CATGGAGAAG CAGCCAGTAA
651 AATCAAACAG TTGGATCAGC ACTACATTAC CCACAAGATT GCCCaTGCCA
701 TAGCGGGCTG TGCGGCTGCG GCGGCGAATA AGGGCAAGTG TCAGGATGGT
751 GCGATAgGTG CGGCTGTGGG CGAGATAGTC GGGGAgGCTT TGACAAACGG
801 CAAAAATCCT GACACTTTGA CAGCTAAAgA ACGCGaACAG ATTTTGGCAT
951 ACAGCAAACT GGTTGCCGGT ACCGTAAGCG GTGTGGTCGG CGGCGATGTA
901 AATGCGGCGG CGAATGCGGC TGAGGTAGCG GTGAAAAATA ATCAGCTTAG
951 CGACAAAtGA

This corresponds to the amino acid sequence <SEQ ID 68; ORF41>:

1   QCRLKSSQFY RRHLLCKYIY RFPIYCPXAC VAEDTPYACY LXQLQVTKDV
51  NWNQVXLAYD KWDYKQEG                                                                        LT GAGAAIIALA VTVVT                                                                    AGAGA GAALGLNGAA
101 AAATDAAFAS LASQASVSLI NNKGNIGNTL KELGRSSTVK NLMVAVATAG
151 VADKIGASAL NNVSDKQWIN NLTVNLANAG SAALINTAVN GGSLKDNLEA
201 NILAALVNTA HGEAASKIKQ LDQHYITHKI AHAIAGCAAA AANKGKCQDG
251 AIGAAVGEIV GEALTNGKNP DTLTAKEREQ ILAYSKLVAG TVSGVVGGDV
301 NAAANAAEVA VKNNQLSDK*

Further work revealed the complete nucleotide sequence <SEQ ID 69>:

1    ATGCAAGTAA ATATTCAGAT TCCCTATATA CTGCCCAGAT GCGTGCGTGC
51   TGAAGACACC CCCTACGCTT GCTATTTGAA ACAGCTCCAA GTCACCAAAG
101  ACGTCAACTG GAACCAGGTA CAACTGGCGT ACGACAAATG GGACTATAAA
151  CAGGAAGGCT TAACCGGAGC CGGAGCAGCG ATTATTGCGC TGGCTGTTAC
201  CGTGGTTACT GCGGGCGCGG GAGCCGGAGC CGCACTGGGC TTAAACGGCG
251  CGGCCGCAGC GGCAACCGAT GCCGCATTCG CCTCGCTGGC CAGCCAGGCT
301  TCCGTATCGC TCATCAACAA CAAAGGCAAT ATCGGTAACA CCCTGAAAGA
351  GCTGGGCAGA AGCAGCACGG TGAAAAATCT GATGGTTGCC GTCGCTACCG
401  CAGGCGTAGC CGACAAAATC GGTGCTTCGG CACTGAACAATGTCAGCGAT
451  AAGCAGTGGA TCAACAACCT GACCGTCAAC CTGGCCAATG CGGGCAGTGC
501  CGCACTGATT AATACCGCTG TCAACGGCGG CAGCCTGAAA GACAATCTGG
551  AAGCGAATAT CCTTGCGGCT TTGGTGAATA CTGCGCATGG AGAAGCAGCC
601  AGTAAAATCA AACAGTTGGA TCAGCACTAC ATTACCCACA AGATTGCCCA
651  TGCCATAGCG GGCTGTGCGG CTGCGGCGGC GAATAAGGGC AAGTGTCAGG
701  ATGGTGCGAT AGGTGCGGCT GTGGGCGAGA TAGTCGGGGA GGCTTTGACA
751  AACGGCAAAA ATCCTGACAC TTTGACAGCT AAAGAACGCG AACAGATTTT
801  GGCATACAGC AAACTGGTTG CCGGTACGGT AAGCGGTGTG GTCGGCGGCG
851  ATGTAAATGC GGCGGCGAAT GCGGCTGAGG TAGCGGTGAA AAATAATCAG
901  CTTAGCGACA AAGAGGGTAG AGAATTTGAT AACGAAATGA CTGCATGCGC
951  CAAACAGAAT AATCCTCAAC TGTGCAGAAA AAATACTGTA AAAAAGTATC
1001 AAAATGTTGC TGATAAAAGA CTTGCTGCTT CGATTGCAAT ATGTACGGAT
1051 ATATCCCGTA GTACTGAATG TAGAACAATC AGAAAACAAC ATTTGATCGA
1101 TAGTAGAAGC CTTCATTCAT CTTGGGAAGC AGGTCTAATT GGTAAAGATG
1151 ATGAATGGTA TAAATTATTC AGCAAATCTT ACACCCAAGC AGATTTGGCT
1201 TTACAGTCTT ATCATTTGAA TACTGCTGCT AAATCTTGGC TTCAATCGGG
1251 CAATACAAAG CCTTTATCCG AATGGATGTC CGACCAAGGT TATACACTTA
1301 TTTCAGGAGT TAATCCTAGA TTCATTCCAA TACCAAGAGG GTTTGTAAAA
1351 CAAAATACAC CTATTACTAA TGTCAAATAC CCGGAAGGCA TCAGTTTCGA
1401 TACAAACCTA AAAAGACATC TGGCAAATGC TGATGGTTTT AGTCAAAAAC
1451 AGGGCATTAA AGGAGCCCAT AACCGCACCA ATTTTATGGC AGAACTAAAT
1501 TCACGAGGAG GACGCGTAAA ATCTGAAACC CAAACTGATA TTGAAGGCAT
1551 TACCCGAATT AAATATGAGA TTCCTACACT AGACAGGACA GGTAAACCTG
1601 ATGGTGGATT TAAGGAAATT TCAAGTATAA AAACTGTTTA TAATCCTAAA
1651 AAATTTTCTG ATGATAAAAT ACTTCAAATG GCTCAAAATG CTGCTTCACA
1701 AGGATATTCA AAAGCCTCTA AAATTGCTCA AAATGAAAGA ACTAAATCAA
1751 TATCGGAAAG AAAAAATGTC ATTCAATTCT CAGAAACCTT TGACGGAATC
1801 AAATTTAGAT CATATTTTGA TGTAAATACA GGAAGAATTA CAAACATTCA
1851 CCCAGAATAA

This corresponds to the amino acid sequence <SEQ ID 70; ORF41-1>:

1   MQVNIQIPYI LPRCVRAEDT PYACYLKQLQ VTKDVNWNQV QLAYDKWDYK
51  QEGLTG                                                                        AGAA IIALAVTVVT AGA                                                                    CAGAALG LNGAAAAATD AAEASLASQA
101 SVSLINNKGN IGNTLKELGR SSTVKNLMVA VATAGVADKI GASALNNVSD
151 KQWTNNLTVN LANAGSAALI NTAVNGGSLK DNLEANILAA LVNTAHGEAA
201 SKIKQLDQHY ITHKIAHAIA GCAAAAANKG KCQDGAIGAA VGEIVGEALT
251 NGKNPDTLTA KEREQILAYS KLVAGTVSGV VGGDVNAAAN AAEVAVKNNQ
301 LSDKEGREFD NEMTACAKQN NPQLCRKNTV KKYQNVADKR LAASIAICTD
351 ISRSTECRTI RKQHLIDSRS LHSSWEAGLI GKDDEWYKLF SKSYTQADLA
401 LQSYHLNTAA KSWLQSGNTK PLSEWMSDQG YTLISGVNPR FIPIPRGFVK
451 QNTPITNVKY PEGISFDTNL KRHLANADGF SQKQGIKGAH NRTNFMAELN
501 SRGGRVKSET QTDIEGITRI KYEIPTLDRT GKPDGGFKEI SSIKTVYNPK
551 KFSDDKILQM AQNAASQGYS KASKIAQNER TKSISERKNV IQFSETFDGI
601 KFRSYFDVNT GRITNIHPE*

Computer analysis of this of this amino acid sequence predicts a transmembrane domain, and homology with an ORF from N.meningitidis (strain A) was also found.

ORF41 (SEQ ID NO:192) shows 92.8% identity over a 279aa overlap with an ORF (ORF41a (SEQ ID NO:193)) from strain A of N.meningitidis :

          10       20         30        40        50        60       69
orf41.pep YPPHLLCKYIYRFPIYCPXACVAEDTPYACYLXQLQVTKDVNWNQVXLAYDKNDYKQEGL
          || ||||:|::||||| ||||:||||||||
orf41a    YLKQLQVAKNINWNQVQLAYDRWDYKQEGL
          10        20        30
          70        80        90       100       110       120      129
orf41.pep TGAGAAIIALAVTVVTAGAGAGAALGLNGAAAAATDAAFASLASQASVSLINNKGNIGNT
          | ||||||||||||||:|||:||:|||||| ||||||||||||||||||:|||||::|:|
ort41a    TEAGAAIIALAVTVVTSGAGTGAVLGLNGAXAAATDAAFASLASQASVSFINNKGDVGKT
          40        50        60        70        80        90
          130       140       150       160       170       180      189
orf41.pep LKELGRSSTVKNLMVAVATAGVADKIGASALNNVSDKQWINNLTVNLANAGSAALINTAV
          |||||||||||||:||:|||||||||||||| ||||||||||||||||||||||||||||
orf41a    LKELGPSSTVKNLVVAAATAGVADKIGASALXNVSDKQWINNLTVNLANAGSAALINTAV
          100       110       120       130       140       150
          190       200       210       220       230       240      249
orf41.pep NGGSLKDNLEANILAALVNTAHGEAASKIKQLDQHYITHKIAHAIAGCAAAAANKGKCQD
          ||||||| |||||||||||||||||||||||||||||:||||||||||||||||||||||
orf41a    NGGSLKDXLEANILAALVNTAHGEAASKIKQLDQHYIVHKIAHAIAGCAAAAANKGKCQD
          160       170       180       190       200       210
          250       260       270       280       290       300      309
orf41.pep GAIGAAVGEIVGEALTNGKNPDTLTAKEREQILAYSKLVAGTVSGVVGGDVNAAANAAEV
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf41a    GAIGAAVGEIVGEALTNGKNPDTLTAKEREQILAYSKLVAGTVSGVVGGDVNAAANAAEV
          220       230       240       250       260       270
          310       320
orf41.pep AVKNNQLSDKX
          |||||||||
orf41a    AVKNNQLSDXEGREFDNEMTACAKQNXPQLCRKNTVKKYQNVADKRLAASIAICTDISRS
          280       290       300       310       320       330

A partial ORF41a nucleotide sequence <SEQ ID 71> is:

1    TATCTGAAAC AGCTCCAAGT AGCGAAAAAC ATCAACTGGA ATCAGGTGCA
51   GCTTGCTTAC GACAGATGGG ACTACAAACA GGAGGGCTTA ACCGAAGCAG
101  GTGCGGCGAT TATCGCACTG GCCGTTACCG TGGTCACCTC AGGCGCAGGA
151  ACCGGAGCCG TATTGGGATT AAACGGTGCG NCCGCCGCCG CAACCGATGC
201  AGCATTCGCC TCTTTGGCCA GCCAGGCTTC CGTATCGTTC ATCAACAACA
251  AAGGCGATGT CGGCAAAACC CTGAAAGAGC TGGGCAGAAG CAGCACGTGC
301  AAAAATCTGG TGGTTGCCGC CGCTACCGCA GGCGTAGCCG ACAAAATCGG
351  CGCTTCGGCA CTGANCAATG TCAGCGATAA GCAGTGGATC AACAACCTGA
401  CCGTCAACCT AGCCAATGCG GGCAGTGCCG CACTGATTAA TACCGCTGTC
451  AACGGCGGCA GCCTGAAAGA CANTCTGGAA GCGAATATCC TTGCGGCTTT
501  GGTCAATACC GCGCATGGAG AAGCAGCCAG TAAAATCAAA CAGTTGGATC
551  AGCACTACAT AGTCCACAAG ATTGCCCATG CCATAGCGGG CTGTGCGGCA
601  GCGGCGGCGA ATAAGGGCAA GTGTCAGGAT GGTGCGATAG GTGCGGCTGT
651  GGGCGAGATA GTCGGGGAGG CTTTGACAAA CGGCAAAAAT CCTGACACTT
701  TGACAGCTAA AGAACGCGAA CAGATTTTGG CATACAGCAA ACTGGTTGCC
751  GGTACGGTAA GCGGTGTGGT CGGCGGCGAT GTAAATGCGG CGGCGAATGC
801  GGCTGAGGTA GCGGTGAAAA ATAATCAGCT TAGCGACNAA GAGGGTAGAG
851  AATTTGATAA CGAAATGACT GCATGCGCCA AACAGAATAN TCCTCAACTG
901  TGCAGAAAAA ATACTGTAAA AAAGTATCAA AATGTTGCTG ATAAAAGACT
951  TGCTGCTTCG ATTGCAATAT GTACGGATAT ATCCCGTAGT ACTGAATGTA
1001 GAACAATCAG AAAACAACAT TTGATCGATA GTAGAAGCCT TCATTCATCT
1051 TGGGAAGCAG GTCTAATTGG TAAAGATGAT GAATGGTATA AATTATTCAG
1101 CAAATCTTAC ACCCAAGCAG ATTTGGCTTT ACAGTCTTAT CATTTGAA7A
1151 CTGCTGCTAA ATCTTGGCTT CAATCGGGCA ATACAAAGCC TTTATCCGAA
1201 TGGATGTCCG ACCAAGGTTA TACACTTATT TCAGGAGTTA ATCCTAGATT
1251 CATTCCAATA CCAAGAGGGT TTGTAAAACA AAATACACCT ATTACTAATG
1301 TCAAATACCC GGAAGGCATC AGTTTCGATA CAAACCTANA AAGACATCTG
1351 GCAAATGCTG ATGGTTTTAG TCAAGAACAG GGCATTAAAG GAGCCCATAA
1401 CCGCACCAAT NTTATGGCAG AACTAAATTC ACGAGGAGGA NGNGTAAAAT
1451 CTGAAACCCA NACTGATATT GAAGGCATTA CCCGAATTAA ATATGAGATT
1501 CCTACACTAG ACAGGACAGG TAAACCTGAT GGTGGATTTA AGGAAATTTC
1551 AAGTATAAAA ACTGTTTATA ATCCTAAAAA NTTTTNNGAT GATAAAATAC
1601 TTCAAATGGC TCAANATGCT GNTTCACAAG GATATTCAAA AGCCTCTAAA
1651 ATTGCTCAAA ATGAAAGAAC TAAATCAATA TCGGAAAGAA AAAATGTCAT
1701 TCAATTCTCA GAAACCTTTG ACGGAATCAA ATTTAGANNN TATNTNGATG
1751 TAAATACAGG AAGAATTACA AACATTCACC CAGAATAA

This encodes a protein having the partial amino acid sequence <SEQ ID 72>:

1   YLKQLQVAKN INWNQVQLAY DRWDYKQEGL TE                                                                        AGAAIIAL AVTVVTSGA                                                                    G
51  TGAVLGLNGA XAAATDAAFA SLASQASVSF INNKGDVGKT LKELGRSSTV
101 KNLVVAAATA GVADKIGASA LXNVSDKQWI NNLTVNLANA GSAALINTAV
151 NGGSLKDXLE ANILAALVNT AHGEAASKIK QLDQRYIVHK IAHAIAGCAA
201 AAANKGKCQD GAIGAAVGEI VGEALTNGKN PDTLTAKERE QILAYSKLVA
251 GTVSGVVGGD VNAAANAAEV AVKNNQLSDX EGREFDNEMT ACAKQNX+190 QL
301 CRKNTVKKYQ NVADKRLAAS IAICTDISRS TECRTIRKQH LIDSRSLHSS
351 WEAGLIGKDD EWYKLFSKSY TQADLALQSY HLNTAAKSWL QSGNTKFLSE
401 WMSDQGYTLI SGVNPRFIPI PRGFVKQNTP ITNVKYPEGI SFDTNLXRHI
451 ANADGFSQEQ GIKGAHNRTN XMAELNSRGG XVKSETXTDI EGITRIKYEI
501 PTLDRTGKPD GGFKEISSIK TVYNPKXFXD DKILQMAQXA XSQGYSKASK
551 IAQNERTKSI SERKNVIQFS ETFDGIKFRX YXDVNTGRIT NIHPE*

ORF41a (SEQ ID NO:72) and ORF41-1 (SEQ ID NO:70) show 94.8% identity in 595 aa overlap:

           10        20        30
orf41a.pep YLKQLQVAKNINWNQVQLAYDRWDYKQEGLTEAGAA
           |||||||:|::||||||||||:|||||||||:||||
orf41-1    MQVNIQIPYILPRCVRAEDTPYACYLKQLQVTKDVNWNQVQLAYDKWDYKQEGLTGAGAA
           10        20        30        40        50        60
           40        50        60        70        80        90
orf41a.pep ITALAVTVVTSGAGTGAVLGLNGAXAAATDAAFASLASQASVSFINNKGDVGKTLKELGR
           ||||||||||:|||:||:|||||| ||||||||||||||||||:|||||::|:|||||||
orf41-1    IIALAVTVVTAGAGAGAALGLNGAAAAATDAAFASLASQASVSLINNKGNIGNTLKELGR
           70        80        90       100       110       120
           100       110       120       130       140       150
orf40a.pep SSTVKNLVVAAATAGVADKIGASALXNVSDKQWINNLTVNLANAGSAALINTAVNGGSLK
           |||||||:||:|||||||||||||| ||||||||||||||||||||||||||||||||||
orf41-1    SSTVKNLMVAVATAGVADKIGASALNNVSDKQWINNLTVNLANAGSAALINTAVNGGSLK
           130       140       150       160       170       180
           160       170       180       190       200       210
orf41a.pep DXLEANILAALVNTAHGEAASKIKQLDQHYIVHKIAHAIAGCAAAAANKGKCQDGAIGAA
           | |||||||||||||||||||||||||||||:||||||||||||||||||||||||||||
orf41-1    DNLEANILAALVNTAHGEAASKIKQLDQHYITHKIAHAIAGCAAAAANKGKCQDGAIGAA
           190       200       210       220       230       240
           220       230       240       250       260       270
orf41a.pep VGEIVGEALTNGKNPDTLTAKEREQILAYSKLVAGTVSGVVGGDVNAAANAAEVAVKNNQ
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf41-1    VGEIVGEALTNGKNPDTLTAKEREQILAYSKLVAGTVSGVVGGDVNAAANAAEVAVKNNQ
           250       260       270       280       290       300
           280       290       300       310       320       330
orf41a.pep LSDXEGREFDNEMTACAKQNXPQLCRKNTVKKYQNVADKRLAASIAICTDISRSTECRTI
           ||| |||||||||||||||| |||||||||||||||||||||||||||||||||||||||
orf41-1    LSDKEGREFDNEMTACAKQNNPQLCRKNTVKKYQNVADKRLAASIAICTDISRSTECRTI
           310       320       330       340       350       360
           340       350       360       370       380       390
orf41a.pep RKQHLIDSRSLHSSWEAGLIGKDDEWYKLFSKSYTQADLALQSYHLNTAAKSWLQSGNTK
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf41-1    RKQHLIDSRSLRSSWEAGLIGKDDEWYKLFSKSYTQADLALQSYHLNTAAKSWLQSGNTK
           370       380       390       400       410       420
           400       410       420       430       440       450
orf41a.pep PLSEWMSDQGYTLISGVNFRFIPIPRGFVKQNTPITNVKYPEGISFDTNLXRHLANADGF
           |||||||||||||||||||||||||||||||||||||||||||||||||| |||||||||
orf41-1    PLSEWMSDQGYTLISGVNPRFIPIPRGFVKQNTPITNVKYPEGISFDTNLKRRLANADGF
           430       440       450       460       470       480
           460       470       480       490       500       510
orf41a.pep SQEQGIKGAHNRTNXMAELNSRGGXVKSETXTDIEGITRIKYEIPTLDRTGKPDGGFKEI
           ||:||||||||||| ||||||||| ||||| |||||||||||||||||||||||||||||
orf41-1    SQKQGIKGAHNRTNFMAELNSRGGRVKSETQTDIEGITRIKYEIFTLDRTGKPDGGFKEI
           490       500       510       520       530       540
           520       530       540       550       560       570
orf41a.pep SSIKTVYNPKXFXDDKILQMAQXAXSQGYSKASKIAQNERTKSISERKNVIQFSETFDGI
           |||||||||| | ||||||||| | |||||||||||||||||||||||||||||||||||
orf41-1    SSIKTVYNPKKFSDDKILQMAQNAASQGYSKASKIAQNERTKSISERKNVIQFSETFDGI
           550       560       570       580       590       600
           580       590
orf41a.pep KFRXYXDVNTGRITNIHPEX
           ||| | ||||||||||||||
orf41-1    KFRSYFDVNTGRITNIHPEX
           610       620

Amino acids 25-619 of ORF41-1 were amplified as described above. FIG. 6 shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF41-1.

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 17

The following DNA sequence was identified in N.meningitidis <SEQ ID 73>

1   ATGGCAATCA TTACATTGTA TTATTCTGTC AATGGTATTT TAAATGTATG
51  TGCAAAAGCA AAAAATATTC AAGTAGTTCC CAATAATAAG AATATGGTTC
101 TTTTTGGGTT TTTGGsmrGC ATCATCGGCG GTTCAACCAA TGCCATGTCT
151 CCCATATTGT TAATATTTTT GCTTAGCGAA ACAGAAAATA AAAATcgTAT
201 CGTAAAATCA AGCAATCTAT GCTATCTTTT GGCGAAAATT GTTCAAATAT
251 ATATGCTAAG AGACCAGTAT TGGTTATTAA ATAAGAGTGA ATACGdTTTA
301 ATATTTTTAC TGTCCGTATT GTCTGTTATT GGATTGTATG TTGGAATTCG
351 GTTAAGGACT AAGATTAGCC CAaATTTTTT TAAAATGTTA ATTTTTATTG
401 tTTTATTGGT ATTGGCtCTG AAAATCGGGC AttCGGGTTT AAtCAAACTT
451 TAA

This corresponds to the amino acid sequence <SEQ ID 74; ORF51>:

1   MAIITLYYSV NGILNVCAKA KNIQVVANNK NMVLFG                                                                        FLXX IIGGSTNAMS
51  PIL                                                                    LIFLLSE TENKNRIVKS SNLCYLLAKI VQIYMLRDQY WLLNKS                                                                        EYXL
101 IFLLSVLSVI GLY                                                                    VGIRLRT KI                                                                        SPNFFKML IFIVLLVLA                                                                    L KIGHSGLIKL
151 *

Further work revealed the complete nucteotide sequence <SEQ ID 75>:

1   ATGCAAGAAA TAATGCAATC TATCGTTTTT GTTGCTGCCG CAATACTGCA
51  CGGAATTACA GGCATGGGAT TTCCGATGCT CGGTACAACC GCATTGGCTT
101 TTATCATGCC ATTGTCTAAG GTTGTTGCCT TGGTGGCATT ACCAAGCCTG
151 TTAATGAGCT TGTTGGTTCT ATGCAGCAAT AACAAAAAGG GTTTTTGGCA
201 AGAGATTGTT TATTATTTAA AAACCTATAA ATTGCTTGCT ATCGGCAGCG
251 TCGTTGGCAG CATTTTGGGG GTGAAGTTGC TTTTGATACT TCCAGTGTCT
301 TGGCTGCTTT TACTGATGGC AATCATTACA TTGTATTATT CTGTCAATGG
351 TATTTTAAAT GTATGTGCAA AAGCAAAAAA TATTCAAGTA GTTGCCAATA
401 ATAAGAATAT GGTTCTTTTT GGGTTTTTGG CAGGCATCAT CGGCGGTTCA
451 ACCAATGCCA TGTCTCCCAT ATTGTTAATA TTTTTGCTTA GCGAAACAGA
501 AAATAAAAAT CGTATCGTAA AATCAAGCAA TCTATGCTAT CTTTTGGCGA
551 AAATTGTTCA AATATATATG CTAAGAGACC AGTATTGGTT ATTAAATAAG
601 AGTGAATACG GTTTAATATT TTTACTGTCC GTATTGTCTG TTATTGGATT
651 GTATGTTGGA ATTCGGTTAA GGACTAAGAT TAGCCCAAAT TTTTTTAAAA
701 TGTTAATTTT TATTGTTTTA TTGGTATTGG CTCTGAAAAT CGGGCATTCG
751 GGTTTAATCA AACTTTAA

This corresponds to the amino acid sequence <SEQ ID 76; ORF51-1>:

1   MQEIMQSIVF VAAAILHGIT GMGFPMLGTT ALA                                                                    FIMPLSK V                                                                        VALVALPSL
51  LMSLLVL                                                                    CSN NKKGFWQEIV YYLKTYKLLA IGSVVGSILG VK                                                                        LLLILPVS
101 WLLLL                                                                                    MAII                                                                                                                          T LYYSVNGILN VCAKAKNIQV VANNKNMVLF GFLAG                                                                        IIGGS
151 TNAMSPILLI FL                                                                    LSETENKN RIVKSSNLCY LLAKIVQIYM LRDQYWLLNK
201 SEYG                                                                        LIFLLS VLSVIGLYVG I                                                                    RLRTKISPN                                                                         FFKNLIFIVL LVLALKI                                                                    GHS
251 GLIKL*

Computer analysis of this amino acid sequence reveals three putative transmembrane domains. A corresponding ORF from strain A of N.meningitidis was also identified:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF51 (SEQ ID NO:74) shows 96.7% identity over a 150aa overlap with an ORF (ORF51a (SEQ ID NO: 194)) from strain A of N.meningitidis :

          10        20        30
orf51.pep MAII                                                                    TLYYSVNGILNVCAKAKNIQVVANNK
          ||||||||||||||||||||||||||||||
orf51a    YKLLAIGSVVGSILGVK                                                                        LLLILPVSWLLLIMAII                                                                    TLYYSVNGILNVCAKAKNIQVVANNK
          80        90       100       110       120       130
          40        50        60        70        80        90
orf51.pep NMVLFGFLXX                                                                        IIGGSTNAMSPILLIFL                                                                    LSETENKNRIVKSSNLCYLLAKIVQIYMLRDQY
          ||||||||  |||||||||||||||||||||||||||:||||||||||||||||||||||
orf51a    NMVLFGFLAG                                                                        IIGGSTNAMSPILLIFL                                                                    LSETENKNRIAKSSNLCYLLAKIVQIYMLRDQY
          140       150       160       170       180       190
          100       110       120       130       140       150
orf51.pep WLLNKSEYX                                                                        LIFLLSVLSVIGLYVGI                                                                    RLRTKISPN                                                                        FFKMLIFIVLLVLALKI                                                                    GHSGLIKL
          |||||||| ||||||||||||||||||||||||||||||||||||||||||||:||||||
orf51a    WLLNKSEYG                                                                        LIFLLSVLSVIGLYVGI                                                                    RLRTKISPN                                                                        FFKMLIFIVLLVLALKI                                                                    GYSGLIKL
          200       210       220       230       240       250

ORF51-1 (SEQ ID NO:76) and ORF51a (SEQ ID NO:78) show 99.2% identity in 255 aa overlap:

orf51a.pep MQEIMQSIVFVAAAILHGITGMGFPMLGTTALAFIMPLSKVVALVALPSLLMSLLVLCSN
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf51-1    MQEIMQSIVFVAAAILHGITGMGFPMLGTTALAFIMPLSKVVALVALPSLLMSLLVLCSN
orf51a.pep NKKGFWQEIVYYLKTYKLLAIGSVVGSILGVKLLLILPVSWLLLLMAIITLYYSVNGILN
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf51-1    NKKGFWQEIVYYLKTYKLLAIGSVVGSILGVKLLLILPVSWLLLLMAIITLYYSVNGILN
orf51a.pep VCAKAKNIQVVANNKNMVLFGFLAGIIGGSTNAMSPILLIFLLSETENKNRIAKSSNLCY
           ||||||||||||||||||||||||||||||||||||||||||||||||||||:|||||||
orf51-1    VCAKAKNIQVVANNKNMVLFGFLAGIIGGSTNAMSPILLIFLLSETENKNRIVKSSNLCY
orf51a.pep LLAKIVQIYMLRDQYWLLNKSEYGLIFLLSVLSVIGLYVGIRLRTKISPNFFKMLIFIVL
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf51-1    LLAKIVQIYMLRDQYWLLNKSEYGLIFLLSVLSVIGLYVGIRLRTKISPNFFKMLIFIVL
ort51a.pep LVLALKIGYSGLIKLX
           ||||||||:|||||||
ort51-1    LVLALKIGHSGLIKLX

The complete length ORF51 a nucleotide sequence <SEQ ID 77> is:

1   ATGCAAGAAA TAATGCAATC TATCGTTTTT GTTGCTGCCG CAATACTGCA
51  CGGAATTACA GGCATGGGAT TTCCGATGCT CGGTACAACC GCATTGGCTT
101 TTATCATGCC ATTGTCTAAG GTTGTTGCCT TGGTGGCATT ACCAAGCCTG
151 TTAATGAGCT TGTTGGTTCT ATGCAGCAAT AACAAAAAGG GTTTTTGGCA
201 AGAGATTGTT TATTATTTAA AAACCTATAA ATTGCTTGCT ATCGGCAGCG
251 TCGTTGGCAG CATTTTGGGG GTGAAGTTGC TTTTGATACT TCCAGTGTCT
301 TGGCTGCTTT TACTGATGGC AATCATTACA TTGTATTATT CTGTCAATGG
351 TATTTTAAAT GTATGTGCAA AAGCAAAAAA TATTCAAGTA GTTGCCAATA
401 ATAAGAATAT GGTTCTTTTT GGGTTTTTGG GAGGCATCAT CGGCGGTTCA
451 ACCAATGCCA TGTCTCCCAT ATTGTTAATA TTTTTGCTTA GCGAAACAGA
501 GAATAAAAAT CGTATCGCAA AATCAAGCAA TCTATGCTAT CTTTTGGCAA
551 AAATTGTTCA AATATATATG CTAAGAGACC AGTATTGGTT ATTAAATAAG
601 AGTGAATACG GTTTAATATT TTTACTGTCC GTATTGTCTG TTATTGGATT
651 GTATGTTGGA ATTCGGTTAA GGACTAAGAT TAGCCCAAAT TTTTTTAAAA
701 TGTTAATTTT TATTGTTTTA TTGGTATTGG CTCTGAAAAT CGGGTATTCA
751 GGTTTAATCA AACTTTAA

This encodes a protein having amino acid sequence <SEQ ID 78>:

1   MQEIMQSIVF VAAAILHGIT GMGFPMLGTT ALA                                                                    FIMPLSK V                                                                        VALVALPSL
51  LMSLLVL                                                                    CSN NKKGFWQEIV YYLKTYKLLA IGSVVCSILG VK                                                                        LLLILPVS
101 WLLLLMAII                                                                    T LYYSVNGILN VCAKAKNIQV VANNKNMVLF GFLAG                                                                        IIGGS
151 TNAMSPILLI FL                                                                    LSETENKN RIAKSSNLCY LLAKIVQIYM LRDQYWLLNK
201 SEYG                                                                        LIFLLS VLSVIGLYVG I                                                                    RLRTKISPN                                                                         FFKMLIFIVL LVLALKI                                                                    GYS
251 GLIKL*

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 18

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 79>

1   ATGAGACATA TGAAAATACA AAATTATTTA CTAGTATTTA TAGTTTTACA
51  TATAGCCTTG ATAGTAATTA ATATAGTGTT TGGTTATTTT GTTTTTCTAT
101 TTGATTTTTT TGCGTTTTTG TTTTTTGCAA ACGTCTTTCT TGCTGTAAAT
151 TTATTATTTT TAGAAAAAAA CATAAAAAAC AAATTATTGT TTTTATTGCC
201 GATTTCTATT ATTATATGGA TGGTAATTCA TATTAGTATG ATAAATATAA
251 AATTTTATAA ATTTGAGCAT CAAATAAAGG AACAAAATAT ATCCTCGATT
301 ACTGGGGTGA TAAAACCACA TGATAGTTAT AATTATGTTT ATGACTCAAA
351 AAACACCTTA TATTGATGTA GTTGCATCTG ATGTTAAAAA TAAATCCATA
401 AAACACCTTA TATTGATGTA GTTGCATCTG ATGTTAAAAA TAAATCCATA
451 AGATTAAGCT TGGTTTGTGG TATTCATTCA TATGCTCCAT GTGCCAATTT
501 TATAAAATTT GTCAGG..

This corresponds to the amino acid sequence <SEQ ID 80; ORF82>:

1   MRHMKIQNYL LVFIVLHIAL IVINIVFGYF VFLFDFFAFL FFANVFLA                                                                    VN
51  LLFLEKNIKN KLLFLLPISI IIWMVIHISM INIKFYKFEH QIKEQNISSI
101 TGVIKPHDSY NYVYDSNGYA KLKDNHRYGR VIRETPYIDV VASDVKNKSI
151 RLSLVCGIHS YAPCANFIKF VR..

Further work revealed the complete nucleotide sequence <SEQ ID 81>:

1   ATGAGACATA TGAAAAATAA AAATTATTTA CTAGTATTTA TAGTTTTACA
51  TATAGCCTTG ATAGTAATTA ATATAGTGTT TGGTTATTTT GTTTTTCTAT
101 TTGATTTTTT TGCGTTTTTG TTTTTTGCAA ACGTCTTTCT TGCTGTAAAT
151 TTATTATTTT TAGAAAAAAA CATAAAAAAC AAATTATTGT TTTTATTGCC
201 GATTTCTATT ATTATATGGA TGGTAATTCA TATTAGTATG ATAAATATAA
251 AATTTTATAA ATTTGAGCAT CAAATAAAGG AACAAAATAT ATCCTCGATT
301 ACTGGGGTGA TAAAACCACA TGATAGTTAT AATTATGTTT ATGACTCAAA
351 TGGATATGCT AAATTAAAAG ATAATCATAG ATATGGTAGG GTAATTAGAG
401 AAACACCTTA TATTGATGTA GTTGCATCTG ATGTTAAAAA TAAATCCATA
451 AGATTAAGCT TGGTTTGTGG TATTCATTCA TATGCTCCAT GTGCCAATTT
501 TATAAAATTT GCAAAAAAAC CTGTTAAAAT TTATTTTTAT AATCAACCTC
551 AAGGAGATTT TATAGATAAT GTAATATTTG AAATTGGTGA TGGAAACAAA
601 AGTTTGTACT TGTTAGATAA GTATAAAACA TTTTTTCTTA TTGAAAACAG
651 TGTTTGTATC GTATTAATTA TTTTATATTT AAAATTTAAT TTGCTTTTAT
701 ATAGGACTTA CTTCAATGAG TTGGAATAG

This corresponds to the amino acid sequence <SEQ ID 82; ORF82-1>:

1   MRHMKNKNYL LVFIVLHIAL IVINIVFGYF VFLFDFFAFL FFANVFLA                                                                    VN
51  LLFLEKNIKN K                                                                        LLFLLPISI IIWMVIHI                                                                    SM INIKFYKFEH QIKEQNISSI
101 TGVIKPHDSY NYVYDSNGYA KLKDNHRYGR VIRETPYIDV VASDVKNKSI
151 RLSLVCGIHS YAPCANFIKF AKKPVKIYFY NQPQGDFIDN VIFEINDGNK
201 SLYLLDKYKT                                                                         FFLIENSVCI VLIILYL                                                                    KFN LLLYRTYFNE LE*

Computer analysis of this amino acid sequence reveals a predicted leader peptide.

A corresponding ORF from strain A of N.meningitidis was also identified:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF82 (SEQ ID NO:80) shows 97.1% identity over a 172aa overlap with an ORF (ORF82a (SEQ ID NO:195)) from strain A of N.meningitidis :

          10        20        30        40        50        60
orf82.pep MRHMKIQNYLLVFIVLHIALIVINIVFGYFVFLFDFFAFLFFANVFLA                                                                    VNLLFLEKNIKN
          ||||| :|||||||||||:|||||||||||||||||||||||||||||||||||||||||
orf82a    MRHMKNKNYLLVFIVLHITLIVINIVFGYFVFLFDFFAFLFFANVFLA                                                                    VNLLFLEKNIKN
          10        20        30        40        50        60
          70        80        90       100       110       120
orf82.pep K                                                                        LLFLLPISIIIWMVIHI                                                                    SMINIKFYKFEHQIKEQNISSITGVIKPHDSYNYVYDSNGYA
          ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf82a    K                                                                        LLFLLPISIIIWMVIHI                                                                    SMINIKFYKFEHQIKEQNISSITGVIKPHDSYNYVYDSNGYA
          70        80        90       100       110       120
          130       140       150       160       170
orf82.pep KLKDNHRYGRVIRETPYIDVVASDVKNKSIRLSLVCGIHSYAPCANFIKFVR
          ||||||||||||||||||||||||||||||||||||||||||||||||||::
orf82a    KLKDNHRYGRVIRETPYIDVVASDVKNKSIRLSLVCGIHSYAPCANFIKFAKKPVKIYFY
          130       140       150       160       170       180

ORF82a (SEQ ID NO:84) and ORF82-1 (SEQ ID NO:82) show 99.2% identity in 242 aa overlap:

orf82a.pep MRHMKNKNYLLVFIVLHITLIVINIVFGYFVFLFDFFAFLFFANVFLAVNLLFLEKNIKN
           ||||||||||||||||||:|||||||||||||||||||||||||||||||||||||||||
orf82-1    MRHMKNKNYLLVFIVLHIALIVINIVFGYFVFLFDFFAFLFFANVFLAVNLLFLEKNIKN
orf82a.pep KLLFLLPISIIIWMVIHISMINIKFYKFEHQIKEQNISSITGVIKPHDSYNYVYDSNGYA
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf82-1    KLLFLLPISIIIWMVIHISMINIKFYKFEHQIKEQNISSITGVIKPHDSYNYVYDSNGYA
orf82a.pep KLKDNHRYGRVIRETPYIDVVASDVKNKSIRLSLVCGIHSYAPCANFIKFAKKPVKIYFY
           ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
orf82-1    KLKDNHRYGRVIRETPYIDVVASDVKNKSIRLSLVCGIHSYAPCANFIKFAKKPVKIYFY
orf82a.pep NQPQGDFIDNVIFEINDGKKSLYLLDKYKTFFLIENSVCIVLIILYLKFNLLLYRTYFNE
           ||||||||||||||||||:|||||||||||||||||||||||||||||||||||||||||
orf82-1    NQPQGDFIDNVIFEINDGNKSLYLLDKYKTFFLIENSVCIVLIILYLKFNLLLYRTYFNE
orf82a.pep LEX
           |||
orf82-1    LEX

The complere length ORF82a nucleotide sequence <SEQ ID 83> is:

1   ATGAGACATA TGAAAAATAA AAATTATTTA CTAGTATTTA TAGTTTTACA
51  TATAACCTTG ATAGTAATTA ATATAGTGTT TGGTTATTTT GTTTTTCTAT
101 TTGATTTTTT TGCGTTTTTG TTTTTTGCAA ACGTCTTTCT TGCTGTAAAT
151 TTATTATTTT TAGAAAAAAA CATAAAAAAC AAATTATTGT TTTTATTGCC
201 GATTTCTATT ATTATATGGA TGGTAATTCA TATTAGTATG ATAAATATAA
251 AATTTTATAA ATTTGAGCAT CAAATAAAGG AACAAAATAT ATCCTCGATT
301 ACTGGGGTGA TAAAACCACA TGATAGTTAT AATTATGTTT ATGACTGAAA
351 TGGATATGCT AAATTAAAAG ATAATCATAG ATATGGTAGG GTAATTAGAG
401 AAACACCTTA TATTGATGTA GTTGCATCTG ATGTTAAAAA TAAATCCATA
451 AGATTAAGCT TGGTTTGTGG TATTCATTCA TATGCTCCAT GTGCCAATTT
501 TATAAAATTT GCAAAAAAAC CTGTTAAAAT TTATTTTTAT AATCAACCTC
551 AAGGAGATTT TATAGATAAT GTAATATTTG AAATTAATGA TGGAAAAAAA
601 AGTTTGTACT TGTTAGATAA GTATAAAACA TTTTTTCTTA TTGAAAACAG
651 TGTTTGTATC GTATTAATTA TTTTATATTT AAAATTTAAT TTGCTTTTAT
701 ATAGGACTTA CTTCAATGAG TTGGAATAG

This encodes a protein having amino acid sequence <SEQ ID 84>:

1   MRHMKNKNYL LVFIVLHITL IVINIVFGYF VFLFDFFAFL FFANVFLA                                                                    VN
51  LLFLEKNIKN K                                                                        LLFLLPISI IIWMVIHI                                                                    SM INIKFYKFEH QIKEQNISSI
101 TGVIKPHDSY NYVYDSNGYA KLKDNHRYGR VIRETPYIDV VASDVKNKSI
151 RLSLVCGIHS YAPCANFIKF AKKPVKIYFY NQPQGDFIDN VIFElNDGKK
201 SLYLLDKYKT                                                                         FFLIENSVCI VLIILYL                                                                    KFN LLLYRTYFNE LE*

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 19

The following partial DNA sequence was identified in N.meningitidis <SEQ ID 85>

1   ACCCCCAACA GCGTGACCGT CTTGCCGTCT TTCGGCGGAT TCGGGCGTAC
51  CGGCGCGACC ATCAATGCAG CAGGCGGGGT CGGCATGACT GCCTTTTCGA
101 CAACCTTAAT TTCCGTAGCC GAGGGCGCGG TTGTAGAGCT GCAGGCCGTG
151 AGAGCCAAAG CCGTCAATGC AACCGCCGCT TGCATTTTTA CGGTCTTGAG
261 TAAGGACATT TTCGATTTCC TTTTTATTTT CCGTTTTCAG ACGGCTGACT
251 TCCGCCTGTA TTTTCGCCAA AGCCATGCCG ACAGCGTGCG CCTTGACTTC
301 ATATTTAAAA GCTTCCGCGC GTGCCAGTTC CAGTTCGCGC GCATAGTTTT
351 GAGCCGACAA CAGCAGGGCT TGCGCCTTGT CGCGCTCCAT CTTGTCGATG
401 ACCGCCTGCA GCTTCGCAAA TGCCGACTTG TAGCCTTGAT GGTGCGACAC
451 AGCCAAGCCC GTGCCGACAA GCGCGATAAT GGCAATCGGT TGCCAGTAAT
501 TCGCCAGCAG TTTCACGAGA TTCATTCTCG ACCTCCTGAC GCTTCACGCT
551 GA

This corresponds to the amino acid sequence <SEQ ID 86; ORF124>:

1   ..TPNSVTVLPS FGGFGRTGAT INAAGGVG                                                                        MT AF                                                                    STTLISVA EGAVVELQAV
51  RAKAVNATAA                                                                         CIFTVLSKDI FDFLFIF                                                                    RFQ TADFRLYFRQ SHADSVRLDF
101 IFKSFRACQF QFARIVLSRQ QQGLRLVALH LVDDRLQLRK CRLVALMVRH
151 SQARADKRDN GNRLPVIRQQ FHEIHSRPPD ASR*

Computer analysis of this amino acid sequence predicts a transmembrane domain.

Further work revealed the complete nucleotide sequence <SEQ ID 87>:

1   ATGACTGCCT TTTCGACAAC CTTAATTTCC GTAGCCGAGG GCGCGGTTGT
51  AGAGCTGCAG GCCGTGAGAG CCAAAGCCGT CAATGCAACC GCCGCTTGCA
161 TTTTTACGGT CTTGAGTAAG GACATTTTCG ATTTCCTTTT TATTTTCCGT
151 TTTCAGACGG CTGACTTCCG CCTGTTTTTT CGCCAAAGCC ATGCCGACAG
201 CGTGCGCCTT GACTTCATAT TTTTTAGCTT CCGCGCGTGC CAGTTCCAGT
251 TCGCGCGCAT AGTTTTGAGC CGACAACAGC AGGGCTTGCG CCTTGTCGCG
301 CTCCATCTTG TCGATGACCG CCTGCTGCTT CGCAAATGCC GACTTGTAGC
351 CTTGATGGTG CGACACAGCC AAGCCCGTGC CGACAAGCGC GATAATGGCA
401 ATCGGTTGCC AGTTATTCGC CAGCAGTTTC ACGAGATTCA TTCTCGACCT
451 CCTGACGCTT CACGCTGA

This corresponds to the amino acid sequence <SEQ ID 88; ORF124-1>:

1   MTAFSTTLIS VAEGAVVELQ AVRAKAVNAT AA                                                                        CIFTVLSK DIFDFLFIF                                                                    R
51  FQTADFRLFF RQSHADSVRL DFIFFSFRAC QFQFARIVLS RQQQGLRLVA
101 LHLVDDRLLL RKCRLVALMV RHSQARADKR DNGNRLPVIR QQFHEIHSRP
151 PDASR*

A corresponding ORF from strain A of N.meningitidis was also identified:

Homology With a Predicted ORF From N.meningitidis (Strain A)

ORF124 (SEQ ID NO:86) shows 87.5% identity over a 152aa overlap with an ORF (ORFI24a (SEQ ID NO:90)) from strain A of N.meningitidis :

           10        20        30        40        50        60
orf124.pep TPNSVTVLPSFGGFGRTGATINAAGGVGMTAFSTTLISVAEGAVVELQAVRAKAVNATAA
           |||||||||||||||:|||||| |||||:|||
orf124a    MTAFSTTLISVAEGALVELQAVMAKAVNTTAA
           10        20        30
           70        80        90       100       110       120
orf124.pep CIFTVLSKDIFDFLFIFRFQTADFRLYFRQSHADSVRLDFIFKSFRACQFQFARIVLSRQ
           ||||||||||||||||||||||||||:|||||||:||||||| |||:  |||| :|||||
orf124a    CIFTVLSKDIFDFLFIFRFQTADFRLFFRQSHADGVRLDFIFFSFRTRLFQFAGVVLSRQ
           40        50        60        70        80        90
           130       140       150       160       170       180
orf124.pep QQGLRLVALHLVDDRLQLRKCRLVALMVRHSQARADKRDNGNRLPVIRQQFHEIHSRPPD
           ||||||||||:::||| ||| ||||||||| |:||||||:||||||||||||||||||||
orf124a    QQGLRLVALHFLNDRLLLRKSRLVALMVRHRQTRADKRDDGNRLPVIRQQFHEIHSRPPD
           100       110       120       130       140       150
orf124.pep ASRX
           :
orf124a    VX

ORF124a (SEQ ID NO:90) and ORF124-1 (SEQ ID NO:88) show 89.5% identity in 152 aa overlap:

orf124-1.pep MTAFSTTLISVAEGAVVELQAVRAKAVNATAACIFTVLSKDIFDFLFIFRFQTADFRLFF
             |||||||||||||||:|||||| |||||:|||||||||||||||||||||||||||||||
orf124a      MTAFSTTLISVAEGALVELQAVMAKAVNTTAACIFTVLSKDIFDFLFIFRFQTADFRLFF
orf124-1.pep RQSHADSVRLDFIFFSFRACQFQFARIVLSRQQQGLRLVALHLVDDRLLLRKCRLVALMV
             ||||||:|||||||||||:  |||| :|||||||||||||||:::||||||| |||||||
orf124a      RQSHADGVRLDFIFFSFRTRLFQFAGVVLSRQQQGLRLVALHFLNDRLLLRKSRLVALMV
orf124-1.pep RHSQARADKRDNGNRLPVIRQQFHEIHSRPPDASRX
             || |:||||||:||||||||||||||||||||:
orf124a      RHRQTRADKRDDGNRLPVIRQQFHEIHSRPPDVX

The complete length ORF124a nucleotide sequence <SEQ ID 89> is:

1   ATGACCGCCT TTTCGACAAC CTTAATTTCC GTAGCCGAGG GCGCGCTTGT
51  AGAGCTGCAA GCCGTGATGG CCAAAGCCGT CAATACAACC GCCGCCTGCA
101 TTTTTACGGT CTTGAGTAAG GACATTTTCG ATTTCCTTTT TATTTTCCGT
151 TTTCAGACGG CTGACTTCCG CCTGTTTTTT CGCCAAAGCC ATGCCGACGG
201 CGTGCGCCTT GACTTCATAT TTTTTAGCTT CCGCACGCGC CTGTTCCAGT
251 TCGCGGGCGT AGTTTTGAGC CGACAACAGC AGGGCTTGCG CCTTGTCGCG
301 CTTCATTTTC TCAATGACCG CCTGCTGCTT CGCAAAAGCC GACTTGTAGC
351 CTTGATGGTG CGACACCGCC AAACCCGTGC CGACAAGCGC GATGATGGCA
401 ATCGGTTGCC AGTTATTCGC CAGCAGTTTC ACGAGATTCA TTCTCGACCT
451 CCTGACGTTT GA
PS

ORF124-1 was amplified as described above. FIG. 7 shows plots of hydrophilicity, antigenic index, and AMPHI regions for ORF 124-1.

Based on this analysis, it is predicted that this protein from N.meningitidis , and its epitopes, could be useful antigens for vaccines or diagnostics.

Example 20

Table III lists several Neisseria strains which were used to assess the conservation of the sequence of ORF 40 among different strains.

	TABLE III
	List of Neisseria Strains Used for Gene Variability
	Study of ORF 40
	Identification
	number	Strains	Source/reference
		Group B
	zn02_1	BZ198		R. Moxon/Seiler et al.,
				1996
	zn03_1	NG3/88		R. Moxon/Seiler et al.,
				1996
	zn04_1	297-0		R. Moxon/Seiler et al.,
				1996
	zn06_1	BZ147		R. Moxon/Seiler et al.,
				1996
	zn07_1	BZ169		R. Moxon/Seiler et al.,
				1996
	zn08_1	528		R. Moxon/Seiler et al.,
				1996
	zn10_1	BZ133		R. Moxon/Seiler et al.,
				1996
	zn11_1ass	NGE31		R. Moxon/Seiler et al.,
				1996
	zn14_1	NGH38		R. Moxon/Seiler et al.,
				1996
	zn16_1	NGH15		R. Moxon/Seiler et al.,
				1996
	zn18_1	BZ232		R. Moxon/Seiler et al.,
				1996
	zn19_1	BZ83		R. Moxon/Seiler et al.,
				1996
	zn20_1	44/76		R. Moxon/Seiler et al.,
				1996
	zn21_1	MC58		R. Moxon
		Group A
	zn22_1	205900		R. Moxon
	zn23_1	F6124		R. Moxon
	z2491_1	Z2491		R. Moxon/Maiden et al.,
				1998
		Group C
	zn24_1	90/18311		R. Moxon
	zn25_1ass	93/4286		R. Moxon
		Others
	zn28_1ass	860800	(group Y)	R. Moxon/Maiden et al.,
				1998
	zn29_1ass	E32	(group Z)	R. Moxon/Maiden et al.,
				1998
	References:
	Seiler A. et al., Mol. Microbiol., 1996, 19(4): 841-856.
	Maiden et al., Proc. Natl. Acad. Sci. USA, 1998, 95: 3140-3145.

The amino acid sequences for each listed strain are as follows:

>Z2491 <SEQ ID 91>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL
ESVQRSVVGSIQASMEGSGELETISLSMTNDSKEFVDPYIVVTLKAGDNLKIKQNTNENT
NASSFTYSLKKDLTGLINVETEKLSFGANGKKVNIISDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSENVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEISRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANKPV
RITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKS
MMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN01_1 <SEQ ID 92>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVATAVLATLLFATVQANATDDDDLYLE
PVQRTAVVLSFRSDKEGTGEKEGTEDSNWAVYFDEKRVLKAGAITLKAGDNLKIKQNTNE
NTNDSSFRYSLKKDLTDLTSVETEKLSFGANGNKVNITSDTKGLNFAKETAGTNGDPTVH
LNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNV
DFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKGKDENG
SSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTA
TVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDE
TVNINAGNNIEITRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNVGSKDTNK
PVRITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPG
KSMMAIGGDTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN03_1 <SEQ ID 93>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVATAVLATLLFATVQASTTDDDDLYLE
PVQRTAPVLSFHADSEGTGEKEVTEDSNWGVYFDKKGVLTAGTITLKAGDNLKIKQNTDE
NTNASSFTYSLKKDLTDLTSVETEKLSFGANGKKVNITSDTKGLNFAKETAGTNGDTTVH
LNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNV
DFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENG
SSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFASGNGTTA
TVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDE
TVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANK
PVRITNVAPGVKEGDVTNVAWLKGVAQNLNNHIDNVDGNARAGIAQAIATAGLVQAYLPG
KSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN04_1 <SEQ ID 94>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVATAVLATLLFATVQANATDDDDLYLE
PVQRTAVVLSFRSDKEGTGEKEGTEDSNWAVYFDEKRVLKAGAITLKAGDNLKIKQNTNE
NTNDSSFRYSLKKDLTDLTSVETEKLSFGANGNKVNITSDTKGLNFAKETAGTNGDPTVH
LNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNV
DFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKGKDENG
SSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTA
TVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDE
TVNINAGNNIEITRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNVGSKDTNK
PVRITNVAPGVKEGDVTNVAWLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPG
KSMMAIGGDTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN06_1 <SEQ ID 95>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVETAVLATLLFATVQASANNEEQEEDL
TLDPVQRTVAVLIVNSDKEGTGEKEKVEENSDWAVYGNEKGVLTAREITLKAGDNLKIKQ
NGTNFTYSLKKDLTDLTSVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDGDALNVGSKKDNKPVR
ITNVAPGVKEGDVTNVAWLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKSM
MAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>XN07_1 <SEQ ID 96>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVKTAVLATLLFATVQASANNEEQEEDL
YLDPVQRTVAVLIVNSDKEGTGEKEKVEENSDWAVYFNEKGVLTAREITLKAGDNLKIKQ
NGTNFTYSLKKDLTDLTSVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKGETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDGDALNVGSKKDNKPVR
ITNVAPGVKEGDVTNVAWLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKSM
MAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN08_1 <SEQ ID 97>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVETAVLATLLFATVQANATDTDEDDEL
EPVVRSALVLQFMIDKEGNGEIESTGDIGWSIYYDDHNTLHGATVTLKAGDNLKIKQNTD
ENTNASSFTYSLKKDLTDLTSVGTEELSFGANGNKVNITSDTKGLNFAKKTAGTNGDTTV
HLNGIGSTLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSEN
VDFVRTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGEN
GSSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTGASGKGTT
ATVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMD
ETVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSKDAN
KPVRITNVAPGVKEGDVTNVAQLKGVAWNLNNHIDNVDGNARAGIAQAIATAGLVQAYLP
GKSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN10_1 <SEQ ID 98>
MNKIYRIIWNSALNAWVAVSELTRNKTKRASATVKTAVLATLLFATVQANATDEDEEEEL
ESVQRSVVGSIQASMEGSGELETISLSMTNDSKEFVDPYIVVTLKAGDNLKIKQNTNENT
NASSFTYSLKKDLTGLINVETEKLSFGANGKKVNIISDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSENVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEISRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANKPV
RITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKS
MMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>XN11_ASS <SEQ ID 99>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVATAVLATLLFATVQASTTDDDDLYLE
PVQRTAPVLSFHADSEGTGEKEVTEDSNWGVYFDKKGVLTAGTITLKAGDNLKIKQNTDE
NTNASSFTYSLKKDLTDLTSVETEKLSFGANGKKVNITSDTKGLNFAKETAGTNGDTTVH
LNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNV
DFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENG
SSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFASGNGTTA
TVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDE
TVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANK
PVRITNVAPGVKEGDVTNVAWLKGVAQNLNNHIDNVDGNARAGIAQAIATASLVQAYLPG
KSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN14_1 <SEQ ID 100>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL
EPVVRSALVLQFMIDKEGNGENESTGNIGWSIYYDNHNTLHGATVTLKAGDNLKIKQNTN
KNTNENTNDSSFTYSLKKDLTDLTSVETEKLSFGANGNKVNITSDTKGLNFAKETAGTNG
DTTVHLNGIGSTLTDTLLNTGATTNVTNDNVTDDKKKRAASVKDVLNAGWNIKGVKPGTT
ASDNVDFVHTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKG
KGENGSSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASG
KGTTATVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSK
GKMDETVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDKGALNVGS
KDANKPVRITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQ
AYLPGKSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>SN16_1 <SEQ ID 101>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVATAVLATLLFATVQANATDDDDLYLE
PVQRTAVVLSFRSDKEGTGEKEGTEDSNWAVYFDEKRVLKAGAITLKAGDNLKIKQNTNE
NTNENTNDSSFTYSLKKDLTDLTSVETEKLSFGANGNKVNITSDTKGLNFAKETAGTNGD
PTVHLNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTA
SDNVDFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKGK
DENGSSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFASGN
GTTATVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKG
KMDETVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSK
DANKPVRITNVAPGVKEGDVTNVAWLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLAQA
YLPGKSMMAIGGGTYRGEAGYAIGYSSISDTGNWVIKGTASGNSRGHFGASASVGTQW*
>SN18_1 <SEQ ID 102>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVATAVLATLLFATVQASTTDDDDLYLE
PVQRTAPVLSFHADSEGTGEKEVTEDSNWGVYFDKKGVLTAGTITLKAGDNLKIKQNTDE
NTNASSFTYSLKKDLTDLTSVETEKLSFGANGKKVNITSDTKGLNFAKETAGTNGDTTVH
LNGIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNV
DFVRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENG
SSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFASGNGTTA
TVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDE
TVNINAGNNIEITRNSKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANK
PVRITNVAPGVKEGDVTNVAWLKGVAQNLNNHIDNVDGNARAGIAQAIATAGLVQAYLPG
KSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHGFASASVGYQW*
>SN19_1 <SEQ ID 103>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVKTAVLATLLFATVQASANNEEQEEDL
YLDPVQRTVAVLIVNSDKEGTGEKEKVEENSDWAVYGNEKGVLTAREITLKAGDNLKIKQ
NGTNFTYSLKKDLTDLTDVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNHLQNSGWDLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDGDALNVGSKKDNKPVR
ITNVAPGVKEGDVTNVAQLKGVAONLNNRIDNVDGNARAGIAQAIATAGLVQAYSPGKSM
MAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN20_1 <SEQ ID NO104>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVKTAVLATLLFATVQASANNEEQEEDL
YLDPVQRTVAVLIVNSDKEGTGEKEKVEENSDWAVYGNEKGVLTAREITLKAGDNLKIKQ
NGTNFTYSLKKDLTDLTSVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDGDALNVGSKKDNKPVR
ITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKSM
MAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN21_1 <SEQ ID 105>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVKTAVLATLLFATVQASANNEEQEEDL
YLDPVQRTVAVLIVNSDKEGTGEKEKVEENSDWAVYFNEKGVLTAREITLKAGDNLKIKQ
NGTNFTYSLKKDLTDLTSVGTEKLSFSANGNKVNITSDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLLNTGATTNVTNDNVTDDEDDRAASVKDVLNAGWNIKGVKPGTTASDNVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKDKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDGDALNVGSKKDNKPVR
ITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKSM
MAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>SN22_1 <SEQ ID 106>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL
ESVQRSVVGSIQASMEGSGELETISLSMTNDSKEFVDPYIVVTLKAGDNLKIKQNTNENT
NASSFTYSLKKDLTGLINVETEKLSFGANGKKVNIISDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSENVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEISRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANKPV
RITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKS
MMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN23_1 <SEQ ID 107>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL
ESVQRSVVGSIQASMEGSGELETISLSMTNDSKEFVDPYIVVTLKAGDNLKIKQNTNENT
NASSFTYSLKKDLTGLINVETEKLSFGANGKKVNIISDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSENVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEISRNGKNIDIATSMAPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANKPV
RITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKS
MMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN24_1 <SEQ ID 108>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVATAVLATLLSATVQANATDTDEDEEL
ESVVRSALVLQFMIDKEGNGEIESTGDIGWSIYYDDHNTLHGATVTLKAGDNLKIKQSGK
DFTYSLKKELKDLTSVETEKLSFGANGNKVNITSDTKGLNFAKETAGTNGDPTVHLNGIG
STLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSENVDFVRT
YDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSSTDE
GEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFASGNGTTATVSKD
DQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETVNIN
AGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANKPVRIT
NVAPGVKEGDVTNVAWLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLAQAYLPGKSMMA
IGGGTYRGEAGYAIGYSSISDTGNWVIKGTASGNSRGHFGTSASVGYQW*
>ZN25_ASS <SEQ ID 109>
MNKIYRIIWNSALNAWVVVSELTRNHTKRASATVATAVLATLLSATVQANATDTDEDEEL
ESVVRSALVLQFMIDKEGNGEIESTGDIGWSIYYDDHNTLHGATVTLKAGDNLKIKQSGK
DFTYSLKKELKDLTSVETEKLSFGANGNKVNITSDTKGLNFAKETAGTNGDPTVHLNGIG
STLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTTGQSENVDFVRT
YDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGKGKGENGSSTDE
GEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFASGNGTTATVSKD
DQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETVNIN
AGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVGSKDANKPVRIT
NVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLAQAYLPGKSMMA
IGGGTYRGEAGYAIGYSSISDTGNWVIKGTASGNSRGHFGTSASVGYQW*
>ZN28_ASS <SEQ ID 110>
MNKIYRIIWNSALNAWVAVSELTRNHTKRASATVKTAVLATLLFATVQANATDEDEEEEL
ESVQRSVVGSIQASMEGSGELETISLSMTNDSKEFVDPYIVVTLKAGDNLKIKQNTNENT
NASSFTYSLKKDLTGLINVETEKLSFGANGKKVNIISDTKGLNFAKETAGTNGDTTVHLN
GIGSTLTDMLLNTGATTNVTNDNVTDDEKKRAASVKDVLNAGWNIKGVKPGTTASDNVDF
VRTYDTVEFLSADTKTTTVNVESKDNGKKTEVKIGAKTSVIKEKDGKLVTGKGKGENGSS
TDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTNVTFASGKGTTATV
SKDDQGNITVMYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPSKGKMDETV
NINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDKGALNVGSKDANKPV
RITNVAPGVKEGDVTNVAQLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLVQAYLPGKS
MMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW*
>ZN29_ASS <SEQ ID 111>
MNKIYRIIWNIALNAWVVVSELTRNHTKRASATVATAVLATLLSATVQANATDEEDNEDL
EPVVRTAPVLSFHSDKEGTGEKEEVGASSNLTVYFDKNRVLKAGTITLKAGDNLKIKQNT
NENTNENTNASSFTYSLKKDLTGLINVETEKLSFGANGKKVNIISDTKGLNFAKETAGTN
GDPTVHLNGIGSTLTDTLAGSSASHVDAGNQSTHYTRAASIKDVLNAGWNIKGVKTGSTT
GQSENVDFVRTYDTVEFLSADTKTTTVNVESKDNGKRTEVKIGAKTSVIKEKDGKLVTGK
GKGENGSSTDEGEGLVTAKEVIDAVNKAGWRMKTTTANGQTGQADKFETVTSGTKVTFAS
GNGTTATVSKDDQGNITVKYDVNVGDALNVNQLQNSGWNLDSKAVAGSSGKVISGNVSPS
KGKMDETVNINAGNNIEITRNGKNIDIATSMTPQFSSVSLGAGADAPTLSVDDEGALNVG
SKDANKPVRITNVAPGVKEGDVTNVAWLKGVAQNLNNRIDNVDGNARAGIAQAIATAGLV
QAYLPGKSMMAIGGGTYRGEAGYAIGYSSISDGGNWIIKGTASGNSRGHFGASASVGYQW
*

FIG. 8 shows the results of aligning the sequences of each of these strains. Dark shading indicates regions of homology, and gray shading indicates the conservation of amino acids with similar characteristics. As is readily discernible, there is significant conservation among the various strains of ORF 40, further confirming its utility as an antigen for both vaccines and diagnostics.

It will be appreciated that the invention has been described by means of example only, and that modifications may be made whilst remaining within the spirit and scope of the invention.

195 1 736 DNA Neisseria meningitidis 1acactgttgt ttgcaacggt tcaggcaagt gctaaccaat gaagagcaag aagaagattt 60atatttagac cccgtacaac gcactgttgc cgtgttgata gtcaattccg ataaagaagg 120cacgggagaa aaagaaaaag tagaagaaaa ttcagattgg gcagtatatt tcaacgagaa 180aggagtacta acagccagag aaatcaccyt caaagccggc gacaacctga aaatcaaaca 240aaacggcaca aacttcacct actcgctgaa aaaagacctc acagatctga ccagtgttgg 300aactgaaaaa ttatcgttta gcgcaaacgg caataaagtc aacatcacaa gcgacaccaa 360aggcttgaat tttgcgaaag aaacggctgg sacgaacggc gacaccacgg ttcatctgaa 420cggtattggt tcgactttga ccgatacgct gctgaatacc ggagcgacca caaacgtaac 480caacgacaac gttaccgatg acgagaaaaa acgtgcggca agcgttaaag acgtattaaa 540cgctggctgg aacattaaag gcgttaaacc cggtacaaca gcttccgata acgttgattt 600cgtccgcact tacgacacag tcgagttctt gagcgcagat acgaaaacaa cgactgttaa 660tgtggaaagc aaagacaacg gcaagaaaac cgaagttaaa atcggtgcga agacttctgt 720tattaaagaa aaagac 736 2 245 PRT Neisseria meningitidis SITE (70) unknown 2Thr Leu Leu Phe Ala Thr Val Gln Ala Ser Ala Asn Gln Glu Glu Gln 1 5 10 15Glu Glu Asp Leu Tyr Leu Asp Pro Val Gln Arg Thr Val Ala Val Leu 20 25 30Ile Val Asn Ser Asp Lys Glu Gly Thr Gly Glu Lys Glu Lys Val Glu 35 40 45Glu Asn Ser Asp Trp Ala Val Tyr Phe Asn Glu Lys Gly Val Leu Thr 50 55 60Ala Arg Glu Ile Thr Xaa Lys Ala Gly Asp Asn Leu Lys Ile Lys Gln 65 70 75 80Asn Gly Thr Asn Phe Thr Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu 85 90 95Thr Ser Val Gly Thr Glu Lys Leu Ser Phe Ser Ala Asn Gly Asn Lys 100 105 110Val Asn Ile Thr Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr 115 120 125Ala Gly Thr Asn Gly Asp Thr Thr Val His Leu Asn Gly Ile Gly Ser 130 135 140Thr Leu Thr Asp Thr Leu Leu Asn Thr Gly Ala Thr Thr Asn Val Thr145 150 155 160Asn Asp Asn Val Thr Asp Asp Glu Lys Lys Arg Ala Ala Ser Val Lys 165 170 175Asp Val Leu Asn Ala Gly Trp Asn Ile Lys Gly Val Lys Pro Gly Thr 180 185 190Thr Ala Ser Asp Asn Val Asp Phe Val Arg Thr Tyr Asp Thr Val Glu 195 200 205Phe Leu Ser Ala Asp Thr Lys Thr Thr Thr Val Asn Val Glu Ser Lys 210 215 220Asp Asn Gly Lys Lys Thr Glu Val Lys Ile Gly Ala Lys Thr Ser Val225 230 235 240Ile Lys Glu Lys Asp 245 3 1776 DNA Neisseria meningitidis 3atgaacaaaa tataccgcat catttggaat agtgccctca atgcctgggt cgtcgtatcc 60gagctcacac gcaaccacac caaacgcgcc tccgcaaccg tgaagaccgc cgtattggcg 120acactgttgt ttgcaacggt tcaggcaagt gctaacaatg aagagcaaga agaagattta 180tatttagacc ccgtacaacg cactgttgcc gtgttgatag tcaattccga taaagaaggc 240acgggagaaa aagaaaaagt agaagaaaat tcagattggg cagtatattt caacgagaaa 300ggagtactaa cagccagaga aatcaccctc aaagccggcg acaacctgaa aatcaaacaa 360aacggcacaa acttcaccta ctcgctgaaa aaagacctca cagatctgac cagtgttgga 420actgaaaaat tatcgtttag cgcaaacggc aataaagtca acatcacaag cgacaccaaa 480ggcttgaatt ttgcgaaaga aacggctggg acgaacggcg acaccacggt tcatctgaac 540ggtattggtt cgactttgac cgatacgctg ctgaataccg gagcgaccac aaacgtaacc 600aacgacaacg ttaccgatga cgagaaaaaa cgtgcggcaa gcgttaaaga cgtattaaac 660gctggctgga acattaaagg cgttaaaccc ggtacaacag cttccgataa cgttgatttc 720gtccgcactt acgacacagt cgagttcttg agcgcagata cgaaaacaac gactgttaat 780gtggaaagca aagacaacgg caagaaaacc gaagttaaaa tcggtgcgaa gacttctgtt 840attaaagaaa aagacggtaa gttggttact ggtaaagaca aaggcgagaa tggttcttct 900acagacgaag gcgaaggctt agtgactgca aaagaagtga ttgatgcagt aaacaaggct 960ggttggagaa tgaaaacaac aaccgctaat ggtcaaacag gtcaagctga caagtttgaa 1020accgttacat caggcacaaa tgtaaccttt gctagtggta aaggtacaac tgcgactgta 1080agtaaagatg atcaaggcaa catcactgtt atgtatgatg taaatgtcgg cgatgcccta 1140aacgtcaatc agctgcaaaa cagcggttgg aatttggatt ccaaagcggt tgcaggttct 1200tcgggcaaag tcatcagcgg caatgtttcg ccgagcaagg gaaagatgga tgaaaccgtc 1260aacattaatg ccggcaacaa catcgagatt acccgcaacg gtaaaaatat cgacatcgcc 1320acttcgatga ccccgcagtt ttccagcgtt tcgctcggcg cgggggcgga tgcgcccact 1380ttgagcgtgg atggggacgc attgaatgtc ggcagcaaga aggacaacaa acccgtccgc 1440attaccaatg tcgccccggg cgttaaagag ggggatgtta caaacgtcgc acaacttaaa 1500ggcgtggcgc aaaacttgaa caaccgcatc gacaatgtgg acggcaacgc gcgtgcgggc 1560atcgcccaag cgattgcaac cgcaggtctg gttcaggcgt atttgcccgg caagagtatg 1620atggcgatcg gcggcggcac ttatcgcggc gaagccggtt acgccatcgg ctactccagt 1680atttccgacg gcggaaattg gattatcaaa ggcacggctt ccggcaattc gcgcggccat 1740ttcggtgctt ccgcatctgt cggttatcag tggtaa 1776 4 591 PRT Neisseria meningitidis 4Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Ala Asn Asn Glu Glu Gln Glu Glu Asp Leu Tyr Leu Asp Pro 50 55 60Val Gln Arg Thr Val Ala Val Leu Ile Val Asn Ser Asp Lys Glu Gly 65 70 75 80Thr Gly Glu Lys Glu Lys Val Glu Glu Asn Ser Asp Trp Ala Val Tyr 85 90 95Phe Asn Glu Lys Gly Val Leu Thr Ala Arg Glu Ile Thr Leu Lys Ala 100 105 110Gly Asp Asn Leu Lys Ile Lys Gln Asn Gly Thr Asn Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr Glu Lys Leu 130 135 140Ser Phe Ser Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Gly Asp Ala Leu Asn Val Gly Ser Lys Lys Asp Asn Lys Pro Val Arg465 470 475 480Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val 485 490 495Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn 500 505 510Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala 515 520 525Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly 530 535 540Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser545 550 555 560Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly Asn 565 570 575Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 5 1779 DNA Neisseria meningitidis misc_feature (48) any nucleotide 5atgaacaaaa tataccgcat catttggaat agtgccctca atgcctgngt cgccgtatcc 60gagctcacac gcaaccacac caaacgcgcc tccgcaaccg tgaagaccgc cgtattggcg 120acactgttgt ttgcaacggt tcaggcgaat gctaccgatg aagatgaaga agaagagtta 180gaatccgtac aacgctctgt cgtagggagc attcaagcca gtatggaagg cagcggcgaa 240ttggaaacga tatcattatc aatgactaac gacagcaagg aatttgtaga cccatacata 300gtagttaccc tcaaagccgg cgacaacctg aaaatcaaac aaaacaccaa tgaaaacacc 360aatgccagta gcttcaccta ctcgctgaaa aaagacctca caggcctgat caatgttgan 420actgaaaaat tatcgtttgg cgcaaacggc aagaaagtca acatcataag cgacaccaaa 480ggcttgaatt tcgcgaaaga aacggctggg acgaacggcg acaccacggt tcatctgaac 540ggtatcggtt cgactttgac cgatacgctt gcgggttctt ctgcttctca cgttgatgcg 600ggtaaccnaa gtacacatta cactcgtgca gcaagtatta aggatgtgtt gaatgcgggt 660tggaatatta agggtgttaa annnggctca acaactggtc aatcagaaaa tgtcgatttc 720gtccgcactt acgacacagt cgagttcttg agcgcagata cgnaaacaac gacngttaat 780gtggaaagca aagacaacgg caagagaacc gaagttaaaa tcggtgcgaa gacttctgtt 840attaaagaaa aagacggtaa gttggttact ggtaaaggca aaggcgagaa tggttcttct 900acagacgaag gcgaaggctt agtgactgca aaagaagtga ttgatgcagt aaacaaggct 960ggttggagaa tgaaaacaac aaccgctaat ggtcaaacag gtcaagctga caagtttgaa 1020accgttacat caggcacaaa tgtaaccttt gctagtggta aaggtacaac tgcgactgta 1080agtaaagatg atcaaggcaa catcactgtt atgtatgatg taaatgtcgg cgatgcccta 1140aacgtcaatc agctgcaaaa cagcggttgg aatttggatt ccaaagcggt tgcaggttct 1200tcgggcaaag tcatcagcgg caatgtttcg ccgagcaagg gaaagatgga tgaaaccgtc 1260aacattaatg ccggcaacaa catcgagatt agccgcaacg gtaaaaatat cgacatcgcc 1320acttcgatgg cgccgcagtt ttccagcgtt tcgctcggcg cgggggcaga tgcgcccact 1380ttaagcgtgg atgacgaggg cgcgttgaat gtcggcagca aggatgccaa caaacccgtc 1440cgcattacca atgtcgcccc gggcgttaaa ganggggatg ttacaaacgt cncacaactt 1500aaaggcgtgg cgcaaaactt gaacaaccgc atcgacaatg tggacggcaa cgcgcgtgcn 1560ggcatcgccc aagcgattgc aaccgcaggt ctggttcagg cgtatctgcc cggcaagagt 1620atgatggcga tcggcggcgg cacttatcgc ggcgaagccg gttacgccat cggctactcc 1680agtatttccg acggcggaaa ttggattatc aaaggcacgg cttccggcaa ttcgcgcggc 1740catttcggtg cttccgcatc tgtcggttat cagtggtaa 1779 6 592 PRT Neisseria meningitidis SITE (16) unknown 6Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Xaa 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Ser Val Gln 50 55 60Arg Ser Val Val Gly Ser Ile Gln Ala Ser Met Glu Gly Ser Gly Glu 65 70 75 80Leu Glu Thr Ile Ser Leu Ser Met Thr Asn Asp Ser Lys Glu Phe Val 85 90 95Asp Pro Tyr Ile Val Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile 100 105 110Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn Val Xaa Thr Glu Lys Leu 130 135 140Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Ile Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly 180 185 190Ser Ser Ala Ser His Val Asp Ala Gly Asn Xaa Ser Thr His Tyr Thr 195 200 205Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 210 215 220Gly Val Lys Xaa Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Xaa Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Ser Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val465 470 475 480Arg Ile Thr Asn Val Ala Pro Gly Val Lys Xaa Gly Asp Val Thr Asn 485 490 495Val Xaa Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp 500 505 510Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr 515 520 525Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile 530 535 540Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser545 550 555 560Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly 565 570 575Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 7 492 DNA Neisseria meningitidis 7atgttacgtt tgactgcttt agccgtatgc accgccctcg ctttgggcgc gtgttcgccg 60caaaattccg actctgcccc acaagccaaa gaacaggcgg tttccgccgc acaaaccgaa 120ggcgcgtccg ttaccgtcaa aaccgcgcgc ggcgacgttc aaataccgca aaaccccgaa 180cgcatcgccg tttacgattt gggtatgctc gacaccttga gcaaactggg cgtgaaaacc 240ggtttgtccg tcgataaaaa ccgcctgccg tatttagagg aatatttcaa aacgacaaaa 300cctgccggca ctttgttcga gccggattac gaaacgctca acgcttacaa accgcagctc 360atcatcatcg gcagccgcgc cgccaaggcg tttgacaaat tgaacgaaat cgcgccgacc 420atcgrmwtga ccgccgatac cgccaacctc aaagaaagtg ccaargaggc atcgacgctg 480gcgcaaatct tc 492 8 164 PRT Neisseria meningitidis SITE (142)..(143) unknown 8Met Leu Arg Leu Thr Ala Leu Ala Val Cys Thr Ala Leu Ala Leu Gly 1 5 10 15Ala Cys Ser Pro Gln Asn Ser Asp Ser Ala Pro Gln Ala Lys Glu Gln 20 25 30Ala Val Ser Ala Ala Gln Thr Glu Gly Ala Ser Val Thr Val Lys Thr 35 40 45Ala Arg Gly Asp Val Gln Ile Pro Gln Asn Pro Glu Arg Ile Ala Val 50 55 60Tyr Asp Leu Gly Met Leu Asp Thr Leu Ser Lys Leu Gly Val Lys Thr 65 70 75 80Gly Leu Ser Val Asp Lys Asn Arg Leu Pro Tyr Leu Glu Glu Tyr Phe 85 90 95Lys Thr Thr Lys Pro Ala Gly Thr Leu Phe Glu Pro Asp Tyr Glu Thr 100 105 110Leu Asn Ala Tyr Lys Pro Gln Leu Ile Ile Ile Gly Ser Arg Ala Ala 115 120 125Lys Ala Phe Asp Lys Leu Asn Glu Ile Ala Pro Thr Ile Xaa Xaa Thr 130 135 140Ala Asp Thr Ala Asn Leu Lys Glu Ser Ala Lys Glu Ala Ser Thr Leu145 150 155 160Ala Gln Ile Phe 9 966 DNA Neisseria meningitidis 9atgttacgtt tgactgcttt agccgtatgc accgccctcg ctttgggcgc gtgttcgccg 60caaaattccg actctgcccc acaagccaaa gaacaggcgg tttccgccgc acaaaccgaa 120ggcgcgtccg ttaccgtcaa aaccgcgcgc ggcgacgttc aaataccgca aaaccccgaa 180cgcatcgccg tttacgattt gggtatgctc gacaccttga gcaaactggg cgtgaaaacc 240ggtttgtccg tcgataaaaa ccgcctgccg tatttagagg aatatttcaa aacgacaaaa 300cctgccggca ctttgttcga gccggattac gaaacgctca acgcttacaa accgcagctc 360atcatcatcg gcagccgcgc cgccaaggcg tttgacaaat tgaacgaaat cgcgccgacc 420atcgaaatga ccgccgatac cgccaacctc aaagaaagtg ccaaagagcg catcgacgcg 480ctggcgcaaa tcttcggcaa acaggcggaa gccgacaagc tgaaggcgga aatcgacgcg 540tcttttgaag ccgcgaaaac tgccgcacaa ggtaagggca aaggtttggt gattttggtc 600aacggcggca agatgtcggc tttcggcccg tcttcacgct tgggcggctg gctgcacaaa 660gacatcggcg ttcccgctgt cgatgaatca attaaagaag gcagccacgg tcagcctatc 720agctttgaat acctgaaaga gaaaaatccc gactggctgt ttgtccttga ccgaagcgcg 780gccatcggcg aagagggtca ggcggcgaaa gacgtgttgg ataatccgct ggttgccgaa 840acaaccgctt ggaaaaaagg acaggtcgtg tacctcgttc ctgaaactta tttggcagcc 900ggtggcgcgc aagagctgct gaatgcaagc aaacaggttg ccgacgcttt taacgcggca 960aaataa 966 10 321 PRT Neisseria meningitidis 10Met Leu Arg Leu Thr Ala Leu Ala Val Cys Thr Ala Leu Ala Leu Gly 1 5 10 15Ala Cys Ser Pro Gln Asn Ser Asp Ser Ala Pro Gln Ala Lys Glu Gln 20 25 30Ala Val Ser Ala Ala Gln Thr Glu Gly Ala Ser Val Thr Val Lys Thr 35 40 45Ala Arg Gly Asp Val Gln Ile Pro Gln Asn Pro Glu Arg Ile Ala Val 50 55 60Tyr Asp Leu Gly Met Leu Asp Thr Leu Ser Lys Leu Gly Val Lys Thr 65 70 75 80Gly Leu Ser Val Asp Lys Asn Arg Leu Pro Tyr Leu Glu Glu Tyr Phe 85 90 95Lys Thr Thr Lys Pro Ala Gly Thr Leu Phe Glu Pro Asp Tyr Glu Thr 100 105 110Leu Asn Ala Tyr Lys Pro Gln Leu Ile Ile Ile Gly Ser Arg Ala Ala 115 120 125Lys Ala Phe Asp Lys Leu Asn Glu Ile Ala Pro Thr Ile Glu Met Thr 130 135 140Ala Asp Thr Ala Asn Leu Lys Glu Ser Ala Lys Glu Arg Ile Asp Ala145 150 155 160Leu Ala Gln Ile Phe Gly Lys Gln Ala Glu Ala Asp Lys Leu Lys Ala 165 170 175Glu Ile Asp Ala Ser Phe Glu Ala Ala Lys Thr Ala Ala Gln Gly Lys 180 185 190Gly Lys Gly Leu Val Ile Leu Val Asn Gly Gly Lys Met Ser Ala Phe 195 200 205Gly Pro Ser Ser Arg Leu Gly Gly Trp Leu His Lys Asp Ile Gly Val 210 215 220Pro Ala Val Asp Glu Ser Ile Lys Glu Gly Ser His Gly Gln Pro Ile225 230 235 240Ser Phe Glu Tyr Leu Lys Glu Lys Asn Pro Asp Trp Leu Phe Val Leu 245 250 255Asp Arg Ser Ala Ala Ile Gly Glu Glu Gly Gln Ala Ala Lys Asp Val 260 265 270Leu Asp Asn Pro Leu Val Ala Glu Thr Thr Ala Trp Lys Lys Gly Gln 275 280 285Val Val Tyr Leu Val Pro Glu Thr Tyr Leu Ala Ala Gly Gly Ala Gln 290 295 300Glu Leu Leu Asn Ala Ser Lys Gln Val Ala Asp Ala Phe Asn Ala Ala305 310 315 320Lys 11 966 DNA Neisseria meningitidis 11atgttacgtt tgactgcttt agccgtatgc accgccctcg ctttgggcgc gtgttcgccg 60caaaattccg actctgcccc acaagccaaa gaacaggcgg tttccgccgc acaatccgaa 120ggcgtgtccg ttaccgtcaa aacggcgcgc ggcgatgttc aaataccgca aaaccccgaa 180cgtatcgccg tttacgattt gggtatgctc gacaccttga gcaaactggg cgtgaaaacc 240ggtttgtccg tcgataaaaa ccgcctgccg tatttagagg aatatttcaa aacgacaaaa 300cctgccggaa ctttgttcga gccggattac gaaacgctca acgcttacaa accgcagctc 360atcatcatcg gcagccgcgc agccaaagcg tttgacaaat tgaacgaaat cgcgccgacc 420atcgaaatga ccgccgatac cgccaacctc aaagaaagtg ccaaagagcg tatcgacgcg 480ctggcgcaaa tcttcggcaa aaaggcggaa gccgacaagc tgaaggcgga aatcgacgcg 540tcttttgaag ccgcgaaaac tgccgcgcaa ggcaaaggca agggtttggt gattttggtc 600aacggcggca agatgtccgc cttcggcccg tcttcacgac tgggcggctg gctgcacaaa 660gacatcggcg ttcccgctgt tgacgaagcc atcaaagaag gcagccacgg tcagcctatc 720agctttgaat acctgaaaga gaaaaatccc gactggctgt ttgtccttga ccgcagcgcg 780gccatcggcg aagagggtca ggcggcgaaa gacgtgttga acaatccgct ggttgccgaa 840acaaccgctt ggaaaaaagg acaagtcgtt taccttgttc ctgaaactta tttggcagcc 900ggtggcgcgc aagagctact gaatgcaagc aaacaggttg ccgacgcttt taacgcggca 960aaataa 966 12 321 PRT Neisseria meningitidis 12Met Leu Arg Leu Thr Ala Leu Ala Val Cys Thr Ala Leu Ala Leu Gly 1 5 10 15Ala Cys Ser Pro Gln Asn Ser Asp Ser Ala Pro Gln Ala Lys Glu Gln 20 25 30Ala Val Ser Ala Ala Gln Ser Glu Gly Val Ser Val Thr Val Lys Thr 35 40 45Ala Arg Gly Asp Val Gln Ile Pro Gln Asn Pro Glu Arg Ile Ala Val 50 55 60Tyr Asp Leu Gly Met Leu Asp Thr Leu Ser Lys Leu Gly Val Lys Thr 65 70 75 80Gly Leu Ser Val Asp Lys Asn Arg Leu Pro Tyr Leu Glu Glu Tyr Phe 85 90 95Lys Thr Thr Lys Pro Ala Gly Thr Leu Phe Glu Pro Asp Tyr Glu Thr 100 105 110Leu Asn Ala Tyr Lys Pro Gln Leu Ile Ile Ile Gly Ser Arg Ala Ala 115 120 125Lys Ala Phe Asp Lys Leu Asn Glu Ile Ala Pro Thr Ile Glu Met Thr 130 135 140Ala Asp Thr Ala Asn Leu Lys Glu Ser Ala Lys Glu Arg Ile Asp Ala145 150 155 160Leu Ala Gln Ile Phe Gly Lys Lys Ala Glu Ala Asp Lys Leu Lys Ala 165 170 175Glu Ile Asp Ala Ser Phe Glu Ala Ala Lys Thr Ala Ala Gln Gly Lys 180 185 190Gly Lys Gly Leu Val Ile Leu Val Asn Gly Gly Lys Met Ser Ala Phe 195 200 205Gly Pro Ser Ser Arg Leu Gly Gly Trp Leu His Lys Asp Ile Gly Val 210 215 220Pro Ala Val Asp Glu Ala Ile Lys Glu Gly Ser His Gly Gln Pro Ile225 230 235 240Ser Phe Glu Tyr Leu Lys Glu Lys Asn Pro Asp Trp Leu Phe Val Leu 245 250 255Asp Arg Ser Ala Ala Ile Gly Glu Glu Gly Gln Ala Ala Lys Asp Val 260 265 270Leu Asn Asn Pro Leu Val Ala Glu Thr Thr Ala Trp Lys Lys Gly Gln 275 280 285Val Val Tyr Leu Val Pro Glu Thr Tyr Leu Ala Ala Gly Gly Ala Gln 290 295 300Glu Leu Leu Asn Ala Ser Lys Gln Val Ala Asp Ala Phe Asn Ala Ala305 310 315 320Lys 13 375 DNA Neisseria meningitidis 13atgaaacttc tgaccaccgc aatcctgtct tccgcaatcg cgctcagcag tatggctgcc 60gccgctggca cggacaaccc cactgttgca aaaaaaaccg tcagctacgt ctgccagcaa 120ggtaaaaaag tcaaagtaac ctacggcttc aacaaacagg gtctgaccac atacgcttcc 180gccgtcatca acggcaaacg cgtgcaaatg cctgtcaatt tggacaaatc cgacaatgtg 240gaaacattct acggcaaaga aggcggttat gttttgggta ccggcgtgat ggatggcaaa 300tcctaccgca aacagcccat tatgattacc gcacctgaca accaaatcgt cttcaaagac 360tgttccccac gttaa 375 14 124 PRT Neisseria meningitidis 14Met Lys Leu Leu Thr Thr Ala Ile Leu Ser Ser Ala Ile Ala Leu Ser 1 5 10 15Ser Met Ala Ala Ala Ala Gly Thr Asp Asn Pro Thr Val Ala Lys Lys 20 25 30Thr Val Ser Tyr Val Cys Gln Gln Gly Lys Lys Val Lys Val Thr Tyr 35 40 45Gly Phe Asn Lys Gln Gly Leu Thr Thr Tyr Ala Ser Ala Val Ile Asn 50 55 60Gly Lys Arg Val Gln Met Pro Val Asn Leu Asp Lys Ser Asp Asn Val 65 70 75 80Glu Thr Phe Tyr Gly Lys Glu Gly Gly Tyr Val Leu Gly Thr Gly Val 85 90 95Met Asp Gly Lys Ser Tyr Arg Lys Gln Pro Ile Met Ile Thr Ala Pro 100 105 110Asp Asn Gln Ile Val Phe Lys Asp Cys Ser Pro Arg 115 120 15 375 DNA Neisseria meningitidis 15atgaaacttc tgaccaccgc aatcctgtct tccgcaatcg cgctcagcag tatggctgct 60gctgccggca cgaacaaccc caccgttgcc aaaaaaaccg tcagctacgt ctgccagcaa 120ggtaaaaaag tcaaagtaac ctacggcttt aacaaacagg gcctgaccac atacgcttcc 180gccgtcatca acggcaaacg tgtgcaaatg cctgtcaatt tggacaaatc cgacaatgtg 240gaaacattct acggcaaaga aggcggttat gttttgggta ccggcgtgat ggatggcaaa 300tcctatcgca aacagcctat tatgattacc gcacctgaca accaaatcgt cttcaaagac 360tgttccccac gttaa 375 16 124 PRT Neisseria meningitidis 16Met Lys Leu Leu Thr Thr Ala Ile Leu Ser Ser Ala Ile Ala Leu Ser 1 5 10 15Ser Met Ala Ala Ala Ala Gly Thr Asn Asn Pro Thr Val Ala Lys Lys 20 25 30Thr Val Ser Tyr Val Cys Gln Gln Gly Lys Lys Val Lys Val Thr Tyr 35 40 45Gly Phe Asn Lys Gln Gly Leu Thr Thr Tyr Ala Ser Ala Val Ile Asn 50 55 60Gly Lys Arg Val Gln Met Pro Val Asn Leu Asp Lys Ser Asp Asn Val 65 70 75 80Glu Thr Phe Tyr Gly Lys Glu Gly Gly Tyr Val Leu Gly Thr Gly Val 85 90 95Met Asp Gly Lys Ser Tyr Arg Lys Gln Pro Ile Met Ile Thr Ala Pro 100 105 110Asp Asn Gln Ile Val Phe Lys Asp Cys Ser Pro Arg 115 120 17 519 DNA Neisseria meningitidis misc_feature (425) any nucleotide 17ggcaccgaat tcaaaaccac cctttccgga gccgacatac aggcaggggt gggtgaaaaa 60gcccgagccg atgcgaaaat tatcctaaaa ggcatcgtta accgcatcca aaccgaagaa 120aagctggaat ccaactcgac cgtatggcaa aagcaggccg gaagcggcag cacggttgaa 180acgctgaagc taccgagctt tgaagggccg gcactgccta agctgaccgc tcccggcggc 240tatatcgccg acatccccaa aggcaacctc aaaaccgaaa tcgaaaagct ggccaaacag 300cccgaatatg cctatctgaa acagcttcag acggtcaagg acgtgaactg gaaccaagta 360cagctcgctt acgacaaatg ggactataaa caggaaggcc taaccggagc cggagccgca 420attancgcac tggccgttac cgtggtcacc tcaggcgcag gaaccggagc cgtattggga 480ttaanacgng tggccgccgc cgcaaccgat gcagcattt 519 18 173 PRT Neisseria meningitidis SITE (142) unknown 18Gly Thr Glu Phe Lys Thr Thr Leu Ser Gly Ala Asp Ile Gln Ala Gly 1 5 10 15Val Gly Glu Lys Ala Arg Ala Asp Ala Lys Ile Ile Leu Lys Gly Ile 20 25 30Val Asn Arg Ile Gln Thr Glu Glu Lys Leu Glu Ser Asn Ser Thr Val 35 40 45Trp Gln Lys Gln Ala Gly Ser Gly Ser Thr Val Glu Thr Leu Lys Leu 50 55 60Pro Ser Phe Glu Gly Pro Ala Leu Pro Lys Leu Thr Ala Pro Gly Gly 65 70 75 80Tyr Ile Ala Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile Glu Lys 85 90 95Leu Ala Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln Thr Val 100 105 110Lys Asp Val Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Lys Trp Asp 115 120 125Tyr Lys Gln Glu Gly Leu Thr Gly Ala Gly Ala Ala Ile Xaa Ala Leu 130 135 140Ala Val Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val Leu Gly145 150 155 160Leu Xaa Arg Val Ala Ala Ala Ala Thr Asp Ala Ala Phe 165 170 19 1923 DNA Neisseria meningitidis 19atgcaactgc tggcagccga aggcattcac caacaccaat tgaatgttca gaaaagtacc 60cgtttcatcg gcatcaaagt gggtaaaagc aattacagca aaaacgagct gaacgaaacc 120aaactgcccg tacgcgttat cgcccaaaca gccaaaaccc gttccggctg ggataccgta 180ctcgaaggca ccgaattcaa aaccaccctt tccggagccg acatacaggc aggggtgggt 240gaaaaagccc gagccgatgc gaaaattatc ctaaaaggca tcgttaaccg catccaaacc 300gaagaaaagc tggaatccaa ctcgaccgta tggcaaaagc aggccggaag cggcagcacg 360gttgaaacgc tgaagctacc gagctttgaa gggccggcac tgcctaagct gaccgctccc 420ggcggctata tcgccgacat ccccaaaggc aacctcaaaa ccgaaatcga aaagctggcc 480aaacagcccg aatatgccta tctgaaacag cttcagacgg tcaaggacgt gaactggaac 540caagtacagc tcgcttacga caaatgggac tataaacagg aaggcctaac cggagccgga 600gccgcaatta tcgcactggc cgttaccgtg gtcacctcag gcgcaggaac cggagccgta 660ttgggattaa acggtgcggc cgccgccgca accgatgcag catttgcctc tttggccagc 720caggcttccg tatcgttcat caacaacaaa ggcaatatcg gtaacaccct gaaagagctg 780ggcagaagca gcacggtgaa aaatctgatg gttgccgtcg ctaccgcagg cgtagccgac 840aaaatcggtg cttcggcact gaacaatgtc agcgataagc agtggatcaa caacctgacc 900gtcaacctgg ccaatgcggg cagtgccgca ctgattaata ccgctgtcaa cggcggcagc 960ctgaaagaca atctggaagc gaatatcctt gcggctttgg tgaatactgc gcatggagag 1020gcagcaagta aaatcaaaca gttggatcag cactacattg cccataagat tgcccatgcc 1080atagcgggct gtgcggcagc ggcggcgaat aagggcaagt gtcaagatgg tgcgatcggt 1140gcggcggtcg gtgaaatcct tggcgaaacc ctactggacg gcagagaccc tggcagcctg 1200aatgtgaagg acagggcaaa aatcattgct aaggcgaagc tggcagcagg ggcggttgcg 1260gcgttgagta agggggatgt gagtacggcg gcgaatgcgg ctgctgtggc ggtagagaat 1320aattctttaa atgatataca ggatcgtttg ttgagtggaa attatgcttt atgtatgagt 1380gcaggaggag cagaaagctt ttgtgagtct tatcgaccac tgggcttgcc acactttgta 1440agtgtttcag gagaaatgaa attacctaat aaattcggga atcgtatggt taatggaaaa 1500ttaattatta acactagaaa tggcaatgta tatttctctg taggtaaaat atggagtact 1560gtaaaatcaa caaaatcaaa tataagtggg gtatctgtcg gttgggtttt aaatgtttcc 1620cctaatgatt atttaaaaga agcatctatg aatgatttca gaaatagtaa tcaaaataaa 1680gcctatgcag aaatgatttc ccagactttg gtaggtgaga gtgttggtgg tagtctttgt 1740ctgacaagag cctgcttttc ggtaagttca acaatatcta aatctaaatc tccttttaaa 1800gattcaaaaa ttattgggga aatcggtttg ggaagtggtg ttgctgcagg agtagaaaaa 1860acaatataca taggtaacat aaaagatatt gataaattta ttagtgcaaa cataaaaaaa 1920tag 1923 20 640 PRT Neisseria meningitidis 20Met Gln Leu Leu Ala Ala Glu Gly Ile His Gln His Gln Leu Asn Val 1 5 10 15Gln Lys Ser Thr Arg Phe Ile Gly Ile Lys Val Gly Lys Ser Asn Tyr 20 25 30Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val Arg Val Ile Ala 35 40 45Gln Thr Ala Lys Thr Arg Ser Gly Trp Asp Thr Val Leu Glu Gly Thr 50 55 60Glu Phe Lys Thr Thr Leu Ser Gly Ala Asp Ile Gln Ala Gly Val Gly 65 70 75 80Glu Lys Ala Arg Ala Asp Ala Lys Ile Ile Leu Lys Gly Ile Val Asn 85 90 95Arg Ile Gln Thr Glu Glu Lys Leu Glu Ser Asn Ser Thr Val Trp Gln 100 105 110Lys Gln Ala Gly Ser Gly Ser Thr Val Glu Thr Leu Lys Leu Pro Ser 115 120 125Phe Glu Gly Pro Ala Leu Pro Lys Leu Thr Ala Pro Gly Gly Tyr Ile 130 135 140Ala Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile Glu Lys Leu Ala145 150 155 160Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln Thr Val Lys Asp 165 170 175Val Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Lys Trp Asp Tyr Lys 180 185 190Gln Glu Gly Leu Thr Gly Ala Gly Ala Ala Ile Ile Ala Leu Ala Val 195 200 205Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val Leu Gly Leu Asn 210 215 220Gly Ala Ala Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser225 230 235 240Gln Ala Ser Val Ser Phe Ile Asn Asn Lys Gly Asn Ile Gly Asn Thr 245 250 255Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Met Val Ala 260 265 270Val Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala Ser Ala Leu Asn 275 280 285Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr Val Asn Leu Ala 290 295 300Asn Ala Gly Ser Ala Ala Leu Ile Asn Thr Ala Val Asn Gly Gly Ser305 310 315 320Leu Lys Asp Asn Leu Glu Ala Asn Ile Leu Ala Ala Leu Val Asn Thr 325 330 335Ala His Gly Glu Ala Ala Ser Lys Ile Lys Gln Leu Asp Gln His Tyr 340 345 350Ile Ala His Lys Ile Ala His Ala Ile Ala Gly Cys Ala Ala Ala Ala 355 360 365Ala Asn Lys Gly Lys Cys Gln Asp Gly Ala Ile Gly Ala Ala Val Gly 370 375 380Glu Ile Leu Gly Glu Thr Leu Leu Asp Gly Arg Asp Pro Gly Ser Leu385 390 395 400Asn Val Lys Asp Arg Ala Lys Ile Ile Ala Lys Ala Lys Leu Ala Ala 405 410 415Gly Ala Val Ala Ala Leu Ser Lys Gly Asp Val Ser Thr Ala Ala Asn 420 425 430Ala Ala Ala Val Ala Val Glu Asn Asn Ser Leu Asn Asp Ile Gln Asp 435 440 445Arg Leu Leu Ser Gly Asn Tyr Ala Leu Cys Met Ser Ala Gly Gly Ala 450 455 460Glu Ser Phe Cys Glu Ser Tyr Arg Pro Leu Gly Leu Pro His Phe Val465 470 475 480Ser Val Ser Gly Glu Met Lys Leu Pro Asn Lys Phe Gly Asn Arg Met 485 490 495Val Asn Gly Lys Leu Ile Ile Asn Thr Arg Asn Gly Asn Val Tyr Phe 500 505 510Ser Val Gly Lys Ile Trp Ser Thr Val Lys Ser Thr Lys Ser Asn Ile 515 520 525Ser Gly Val Ser Val Gly Trp Val Leu Asn Val Ser Pro Asn Asp Tyr 530 535 540Leu Lys Glu Ala Ser Met Asn Asp Phe Arg Asn Ser Asn Gln Asn Lys545 550 555 560Ala Tyr Ala Glu Met Ile Ser Gln Thr Leu Val Gly Glu Ser Val Gly 565 570 575Gly Ser Leu Cys Leu Thr Arg Ala Cys Phe Ser Val Ser Ser Thr Ile 580 585 590Ser Lys Ser Lys Ser Pro Phe Lys Asp Ser Lys Ile Ile Gly Glu Ile 595 600 605Gly Leu Gly Ser Gly Val Ala Ala Gly Val Glu Lys Thr Ile Tyr Ile 610 615 620Gly Asn Ile Lys Asp Ile Asp Lys Phe Ile Ser Ala Asn Ile Lys Lys625 630 635 640 21 2291 DNA Neisseria meningitidis misc_feature (1) any nucleotide 21ntgcaactgc tggcagaaga aggcatccac aagcacgagt tggatgtcca aaaaagccgc 60cgctttatcg gcatcaaggt aggtnagagc aattacagta aaaacgaact gaacgaaacc 120aaattgcctg tccgcgtcgt cgcccaaant gcagccaccc gttcaggctg ggataccgtg 180ctcgaaggta ccgaattcaa aaccacgctg gccggtgccg acattcaggc aggtgtangc 240gaaaaagccc gtgtcgatgc gaaaattatc ctcaaaggca ttgtgaaccg tatccagtcg 300gaagaaaaat tagaaaccaa ctcaaccgta tggcagaaac aggccggacg cggcagcact 360atcgaaacgc taaaactgcc cagcttcgaa agccctactc cgcccaaatt gtccgcaccc 420ggcggntata tcgtcgacat tccgaaaggc aatctgaaaa ccgaaatcga aaagctgtcc 480aaacagcccg agtatgccta tctgaaacag ctccaagtag cgaaaaacat caactggaat 540caggtgcagc ttgcttacga cagatgggac tacaaacagg agggcttaac cgaagcaggt 600gcggcgatta tcgcactggc cgttaccgtg gtcacctcag gcgcaggaac cggagccgta 660ttgggattaa acggtgcgnc cgccgccgca accgatgcag cattcgcctc tttggccagc 720caggcttccg tatcgttcat caacaacaaa ggcgatgtcg gcaaaaccct gaaagagctg 780ggcagaagca gcacggtgaa aaatctggtg gttgccgccg ctaccgcagg cgtagccgac 840aaaatcggcg cttcggcact gancaatgtc agcgataagc agtggatcaa caacctgacc 900gtcaacctag ccaatgcggg cagtgccgca ctgattaata ccgctgtcaa cggcggcagc 960ctgaaagaca ntctggaagc gaatatcctt gcggctttgg tcaataccgc gcatggagaa 1020gcagccagta aaatcaaaca gttggatcag cactacatag tccacaagat tgcccatgcc 1080atagcgggct gtgcggcagc ggcggcgaat aagggcaagt gtcaggatgg tgcgataggt 1140gcggctgtgg gcgagatagt cggggaggct ttgacaaacg gcaaaaatcc tgacactttg 1200acagctaaag aacgcgaaca gattttggca tacagcaaac tggttgccgg tacggtaagc 1260ggtgtggtcg gcggcgatgt aaatgcggcg gcgaatgcgg ctgaggtagc ggtgaaaaat 1320aatcagctta gcgacnaaga gggtagagaa tttgataacg aaatgactgc atgcgccaaa 1380cagaatantc ctcaactgtg cagaaaaaat actgtaaaaa agtatcaaaa tgttgctgat 1440aaaagacttg ctgcttcgat tgcaatatgt acggatatat cccgtagtac tgaatgtaga 1500acaatcagaa aacaacattt gatcgatagt agaagccttc attcatcttg ggaagcaggt 1560ctaattggta aagatgatga atggtataaa ttattcagca aatcttacac ccaagcagat 1620ttggctttac agtcttatca tttgaatact gctgctaaat cttggcttca atcgggcaat 1680acaaagcctt tatccgaatg gatgtccgac caaggttata cacttatttc aggagttaat 1740cctagattca ttccaatacc aagagggttt gtaaaacaaa atacacctat tactaatgtc 1800aaatacccgg aaggcatcag tttcgataca aacctanaaa gacatctggc aaatgctgat 1860ggttttagtc aagaacaggg cattaaagga gcccataacc gcaccaatnt tatggcagaa 1920ctaaattcac gaggaggang ngtaaaatct gaaacccana ctgatattga aggcattacc 1980cgaattaaat atgagattcc tacactagac aggacaggta aacctgatgg tggatttaag 2040gaaatttcaa gtataaaaac tgtttataat cctaaaaant tttnngatga taaaatactt 2100caaatggctc aanatgctgn ttcacaagga tattcaaaag cctctaaaat tgctcaaaat 2160gaaagaacta aatcaatatc ggaaagaaaa aatgtcattc aattctcaga aacctttgac 2220ggaatcaaat ttagannnta tntngatgta aatacaggaa gaattacaaa cattcaccca 2280gaataattta a 2291 22 761 PRT Neisseria meningitidis SITE (1) unknown 22Xaa Gln Leu Leu Ala Glu Glu Gly Ile His Lys His Glu Leu Asp Val 1 5 10 15Gln Lys Ser Arg Arg Phe Ile Gly Ile Lys Val Gly Xaa Ser Asn Tyr 20 25 30Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val Arg Val Val Ala 35 40 45Gln Xaa Ala Ala Thr Arg Ser Gly Trp Asp Thr Val Leu Glu Gly Thr 50 55 60Glu Phe Lys Thr Thr Leu Ala Gly Ala Asp Ile Gln Ala Gly Val Xaa 65 70 75 80Glu Lys Ala Arg Val Asp Ala Lys Ile Ile Leu Lys Gly Ile Val Asn 85 90 95Arg Ile Gln Ser Glu Glu Lys Leu Glu Thr Asn Ser Thr Val Trp Gln 100 105 110Lys Gln Ala Gly Arg Gly Ser Thr Ile Glu Thr Leu Lys Leu Pro Ser 115 120 125Phe Glu Ser Pro Thr Pro Pro Lys Leu Ser Ala Pro Gly Gly Tyr Ile 130 135 140Val Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile Glu Lys Leu Ser145 150 155 160Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln Val Ala Lys Asn 165 170 175Ile Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Arg Trp Asp Tyr Lys 180 185 190Gln Glu Gly Leu Thr Glu Ala Gly Ala Ala Ile Ile Ala Leu Ala Val 195 200 205Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val Leu Gly Leu Asn 210 215 220Gly Ala Xaa Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser225 230 235 240Gln Ala Ser Val Ser Phe Ile Asn Asn Lys Gly Asp Val Gly Lys Thr 245 250 255Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Val Val Ala 260 265 270Ala Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala Ser Ala Leu Xaa 275 280 285Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr Val Asn Leu Ala 290 295 300Asn Ala Gly Ser Ala Ala Leu Ile Asn Thr Ala Val Asn Gly Gly Ser305 310 315 320Leu Lys Asp Xaa Leu Glu Ala Asn Ile Leu Ala Ala Leu Val Asn Thr 325 330 335Ala His Gly Glu Ala Ala Ser Lys Ile Lys Gln Leu Asp Gln His Tyr 340 345 350Ile Val His Lys Ile Ala His Ala Ile Ala Gly Cys Ala Ala Ala Ala 355 360 365Ala Asn Lys Gly Lys Cys Gln Asp Gly Ala Ile Gly Ala Ala Val Gly 370 375 380Glu Ile Val Gly Glu Ala Leu Thr Asn Gly Lys Asn Pro Asp Thr Leu385 390 395 400Thr Ala Lys Glu Arg Glu Gln Ile Leu Ala Tyr Ser Lys Leu Val Ala 405 410 415Gly Thr Val Ser Gly Val Val Gly Gly Asp Val Asn Ala Ala Ala Asn 420 425 430Ala Ala Glu Val Ala Val Lys Asn Asn Gln Leu Ser Asp Xaa Glu Gly 435 440 445Arg Glu Phe Asp Asn Glu Met Thr Ala Cys Ala Lys Gln Asn Xaa Pro 450 455 460Gln Leu Cys Arg Lys Asn Thr Val Lys Lys Tyr Gln Asn Val Ala Asp465 470 475 480Lys Arg Leu Ala Ala Ser Ile Ala Ile Cys Thr Asp Ile Ser Arg Ser 485 490 495Thr Glu Cys Arg Thr Ile Arg Lys Gln His Leu Ile Asp Ser Arg Ser 500 505 510Leu His Ser Ser Trp Glu Ala Gly Leu Ile Gly Lys Asp Asp Glu Trp 515 520 525Tyr Lys Leu Phe Ser Lys Ser Tyr Thr Gln Ala Asp Leu Ala Leu Gln 530 535 540Ser Tyr His Leu Asn Thr Ala Ala Lys Ser Trp Leu Gln Ser Gly Asn545 550 555 560Thr Lys Pro Leu Ser Glu Trp Met Ser Asp Gln Gly Tyr Thr Leu Ile 565 570 575Ser Gly Val Asn Pro Arg Phe Ile Pro Ile Pro Arg Gly Phe Val Lys 580 585 590Gln Asn Thr Pro Ile Thr Asn Val Lys Tyr Pro Glu Gly Ile Ser Phe 595 600 605Asp Thr Asn Leu Xaa Arg His Leu Ala Asn Ala Asp Gly Phe Ser Gln 610 615 620Glu Gln Gly Ile Lys Gly Ala His Asn Arg Thr Asn Xaa Met Ala Glu625 630 635 640Leu Asn Ser Arg Gly Gly Xaa Val Lys Ser Glu Thr Xaa Thr Asp Ile 645 650 655Glu Gly Ile Thr Arg Ile Lys Tyr Glu Ile Pro Thr Leu Asp Arg Thr 660 665 670Gly Lys Pro Asp Gly Gly Phe Lys Glu Ile Ser Ser Ile Lys Thr Val 675 680 685Tyr Asn Pro Lys Xaa Phe Xaa Asp Asp Lys Ile Leu Gln Met Ala Gln 690 695 700Xaa Ala Xaa Ser Gln Gly Tyr Ser Lys Ala Ser Lys Ile Ala Gln Asn705 710 715 720Glu Arg Thr Lys Ser Ile Ser Glu Arg Lys Asn Val Ile Gln Phe Ser 725 730 735Glu Thr Phe Asp Gly Ile Lys Phe Arg Xaa Tyr Xaa Asp Val Asn Thr 740 745 750Gly Arg Ile Thr Asn Ile His Pro Glu 755 760 23 336 DNA Neisseria meningitidis 23cggatcgttg taggtttgcg gatttcttgc gccgtagtca ccgtagtccc aagtataacc 60caaggctttg tcttcgcctt tcattccgat aagggatatg acgctttggt cggtatagcc 120gtcttgggaa cctttgtcca cccaacgcat atctgcctgc ggattctcat tgccgcttct 180tggctgctga tttttctgcc ttcgcgtttt tcaacttcgc gcttgagggc ttcggcatat 240ttgtcggcca acgccatttc tttcggatgc agctgcctat tgttccaatc tacattcgca 300cccaccacag caccaccact accaccagtt gcatag 336 24 111 PRT Neisseria meningitidis 24Arg Ile Val Val Gly Leu Arg Ile Ser Cys Ala Val Val Thr Val Val 1 5 10 15Pro Ser Ile Thr Gln Gly Phe Val Phe Ala Phe His Ser Asp Lys Gly 20 25 30Tyr Asp Ala Leu Val Gly Ile Ala Val Leu Gly Thr Phe Val His Pro 35 40 45Thr His Ile Cys Leu Arg Ile Leu Ile Ala Ala Ser Trp Leu Leu Ile 50 55 60Phe Leu Pro Ser Arg Phe Ser Thr Ser Arg Leu Arg Ala Ser Ala Tyr 65 70 75 80Leu Ser Ala Asn Ala Ile Ser Phe Gly Cys Ser Cys Leu Leu Phe Gln 85 90 95Ser Thr Phe Ala Pro Thr Thr Ala Pro Pro Leu Pro Pro Val Ala 100 105 110 25 1716 DNA Neisseria meningitidis unsure (496)..(1542) N = Unknown 25aagtttgact ttacctggtt tattccggcg gtaatcaaat accgccggtt gttttttgaa 60gtattggtgg tgtcggtggt gttgcagctg tttgcgctga ttacgcctct gtttttccaa 120gtggtgatgg acaaggtgct ggtacatcgg ggattctcta ctttggatgt ggtgtcggtg 180gctttgttgg tggtgtcgct gtttgagatt gtgttgggcg gtttgcggac gtatctgttt 240gcacatacga cttcacgtat tgatgtggaa ttgggcgcgc gtttgttccg gcatctgctt 300tccctgcctt tatcctattt cgagcacaga cgagtgggtg atacggtggc tcgggtgcgg 360gaattggagc agattcgcaa tttcttgacc ggtcaggcgc tgacttcggt gttggatttg 420gcgttttcgt ttatctttct ggcggtgatg tggtattaca gctccactct gacttgggtg 480gtattggctt cgttgnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 540nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 600nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 660nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 720nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 780nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 840nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 900nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 960nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1020nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1080nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1140nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1200nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1260nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1320nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1380nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1440nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1500nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnatttgcgc caaccggacg 1560gtgctgatta tcgcccaccg tctgtccact gttaaaacgg cacaccggat cattgccatg 1620gataaaggca ggattgtgga agcgggaaca cagcaggaat tgctggcgaa cgnnaacgga 1680tattaccgct atctgtatga tttacagaac gggtag 1716 26 571 PRT Neisseria meningitidis UNSURE (166)..(514) x = Unknown 26Lys Phe Asp Phe Thr Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Arg 1 5 10 15Leu Phe Phe Glu Val Leu Val Val Ser Val Val Leu Gln Leu Phe Ala 20 25 30Leu Ile Thr Pro Leu Phe Phe Gln Val Val Met Asp Lys Val Leu Val 35 40 45His Arg Gly Phe Ser Thr Leu Asp Val Val Ser Val Ala Leu Leu Val 50 55 60Val Ser Leu Phe Glu Ile Val Leu Gly Gly Leu Arg Thr Tyr Leu Phe 65 70 75 80Ala His Thr Thr Ser Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe 85 90 95Arg His Leu Leu Ser Leu Pro Leu Ser Tyr Phe Glu His Arg Arg Val 100 105 110Gly Asp Thr Val Ala Arg Val Arg Glu Leu Glu Gln Ile Arg Asn Phe 115 120 125Leu Thr Gly Gln Ala Leu Thr Ser Val Leu Asp Leu Ala Phe Ser Phe 130 135 140Ile Phe Leu Ala Val Met Trp Tyr Tyr Ser Ser Thr Leu Thr Trp Val145 150 155 160Val Leu Ala Ser Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165 170 175Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 180 185 190Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 195 200 205Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 210 215 220Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa225 230 235 240Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 245 250 255Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 260 265 270Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 275 280 285Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 290 295 300Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa305 310 315 320Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 325 330 335Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 340 345 350Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 355 360 365Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 370 375 380Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa385 390 395 400Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 405 410 415Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 420 425 430Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 435 440 445Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 450 455 460Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa465 470 475 480Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 485 490 495Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 500 505 510Xaa Xaa Ile Cys Ala Asn Arg Thr Val Leu Ile Ile Ala His Arg Leu 515 520 525Ser Thr Val Lys Thr Ala His Arg Ile Ile Ala Met Asp Lys Gly Arg 530 535 540Ile Val Glu Ala Gly Thr Gln Gln Glu Leu Leu Ala Asn Xaa Asn Gly545 550 555 560Tyr Tyr Arg Tyr Leu Tyr Asp Leu Gln Asn Gly 565 570 27 2133 DNA Neisseria meningitidis 27atgtctatcg tatccgcacc gctccccgcc ctttccgccc tcatcatcct cgcccattac 60cacggcattg ccgccaatcc tgccgatata cagcatgaat tttgtacttc cgcacagagc 120gatttaaatg aaacgcaatg gctgttagcc gccaaatctt tgggattgaa ggcaaaggta 180gtccgccagc ctattaaacg tttggctatg gcgactttac ccgcattggt atggtgtgat 240gacggcaacc atttcatttt ggccaaaaca gacggtgagg gtgagcatgc ccaatttttg 300atacaggatt tggttacgaa taagtctgcg gtattgtctt ttgccgaatt ttctaacaga 360tattcgggca aactgatatt ggttgcttcc cgcgcttcgg tattgggcag tttggcaaag 420tttgacttta cctggtttat tccggcggta atcaaatacc gccggttgtt ttttgaagta 480ttggtggtgt cggtggtgtt gcagctgttt gcgctgatta cgcctctgtt tttccaagtg 540gtgatggaca aggtgctggt acatcgggga ttctctactt tggatgtggt gtcggtggct 600ttgttggtgg tgtcgctgtt tgagattgtg ttgggcggtt tgcggacgta tctgtttgca 660catacgactt cacgtattga tgtggaattg ggcgcgcgtt tgttccggca tctgctttcc 720ctgcctttat cctatttcga gcacagacga gtgggtgata cggtggctcg ggtgcgggaa 780ttggagcaga ttcgcaattt cttgaccggt caggcgctga cttcggtgtt ggatttggcg 840ttttcgttta tctttctggc ggtgatgtgg tattacagct ccactctgac ttgggtggta 900ttggcttcgt tgcctgccta tgcgttttgg tcggcattta tcagtccgat actgcggacg 960cgtctgaacg ataagttcgc gcgcaatgca gacaaccagt cgtttttagt agaaagcatc 1020actgcggtgg gtacggtaaa ggcgatggcg gtggagccgc agatgacgca gcgttgggac 1080aatcagttgg cggcttatgt ggcttcggga tttcgggtaa cgaagttggc ggtggtcggc 1140cagcaggggg tgcagctgat tcagaagctg gtgacggtgg cgacgttgtg gattggcgca 1200cggctggtaa ttgagagcaa gctgacggtg gggcagctga ttgcgtttaa tatgctctcg 1260ggacaggtgg cggcgcctgt tatccgtttg gcgcagttgt ggcaggattt ccagcaggtg 1320gggatttcgg tggcgcgttt gggggatatt ctgaatgcgc cgaccgagaa tgcgtcttcg 1380catttggctt tgcccgatat ccggggggag attacgttcg aacatgtcga tttccgctat 1440aaggcggacg gcaggctgat tttgcaggat ttgaacctgc ggattcgggc gggggaagtg 1500ctggggattg tgggacgttc ggggtcgggc aaatccacac tcaccaaatt ggtgcagcgt 1560ctgtatgtac cggagcaggg acgggtgttg gtggacggca acgatttggc tttggccgct 1620cctgcctggc tgcggcggca ggtcggcgtg gtcttgcagg agaatgtgct gctcaaccgc 1680agcatacgcg acaatatcgc gctgacggat acgggtatgc cgctggaacg cattatcgaa 1740gcagccaaac tggcgggcgc acacgagttt attatggagc tgccggaagg ctacggcacc 1800gtggtgggcg aacaaggggc cggcttgtcg ggcggacagc ggcagcgtat tgcgattgcc 1860cgcgcgttaa tcaccaatcc gcgcattctg atttttgatg aagccaccag cgcgctggat 1920tatgaaagtg aacgagcgat tatgcagaac atgcaggcca tttgcgccaa ccggacggtg 1980ctgattatcg cccaccgtct gtccactgtt aaaacggcac accggatcat tgccatggat 2040aaaggcagga ttgtggaagc gggaacacag caggaattgc tggcgaagcc gaacggatat 2100taccgctatc tgtatgattt acagaacggg tag 2133 28 710 PRT Neisseria meningitidis 28Met Ser Ile Val Ser Ala Pro Leu Pro Ala Leu Ser Ala Leu Ile Ile 1 5 10 15Leu Ala His Tyr His Gly Ile Ala Ala Asn Pro Ala Asp Ile Gln His 20 25 30Glu Phe Cys Thr Ser Ala Gln Ser Asp Leu Asn Glu Thr Gln Trp Leu 35 40 45Leu Ala Ala Lys Ser Leu Gly Leu Lys Ala Lys Val Val Arg Gln Pro 50 55 60Ile Lys Arg Leu Ala Met Ala Thr Leu Pro Ala Leu Val Trp Cys Asp 65 70 75 80Asp Gly Asn His Phe Ile Leu Ala Lys Thr Asp Gly Glu Gly Glu His 85 90 95Ala Gln Phe Leu Ile Gln Asp Leu Val Thr Asn Lys Ser Ala Val Leu 100 105 110Ser Phe Ala Glu Phe Ser Asn Arg Tyr Ser Gly Lys Leu Ile Leu Val 115 120 125Ala Ser Arg Ala Ser Val Leu Gly Ser Leu Ala Lys Phe Asp Phe Thr 130 135 140Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Arg Leu Phe Phe Glu Val145 150 155 160Leu Val Val Ser Val Val Leu Gln Leu Phe Ala Leu Ile Thr Pro Leu 165 170 175Phe Phe Gln Val Val Met Asp Lys Val Leu Val His Arg Gly Phe Ser 180 185 190Thr Leu Asp Val Val Ser Val Ala Leu Leu Val Val Ser Leu Phe Glu 195 200 205Ile Val Leu Gly Gly Leu Arg Thr Tyr Leu Phe Ala His Thr Thr Ser 210 215 220Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe Arg His Leu Leu Ser225 230 235 240Leu Pro Leu Ser Tyr Phe Glu His Arg Arg Val Gly Asp Thr Val Ala 245 250 255Arg Val Arg Glu Leu Glu Gln Ile Arg Asn Phe Leu Thr Gly Gln Ala 260 265 270Leu Thr Ser Val Leu Asp Leu Ala Phe Ser Phe Ile Phe Leu Ala Val 275 280 285Met Trp Tyr Tyr Ser Ser Thr Leu Thr Trp Val Val Leu Ala Ser Leu 290 295 300Pro Ala Tyr Ala Phe Trp Ser Ala Phe Ile Ser Pro Ile Leu Arg Thr305 310 315 320Arg Leu Asn Asp Lys Phe Ala Arg Asn Ala Asp Asn Gln Ser Phe Leu 325 330 335Val Glu Ser Ile Thr Ala Val Gly Thr Val Lys Ala Met Ala Val Glu 340 345 350Pro Gln Met Thr Gln Arg Trp Asp Asn Gln Leu Ala Ala Tyr Val Ala 355 360 365Ser Gly Phe Arg Val Thr Lys Leu Ala Val Val Gly Gln Gln Gly Val 370 375 380Gln Leu Ile Gln Lys Leu Val Thr Val Ala Thr Leu Trp Ile Gly Ala385 390 395 400Arg Leu Val Ile Glu Ser Lys Leu Thr Val Gly Gln Leu Ile Ala Phe 405 410 415Asn Met Leu Ser Gly Gln Val Ala Ala Pro Val Ile Arg Leu Ala Gln 420 425 430Leu Trp Gln Asp Phe Gln Gln Val Gly Ile Ser Val Ala Arg Leu Gly 435 440 445Asp Ile Leu Asn Ala Pro Thr Glu Asn Ala Ser Ser His Leu Ala Leu 450 455 460Pro Asp Ile Arg Gly Glu Ile Thr Phe Glu His Val Asp Phe Arg Tyr465 470 475 480Lys Ala Asp Gly Arg Leu Ile Leu Gln Asp Leu Asn Leu Arg Ile Arg 485 490 495Ala Gly Glu Val Leu Gly Ile Val Gly Arg Ser Gly Ser Gly Lys Ser 500 505 510Thr Leu Thr Lys Leu Val Gln Arg Leu Tyr Val Pro Glu Gln Gly Arg 515 520 525Val Leu Val Asp Gly Asn Asp Leu Ala Leu Ala Ala Pro Ala Trp Leu 530 535 540Arg Arg Gln Val Gly Val Val Leu Gln Glu Asn Val Leu Leu Asn Arg545 550 555 560Ser Ile Arg Asp Asn Ile Ala Leu Thr Asp Thr Gly Met Pro Leu Glu 565 570 575Arg Ile Ile Glu Ala Ala Lys Leu Ala Gly Ala His Glu Phe Ile Met 580 585 590Glu Leu Pro Glu Gly Tyr Gly Thr Val Val Gly Glu Gln Gly Ala Gly 595 600 605Leu Ser Gly Gly Gln Arg Gln Arg Ile Ala Ile Ala Arg Ala Leu Ile 610 615 620Thr Asn Pro Arg Ile Leu Ile Phe Asp Glu Ala Thr Ser Ala Leu Asp625 630 635 640Tyr Glu Ser Glu Arg Ala Ile Met Gln Asn Met Gln Ala Ile Cys Ala 645 650 655Asn Arg Thr Val Leu Ile Ile Ala His Arg Leu Ser Thr Val Lys Thr 660 665 670Ala His Arg Ile Ile Ala Met Asp Lys Gly Arg Ile Val Glu Ala Gly 675 680 685Thr Gln Gln Glu Leu Leu Ala Lys Pro Asn Gly Tyr Tyr Arg Tyr Leu 690 695 700Tyr Asp Leu Gln Asn Gly705 710 29 2133 DNA Neisseria meningitidis 29atgtctatcg tatccgcacc gctccccgcc ctttccgccc tcatcatcct cgcccattac 60cacggcattg ccgccaatcc tgccgatata cagcatgaat tttgtacttc cgcacagagc 120gatttaaatg aaacgcaatg gctgttagcc gccaaatctt tgggattgaa ggcaaaggta 180gtccgccagc ctattaaacg tttggctatg gcgactttac ccgcattggt atggtgtgat 240gacggcaacc attttatttt ggctaaaaca gacggtgggg gtgagcatgc ccaatatcta 300atacaggatt taactacgaa taagtctgcg gtattgtctt ttgccgaatt ttctaacaga 360tattcgggca aactgatatt ggttgcttcc cgcgcttcgg tattgggcag tttggcaaag 420tttgacttta cctggtttat tccggcggta atcaaatacc gccggttgtt ttttgaagta 480ttggtggtgt cggtggtgtt gcagctgttt gcgctgatta cgcctctgtt tttccaagtg 540gtgatggaca aggtgctggt acatcgggga ttctctactt tggatgtggt gtcggtggct 600ttgttggtgg tgtcgctgtt tgagattgtg ttgggcggtt tgcggacgta tctgtttgca 660catacgactt cacgtattga tgtggaattg ggcgcgcgtt tgttccggca tctgctttcc 720ctgcctttat cctatttcga gcacagacga gtgggtgata cggtggctcg ggtgcgggaa 780ttggagcaga ttcgcaattt cttgaccggt caggcgctga cttcggtgtt ggatttggcg 840ttttcgttta tctttctggc ggtgatgtgg tattacagct ccactctgac ttgggtggta 900ttggcttcgt tgcctgccta tgcgttttgg tcggcattta tcagtccgat actgcggacg 960cgtctgaacg ataagttcgc gcgcaatgca gacaaccagt cgtttttagt agaaagcatc 1020actgcggtgg gtacggtaaa ggcgatggcg gtggagccgc agatgacgca gcgttgggac 1080aatcagttgg cggcttatgt ggcttcggga tttcgggtaa cgaagttggc ggtggtcggc 1140cagcaggggg tgcagctgat tcagaagctg gtgacggtgg cgacgttgtg gattggcgca 1200cggctggtaa ttgagagcaa gctgacggtg gggcagctga ttgcgtttaa tatgctctcg 1260ggacaggtgg cggcgcctgt tatccgtttg gcgcagttgt ggcaggattt ccagcaggtg 1320gggatttcgg tggcgcgttt gggggatatt ctgaatgcgc cgaccgagaa tgcgtcttcg 1380catttggctt tgcccgatat ccggggggag attacgttcg aacatgtcga tttccgctat 1440aaggcggacg gcaggctgat tttgcaggat ttgaacctgc ggattcgggc gggggaagtg 1500ctggggattg tgggacgttc ggggtcgggc aaatccacac tcaccaaatt ggtgcagcgt 1560ctgtatgtac cggcgcaggg acgggtgttg gtggacggca acgatttggc tttggccgct 1620cctgcttggc tgcggcggca ggtcggcgtg gtcttgcagg agaatgtgct gctcaaccgc 1680agcatacgcg acaatatcgc gctgacggat acgggtatgc cgctggaacg cattatcgaa 1740gcagccaaac tggcgggcgc acacgagttt attatggagc tgccggaagg ctacggcacc 1800gtggtgggcg aacaaggggc cggcttgtcg ggcggacagc ggcagcgtat tgcgattgcc 1860cgcgcgttaa tcaccaatcc gcgcattctg atttttgatg aagccaccag cgcgctggat 1920tatgaaagtg aacgagcgat tatgcagaac atgcaggcca tttgcgccaa ccggacggtg 1980ctgattatcg cccaccgtct gtccactgtt aaaacggcac accggatcat tgccatggat 2040aaaggcagga ttgtggaagc gggaacacag caggaattgc tggcgaagcc gaacggatat 2100taccgctatc tgtatgattt acagaacggg tag 2133 30 710 PRT Neisseria meningitidis 30Met Ser Ile Val Ser Ala Pro Leu Pro Ala Leu Ser Ala Leu Ile Ile 1 5 10 15Leu Ala His Tyr His Gly Ile Ala Ala Asn Pro Ala Asp Ile Gln His 20 25 30Glu Phe Cys Thr Ser Ala Gln Ser Asp Leu Asn Glu Thr Gln Trp Leu 35 40 45Leu Ala Ala Lys Ser Leu Gly Leu Lys Ala Lys Val Val Arg Gln Pro 50 55 60Ile Lys Arg Leu Ala Met Ala Thr Leu Pro Ala Leu Val Trp Cys Asp 65 70 75 80Asp Gly Asn His Phe Ile Leu Ala Lys Thr Asp Gly Gly Gly Glu His 85 90 95Ala Gln Tyr Leu Ile Gln Asp Leu Thr Thr Asn Lys Ser Ala Val Leu 100 105 110Ser Phe Ala Glu Phe Ser Asn Arg Tyr Ser Gly Lys Leu Ile Leu Val 115 120 125Ala Ser Arg Ala Ser Val Leu Gly Ser Leu Ala Lys Phe Asp Phe Thr 130 135 140Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Arg Leu Phe Phe Glu Val145 150 155 160Leu Val Val Ser Val Val Leu Gln Leu Phe Ala Leu Ile Thr Pro Leu 165 170 175Phe Phe Gln Val Val Met Asp Lys Val Leu Val His Arg Gly Phe Ser 180 185 190Thr Leu Asp Val Val Ser Val Ala Leu Leu Val Val Ser Leu Phe Glu 195 200 205Ile Val Leu Gly Gly Leu Arg Thr Tyr Leu Phe Ala His Thr Thr Ser 210 215 220Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe Arg His Leu Leu Ser225 230 235 240Leu Pro Leu Ser Tyr Phe Glu His Arg Arg Val Gly Asp Thr Val Ala 245 250 255Arg Val Arg Glu Leu Glu Gln Ile Arg Asn Phe Leu Thr Gly Gln Ala 260 265 270Leu Thr Ser Val Leu Asp Leu Ala Phe Ser Phe Ile Phe Leu Ala Val 275 280 285Met Trp Tyr Tyr Ser Ser Thr Leu Thr Trp Val Val Leu Ala Ser Leu 290 295 300Pro Ala Tyr Ala Phe Trp Ser Ala Phe Ile Ser Pro Ile Leu Arg Thr305 310 315 320Arg Leu Asn Asp Lys Phe Ala Arg Asn Ala Asp Asn Gln Ser Phe Leu 325 330 335Val Glu Ser Ile Thr Ala Val Gly Thr Val Lys Ala Met Ala Val Glu 340 345 350Pro Gln Met Thr Gln Arg Trp Asp Asn Gln Leu Ala Ala Tyr Val Ala 355 360 365Ser Gly Phe Arg Val Thr Lys Leu Ala Val Val Gly Gln Gln Gly Val 370 375 380Gln Leu Ile Gln Lys Leu Val Thr Val Ala Thr Leu Trp Ile Gly Ala385 390 395 400Arg Leu Val Ile Glu Ser Lys Leu Thr Val Gly Gln Leu Ile Ala Phe 405 410 415Asn Met Leu Ser Gly Gln Val Ala Ala Pro Val Ile Arg Leu Ala Gln 420 425 430Leu Trp Gln Asp Phe Gln Gln Val Gly Ile Ser Val Ala Arg Leu Gly 435 440 445Asp Ile Leu Asn Ala Pro Thr Glu Asn Ala Ser Ser His Leu Ala Leu 450 455 460Pro Asp Ile Arg Gly Glu Ile Thr Phe Glu His Val Asp Phe Arg Tyr465 470 475 480Lys Ala Asp Gly Arg Leu Ile Leu Gln Asp Leu Asn Leu Arg Ile Arg 485 490 495Ala Gly Glu Val Leu Gly Ile Val Gly Arg Ser Gly Ser Gly Lys Ser 500 505 510Thr Leu Thr Lys Leu Val Gln Arg Leu Tyr Val Pro Ala Gln Gly Arg 515 520 525Val Leu Val Asp Gly Asn Asp Leu Ala Leu Ala Ala Pro Ala Trp Leu 530 535 540Arg Arg Gln Val Gly Val Val Leu Gln Glu Asn Val Leu Leu Asn Arg545 550 555 560Ser Ile Arg Asp Asn Ile Ala Leu Thr Asp Thr Gly Met Pro Leu Glu 565 570 575Arg Ile Ile Glu Ala Ala Lys Leu Ala Gly Ala His Glu Phe Ile Met 580 585 590Glu Leu Pro Glu Gly Tyr Gly Thr Val Val Gly Glu Gln Gly Ala Gly 595 600 605Leu Ser Gly Gly Gln Arg Gln Arg Ile Ala Ile Ala Arg Ala Leu Ile 610 615 620Thr Asn Pro Arg Ile Leu Ile Phe Asp Glu Ala Thr Ser Ala Leu Asp625 630 635 640Tyr Glu Ser Glu Arg Ala Ile Met Gln Asn Met Gln Ala Ile Cys Ala 645 650 655Asn Arg Thr Val Leu Ile Ile Ala His Arg Leu Ser Thr Val Lys Thr 660 665 670Ala His Arg Ile Ile Ala Met Asp Lys Gly Arg Ile Val Glu Ala Gly 675 680 685Thr Gln Gln Glu Leu Leu Ala Lys Pro Asn Gly Tyr Tyr Arg Tyr Leu 690 695 700Tyr Asp Leu Gln Asn Gly705 710 31 186 DNA Neisseria meningitidis 31atgaaatact tgatccgcac cgccttactc gcagtcgcag ccgccggcat ctacgcctgc 60caaccgcaat ccgaagccgc agtgcaagtc aaggctgaaa acagcctgac cgctatgcgc 120ttagccgtcg ccgacaaaca ggcagagatt gacgggttga acgcccaaak sgacgccgaa 180atcaga 186 32 62 PRT Neisseria meningitidis SITE (57) unknown 32Met Lys Tyr Leu Ile Arg Thr Ala Leu Leu Ala Val Ala Ala Ala Gly 1 5 10 15Ile Tyr Ala Cys Gln Pro Gln Ser Glu Ala Ala Val Gln Val Lys Ala 20 25 30Glu Asn Ser Leu Thr Ala Met Arg Leu Ala Val Ala Asp Lys Gln Ala 35 40 45Glu Ile Asp Gly Leu Asn Ala Gln Xaa Asp Ala Glu Ile Arg 50 55 60 33 261 DNA Neisseria meningitidis 33atgaaatact tgatccgcac cgccttactc gcagtcgcag ccgccggcat ctacgcctgc 60caaccgcaat ccgaagccgc agtgcaagtc aaggctgaaa acagcctgac cgctatgcgc 120ttagccgtcg ccgacaaaca ggcagagatt gacgggttga acgcccaaat cgacgccgaa 180atcagacaac gcgaagccga agaattgaaa gactaccgat ggatacacgg cgacgcggaa 240gtgccggagc tggaaaaatg a 261 34 86 PRT Neisseria meningitidis 34Met Lys Tyr Leu Ile Arg Thr Ala Leu Leu Ala Val Ala Ala Ala Gly 1 5 10 15Ile Tyr Ala Cys Gln Pro Gln Ser Glu Ala Ala Val Gln Val Lys Ala 20 25 30Glu Asn Ser Leu Thr Ala Met Arg Leu Ala Val Ala Asp Lys Gln Ala 35 40 45Glu Ile Asp Gly Leu Asn Ala Gln Ile Asp Ala Glu Ile Arg Gln Arg 50 55 60Glu Ala Glu Glu Leu Lys Asp Tyr Arg Trp Ile His Gly Asp Ala Glu 65 70 75 80Val Pro Glu Leu Glu Lys 85 35 279 DNA Neisseria meningitidis 35atggttatcg gaatattact cgcatcaagc aagcatgctc ttgtcattac tctattgtta 60aatcccgtct tccatgcatc cagttgcgta tcgcgttsgg caatacggaa taaaatctgc 120tgttctgctt tggctaaatt tgccaaattg tttattgttt ctttaggagc agcttgctta 180gccgccttcg ctttcgacaa cgcccccaca ggcgcttccc aagcgttgcc taccgttacc 240gcacccgtgg cgattcccgc gcccgcttcg gcagcctga 279 36 92 PRT Neisseria meningitidis SITE (33) unknown 36Met Val Ile Gly Ile Leu Leu Ala Ser Ser Lys His Ala Leu Val Ile 1 5 10 15Thr Leu Leu Leu Asn Pro Val Phe His Ala Ser Ser Cys Val Ser Arg 20 25 30Xaa Ala Ile Arg Asn Lys Ile Cys Cys Ser Ala Leu Ala Lys Phe Ala 35 40 45Lys Leu Phe Ile Val Ser Leu Gly Ala Ala Cys Leu Ala Ala Phe Ala 50 55 60Phe Asp Asn Ala Pro Thr Gly Ala Ser Gln Ala Leu Pro Thr Val Thr 65 70 75 80Ala Pro Val Ala Ile Pro Ala Pro Ala Ser Ala Ala 85 90 37 312 DNA Neisseria meningitidis 37atggcttgta caggtttgat ggtttttccg ttaatggtta tcggaatatt acttgcatca 60agcaagcctg ctcctttcct tactctattg ttaaatcccg tcttccatgc atccagttgc 120gtatcgcgtt gggcaatacg gaataaaatc tgctgttctg ctttggctaa atttgccaaa 180ttgtttattg tttctttagg agcagcttgc ttagccgcct tcgctttcga caacgccccc 240acaggcgctt cccaagcgtt gcctaccgtt accgcacccg tggcgattcc cgcgcccgct 300tcggcagcct ga 312 38 103 PRT Neisseria meningitidis 38Met Ala Cys Thr Gly Leu Met Val Phe Pro Leu Met Val Ile Gly Ile 1 5 10 15Leu Leu Ala Ser Ser Lys Pro Ala Pro Phe Leu Thr Leu Leu Leu Asn 20 25 30Pro Val Phe His Ala Ser Ser Cys Val Ser Arg Trp Ala Ile Arg Asn 35 40 45Lys Ile Cys Cys Ser Ala Leu Ala Lys Phe Ala Lys Leu Phe Ile Val 50 55 60Ser Leu Gly Ala Ala Cys Leu Ala Ala Phe Ala Phe Asp Asn Ala Pro 65 70 75 80Thr Gly Ala Ser Gln Ala Leu Pro Thr Val Thr Ala Pro Val Ala Ile 85 90 95Pro Ala Pro Ala Ser Ala Ala 100 39 255 DNA Neisseria meningitidis 39atgttcagta ttttaaatgt gtttcttcat tgtattctgg cttgtgtagt ctctggtgag 60acgcctacta tatttggtat ccttgctctt ttttacttat tgtatctttc ttatcttgct 120gtttttaaga ttttcttttc ttttttctta gacagagttt cactccggtc tcccaggctg 180gagtgcaaat ggcatgaccc tttggctcac tggctcacgg ccacttctgc tattctgccg 240cctcagcctc caggg 255 40 85 PRT Neisseria meningitidis 40Met Phe Ser Ile Leu Asn Val Phe Leu His Cys Ile Leu Ala Cys Val 1 5 10 15Val Ser Gly Glu Thr Pro Thr Ile Phe Gly Ile Leu Ala Leu Phe Tyr 20 25 30Leu Leu Tyr Leu Ser Tyr Leu Ala Val Phe Lys Ile Phe Phe Ser Phe 35 40 45Phe Leu Asp Arg Val Ser Leu Arg Ser Pro Arg Leu Glu Cys Lys Trp 50 55 60His Asp Pro Leu Ala His Trp Leu Thr Ala Thr Ser Ala Ile Leu Pro 65 70 75 80Pro Gln Pro Pro Gly 85 41 237 DNA Neisseria meningitidis 41gtgcggacgt ggttggtttt ttggttgcag cgtttgaaat acccgttgtt gctttggatt 60gcggatatgt tgctgtaccg gttgttgggc ggcgcggaaa tcgaatgcgg ccgttgccct 120gtgccgccga tgacggattg gcagcatttt ttgccggcga tgggaacggt gtcggcttgg 180gtggcggtga tttgggcata cctgatgatt gaaagtgaaa aaaacggaag atattga 237 42 78 PRT Neisseria meningitidis 42Val Arg Thr Trp Leu Val Phe Trp Leu Gln Arg Leu Lys Tyr Pro Leu 1 5 10 15Leu Leu Trp Ile Ala Asp Met Leu Leu Tyr Arg Leu Leu Gly Gly Ala 20 25 30Glu Ile Glu Cys Gly Arg Cys Pro Val Pro Pro Met Thr Asp Trp Gln 35 40 45His Phe Leu Pro Ala Met Gly Thr Val Ser Ala Trp Val Ala Val Ile 50 55 60Trp Ala Tyr Leu Met Ile Glu Ser Glu Lys Asn Gly Arg Tyr 65 70 75 43 237 DNA Neisseria meningitidis 43gtgcggacgt ggttggtttt ttggttgcag cgtttgaaat acccgttgtt gctttgtatt 60gcggatatgc tgctgtaccg gttgttgggc ggcgcggaaa tcgaatgcgg ccgttgccct 120gtaccgccga tgacggattg gcagcatttt ttgccgacga tgggaacggt ggcggcttgg 180gtggcggtga tttgggcata cctgatgatt gaaagtgaaa aaaacggaag atattga 237 44 78 PRT Neisseria meningitidis 44Val Arg Thr Trp Leu Val Phe Trp Leu Gln Arg Leu Lys Tyr Pro Leu 1 5 10 15Leu Leu Cys Ile Ala Asp Met Leu Leu Tyr Arg Leu Leu Gly Gly Ala 20 25 30Glu Ile Glu Cys Gly Arg Cys Pro Val Pro Pro Met Thr Asp Trp Gln 35 40 45His Phe Leu Pro Thr Met Gly Thr Val Ala Ala Trp Val Ala Val Ile 50 55 60Trp Ala Tyr Leu Met Ile Glu Ser Glu Lys Asn Gly Arg Tyr 65 70 75 45 660 DNA Neisseria meningitidis 45atgtttcaaa attttgattt gggcgtgttc ctgcttgccg tcctccccgt gctgccctcc 60attaccgtct cgcacgtggc gcgcggctat acggcgcgct actggggaga caacactgcc 120gaacaatacg gcaggctgac actgaacccc ctgccccata tcgatttggt cggcacaatc 180atcgtaccgc tgcttacttt gatgttcacg cccttcctgt tcggctgggc gcgtccgatt 240cctatcgatt cgcgcaactt ccgcaacccg cgccttgcct ggcgttgcgt tgccgcgtcc 300ggcccgctgt cgaatctagc gatggctgtw ctgtggggcg tggttttggt gctgactccg 360tatgtcggcg gggcgtatca gatgccgttg gctcaaatgg caaactacgg tattctgatc 420aatgcgattc tgttcgcgct caacatcatc cccatcctgc cttgggacgg cggcattttc 480atcgacacct tcctgtcggc gaaatattcg caagcgttcc gcaaaatcga accttatggg 540acgtggatta tcctactgct gatgctgacc sgggttttgg gtgcgtttat wgcaccgatt 600stgcggmtgc gtgattgcrt ttgtgcagat gtwcgtctga ctggctttca gacggcataa 660 46 219 PRT Neisseria meningitidis SITE (191) unknown 46Met Phe Gln Asn Phe Asp Leu Gly Val Phe Leu Leu Ala Val Leu Pro 1 5 10 15Val Leu Pro Ser Ile Thr Val Ser His Val Ala Arg Gly Tyr Thr Ala 20 25 30Arg Tyr Trp Gly Asp Asn Thr Ala Glu Gln Tyr Gly Arg Leu Thr Leu 35 40 45Asn Pro Leu Pro His Ile Asp Leu Val Gly Thr Ile Ile Val Pro Leu 50 55 60Leu Thr Leu Met Phe Thr Pro Phe Leu Phe Gly Trp Ala Arg Pro Ile 65 70 75 80Pro Ile Asp Ser Arg Asn Phe Arg Asn Pro Arg Leu Ala Trp Arg Cys 85 90 95Val Ala Ala Ser Gly Pro Leu Ser Asn Leu Ala Met Ala Val Leu Trp 100 105 110Gly Val Val Leu Val Leu Thr Pro Tyr Val Gly Gly Ala Tyr Gln Met 115 120 125Pro Leu Ala Gln Met Ala Asn Tyr Gly Ile Leu Ile Asn Ala Ile Leu 130 135 140Phe Ala Leu Asn Ile Ile Pro Ile Leu Pro Trp Asp Gly Gly Ile Phe145 150 155 160Ile Asp Thr Phe Leu Ser Ala Lys Tyr Ser Gln Ala Phe Arg Lys Ile 165 170 175Glu Pro Tyr Gly Thr Trp Ile Ile Leu Leu Leu Met Leu Thr Xaa Val 180 185 190Leu Gly Ala Phe Ile Ala Pro Ile Xaa Arg Xaa Arg Asp Cys Xaa Cys 195 200 205Ala Asp Val Arg Leu Thr Gly Phe Gln Thr Ala 210 215 47 639 DNA Neisseria meningitidis 47atgtttcaaa attttgattt gggcgtgttt ctgcttgccg tcctgcccgt gctgctctcc 60attaccgtca gggaggtggc gcgcggctat acggcgcgct actggggaga caacactgcc 120gaacaatacg gcaggctgac actgaacccc ctgccccata tcgatttggt cggcacaatc 180atcgtaccgc tgcttacttt gatgttcacg cccttcctgt tcggctgggc gcgtccgatt 240cctatcgatt cgcgcaactt ccgcaacccg cgccttgcct ggcgttgcgt tgccgcgtcc 300ggcccgctgt cgaatctagc gatggctgtt ctgtggggcg tggttttggt gctgactccg 360tatgtcggcg gggcgtatca gatgccgttg gctcaaatgg caaactacgg tattctgatc 420aatgcgattc tgttcgcgct caacatcatc cccatcctgc cttgggacgg cggcattttc 480atcgacacct tcctgtcggc gaaatattcg caagcgttcc gcaaaatcga accttatggg 540acgtggatta tcctactgct gatgctgacc ggggttttgg gtgcgtttat tgcaccgatt 600gtgcggctgg tgattgcgtt tgtgcagatg ttcgtctga 639 48 212 PRT Neisseria meningitidis 48Met Phe Gln Asn Phe Asp Leu Gly Val Phe Leu Leu Ala Val Leu Pro 1 5 10 15Val Leu Leu Ser Ile Thr Val Arg Glu Val Ala Arg Gly Tyr Thr Ala 20 25 30Arg Tyr Trp Gly Asp Asn Thr Ala Glu Gln Tyr Gly Arg Leu Thr Leu 35 40 45Asn Pro Leu Pro His Ile Asp Leu Val Gly Thr Ile Ile Val Pro Leu 50 55 60Leu Thr Leu Met Phe Thr Pro Phe Leu Phe Gly Trp Ala Arg Pro Ile 65 70 75 80Pro Ile Asp Ser Arg Asn Phe Arg Asn Pro Arg Leu Ala Trp Arg Cys 85 90 95Val Ala Ala Ser Gly Pro Leu Ser Asn Leu Ala Met Ala Val Leu Trp 100 105 110Gly Val Val Leu Val Leu Thr Pro Tyr Val Gly Gly Ala Tyr Gln Met 115 120 125Pro Leu Ala Gln Met Ala Asn Tyr Gly Ile Leu Ile Asn Ala Ile Leu 130 135 140Phe Ala Leu Asn Ile Ile Pro Ile Leu Pro Trp Asp Gly Gly Ile Phe145 150 155 160Ile Asp Thr Phe Leu Ser Ala Lys Tyr Ser Gln Ala Phe Arg Lys Ile 165 170 175Glu Pro Tyr Gly Thr Trp Ile Ile Leu Leu Leu Met Leu Thr Gly Val 180 185 190Leu Gly Ala Phe Ile Ala Pro Ile Val Arg Leu Val Ile Ala Phe Val 195 200 205Gln Met Phe Val 210 49 558 DNA Neisseria meningitidis misc_feature (312) any nucleotide 49cgcggctata cagcgcgcta ctggggtgac aacactgccg aacaatacgg caggctgaca 60ctgaaccccc tgccccatat cgatttggtc ggcacaatca tcgtaccgct gcttactttg 120atgtttacgc ccttcctgtt cggctgggcg cgtccgattc ctatcgattc gcgcaacttc 180cgcaacccgc gccttgcctg gcgttgcgtt gccgcgtccg gcccgctgtc gaatctggcg 240atggctgttc tgtggggcgt ggttttggtg ctgactccgt atgtcggtgg ggcgtatcag 300atgccgttgg cncaaatggc aaactacnnn attctgatca atgcgattct gtncgcgctc 360aacatcatcc ccatcctgcc ttgggacggc ggcattttca tcgacacctt cctgtcggcn 420aaatantcgc aagcgttccg caaaatcgaa ccttatggga cgtggattat ccngctgctt 480atgctgaccg gggttttggg tgcgtntatt gcaccgattg tgcagctggt gattgcgttt 540gtgcagatgt tcgtctga 558 50 185 PRT Neisseria meningitidis SITE (110) unknown 50Arg Gly Tyr Thr Ala Arg Tyr Trp Gly Asp Asn Thr Ala Glu Gln Tyr 1 5 10 15Gly Arg Leu Thr Leu Asn Pro Leu Pro His Ile Asp Leu Val Gly Thr 20 25 30Ile Ile Val Pro Leu Leu Thr Leu Met Phe Thr Pro Phe Leu Phe Gly 35 40 45Trp Ala Arg Pro Ile Pro Ile Asp Ser Arg Asn Phe Arg Asn Pro Arg 50 55 60Leu Ala Trp Arg Cys Val Ala Ala Ser Gly Pro Leu Ser Asn Leu Ala 65 70 75 80Met Ala Val Leu Trp Gly Val Val Leu Val Leu Thr Pro Tyr Val Gly 85 90 95Gly Ala Tyr Gln Met Pro Leu Ala Gln Met Ala Asn Tyr Xaa Ile Leu 100 105 110Ile Asn Ala Ile Leu Xaa Ala Leu Asn Ile Ile Pro Ile Leu Pro Trp 115 120 125Asp Gly Gly Ile Phe Ile Asp Thr Phe Leu Ser Ala Lys Xaa Ser Gln 130 135 140Ala Phe Arg Lys Ile Glu Pro Tyr Gly Thr Trp Ile Ile Xaa Leu Leu145 150 155 160Met Leu Thr Gly Val Leu Gly Ala Xaa Ile Ala Pro Ile Val Gln Leu 165 170 175Val Ile Ala Phe Val Gln Met Phe Val 180 185 51 498 DNA Neisseria meningitidis 51atgaacctga tttcacgtta catcatccgt caaatggcgg ttatggcggt ttacgcgctc 60cttgccttcc tcgctttgta cagctttttt gaaatcctgt acgaaaccgg caacctcggc 120aaaggcagtt acggcatatg ggaaatgctg ggctacaccg ccctcaaaat gcccgcccgc 180gcctacgaac tgattcccct cgccgtcctt atcggcggac tggtctccct cagccagctt 240gccgccggca gcgaactgac cgtcatcaaa gccagcggca tgagcaccaa aaagctgctg 300ttgattctgt cgcagttcgg ttttattttt gctattgcca ccgtcgcgct cggcgaatgg 360gttgcgccca cactgagcca aaaagccgaa aacatcaaag ccgccgccat caacggcaaa 420atcagcaccg gcaataccgg cctttggctg aaagaaaaaa acagcgtgat caatgtgcgc 480gaaatgttgc ccgaccat 498 52 166 PRT Neisseria meningitidis 52Met Asn Leu Ile Ser Arg Tyr Ile Ile Arg Gln Met Ala Val Met Ala 1 5 10 15Val Tyr Ala Leu Leu Ala Phe Leu Ala Leu Tyr Ser Phe Phe Glu Ile 20 25 30Leu Tyr Glu Thr Gly Asn Leu Gly Lys Gly Ser Tyr Gly Ile Trp Glu 35 40 45Met Leu Gly Tyr Thr Ala Leu Lys Met Pro Ala Arg Ala Tyr Glu Leu 50 55 60Ile Pro Leu Ala Val Leu Ile Gly Gly Leu Val Ser Leu Ser Gln Leu 65 70 75 80Ala Ala Gly Ser Glu Leu Thr Val Ile Lys Ala Ser Gly Met Ser Thr 85 90 95Lys Lys Leu Leu Leu Ile Leu Ser Gln Phe Gly Phe Ile Phe Ala Ile 100 105 110Ala Thr Val Ala Leu Gly Glu Trp Val Ala Pro Thr Leu Ser Gln Lys 115 120 125Ala Glu Asn Ile Lys Ala Ala Ala Ile Asn Gly Lys Ile Ser Thr Gly 130 135 140Asn Thr Gly Leu Trp Leu Lys Glu Lys Asn Ser Val Ile Asn Val Arg145 150 155 160Glu Met Leu Pro Asp His 165 53 980 DNA Neisseria meningitidis 53atgaacctga tttcacgtta catcatccgt caaatggcgg ttatggcggt ttacgcgctc 60cttgccttcc tcgctttgta cagctttttt gaaatcctgt acgaaaccgg caacctcggc 120aaaggcagtt acggcatatg ggaaatgctg ggctacaccg ccctcaaaat gcccgcccgc 180gcctacgaac tgattcccct cgccgtcctt atcggcggac tggtctccct cagccagctt 240gccgccggca gcgaactgac cgtcatcaaa gccagcggca tgagcaccaa aaagctgctg 300ttgattctgt cgcagttcgg ttttattttt gctattgcca ccgtcgcgct cggcgaatgg 360gttgcgccca cactgagcca aaaagccgaa aacatcaaag ccgccgccat caacggcaaa 420atcagcaccg gcaataccgg cctttggctg aaagaaaaaa acagcrtkat caatgtgcgc 480gaaatgttgc ccgaccatac gcttttgggc atcaaaattt gggcgcgcaa cgataaaaac 540gaattggcag aggcagtgga agccgattcc gccgttttga acagcgacgg cagttggcag 600ttgaaaaaca tccgccgcag cacgcttggc gaagacaaag tcgaggtctc tattgcggct 660gaagaaaact ggccgatttc cgtcaaacgc aacctgatgg acgtattgct cgtcaaaccc 720gaccaaatgt ccgtcggcga actgaccacc tacatccgcc acctccaaaa caacagccaa 780aacacccgaa tctacgccat cgcatggtgg cgcaaattgg tttaccccgc cgcagcctgg 840gtgatggcgc tcgtcgcctt tgcctttacc ccgcaaacca cccgccacgg caatatgggc 900ttaaaactct tcggcggcat ctgtstcgga ttgctgttcc accttgccgg acggctcttt 960gggtttacca gccaactcgg 980 54 326 PRT Neisseria meningitidis SITE (156) unknown 54Met Asn Leu Ile Ser Arg Tyr Ile Ile Arg Gln Met Ala Val Met Ala 1 5 10 15Val Tyr Ala Leu Leu Ala Phe Leu Ala Leu Tyr Ser Phe Phe Glu Ile 20 25 30Leu Tyr Glu Thr Gly Asn Leu Gly Lys Gly Ser Tyr Gly Ile Trp Glu 35 40 45Met Leu Gly Tyr Thr Ala Leu Lys Met Pro Ala Arg Ala Tyr Glu Leu 50 55 60Ile Pro Leu Ala Val Leu Ile Gly Gly Leu Val Ser Leu Ser Gln Leu 65 70 75 80Ala Ala Gly Ser Glu Leu Thr Val Ile Lys Ala Ser Gly Met Ser Thr 85 90 95Lys Lys Leu Leu Leu Ile Leu Ser Gln Phe Gly Phe Ile Phe Ala Ile 100 105 110Ala Thr Val Ala Leu Gly Glu Trp Val Ala Pro Thr Leu Ser Gln Lys 115 120 125Ala Glu Asn Ile Lys Ala Ala Ala Ile Asn Gly Lys Ile Ser Thr Gly 130 135 140Asn Thr Gly Leu Trp Leu Lys Glu Lys Asn Ser Xaa Ile Asn Val Arg145 150 155 160Glu Met Leu Pro Asp His Thr Leu Leu Gly Ile Lys Ile Trp Ala Arg 165 170 175Asn Asp Lys Asn Glu Leu Ala Glu Ala Val Glu Ala Asp Ser Ala Val 180 185 190Leu Asn Ser Asp Gly Ser Trp Gln Leu Lys Asn Ile Arg Arg Ser Thr 195 200 205Leu Gly Glu Asp Lys Val Glu Val Ser Ile Ala Ala Glu Glu Asn Trp 210 215 220Pro Ile Ser Val Lys Arg Asn Leu Met Asp Val Leu Leu Val Lys Pro225 230 235 240Asp Gln Met Ser Val Gly Glu Leu Thr Thr Tyr Ile Arg His Leu Gln 245 250 255Asn Asn Ser Gln Asn Thr Arg Ile Tyr Ala Ile Ala Trp Trp Arg Lys 260 265 270Leu Val Tyr Pro Ala Ala Ala Trp Val Met Ala Leu Val Ala Phe Ala 275 280 285Phe Thr Pro Gln Thr Thr Arg His Gly Asn Met Gly Leu Lys Leu Phe 290 295 300Gly Gly Ile Cys Xaa Gly Leu Leu Phe His Leu Ala Gly Arg Leu Phe305 310 315 320Gly Phe Thr Ser Gln Leu 325 55 1071 DNA Neisseria meningitidis misc_feature (148) any nucleotide 55atgaacctga tttcacgtta catcatccgt caaatggcgg ttatggcggt ttacgcgctc 60cttgccttcc tcgctttgta cagctttttt gaaatcctgt acgaaaccgg caacctcggc 120aaaggcagtt acggcatatg ggaaatgntg ggntacaccg ccctcaaaat gnccgcccgc 180gcctacgaac tgatgcccct cgccgtcctt atcggcggac tggtctctnt cagccagctt 240gccgccggca gcgaactgan cgtcatcaaa gccagcggca tgagcaccaa aaagctgctg 300ttgattctgt cgcagttcgg ttttattttt gctattgcca ccgtcgcgct cggcgaatgg 360gttgcgccca cactgagcca aaaagccgaa aacatcaaag ccgcggccat caacggcaaa 420atcagtaccg gcaataccgg cctttggctg aaagaaaaaa acagcattat caatgtgcgc 480gaaatgttgc ccgaccatac cctgctgggc attaaaatct gggcccgcaa cgataaaaac 540gaactggcag aggcagtgga agccgattcc gccgttttga acagcgacgg cagttggcag 600ttgaaaaaca tccgccgcag cacgcttggc gaagacaaag tcgaggtctc tattgcggct 660gaagaaaant ggccgatttc cgtcaaacgc aacctgatgg acgtattgct cgtcaaaccc 720gaccaaatgt ccgtcggcga actgaccacc tacatccgcc acctccaaan nnacagccaa 780aacacccgaa tctacgccat cgcatggtgg cgcaaattgg tttaccccgc cgcagcctgg 840gtgatggcgc tcgtcgcctt tgcctttacc ccgcaaacca cccgccacgg caatatgggc 900ttaaaantct tcggcggcat ctgtctcgga ttgctgttcc accttgccgg ncggctcttc 960nggtttacca gccaactcta cggcatcccg cccttcctcg ncggcgcact acctaccata 1020gccttcgcct tgctcgccgt ttggctgata cgcaaacagg aaaaacgcta a 1071 56 356 PRT Neisseria meningitidis SITE (50) unknown 56Met Asn Leu Ile Ser Arg Tyr Ile Ile Arg Gln Met Ala Val Met Ala 1 5 10 15Val Tyr Ala Leu Leu Ala Phe Leu Ala Leu Tyr Ser Phe Phe Glu Ile 20 25 30Leu Tyr Glu Thr Gly Asn Leu Gly Lys Gly Ser Tyr Gly Ile Trp Glu 35 40 45Met Xaa Gly Tyr Thr Ala Leu Lys Met Xaa Ala Arg Ala Tyr Glu Leu 50 55 60Met Pro Leu Ala Val Leu Ile Gly Gly Leu Val Ser Xaa Ser Gln Leu 65 70 75 80Ala Ala Gly Ser Glu Leu Xaa Val Ile Lys Ala Ser Gly Met Ser Thr 85 90 95Lys Lys Leu Leu Leu Ile Leu Ser Gln Phe Gly Phe Ile Phe Ala Ile 100 105 110Ala Thr Val Ala Leu Gly Glu Trp Val Ala Pro Thr Leu Ser Gln Lys 115 120 125Ala Glu Asn Ile Lys Ala Ala Ala Ile Asn Gly Lys Ile Ser Thr Gly 130 135 140Asn Thr Gly Leu Trp Leu Lys Glu Lys Asn Ser Ile Ile Asn Val Arg145 150 155 160Glu Met Leu Pro Asp His Thr Leu Leu Gly Ile Lys Ile Trp Ala Arg 165 170 175Asn Asp Lys Asn Glu Leu Ala Glu Ala Val Glu Ala Asp Ser Ala Val 180 185 190Leu Asn Ser Asp Gly Ser Trp Gln Leu Lys Asn Ile Arg Arg Ser Thr 195 200 205Leu Gly Glu Asp Lys Val Glu Val Ser Ile Ala Ala Glu Glu Xaa Trp 210 215 220Pro Ile Ser Val Lys Arg Asn Leu Met Asp Val Leu Leu Val Lys Pro225 230 235 240Asp Gln Met Ser Val Gly Glu Leu Thr Thr Tyr Ile Arg His Leu Gln 245 250 255Xaa Xaa Ser Gln Asn Thr Arg Ile Tyr Ala Ile Ala Trp Trp Arg Lys 260 265 270Leu Val Tyr Pro Ala Ala Ala Trp Val Met Ala Leu Val Ala Phe Ala 275 280 285Phe Thr Pro Gln Thr Thr Arg His Gly Asn Met Gly Leu Lys Xaa Phe 290 295 300Gly Gly Ile Cys Leu Gly Leu Leu Phe His Leu Ala Gly Arg Leu Phe305 310 315 320Xaa Phe Thr Ser Gln Leu Tyr Gly Ile Pro Pro Phe Leu Xaa Gly Ala 325 330 335Leu Pro Thr Ile Ala Phe Ala Leu Leu Ala Val Trp Leu Ile Arg Lys 340 345 350Gln Glu Lys Arg 355 57 854 DNA Neisseria meningitidis 57gcagtagccg aaactgccaa cagccagggc aaaggtaaac aggcaggcag ttcggtttct 60gtttcactga aaacttcagg cgacctttgc ggcaaactca aaaccaccct taaaactttg 120gtctgctctt tggtttccct gagtatggta ttgcctgccc atgcccaaat taccaccgac 180aaatcagcac ctaaaaacca gcaggtcgtt atccttaaaa ccaacactgg tgcccccttg 240gtgaatatcc aaactccgaa tggacgcgga ttgagccaca accgctatac gcatttgatg 300ttgacaacaa aggggcagtg ttaaacaacg accgtaacaa taatccgttt gtggtcaaag 360gcagtgcgca attgattttg aacgaggtac gcggtacggc tagcaaactc aacggcatcg 420ttaccgtagg cggtcaaaag gccgacgtga ttattgccaa ccccaacggc attaccgtta 480atggcggcgg ctttaaaaat gtcggtcggg gcatcttaac taccggtgcg ccccaaatcg 540gcaaagacgg tgcactgaca ggatttgatg tgcgtcaagg cacattggac cgtagragca 600gcaggttgga atgataaagg cggagcmrmy tacaccgggg tacttgctcg tgcagttgct 660ttgcagggga aattwmmggg taaaaactgg cggtttctac cggtcctcag aaagtagatt 720acgccagcgg cgaaatcagt gcaggtacgg cagcgggtac gaaaccgact attgcccttg 780atactgccgc actgggcggt atgtacgccg acagcatcac actgattgcc aatgaaaaag 840gcgtaggcgt ctaa 854 58 284 PRT Neisseria meningitidis SITE (96) unknown 58Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln Ala Gly 1 5 10 15Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Leu Cys Gly Lys 20 25 30Leu Lys Thr Thr Leu Lys Thr Leu Val Cys Ser Leu Val Ser Leu Ser 35 40 45Met Val Leu Pro Ala His Ala Gln Ile Thr Thr Asp Lys Ser Ala Pro 50 55 60Lys Asn Gln Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala Pro Leu 65 70 75 80Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn Arg Xaa 85 90 95Tyr Ala Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn Asp Arg 100 105 110Asn Asn Asn Pro Phe Val Val Lys Gly Ser Ala Gln Leu Ile Leu Asn 115 120 125Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr Val Gly 130 135 140Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile Thr Val145 150 155 160Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr Thr Gly 165 170 175Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp Val Val 180 185 190Lys Ala His Trp Thr Val Xaa Ala Ala Gly Trp Asn Asp Lys Gly Gly 195 200 205Ala Xaa Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln Gly Lys 210 215 220Xaa Xaa Gly Lys Xaa Leu Ala Val Ser Thr Gly Pro Gln Lys Val Asp225 230 235 240Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr Lys Pro 245 250 255Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala Asp Ser 260 265 270Ile Thr Leu Ile Ala Asn Glu Lys Gly Val Gly Val 275 280 59 5937 DNA Neisseria meningitidis 59atgaataaag gtttacatcg cattatcttt agtaaaaagc acagcaccat ggttgcagta 60gccgaaactg ccaacagcca gggcaaaggt aaacaggcag gcagttcggt ttctgtttca 120ctgaaaactt caggcgacct ttgcggcaaa ctcaaaacca cccttaaaac tttggtctgc 180tctttggttt ccctgagtat ggtattgcct gcccatgccc aaattaccac cgacaaatca 240gcacctaaaa accagcaggt cgttatcctt aaaaccaaca ctggtgcccc cttggtgaat 300atccaaactc cgaatggacg cggattgagc cacaaccgct atacgcagtt tgatgttgac 360aacaaagggg cagtgttaaa caacgaccgt aacaataatc cgtttgtggt caaaggcagt 420gcgcaattga ttttgaacga ggtacgcggt acggctagca aactcaacgg catcgttacc 480gtaggcggtc aaaaggccga cgtgattatt gccaacccca acggcattac cgttaatggc 540ggcggcttta aaaatgtcgg tcggggcatc ttaactaccg gtgcgcccca aatcggcaaa 600gacggtgcac tgacaggatt tgatgtgcgt caaggcacat tgaccgtagg agcagcaggt 660tggaatgata aaggcggagc cgactacacc ggggtacttg ctcgtgcagt tgctttgcag 720gggaaattac agggtaaaaa cctggcggtt tctaccggtc ctcagaaagt agattacgcc 780agcggcgaaa tcagtgcagg tacggcagcg ggtacgaaac cgactattgc ccttgatact 840gccgcactgg gcggtatgta cgccgacagc atcacactga ttgccaatga aaaaggcgta 900ggcgtcaaaa atgccggcac actcgaagcg gccaagcaat tgattgtgac ttcgtcaggc 960cgcattgaaa acagcggccg catcgccacc actgccgacg gcaccgaagc ttcaccgact 1020tatctctcca tcgaaaccac cgaaaaagga gcggcaggca catttatctc caatggtggt 1080cggatcgaga gcaaaggctt attggttatt gagacgggag aagatatcag cttgcgtaac 1140ggagccgtgg tgcagaataa cggcagtcgc ccagctacca cggtattaaa tgctggtcat 1200aatttggtga ttgagagcaa aactaatgtg aacaatgcca aaggcccggc tactctgtcg 1260gccgacggcc gtaccgtcat caaggaggcc agtattcaga ctggcactac cgtatacagt 1320tccagcaaag gcaacgccga attaggcaat aacacacgca ttaccggggc agatgttacc 1380gtattatcca acggcaccat cagcagttcc gccgtaatag atgccaaaga caccgcacac 1440atcgaagcag gcaaaccgct ttctttggaa gcttcaacag ttacctccga tatccgctta 1500aacggaggca gtatcaaggg cggcaagcag cttgctttac tggcagacga taacattact 1560gccaaaacta ccaatctgaa tactcccggc aatctgtatg ttcatacagg taaagatctg 1620aatttgaatg ttgataaaga tttgtctgcc gccagcatcc atttgaaatc ggataacgct 1680gcccatatta ccggcaccag taaaaccctc actgcctcaa aagacatggg tgtggaggca 1740ggctcgctga atgttaccaa taccaatctg cgtaccaact cgggtaatct gcacattcag 1800gcagccaaag gcaatattca gcttcgcaat accaagctga acgcagccaa ggctctcgaa 1860accaccgcat tgcagggcaa tatcgtttca gacggccttc atgctgtttc tgcagacggt 1920catgtatcct tattggccaa cggtaatgcc gactttaccg gtcacaatac cctgacagcc 1980aaggccgatg tcaatgcagg atcggttggt aaaggccgtc tgaaagcaga caataccaat 2040atcacttcat cttcaggaga tattacgttg gttgccggca acggtattca gcttggtgac 2100ggaaaacaac gcaattcaat caacggaaaa cacatcagca tcaaaaacaa cggtggtaat 2160gccgacttaa aaaaccttaa cgtccatgcc aaaagcgggg cattgaacat tcattccgac 2220cgggcattga gcatagaaaa taccaagctg gagtctaccc ataatacgca tcttaatgca 2280caacacgagc gggtaacgct caaccaagta gatgcctacg cacaccgtca tctaagcatt 2340accggcagcc agatttggca aaacgacaaa ctgccttctg ccaacaagct ggtggctaac 2400ggtgtattgg cactcaatgc gcgctattcc caaattgccg acaacaccac gctgagagcg 2460ggtgcaatca accttactgc cggtaccgcc ctagtcaagc gcggcaacat caattggagt 2520accgtttcga ccaaaacttt ggaagataat gccgaattaa aaccattggc cggacggctg 2580aatattgaag caggtagcgg cacattaacc atcgaacctg ccaaccgcat cagtgcgcat 2640accgacctga gcatcaaaac aggcggaaaa ttgctgttgt ctgcaaaagg aggaaatgca 2700ggtgcgccta gtgctcaagt ttcctcattg gaagcaaaag gcaatatccg tctggttaca 2760ggagaaacag atttaagagg ttctaaaatt acagccggta aaaacttggt tgtcgccacc 2820accaaaggca agttgaatat cgaagccgta aacaactcat tcagcaatta ttttcctaca 2880caaaaagcgg ctgaactcaa ccaaaaatcc aaagaattgg aacagcagat tgcgcagttg 2940aaaaaaagct cgcctaaaag caagctgatt ccaaccctgc aagaagaacg cgaccgtctc 3000gctttctata ttcaagccat caacaaggaa gttaaaggta aaaaacccaa aggcaaagaa 3060tacctgcaag ccaagctttc tgcacaaaat attgacttga tttccgcaca aggcatcgaa 3120atcagcggtt ccgatattac cgcttccaaa aaactgaacc ttcacgccgc aggcgtattg 3180ccaaaggcag cagattcaga ggcggctgct attctgattg acggcataac cgaccaatat 3240gaaattggca agcccaccta caagagtcac tacgacaaag ctgctctgaa caagccttca 3300cgtttgaccg gacgtacagg ggtaagtatt catgcagctg cggcactcga tgatgcacgt 3360attattatcg gtgcatccga aatcaaagct ccctcaggca gcatagacat caaagcccat 3420agtgatattg tactggaggc tggacaaaac gatgcctata ccttcttaaa aaccaaaggt 3480aaaagcggca aaatcatcag aaaaaccaag tttaccagca cccgcgacca cctgattatg 3540ccagcccccg tcgagctgac cgccaacggc ataacgcttc aggcaggcgg caacatcgaa 3600gctaatacca cccgcttcaa tgcccctgca ggtaaagtta ccctggttgc gggtgaagag 3660ctgcaactgc tggcagaaga aggcatccac aagcacgagt tggatgtcca aaaaagccgc 3720cgctttatcg gcatcaaggt aggcaagagc aattacagta aaaacgaact gaacgaaacc 3780aaattgcctg tccgcgtcgt cgcccaaact gcagccaccc gttcaggctg ggataccgtg 3840ctcgaaggta ccgaattcaa aaccacgctg gccggtgcgg acattcaggc aggtgtaggc 3900gaaaaagccc gtgccgatgc gaaaattatc ctcaaaggca ttgtgaaccg tatccagtcg 3960gaagaaaaat tagaaaccaa ctcaaccgta tggcagaaac aggccggacg cggcagcact 4020atcgaaacgc tgaaactgcc cagcttcgaa agccctactc cgcccaaact gaccgccccc 4080ggtggctata tcgtcgacat tccgaaaggc aatttgaaaa ccgaaatcga aaagctggcc 4140aaacagcccg agtatgccta tctgaaacag ctccaagtag cgaaaaacgt caactggaac 4200caggtgcaac tggcttacga taaatgggac tataagcagg aaggcttaac cagagccggt 4260gcagcgattg ttaccataat cgtaaccgca ctgacttatg gatacggcgc aaccgcagcg 4320ggcggtgtag ccgcttcagg aagtagtaca gccgcagctg ccggaacagc cgccacaacg 4380acagcagcag ctactaccgt ttctacagcg actgccatgc aaaccgctgc tttagcctcc 4440ttgtatagcc aagcagctgt atccatcatc aataataaag gtgatgtcgg caaagcgttg 4500aaagatctcg gcaccagtga tacggtcaag cagattgtca cttctgccct gacggcgggt 4560gcattaaatc agatgggcgc agatattgcc caattgaaca gcaaggtaag aaccgaactg 4620ttcagcagta cgggcaatca aactattgcc aaccttggag gcagactggc taccaatctc 4680agtaatgcag gtatctcagc tggtatcaat accgccgtca acggcggcag cctgaaagac 4740aacttaggca atgccgcatt aggagcattg gttaatagct tccaaggaga agccgccagc 4800aaaatcaaaa caaccttcag cgacgattat gttgccaaac agttcgccca cgctttggct 4860gggtgtgtta gcggattggt acaaggaaaa tgtaaagacg gggcaattgg cgcagcagtt 4920ggggaaatcg tagccgactc catgcttggc ggcagaaacc ctgctacact cagcgatgcg 4980gaaaagcata aggttatcag ttactcgaag attattgccg gcagcgtggc ggcactcaac 5040ggcggcgatg tgaatactgc ggcgaatgcg gctgaggtgg cggtagtgaa taatgctttg 5100aattttgaca gtacccctac caatgcgaaa aagcatcaac cgcagaagcc cgacaaaacc 5160gcactggaaa aaattatcca aggtattatg cctgcacatg cagcaggtgc gatgactaat 5220ccgcaggata aggatgctgc catttggata agcaatatcc gtaatggcat cacaggcccg 5280attgtgatta ccagctatgg ggtttatgct gcaggttgga cagctccgct gatcggtaca 5340gcgggtaaat tagctatcag cacctgcatg gctaatcctt ctggttgtac tgtcatggtc 5400actcaggctg ccgaagcggg cgcgggaatc gccacgggtg cggtaacggt aggcaacgct 5460tgggaagcgc ctgtgggggc gttgtcgaaa gcgaaggcgg ccaagcaggc tataccaacc 5520cagacagtta aagaacttga tggcttacta caagaatcaa aaaatatagg tgctgtaaat 5580acacgaatta atatagcgaa tagtactact cgatatacac caatgagaca aacgggacaa 5640ccggtatctg ctggctttga gcatgttctt gaggggcact tccataggcc tattgcgaat 5700aaccgttcag tttttaccat ctccccaaat gaattgaagg ttatacttca aagtaataaa 5760gtagtttctt ctcccgtatc gatgactcct gatggccaat atatgcggac tgtcgatgta 5820ggaaaagtta ttggtactac ttctattaaa gaaggtggac aacccacaac tacaattaaa 5880gtatttacag ataagtcagg aaatttgatt actacatacc cagtaaaagg aaactaa 5937 60 1978 PRT Neisseria meningitidis 60Met Asn Lys Gly Leu His Arg Ile Ile Phe Ser Lys Lys His Ser Thr 1 5 10 15Met Val Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln 20 25 30Ala Gly Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Leu Cys 35 40 45Gly Lys Leu Lys Thr Thr Leu Lys Thr Leu Val Cys Ser Leu Val Ser 50 55 60Leu Ser Met Val Leu Pro Ala His Ala Gln Ile Thr Thr Asp Lys Ser 65 70 75 80Ala Pro Lys Asn Gln Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala 85 90 95Pro Leu Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn 100 105 110Arg Tyr Thr Gln Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn 115 120 125Asp Arg Asn Asn Asn Pro Phe Val Val Lys Gly Ser Ala Gln Leu Ile 130 135 140Leu Asn Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr145 150 155 160Val Gly Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile 165 170 175Thr Val Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr 180 185 190Thr Gly Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp 195 200 205Val Arg Gln Gly Thr Leu Thr Val Gly Ala Ala Gly Trp Asn Asp Lys 210 215 220Gly Gly Ala Asp Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln225 230 235 240Gly Lys Leu Gln Gly Lys Asn Leu Ala Val Ser Thr Gly Pro Gln Lys 245 250 255Val Asp Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr 260 265 270Lys Pro Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala 275 280 285Asp Ser Ile Thr Leu Ile Ala Asn Glu Lys Gly Val Gly Val Lys Asn 290 295 300Ala Gly Thr Leu Glu Ala Ala Lys Gln Leu Ile Val Thr Ser Ser Gly305 310 315 320Arg Ile Glu Asn Ser Gly Arg Ile Ala Thr Thr Ala Asp Gly Thr Glu 325 330 335Ala Ser Pro Thr Tyr Leu Ser Ile Glu Thr Thr Glu Lys Gly Ala Ala 340 345 350Gly Thr Phe Ile Ser Asn Gly Gly Arg Ile Glu Ser Lys Gly Leu Leu 355 360 365Val Ile Glu Thr Gly Glu Asp Ile Ser Leu Arg Asn Gly Ala Val Val 370 375 380Gln Asn Asn Gly Ser Arg Pro Ala Thr Thr Val Leu Asn Ala Gly His385 390 395 400Asn Leu Val Ile Glu Ser Lys Thr Asn Val Asn Asn Ala Lys Gly Pro 405 410 415Ala Thr Leu Ser Ala Asp Gly Arg Thr Val Ile Lys Glu Ala Ser Ile 420 425 430Gln Thr Gly Thr Thr Val Tyr Ser Ser Ser Lys Gly Asn Ala Glu Leu 435 440 445Gly Asn Asn Thr Arg Ile Thr Gly Ala Asp Val Thr Val Leu Ser Asn 450 455 460Gly Thr Ile Ser Ser Ser Ala Val Ile Asp Ala Lys Asp Thr Ala His465 470 475 480Ile Glu Ala Gly Lys Pro Leu Ser Leu Glu Ala Ser Thr Val Thr Ser 485 490 495Asp Ile Arg Leu Asn Gly Gly Ser Ile Lys Gly Gly Lys Gln Leu Ala 500 505 510Leu Leu Ala Asp Asp Asn Ile Thr Ala Lys Thr Thr Asn Leu Asn Thr 515 520 525Pro Gly Asn Leu Tyr Val His Thr Gly Lys Asp Leu Asn Leu Asn Val 530 535 540Asp Lys Asp Leu Ser Ala Ala Ser Ile His Leu Lys Ser Asp Asn Ala545 550 555 560Ala His Ile Thr Gly Thr Ser Lys Thr Leu Thr Ala Ser Lys Asp Met 565 570 575Gly Val Glu Ala Gly Ser Leu Asn Val Thr Asn Thr Asn Leu Arg Thr 580 585 590Asn Ser Gly Asn Leu His Ile Gln Ala Ala Lys Gly Asn Ile Gln Leu 595 600 605Arg Asn Thr Lys Leu Asn Ala Ala Lys Ala Leu Glu Thr Thr Ala Leu 610 615 620Gln Gly Asn Ile Val Ser Asp Gly Leu His Ala Val Ser Ala Asp Gly625 630 635 640His Val Ser Leu Leu Ala Asn Gly Asn Ala Asp Phe Thr Gly His Asn 645 650 655Thr Leu Thr Ala Lys Ala Asp Val Asn Ala Gly Ser Val Gly Lys Gly 660 665 670Arg Leu Lys Ala Asp Asn Thr Asn Ile Thr Ser Ser Ser Gly Asp Ile 675 680 685Thr Leu Val Ala Gly Asn Gly Ile Gln Leu Gly Asp Gly Lys Gln Arg 690 695 700Asn Ser Ile Asn Gly Lys His Ile Ser Ile Lys Asn Asn Gly Gly Asn705 710 715 720Ala Asp Leu Lys Asn Leu Asn Val His Ala Lys Ser Gly Ala Leu Asn 725 730 735Ile His Ser Asp Arg Ala Leu Ser Ile Glu Asn Thr Lys Leu Glu Ser 740 745 750Thr His Asn Thr His Leu Asn Ala Gln His Glu Arg Val Thr Leu Asn 755 760 765Gln Val Asp Ala Tyr Ala His Arg His Leu Ser Ile Thr Gly Ser Gln 770 775 780Ile Trp Gln Asn Asp Lys Leu Pro Ser Ala Asn Lys Leu Val Ala Asn785 790 795 800Gly Val Leu Ala Leu Asn Ala Arg Tyr Ser Gln Ile Ala Asp Asn Thr 805 810 815Thr Leu Arg Ala Gly Ala Ile Asn Leu Thr Ala Gly Thr Ala Leu Val 820 825 830Lys Arg Gly Asn Ile Asn Trp Ser Thr Val Ser Thr Lys Thr Leu Glu 835 840 845Asp Asn Ala Glu Leu Lys Pro Leu Ala Gly Arg Leu Asn Ile Glu Ala 850 855 860Gly Ser Gly Thr Leu Thr Ile Glu Pro Ala Asn Arg Ile Ser Ala His865 870 875 880Thr Asp Leu Ser Ile Lys Thr Gly Gly Lys Leu Leu Leu Ser Ala Lys 885 890 895Gly Gly Asn Ala Gly Ala Pro Ser Ala Gln Val Ser Ser Leu Glu Ala 900 905 910Lys Gly Asn Ile Arg Leu Val Thr Gly Glu Thr Asp Leu Arg Gly Ser 915 920 925Lys Ile Thr Ala Gly Lys Asn Leu Val Val Ala Thr Thr Lys Gly Lys 930 935 940Leu Asn Ile Glu Ala Val Asn Asn Ser Phe Ser Asn Tyr Phe Pro Thr945 950 955 960Gln Lys Ala Ala Glu Leu Asn Gln Lys Ser Lys Glu Leu Glu Gln Gln 965 970 975Ile Ala Gln Leu Lys Lys Ser Ser Pro Lys Ser Lys Leu Ile Pro Thr 980 985 990Leu Gln Glu Glu Arg Asp Arg Leu Ala Phe Tyr Ile Gln Ala Ile Asn 995 1000 1005Lys Glu Val Lys Gly Lys Lys Pro Lys Gly Lys Glu Tyr Leu Gln Ala 1010 1015 1020Lys Leu Ser Ala Gln Asn Ile Asp Leu Ile Ser Ala Gln Gly Ile Glu1025 1030 1035 1040Ile Ser Gly Ser Asp Ile Thr Ala Ser Lys Lys Leu Asn Leu His Ala 1045 1050 1055Ala Gly Val Leu Pro Lys Ala Ala Asp Ser Glu Ala Ala Ala Ile Leu 1060 1065 1070Ile Asp Gly Ile Thr Asp Gln Tyr Glu Ile Gly Lys Pro Thr Tyr Lys 1075 1080 1085Ser His Tyr Asp Lys Ala Ala Leu Asn Lys Pro Ser Arg Leu Thr Gly 1090 1095 1100Arg Thr Gly Val Ser Ile His Ala Ala Ala Ala Leu Asp Asp Ala Arg1105 1110 1115 1120Ile Ile Ile Gly Ala Ser Glu Ile Lys Ala Pro Ser Gly Ser Ile Asp 1125 1130 1135Ile Lys Ala His Ser Asp Ile Val Leu Glu Ala Gly Gln Asn Asp Ala 1140 1145 1150Tyr Thr Phe Leu Lys Thr Lys Gly Lys Ser Gly Lys Ile Ile Arg Lys 1155 1160 1165Thr Lys Phe Thr Ser Thr Arg Asp His Leu Ile Met Pro Ala Pro Val 1170 1175 1180Glu Leu Thr Ala Asn Gly Ile Thr Leu Gln Ala Gly Gly Asn Ile Glu1185 1190 1195 1200Ala Asn Thr Thr Arg Phe Asn Ala Pro Ala Gly Lys Val Thr Leu Val 1205 1210 1215Ala Gly Glu Glu Leu Gln Leu Leu Ala Glu Glu Gly Ile His Lys His 1220 1225 1230Glu Leu Asp Val Gln Lys Ser Arg Arg Phe Ile Gly Ile Lys Val Gly 1235 1240 1245Lys Ser Asn Tyr Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val 1250 1255 1260Arg Val Val Ala Gln Thr Ala Ala Thr Arg Ser Gly Trp Asp Thr Val1265 1270 1275 1280Leu Glu Gly Thr Glu Phe Lys Thr Thr Leu Ala Gly Ala Asp Ile Gln 1285 1290 1295Ala Gly Val Gly Glu Lys Ala Arg Ala Asp Ala Lys Ile Ile Leu Lys 1300 1305 1310Gly Ile Val Asn Arg Ile Gln Ser Glu Glu Lys Leu Glu Thr Asn Ser 1315 1320 1325Thr Val Trp Gln Lys Gln Ala Gly Arg Gly Ser Thr Ile Glu Thr Leu 1330 1335 1340Lys Leu Pro Ser Phe Glu Ser Pro Thr Pro Pro Lys Leu Thr Ala Pro1345 1350 1355 1360Gly Gly Tyr Ile Val Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile 1365 1370 1375Glu Lys Leu Ala Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln 1380 1385 1390Val Ala Lys Asn Val Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Lys 1395 1400 1405Trp Asp Tyr Lys Gln Glu Gly Leu Thr Arg Ala Gly Ala Ala Ile Val 1410 1415 1420Thr Ile Ile Val Thr Ala Leu Thr Tyr Gly Tyr Gly Ala Thr Ala Ala1425 1430 1435 1440Gly Gly Val Ala Ala Ser Gly Ser Ser Thr Ala Ala Ala Ala Gly Thr 1445 1450 1455Ala Ala Thr Thr Thr Ala Ala Ala Thr Thr Val Ser Thr Ala Thr Ala 1460 1465 1470Met Gln Thr Ala Ala Leu Ala Ser Leu Tyr Ser Gln Ala Ala Val Ser 1475 1480 1485Ile Ile Asn Asn Lys Gly Asp Val Gly Lys Ala Leu Lys Asp Leu Gly 1490 1495 1500Thr Ser Asp Thr Val Lys Gln Ile Val Thr Ser Ala Leu Thr Ala Gly1505 1510 1515 1520Ala Leu Asn Gln Met Gly Ala Asp Ile Ala Gln Leu Asn Ser Lys Val 1525 1530 1535Arg Thr Glu Leu Phe Ser Ser Thr Gly Asn Gln Thr Ile Ala Asn Leu 1540 1545 1550Gly Gly Arg Leu Ala Thr Asn Leu Ser Asn Ala Gly Ile Ser Ala Gly 1555 1560 1565Ile Asn Thr Ala Val Asn Gly Gly Ser Leu Lys Asp Asn Leu Gly Asn 1570 1575 1580Ala Ala Leu Gly Ala Leu Val Asn Ser Phe Gln Gly Glu Ala Ala Ser1585 1590 1595 1600Lys Ile Lys Thr Thr Phe Ser Asp Asp Tyr Val Ala Lys Gln Phe Ala 1605 1610 1615His Ala Leu Ala Gly Cys Val Ser Gly Leu Val Gln Gly Lys Cys Lys 1620 1625 1630Asp Gly Ala Ile Gly Ala Ala Val Gly Glu Ile Val Ala Asp Ser Met 1635 1640 1645Leu Gly Gly Arg Asn Pro Ala Thr Leu Ser Asp Ala Glu Lys His Lys 1650 1655 1660Val Ile Ser Tyr Ser Lys Ile Ile Ala Gly Ser Val Ala Ala Leu Asn1665 1670 1675 1680Gly Gly Asp Val Asn Thr Ala Ala Asn Ala Ala Glu Val Ala Val Val 1685 1690 1695Asn Asn Ala Leu Asn Phe Asp Ser Thr Pro Thr Asn Ala Lys Lys His 1700 1705 1710Gln Pro Gln Lys Pro Asp Lys Thr Ala Leu Glu Lys Ile Ile Gln Gly 1715 1720 1725Ile Met Pro Ala His Ala Ala Gly Ala Met Thr Asn Pro Gln Asp Lys 1730 1735 1740Asp Ala Ala Ile Trp Ile Ser Asn Ile Arg Asn Gly Ile Thr Gly Pro1745 1750 1755 1760Ile Val Ile Thr Ser Tyr Gly Val Tyr Ala Ala Gly Trp Thr Ala Pro 1765 1770 1775Leu Ile Gly Thr Ala Gly Lys Leu Ala Ile Ser Thr Cys Met Ala Asn 1780 1785 1790Pro Ser Gly Cys Thr Val Met Val Thr Gln Ala Ala Glu Ala Gly Ala 1795 1800 1805Gly Ile Ala Thr Gly Ala Val Thr Val Gly Asn Ala Trp Glu Ala Pro 1810 1815 1820Val Gly Ala Leu Ser Lys Ala Lys Ala Ala Lys Gln Ala Ile Pro Thr1825 1830 1835 1840Gln Thr Val Lys Glu Leu Asp Gly Leu Leu Gln Glu Ser Lys Asn Ile 1845 1850 1855Gly Ala Val Asn Thr Arg Ile Asn Ile Ala Asn Ser Thr Thr Arg Tyr 1860 1865 1870Thr Pro Met Arg Gln Thr Gly Gln Pro Val Ser Ala Gly Phe Glu His 1875 1880 1885Val Leu Glu Gly His Phe His Arg Pro Ile Ala Asn Asn Arg Ser Val 1890 1895 1900Phe Thr Ile Ser Pro Asn Glu Leu Lys Val Ile Leu Gln Ser Asn Lys1905 1910 1915 1920Val Val Ser Ser Pro Val Ser Met Thr Pro Asp Gly Gln Tyr Met Arg 1925 1930 1935Thr Val Asp Val Gly Lys Val Ile Gly Thr Thr Ser Ile Lys Glu Gly 1940 1945 1950Gly Gln Pro Thr Thr Thr Ile Lys Val Phe Thr Asp Lys Ser Gly Asn 1955 1960 1965Leu Ile Thr Thr Tyr Pro Val Lys Gly Asn 1970 1975 61 4599 DNA Neisseria meningitidis misc_feature (203) any nucleotide 61atgaataaag gtttacatcg cattatcttt agtaaaaagc acagcaccat ggttgcagta 60gccgaaactg ccaacagcca gggcaaaggt aaacaggcag gcagttcggt ttctgtttca 120ctgaaaactt caggcgacct ttgcggcaaa ctcaaaacca cccttaaaac cttggtctgc 180tctttggttt ccctgagtat ggnattncnn nncnntnccc aaattaccac cgacaaatca 240gcacctaaaa accancaggt cgttatcctt aaaaccaaca ctggtgcccc cttggtgaat 300atccaaactc cgaatggacg cggattgagc cacaaccgct atacgcagtt tgatgttgac 360aacaaagggg cagtgttaaa caacgaccgt aacaataatc cgtttctggt caaaggcagt 420gcgcaattga ttttgaacga ggtacgcggt acggctagca aactcaacgg catcgttacc 480gtaggcggtc aaaaggccga cgtgattatt gccaacccca acggcattac cgttaatggc 540ggcggcttta aaaatgtcgg tcggggcatc ttaactatcg gtgcgcccca aatcggcaaa 600gacggtgcac tgacaggatt tgatgtgcgt caaggcacat tgaccgtagg agcagcaggt 660tggaatgata aaggcggagc cgactacacc ggggtacttg ctcgtgcagt tgctttgcag 720gggaaattac agggtaaaaa cctggcggtt tctaccggtc ctcagaaagt agattacgcc 780agcggcgaaa tcagtgcagg tacggcagcg ggtacgaaac cgactattgc ccttgatact 840gccgcactgg gcggtatgta cgccgacagc atcacactga ttgccantga aaaaggcgta 900ggcgtcaaaa atgccggcac actcgaagcg gccaagcaat tgattgtgac ttcgtcaggc 960cgcattgaaa acagcggccg catcgccacc actgccgacg gcaccgaagc ttcaccgact 1020tatctnncna tcgaaaccac cgaaaaagga gcnncaggca catttatctc caatggtggt 1080cggatcgaga gcaaaggctt attggttatt gagacgggag aagatatcan cttgcgtaac 1140ggagccgtgg tgcagaataa cggcagtcgc ccagctacca cggtattaaa tgctggtcat 1200aatttggtga ttgagagtaa aactaatgtg aacaatgcca aaggctcgnc taatctgtcg 1260gccggcggtc gtactacgat caatgatgct actattcaag cgggcagttc cgtgtacagc 1320tccaccaaag gcgatactga nttgggtgaa aatacccgta ttattgctga aaacgtaacc 1380gtattatcta acggtagtat tggcagtgct gctgtaattg aggctaaaga cactgcacac 1440attgaatcgg gcaaaccgct ttctttagaa acctcgaccg ttgcctccaa catccgtttg 1500aacaacggta acattaaagg cggaaagcag cttgctttac tggcagacga taacattact 1560gccaaaacta ccaatctgaa tactcccggc aatctgtatg ttcatacagg taaagatctg 1620aatttgaatg ttgataaaga tttgtctgcc gccagcatcc atttgaaatc ggataacgct 1680gcccatatta ccggcaccag taaaaccctc actgcctcaa aagacatggg tgtggaggca 1740ggcttgctga atgttaccaa taccaatctg cgtaccaact cgggtaatct gcacattcag 1800gcagccaaag gcaatattca gcttcgcaat accaagctga acgcagccaa ggctctcgaa 1860accaccgcat tgcagggcaa tatcgtttca gacggccttc atgctgtttc tgcagacggt 1920catgtatcct tattggccaa cggtaatgcc gactttaccg gtcacaatac cctgacagcc 1980aaggccgatg tcnatgcagg atcggttggt aaaggccgtc tgaaagcaga caataccaat 2040atcacttcat cttcaggaga tattacgttg gttgccgnnn ncggtattca gcttggtgac 2100ggaaaacaac gcaattcaat caacggaaaa cacatcagca tcaaaaacaa cggtggtaat 2160gccgacttaa aaaaccttaa cgtccatgcc aaaagcgggg cattgaacat tcattccgac 2220cgggcattga gcatagaaaa tacnaagctg gagtctaccc ataatacgca tcttaatgca 2280caacacgagc gggtaacgct caaccaagta gatgcctacg cacaccgtca tctaagcatt 2340ancggcagcc agatttggca aaacgacaaa ctgccttctg ccaacaagct ggtggctaac 2400ggtgtattgg cantcaatgc gcgctattcc caaattgccg acaacaccac gctgagagcg 2460ggtgcaatca accttactgc cggtaccgcc ctagtcaagc gcggcaacat caattggagt 2520accgtttcga ccaagacttt ggaagataat gccgaattaa aaccattggc cggacggctg 2580aatattgaag caggtagcgg cacattaacc atcgaacctg ccaaccgcat cagtgcgcat 2640accgacctga gcatcaaaac aggcggaaaa ttgctgttgt ctgcaaaagg aggaaatgca 2700ggtgcgcnta gtgctcaagt ttcctcattg gaagcaaaag gcaatatccg tctggttaca 2760ggagnaacag atttaagagg ttctaaaatt acagccggta aaaacttggt tgtcgccacc 2820accaaaggca agttgaatat cgaagccgta aacaactcat tcagcaatta ttttcntaca 2880caaaaagngn nngnnctcaa ccaaaaatcc aaagaattgg aacagcagat tgcgcagttg 2940aaaaaaagct cgcntaaaag caagctgatt ccaaccctgc aagaagaacg cgaccgtctc 3000gctttctata ttcaagccat caacaaggaa gttaaaggta aaaaacccaa aggcaaagaa 3060tacctgcaag ccaagctttc tgcacaaaat attgacttga tttccgcaca aggcatcgaa 3120atcagcggtt ccgatattac cgcttccaaa aaactgaacc ttcacgccgc aggcgtattg 3180ccaaaggcag cagattcaga ggcggctgct attctgattg acggcataac cgaccaatat 3240gaaattggca agcccaccta caagagtcac tacgacaaag ctgctctgaa caagccttca 3300cgtttgaccg gacgtacggg ggtaagtatt catgcagctg cggcactcga tgatgcacgt 3360attattatcg gtgcatccga aatcaaagct ccctcaggca gcatagacat caaagcccat 3420agtgatattg tactggaggc tggacaaaac gatgcctata ccttcttana aaccaaaggt 3480aaaagcggca naatnatcag aaaaacnaag tttaccagca ccngcganca cctgattatg 3540ccagccccng tcgagctgac cgccaacggt atcacgcttc aggcaggcgg caacatcgaa 3600gctaatacca cccgcttcaa tgcccctgca ggtaaagtta ccctggttgc gggtgaanag 3660ntgcaactgc tggcagaaga aggcatccac aagcacgagt tggatgtcca aaaaagccgc 3720cgctttatcg gcatcaaggt aggtnagagc aattacagta aaaacgaact gaacgaaacc 3780aaattgcctg tccgcgtcgt cgcccaaant gcagccaccc gttcaggctg ggataccgtg 3840ctcgaaggta ccgaattcaa aaccacgctg gccggtgccg acattcaggc aggtgtangc 3900gaaaaagccc gtgtcgatgc gaaaattatc ctcaaaggca ttgtgaaccg tatccagtcg 3960gaagaaaaat tagaaaccaa ctcaaccgta tggcagaaac aggccggacg cggcagcact 4020atcgaaacgc taaaactgcc cagcttcgaa agccctactc cgcccaaatt gtccgcaccc 4080ggcggntata tcgtcgacat tccgaaaggc aatctgaaaa ccgaaatcga aaagctgtcc 4140aaacagcccg agtatgccta tctgaaacag ctccaagtag cgaaaaacat caactggaat 4200caggtgcagc ttgcttacga cagatgggac tacaaacagg agggcttaac cgaagcaggt 4260gcggcgatta tcgcactggc cgttaccgtg gtcacctcag gcgcaggaac cggagccgta 4320ttgggattaa acggtgcgnc cgccgccgca accgatgcag cattcgcctc tttggccagc 4380caggcttccg tatcgttcat caacaacaaa ggcgatgtcg gcaaaaccct gaaagagctg 4440ggcagaagca gcacggtgaa aaatctggtg gttgccgccg ctaccgcagg cgtagccgac 4500aaaatcggcg cttcggcact gancaatgtc agcgataagc agtggatcaa caacctgacc 4560gtcaacctag ccaatgncgg gcagtgccgc actgattaa 4599 62 1532 PRT Neisseria meningitidis SITE (68)..(73) unknown 62Met Asn Lys Gly Leu His Arg Ile Ile Phe Ser Lys Lys His Ser Thr 1 5 10 15Met Val Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln 20 25 30Ala Gly Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Leu Cys 35 40 45Gly Lys Leu Lys Thr Thr Leu Lys Thr Leu Val Cys Ser Leu Val Ser 50 55 60Leu Ser Met Xaa Xaa Xaa Xaa Xaa Xaa Gln Ile Thr Thr Asp Lys Ser 65 70 75 80Ala Pro Lys Asn Xaa Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala 85 90 95Pro Leu Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn 100 105 110Arg Tyr Thr Gln Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn 115 120 125Asp Arg Asn Asn Asn Pro Phe Leu Val Lys Gly Ser Ala Gln Leu Ile 130 135 140Leu Asn Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr145 150 155 160Val Gly Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile 165 170 175Thr Val Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr 180 185 190Ile Gly Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp 195 200 205Val Arg Gln Gly Thr Leu Thr Val Gly Ala Ala Gly Trp Asn Asp Lys 210 215 220Gly Gly Ala Asp Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln225 230 235 240Gly Lys Leu Gln Gly Lys Asn Leu Ala Val Ser Thr Gly Pro Gln Lys 245 250 255Val Asp Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr 260 265 270Lys Pro Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala 275 280 285Asp Ser Ile Thr Leu Ile Ala Xaa Glu Lys Gly Val Gly Val Lys Asn 290 295 300Ala Gly Thr Leu Glu Ala Ala Lys Gln Leu Ile Val Thr Ser Ser Gly305 310 315 320Arg Ile Glu Asn Ser Gly Arg Ile Ala Thr Thr Ala Asp Gly Thr Glu 325 330 335Ala Ser Pro Thr Tyr Leu Xaa Ile Glu Thr Thr Glu Lys Gly Ala Xaa 340 345 350Gly Thr Phe Ile Ser Asn Gly Gly Arg Ile Glu Ser Lys Gly Leu Leu 355 360 365Val Ile Glu Thr Gly Glu Asp Ile Xaa Leu Arg Asn Gly Ala Val Val 370 375 380Gln Asn Asn Gly Ser Arg Pro Ala Thr Thr Val Leu Asn Ala Gly His385 390 395 400Asn Leu Val Ile Glu Ser Lys Thr Asn Val Asn Asn Ala Lys Gly Ser 405 410 415Xaa Asn Leu Ser Ala Gly Gly Arg Thr Thr Ile Asn Asp Ala Thr Ile 420 425 430Gln Ala Gly Ser Ser Val Tyr Ser Ser Thr Lys Gly Asp Thr Xaa Leu 435 440 445Gly Glu Asn Thr Arg Ile Ile Ala Glu Asn Val Thr Val Leu Ser Asn 450 455 460Gly Ser Ile Gly Ser Ala Ala Val Ile Glu Ala Lys Asp Thr Ala His465 470 475 480Ile Glu Ser Gly Lys Pro Leu Ser Leu Glu Thr Ser Thr Val Ala Ser 485 490 495Asn Ile Arg Leu Asn Asn Gly Asn Ile Lys Gly Gly Lys Gln Leu Ala 500 505 510Leu Leu Ala Asp Asp Asn Ile Thr Ala Lys Thr Thr Asn Leu Asn Thr 515 520 525Pro Gly Asn Leu Tyr Val His Thr Gly Lys Asp Leu Asn Leu Asn Val 530 535 540Asp Lys Asp Leu Ser Ala Ala Ser Ile His Leu Lys Ser Asp Asn Ala545 550 555 560Ala His Ile Thr Gly Thr Ser Lys Thr Leu Thr Ala Ser Lys Asp Met 565 570 575Gly Val Glu Ala Gly Leu Leu Asn Val Thr Asn Thr Asn Leu Arg Thr 580 585 590Asn Ser Gly Asn Leu His Ile Gln Ala Ala Lys Gly Asn Ile Gln Leu 595 600 605Arg Asn Thr Lys Leu Asn Ala Ala Lys Ala Leu Glu Thr Thr Ala Leu 610 615 620Gln Gly Asn Ile Val Ser Asp Gly Leu His Ala Val Ser Ala Asp Gly625 630 635 640His Val Ser Leu Leu Ala Asn Gly Asn Ala Asp Phe Thr Gly His Asn 645 650 655Thr Leu Thr Ala Lys Ala Asp Val Xaa Ala Gly Ser Val Gly Lys Gly 660 665 670Arg Leu Lys Ala Asp Asn Thr Asn Ile Thr Ser Ser Ser Gly Asp Ile 675 680 685Thr Leu Val Ala Xaa Xaa Gly Ile Gln Leu Gly Asp Gly Lys Gln Arg 690 695 700Asn Ser Ile Asn Gly Lys His Ile Ser Ile Lys Asn Asn Gly Gly Asn705 710 715 720Ala Asp Leu Lys Asn Leu Asn Val His Ala Lys Ser Gly Ala Leu Asn 725 730 735Ile His Ser Asp Arg Ala Leu Ser Ile Glu Asn Thr Lys Leu Glu Ser 740 745 750Thr His Asn Thr His Leu Asn Ala Gln His Glu Arg Val Thr Leu Asn 755 760 765Gln Val Asp Ala Tyr Ala His Arg His Leu Ser Ile Xaa Gly Ser Gln 770 775 780Ile Trp Gln Asn Asp Lys Leu Pro Ser Ala Asn Lys Leu Val Ala Asn785 790 795 800Gly Val Leu Ala Xaa Asn Ala Arg Tyr Ser Gln Ile Ala Asp Asn Thr 805 810 815Thr Leu Arg Ala Gly Ala Ile Asn Leu Thr Ala Gly Thr Ala Leu Val 820 825 830Lys Arg Gly Asn Ile Asn Trp Ser Thr Val Ser Thr Lys Thr Leu Glu 835 840 845Asp Asn Ala Glu Leu Lys Pro Leu Ala Gly Arg Leu Asn Ile Glu Ala 850 855 860Gly Ser Gly Thr Leu Thr Ile Glu Pro Ala Asn Arg Ile Ser Ala His865 870 875 880Thr Asp Leu Ser Ile Lys Thr Gly Gly Lys Leu Leu Leu Ser Ala Lys 885 890 895Gly Gly Asn Ala Gly Ala Xaa Ser Ala Gln Val Ser Ser Leu Glu Ala 900 905 910Lys Gly Asn Ile Arg Leu Val Thr Gly Xaa Thr Asp Leu Arg Gly Ser 915 920 925Lys Ile Thr Ala Gly Lys Asn Leu Val Val Ala Thr Thr Lys Gly Lys 930 935 940Leu Asn Ile Glu Ala Val Asn Asn Ser Phe Ser Asn Tyr Phe Xaa Thr945 950 955 960Gln Lys Xaa Xaa Xaa Leu Asn Gln Lys Ser Lys Glu Leu Glu Gln Gln 965 970 975Ile Ala Gln Leu Lys Lys Ser Ser Xaa Lys Ser Lys Leu Ile Pro Thr 980 985 990Leu Gln Glu Glu Arg Asp Arg Leu Ala Phe Tyr Ile Gln Ala Ile Asn 995 1000 1005Lys Glu Val Lys Gly Lys Lys Pro Lys Gly Lys Glu Tyr Leu Gln Ala 1010 1015 1020Lys Leu Ser Ala Gln Asn Ile Asp Leu Ile Ser Ala Gln Gly Ile Glu1025 1030 1035 1040Ile Ser Gly Ser Asp Ile Thr Ala Ser Lys Lys Leu Asn Leu His Ala 1045 1050 1055Ala Gly Val Leu Pro Lys Ala Ala Asp Ser Glu Ala Ala Ala Ile Leu 1060 1065 1070Ile Asp Gly Ile Thr Asp Gln Tyr Glu Ile Gly Lys Pro Thr Tyr Lys 1075 1080 1085Ser His Tyr Asp Lys Ala Ala Leu Asn Lys Pro Ser Arg Leu Thr Gly 1090 1095 1100Arg Thr Gly Val Ser Ile His Ala Ala Ala Ala Leu Asp Asp Ala Arg1105 1110 1115 1120Ile Ile Ile Gly Ala Ser Glu Ile Lys Ala Pro Ser Gly Ser Ile Asp 1125 1130 1135Ile Lys Ala His Ser Asp Ile Val Leu Glu Ala Gly Gln Asn Asp Ala 1140 1145 1150Tyr Thr Phe Leu Xaa Thr Lys Gly Lys Ser Gly Xaa Xaa Ile Arg Lys 1155 1160 1165Thr Lys Phe Thr Ser Thr Xaa Xaa His Leu Ile Met Pro Ala Pro Val 1170 1175 1180Glu Leu Thr Ala Asn Gly Ile Thr Leu Gln Ala Gly Gly Asn Ile Glu1185 1190 1195 1200Ala Asn Thr Thr Arg Phe Asn Ala Pro Ala Gly Lys Val Thr Leu Val 1205 1210 1215Ala Gly Glu Xaa Xaa Gln Leu Leu Ala Glu Glu Gly Ile His Lys His 1220 1225 1230Glu Leu Asp Val Gln Lys Ser Arg Arg Phe Ile Gly Ile Lys Val Gly 1235 1240 1245Xaa Ser Asn Tyr Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val 1250 1255 1260Arg Val Val Ala Gln Xaa Ala Ala Thr Arg Ser Gly Trp Asp Thr Val1265 1270 1275 1280Leu Glu Gly Thr Glu Phe Lys Thr Thr Leu Ala Gly Ala Asp Ile Gln 1285 1290 1295Ala Gly Val Xaa Glu Lys Ala Arg Val Asp Ala Lys Ile Ile Leu Lys 1300 1305 1310Gly Ile Val Asn Arg Ile Gln Ser Glu Glu Lys Leu Glu Thr Asn Ser 1315 1320 1325Thr Val Trp Gln Lys Gln Ala Gly Arg Gly Ser Thr Ile Glu Thr Leu 1330 1335 1340Lys Leu Pro Ser Phe Glu Ser Pro Thr Pro Pro Lys Leu Ser Ala Pro1345 1350 1355 1360Gly Gly Tyr Ile Val Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile 1365 1370 1375Glu Lys Leu Ser Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln 1380 1385 1390Val Ala Lys Asn Ile Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Arg 1395 1400 1405Trp Asp Tyr Lys Gln Glu Gly Leu Thr Glu Ala Gly Ala Ala Ile Ile 1410 1415 1420Ala Leu Ala Val Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val1425 1430 1435 1440Leu Gly Leu Asn Gly Ala Xaa Ala Ala Ala Thr Asp Ala Ala Phe Ala 1445 1450 1455Ser Leu Ala Ser Gln Ala Ser Val Ser Phe Ile Asn Asn Lys Gly Asp 1460 1465 1470Val Gly Lys Thr Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn 1475 1480 1485Leu Val Val Ala Ala Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala 1490 1495 1500Ser Ala Leu Xaa Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr1505 1510 1515 1520Val Asn Leu Ala Asn Xaa Gly Gln Cys Arg Thr Asp 1525 1530 63 1782 DNA Neisseria meningitidis misc_feature (89) any nucleotide 63cgcttcattc atgatgaagc agtcggcagc aacatcggcg gcggcaaaat gattgttgca 60gccgggcagg atatcaatgt acgcggcana agccttattt ctgataaggg cattgtttta 120aaagcaggac acgacatcga tatttctact gcccataatc gctataccgg caatgaatac 180cacgagagca waaawtcagg cgtcatgggt actggcggat tgggctttac tatcggtaac 240cggaaaacta ccgatgacac tgatcgtacc aatattgtsc atacaggcag cattataggc 300agcctgaatg gagacaccgt tacagttgca ggaaaccgct accgacaaac cggcagtacc 360gtctccagcc ccgaggggcg caataccgtc acagccaaaw gcatagatgt agagttcgca 420aacaaccggt atgccactga ctacgcccat acccagggaa caaaaaggcc ttaccgtcgc 480cctcaatgtc ccggttgtcc aagctgcaca aaacttcata caagcagccc aaaatgtggg 540caaaagtaaa aataaacgcg ttaatgccat ggctgcagcc aatgctgcat ggcagagtta 600tcaagcaacc caacaaatgc aacaatttgc tccaagcagc agtgcgggac aaggtcaaaa 660ctacaatcaa agccccagta tcagtgtgtc cattacntac ggcgaacaga aaagtcgtaa 720cgagcaaaaa agacattaca ccgaagcggc agcaagtcaa attatcggca aagggcaaac 780cacacttgcg gcaacaggaa gtggggagca gtccaatatc aatattacag gttccgatgt 840catcggccat gcaggtactc cnctcattgc cgacaaccat atcagactcc aatctgccaa 900acaggacggc agcgagcaaa gcaaaaacaa aagcagtggt tggaatgcag gcgtacgtnn 960caaaataggc aacggcatca ggtttggaat taccgccgga ggaaatatcg gtaaaggtaa 1020agagcaaggg ggaagtacta cccaccgcca cacccatgtc ggcagcacaa ccggcaaaac 1080taccatccga agcggcgggg gataccaccc tcaaaggtgt gcagctcatc ggcaaaggca 1140tacaggcaga tacgcgcaac ctgcatatag aaagtgttca agatactgaa acctatcaga 1200gcaaacagca aaacggcaat gtccaagttt actgtcggtt acggattcag tgcaagcggc 1260agttaccgcc aaagcaaagt caaagcagac catgcctccg taaccgggca aagcggtatt 1320tatgccggag aagacggcta tcaaatyaaa gtyagagaca acacagacct yaagggcggt 1380atcatcacgt ctagccaaag cgcagaagat aagggcaaaa acctttttca gacggccacc 1440cttactgcca gcgacattca aaaccacagc cgctacgaag gcagaagctt cggcataggc 1500ggcagtttcg acctgaacgg cggctgggac ggcacggtta ccgacaaaca aggcaggcct 1560accgacagga taagcccggc agccggctac ggcagcgacg gagacagcaa aaacagcacc 1620acccgcagcg gcgtcaacac ccacaacata cacatcaccg acgaagcggg acaacttgcc 1680cgaacaggca ggactgcaaa agaaaccgaa gcgcgtatct acaccggcat cgacaccgaa 1740actgcggatc aacactcagg ccatctgaaa aacagcttcg ac 1782 64 593 PRT Neisseria meningitidis SITE (30) unknown 64Arg Phe Ile His Asp Glu Ala Val Gly Ser Asn Ile Gly Gly Gly Lys 1 5 10 15Met Ile Val Ala Ala Gly Gln Asp Ile Asn Val Arg Gly Xaa Ser Leu 20 25 30Ile Ser Asp Lys Gly Ile Val Leu Lys Ala Gly His Asp Ile Asp Ile 35 40 45Ser Thr Ala His Asn Arg Tyr Thr Gly Asn Glu Tyr His Glu Ser Xaa 50 55 60Xaa Ser Gly Val Met Gly Thr Gly Gly Leu Gly Phe Thr Ile Gly Asn 65 70 75 80Arg Lys Thr Thr Asp Asp Thr Asp Arg Thr Asn Ile Val His Thr Gly 85 90 95Ser Ile Ile Gly Ser Leu Asn Gly Asp Thr Val Thr Val Ala Gly Asn 100 105 110Arg Tyr Arg Gln Thr Gly Ser Thr Val Ser Ser Pro Glu Gly Arg Asn 115 120 125Thr Val Thr Ala Lys Xaa Ile Asp Val Glu Phe Ala Asn Asn Arg Tyr 130 135 140Ala Thr Asp Tyr Ala His Thr Gln Glu Gln Lys Gly Leu Thr Val Ala145 150 155 160Leu Asn Val Pro Val Val Gln Ala Ala Gln Asn Phe Ile Gln Ala Ala 165 170 175Gln Asn Val Gly Lys Ser Lys Asn Lys Arg Val Asn Ala Met Ala Ala 180 185 190Ala Asn Ala Ala Trp Gln Ser Tyr Gln Ala Thr Gln Gln Met Gln Gln 195 200 205Phe Ala Pro Ser Ser Ser Ala Gly Gln Gly Gln Asn Tyr Asn Gln Ser 210 215 220Pro Ser Ile Ser Val Ser Ile Xaa Tyr Gly Glu Gln Lys Ser Arg Asn225 230 235 240Glu Gln Lys Arg His Tyr Thr Glu Ala Ala Ala Ser Gln Ile Ile Gly 245 250 255Lys Gly Gln Thr Thr Leu Ala Ala Thr Gly Ser Gly Glu Gln Ser Asn 260 265 270Ile Asn Ile Thr Gly Ser Asp Val Ile Gly His Ala Gly Thr Xaa Leu 275 280 285Ile Ala Asp Asn His Ile Arg Leu Gln Ser Ala Lys Gln Asp Gly Ser 290 295 300Glu Gln Ser Lys Asn Lys Ser Ser Gly Trp Asn Ala Gly Val Arg Xaa305 310 315 320Lys Ile Gly Asn Gly Ile Arg Phe Gly Ile Thr Ala Gly Gly Asn Ile 325 330 335Gly Lys Gly Lys Glu Gln Gly Gly Ser Thr Thr His Arg His Thr His 340 345 350Val Gly Ser Thr Thr Gly Lys Thr Thr Ile Arg Ser Gly Gly Asp Thr 355 360 365Thr Leu Lys Gly Val Gln Leu Ile Gly Lys Gly Ile Gln Ala Asp Thr 370 375 380Arg Asn Leu His Ile Glu Ser Val Gln Asp Thr Glu Thr Tyr Gln Ser385 390 395 400Lys Gln Gln Asn Gly Asn Val Gln Val Thr Val Gly Tyr Gly Phe Ser 405 410 415Ala Ser Gly Ser Tyr Arg Gln Ser Lys Val Lys Ala Asp His Ala Ser 420 425 430Val Thr Gly Gln Ser Gly Ile Tyr Ala Gly Glu Asp Gly Tyr Gln Ile 435 440 445Lys Val Arg Asp Asn Thr Asp Leu Lys Gly Gly Ile Ile Thr Ser Ser 450 455 460Gln Ser Ala Glu Asp Lys Gly Lys Asn Leu Phe Gln Thr Ala Thr Leu465 470 475 480Thr Ala Ser Asp Ile Gln Asn His Ser Arg Tyr Glu Gly Arg Ser Phe 485 490 495Gly Ile Gly Gly Ser Phe Asp Leu Asn Gly Gly Trp Asp Gly Thr Val 500 505 510Thr Asp Lys Gln Gly Arg Pro Thr Asp Arg Ile Ser Pro Ala Ala Gly 515 520 525Tyr Gly Ser Asp Gly Asp Ser Lys Asn Ser Thr Thr Arg Ser Gly Val 530 535 540Asn Thr His Asn Ile His Ile Thr Asp Glu Ala Gly Gln Leu Ala Arg545 550 555 560Thr Gly Arg Thr Ala Lys Glu Thr Glu Ala Arg Ile Tyr Thr Gly Ile 565 570 575Asp Thr Glu Thr Ala Asp Gln His Ser Gly His Leu Lys Asn Ser Phe 580 585 590Asp 65 390 DNA Neisseria meningitidis 65acgaccggca gcctcggcgg catactggcc ggcggcggca cttcccttgc cgcaccgtat 60ttggacaaag cggcggaaaa cctcggtccg gcgggcaaag cggcggtcaa cgcactgggc 120ggtgcggcca tcggctatgc aactggtggt agtggtggtg ctgtggtggg tgcgaatgta 180gattggaaca ataggcagct gcatccgaaa gaaatggcgt tggccgacaa atatgccgaa 240gccctcaagc gcgaagttga aaaacgcgaa ggcagaaaaa tcagcagcca agaagcggca 300atgagaatcc gcaggcagat atgcgttggg tggacaaagg ttcccaagac ggctataccg 360accaaagcgt catatccctt atcggaatga 390 66 129 PRT Neisseria meningitidis 66Thr Thr Gly Ser Leu Gly Gly Ile Leu Ala Gly Gly Gly Thr Ser Leu 1 5 10 15Ala Ala Pro Tyr Leu Asp Lys Ala Ala Glu Asn Leu Gly Pro Ala Gly 20 25 30Lys Ala Ala Val Asn Ala Leu Gly Gly Ala Ala Ile Gly Tyr Ala Thr 35 40 45Gly Gly Ser Gly Gly Ala Val Val Gly Ala Asn Val Asp Trp Asn Asn 50 55 60Arg Gln Leu His Pro Lys Glu Met Ala Leu Ala Asp Lys Tyr Ala Glu 65 70 75 80Ala Leu Lys Arg Glu Val Glu Lys Arg Glu Gly Arg Lys Ile Ser Ser 85 90 95Gln Glu Ala Ala Met Arg Ile Arg Arg Gln Ile Cys Val Gly Trp Thr 100 105 110Lys Val Pro Lys Thr Ala Ile Pro Thr Lys Ala Ser Tyr Pro Leu Ser 115 120 125Glu 67 960 DNA Neisseria meningitidis 67caatgccgtc tgaaaagctc acaattttac agacggcatt tgttatgcaa gtacatatac 60agattcccta tatactgccc agrkgcgtgc gtggctgaag acacccccta cgcttgctat 120ttgraacagc tccaagtcac caaagacgtc aactggaacc aggtacwact ggcgtacgac 180aaatgggact ataaacagga aggcttaacc ggagccggag cagcgattat tgcgctggct 240gttaccgtgg ttactgcggg cgcgggagcc ggagccgcac tgggcttaaa cggcgcggcc 300gcagcggcaa ccgatgccgc attcgcctcg ctggccagcc aggcttccgt atcgctcatc 360aacaacaaag gcaatatcgg taacaccctg aaagagctgg gcagaagcag cacggtgaaa 420aatctgatgg ttgccgtcgc taccgcaggc gtagccgaca aaatcggtgc ttcggcactg 480aacaatgtca gcgataagca gtggatcaac aacctgaccg tcaacctggc caatgcgggc 540agtgccgcac tgattaatac cgctgtcaac ggcggcagcc tgaaagacaa tctggaagcg 600aatatccttg cggctttggt gaatactgcg catggagaag cagccagtaa aatcaaacag 660ttggatcagc actacattac ccacaagatt gcccatgcca tagcgggctg tgcggctgcg 720gcggcgaata agggcaagtg tcaggatggt gcgataggtg cggctgtggg cgagatagtc 780ggggaggctt tgacaaacgg caaaaatcct gacactttga cagctaaaga acgcgaacag 840attttggcat acagcaaact ggttgccggt acggtaagcg gtgtggtcgg cggcgatgta 900aatgcggcgg cgaatgcggc tgaggtagcg gtgaaaaata atcagcttag cgacaaatga 960 68 319 PRT Neisseria meningitidis SITE (28) unknown 68Gln Cys Arg Leu Lys Ser Ser Gln Phe Tyr Arg Arg His Leu Leu Cys 1 5 10 15Lys Tyr Ile Tyr Arg Phe Pro Ile Tyr Cys Pro Xaa Ala Cys Val Ala 20 25 30Glu Asp Thr Pro Tyr Ala Cys Tyr Leu Xaa Gln Leu Gln Val Thr Lys 35 40 45Asp Val Asn Trp Asn Gln Val Xaa Leu Ala Tyr Asp Lys Trp Asp Tyr 50 55 60Lys Gln Glu Gly Leu Thr Gly Ala Gly Ala Ala Ile Ile Ala Leu Ala 65 70 75 80Val Thr Val Val Thr Ala Gly Ala Gly Ala Gly Ala Ala Leu Gly Leu 85 90 95Asn Gly Ala Ala Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu Ala 100 105 110Ser Gln Ala Ser Val Ser Leu Ile Asn Asn Lys Gly Asn Ile Gly Asn 115 120 125Thr Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Met Val 130 135 140Ala Val Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala Ser Ala Leu145 150 155 160Asn Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr Val Asn Leu 165 170 175Ala Asn Ala Gly Ser Ala Ala Leu Ile Asn Thr Ala Val Asn Gly Gly 180 185 190Ser Leu Lys Asp Asn Leu Glu Ala Asn Ile Leu Ala Ala Leu Val Asn 195 200 205Thr Ala His Gly Glu Ala Ala Ser Lys Ile Lys Gln Leu Asp Gln His 210 215 220Tyr Ile Thr His Lys Ile Ala His Ala Ile Ala Gly Cys Ala Ala Ala225 230 235 240Ala Ala Asn Lys Gly Lys Cys Gln Asp Gly Ala Ile Gly Ala Ala Val 245 250 255Gly Glu Ile Val Gly Glu Ala Leu Thr Asn Gly Lys Asn Pro Asp Thr 260 265 270Leu Thr Ala Lys Glu Arg Glu Gln Ile Leu Ala Tyr Ser Lys Leu Val 275 280 285Ala Gly Thr Val Ser Gly Val Val Gly Gly Asp Val Asn Ala Ala Ala 290 295 300Asn Ala Ala Glu Val Ala Val Lys Asn Asn Gln Leu Ser Asp Lys305 310 315 69 1860 DNA Neisseria meningitidis 69atgcaagtaa atattcagat tccctatata ctgcccagat gcgtgcgtgc tgaagacacc 60ccctacgctt gctatttgaa acagctccaa gtcaccaaag acgtcaactg gaaccaggta 120caactggcgt acgacaaatg ggactataaa caggaaggct taaccggagc cggagcagcg 180attattgcgc tggctgttac cgtggttact gcgggcgcgg gagccggagc cgcactgggc 240ttaaacggcg cggccgcagc ggcaaccgat gccgcattcg cctcgctggc cagccaggct 300tccgtatcgc tcatcaacaa caaaggcaat atcggtaaca ccctgaaaga gctgggcaga 360agcagcacgg tgaaaaatct gatggttgcc gtcgctaccg caggcgtagc cgacaaaatc 420ggtgcttcgg cactgaacaa tgtcagcgat aagcagtgga tcaacaacct gaccgtcaac 480ctggccaatg cgggcagtgc cgcactgatt aataccgctg tcaacggcgg cagcctgaaa 540gacaatctgg aagcgaatat ccttgcggct ttggtgaata ctgcgcatgg agaagcagcc 600agtaaaatca aacagttgga tcagcactac attacccaca agattgccca tgccatagcg 660ggctgtgcgg ctgcggcggc gaataagggc aagtgtcagg atggtgcgat aggtgcggct 720gtgggcgaga tagtcgggga ggctttgaca aacggcaaaa atcctgacac tttgacagct 780aaagaacgcg aacagatttt ggcatacagc aaactggttg ccggtacggt aagcggtgtg 840gtcggcggcg atgtaaatgc ggcggcgaat gcggctgagg tagcggtgaa aaataatcag 900cttagcgaca aagagggtag agaatttgat aacgaaatga ctgcatgcgc caaacagaat 960aatcctcaac tgtgcagaaa aaatactgta aaaaagtatc aaaatgttgc tgataaaaga 1020cttgctgctt cgattgcaat atgtacggat atatcccgta gtactgaatg tagaacaatc 1080agaaaacaac atttgatcga tagtagaagc cttcattcat cttgggaagc aggtctaatt 1140ggtaaagatg atgaatggta taaattattc agcaaatctt acacccaagc agatttggct 1200ttacagtctt atcatttgaa tactgctgct aaatcttggc ttcaatcggg caatacaaag 1260cctttatccg aatggatgtc cgaccaaggt tatacactta tttcaggagt taatcctaga 1320ttcattccaa taccaagagg gtttgtaaaa caaaatacac ctattactaa tgtcaaatac 1380ccggaaggca tcagtttcga tacaaaccta aaaagacatc tggcaaatgc tgatggtttt 1440agtcaaaaac agggcattaa aggagcccat aaccgcacca attttatggc agaactaaat 1500tcacgaggag gacgcgtaaa atctgaaacc caaactgata ttgaaggcat tacccgaatt 1560aaatatgaga ttcctacact agacaggaca ggtaaacctg atggtggatt taaggaaatt 1620tcaagtataa aaactgttta taatcctaaa aaattttctg atgataaaat acttcaaatg 1680gctcaaaatg ctgcttcaca aggatattca aaagcctcta aaattgctca aaatgaaaga 1740actaaatcaa tatcggaaag aaaaaatgtc attcaattct cagaaacctt tgacggaatc 1800aaatttagat catattttga tgtaaataca ggaagaatta caaacattca cccagaataa 1860 70 619 PRT Neisseria meningitidis 70Met Gln Val Asn Ile Gln Ile Pro Tyr Ile Leu Pro Arg Cys Val Arg 1 5 10 15Ala Glu Asp Thr Pro Tyr Ala Cys Tyr Leu Lys Gln Leu Gln Val Thr 20 25 30Lys Asp Val Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Lys Trp Asp 35 40 45Tyr Lys Gln Glu Gly Leu Thr Gly Ala Gly Ala Ala Ile Ile Ala Leu 50 55 60Ala Val Thr Val Val Thr Ala Gly Ala Gly Ala Gly Ala Ala Leu Gly 65 70 75 80Leu Asn Gly Ala Ala Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu 85 90 95Ala Ser Gln Ala Ser Val Ser Leu Ile Asn Asn Lys Gly Asn Ile Gly 100 105 110Asn Thr Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Met 115 120 125Val Ala Val Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala Ser Ala 130 135 140Leu Asn Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr Val Asn145 150 155 160Leu Ala Asn Ala Gly Ser Ala Ala Leu Ile Asn Thr Ala Val Asn Gly 165 170 175Gly Ser Leu Lys Asp Asn Leu Glu Ala Asn Ile Leu Ala Ala Leu Val 180 185 190Asn Thr Ala His Gly Glu Ala Ala Ser Lys Ile Lys Gln Leu Asp Gln 195 200 205His Tyr Ile Thr His Lys Ile Ala His Ala Ile Ala Gly Cys Ala Ala 210 215 220Ala Ala Ala Asn Lys Gly Lys Cys Gln Asp Gly Ala Ile Gly Ala Ala225 230 235 240Val Gly Glu Ile Val Gly Glu Ala Leu Thr Asn Gly Lys Asn Pro Asp 245 250 255Thr Leu Thr Ala Lys Glu Arg Glu Gln Ile Leu Ala Tyr Ser Lys Leu 260 265 270Val Ala Gly Thr Val Ser Gly Val Val Gly Gly Asp Val Asn Ala Ala 275 280 285Ala Asn Ala Ala Glu Val Ala Val Lys Asn Asn Gln Leu Ser Asp Lys 290 295 300Glu Gly Arg Glu Phe Asp Asn Glu Met Thr Ala Cys Ala Lys Gln Asn305 310 315 320Asn Pro Gln Leu Cys Arg Lys Asn Thr Val Lys Lys Tyr Gln Asn Val 325 330 335Ala Asp Lys Arg Leu Ala Ala Ser Ile Ala Ile Cys Thr Asp Ile Ser 340 345 350Arg Ser Thr Glu Cys Arg Thr Ile Arg Lys Gln His Leu Ile Asp Ser 355 360 365Arg Ser Leu His Ser Ser Trp Glu Ala Gly Leu Ile Gly Lys Asp Asp 370 375 380Glu Trp Tyr Lys Leu Phe Ser Lys Ser Tyr Thr Gln Ala Asp Leu Ala385 390 395 400Leu Gln Ser Tyr His Leu Asn Thr Ala Ala Lys Ser Trp Leu Gln Ser 405 410 415Gly Asn Thr Lys Pro Leu Ser Glu Trp Met Ser Asp Gln Gly Tyr Thr 420 425 430Leu Ile Ser Gly Val Asn Pro Arg Phe Ile Pro Ile Pro Arg Gly Phe 435 440 445Val Lys Gln Asn Thr Pro Ile Thr Asn Val Lys Tyr Pro Glu Gly Ile 450 455 460Ser Phe Asp Thr Asn Leu Lys Arg His Leu Ala Asn Ala Asp Gly Phe465 470 475 480Ser Gln Lys Gln Gly Ile Lys Gly Ala His Asn Arg Thr Asn Phe Met 485 490 495Ala Glu Leu Asn Ser Arg Gly Gly Arg Val Lys Ser Glu Thr Gln Thr 500 505 510Asp Ile Glu Gly Ile Thr Arg Ile Lys Tyr Glu Ile Pro Thr Leu Asp 515 520 525Arg Thr Gly Lys Pro Asp Gly Gly Phe Lys Glu Ile Ser Ser Ile Lys 530 535 540Thr Val Tyr Asn Pro Lys Lys Phe Ser Asp Asp Lys Ile Leu Gln Met545 550 555 560Ala Gln Asn Ala Ala Ser Gln Gly Tyr Ser Lys Ala Ser Lys Ile Ala 565 570 575Gln Asn Glu Arg Thr Lys Ser Ile Ser Glu Arg Lys Asn Val Ile Gln 580 585 590Phe Ser Glu Thr Phe Asp Gly Ile Lys Phe Arg Ser Tyr Phe Asp Val 595 600 605Asn Thr Gly Arg Ile Thr Asn Ile His Pro Glu 610 615 71 1788 DNA Neisseria meningitidis misc_feature (181) any nucleotide 71tatctgaaac agctccaagt agcgaaaaac atcaactgga atcaggtgca gcttgcttac 60gacagatggg actacaaaca ggagggctta accgaagcag gtgcggcgat tatcgcactg 120gccgttaccg tggtcacctc aggcgcagga accggagccg tattgggatt aaacggtgcg 180nccgccgccg caaccgatgc agcattcgcc tctttggcca gccaggcttc cgtatcgttc 240atcaacaaca aaggcgatgt cggcaaaacc ctgaaagagc tgggcagaag cagcacggtg 300aaaaatctgg tggttgccgc cgctaccgca ggcgtagccg acaaaatcgg cgcttcggca 360ctgancaatg tcagcgataa gcagtggatc aacaacctga ccgtcaacct agccaatgcg 420ggcagtgccg cactgattaa taccgctgtc aacggcggca gcctgaaaga cantctggaa 480gcgaatatcc ttgcggcttt ggtcaatacc gcgcatggag aagcagccag taaaatcaaa 540cagttggatc agcactacat agtccacaag attgcccatg ccatagcggg ctgtgcggca 600gcggcggcga ataagggcaa gtgtcaggat ggtgcgatag gtgcggctgt gggcgagata 660gtcggggagg ctttgacaaa cggcaaaaat cctgacactt tgacagctaa agaacgcgaa 720cagattttgg catacagcaa actggttgcc ggtacggtaa gcggtgtggt cggcggcgat 780gtaaatgcgg cggcgaatgc ggctgaggta gcggtgaaaa ataatcagct tagcgacnaa 840gagggtagag aatttgataa cgaaatgact gcatgcgcca aacagaatan tcctcaactg 900tgcagaaaaa atactgtaaa aaagtatcaa aatgttgctg ataaaagact tgctgcttcg 960attgcaatat gtacggatat atcccgtagt actgaatgta gaacaatcag aaaacaacat 1020ttgatcgata gtagaagcct tcattcatct tgggaagcag gtctaattgg taaagatgat 1080gaatggtata aattattcag caaatcttac acccaagcag atttggcttt acagtcttat 1140catttgaata ctgctgctaa atcttggctt caatcgggca atacaaagcc tttatccgaa 1200tggatgtccg accaaggtta tacacttatt tcaggagtta atcctagatt cattccaata 1260ccaagagggt ttgtaaaaca aaatacacct attactaatg tcaaataccc ggaaggcatc 1320agtttcgata caaacctana aagacatctg gcaaatgctg atggttttag tcaagaacag 1380ggcattaaag gagcccataa ccgcaccaat nttatggcag aactaaattc acgaggagga 1440ngngtaaaat ctgaaaccca nactgatatt gaaggcatta cccgaattaa atatgagatt 1500cctacactag acaggacagg taaacctgat ggtggattta aggaaatttc aagtataaaa 1560actgtttata atcctaaaaa nttttnngat gataaaatac ttcaaatggc tcaanatgct 1620gnttcacaag gatattcaaa agcctctaaa attgctcaaa atgaaagaac taaatcaata 1680tcggaaagaa aaaatgtcat tcaattctca gaaacctttg acggaatcaa atttagannn 1740tatntngatg taaatacagg aagaattaca aacattcacc cagaataa 1788 72 595 PRT Neisseria meningitidis SITE (61) unknown 72Tyr Leu Lys Gln Leu Gln Val Ala Lys Asn Ile Asn Trp Asn Gln Val 1 5 10 15Gln Leu Ala Tyr Asp Arg Trp Asp Tyr Lys Gln Glu Gly Leu Thr Glu 20 25 30Ala Gly Ala Ala Ile Ile Ala Leu Ala Val Thr Val Val Thr Ser Gly 35 40 45Ala Gly Thr Gly Ala Val Leu Gly Leu Asn Gly Ala Xaa Ala Ala Ala 50 55 60Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser Gln Ala Ser Val Ser Phe 65 70 75 80Ile Asn Asn Lys Gly Asp Val Gly Lys Thr Leu Lys Glu Leu Gly Arg 85 90 95Ser Ser Thr Val Lys Asn Leu Val Val Ala Ala Ala Thr Ala Gly Val 100 105 110Ala Asp Lys Ile Gly Ala Ser Ala Leu Xaa Asn Val Ser Asp Lys Gln 115 120 125Trp Ile Asn Asn Leu Thr Val Asn Leu Ala Asn Ala Gly Ser Ala Ala 130 135 140Leu Ile Asn Thr Ala Val Asn Gly Gly Ser Leu Lys Asp Xaa Leu Glu145 150 155 160Ala Asn Ile Leu Ala Ala Leu Val Asn Thr Ala His Gly Glu Ala Ala 165 170 175Ser Lys Ile Lys Gln Leu Asp Gln His Tyr Ile Val His Lys Ile Ala 180 185 190His Ala Ile Ala Gly Cys Ala Ala Ala Ala Ala Asn Lys Gly Lys Cys 195 200 205Gln Asp Gly Ala Ile Gly Ala Ala Val Gly Glu Ile Val Gly Glu Ala 210 215 220Leu Thr Asn Gly Lys Asn Pro Asp Thr Leu Thr Ala Lys Glu Arg Glu225 230 235 240Gln Ile Leu Ala Tyr Ser Lys Leu Val Ala Gly Thr Val Ser Gly Val 245 250 255Val Gly Gly Asp Val Asn Ala Ala Ala Asn Ala Ala Glu Val Ala Val 260 265 270Lys Asn Asn Gln Leu Ser Asp Xaa Glu Gly Arg Glu Phe Asp Asn Glu 275 280 285Met Thr Ala Cys Ala Lys Gln Asn Xaa Pro Gln Leu Cys Arg Lys Asn 290 295 300Thr Val Lys Lys Tyr Gln Asn Val Ala Asp Lys Arg Leu Ala Ala Ser305 310 315 320Ile Ala Ile Cys Thr Asp Ile Ser Arg Ser Thr Glu Cys Arg Thr Ile 325 330 335Arg Lys Gln His Leu Ile Asp Ser Arg Ser Leu His Ser Ser Trp Glu 340 345 350 73 453 DNA Neisseria meningitidis 73atggcaatca ttacattgta ttattctgtc aatggtattt taaatgtatg tgcaaaagca 60aaaaatattc aagtagttgc caataataag aatatggttc tttttgggtt tttggsmrgc 120atcatcggcg gttcaaccaa tgccatgtct cccatattgt taatattttt gcttagcgaa 180acagaaaata aaaatcgtat cgtaaaatca agcaatctat gctatctttt ggcgaaaatt 240gttcaaatat atatgctaag agaccagtat tggttattaa ataagagtga atacgdttta 300atatttttac tgtccgtatt gtctgttatt ggattgtatg ttggaattcg gttaaggact 360aagattagcc caaatttttt taaaatgtta atttttattg ttttattggt attggctctg 420aaaatcgggc attcgggttt aatcaaactt taa 453 74 150 PRT Neisseria meningitidis SITE (39)..(40) unknown 74Met Ala Ile Ile Thr Leu Tyr Tyr Ser Val Asn Gly Ile Leu Asn Val 1 5 10 15Cys Ala Lys Ala Lys Asn Ile Gln Val Val Ala Asn Asn Lys Asn Met 20 25 30Val Leu Phe Gly Phe Leu Xaa Xaa Ile Ile Gly Gly Ser Thr Asn Ala 35 40 45Met Ser Pro Ile Leu Leu Ile Phe Leu Leu Ser Glu Thr Glu Asn Lys 50 55 60Asn Arg Ile Val Lys Ser Ser Asn Leu Cys Tyr Leu Leu Ala Lys Ile 65 70 75 80Val Gln Ile Tyr Met Leu Arg Asp Gln Tyr Trp Leu Leu Asn Lys Ser 85 90 95Glu Tyr Xaa Leu Ile Phe Leu Leu Ser Val Leu Ser Val Ile Gly Leu 100 105 110Tyr Val Gly Ile Arg Leu Arg Thr Lys Ile Ser Pro Asn Phe Phe Lys 115 120 125Met Leu Ile Phe Ile Val Leu Leu Val Leu Ala Leu Lys Ile Gly His 130 135 140Ser Gly Leu Ile Lys Leu145 150 75 768 DNA Neisseria meningitidis 75atgcaagaaa taatgcaatc tatcgttttt gttgctgccg caatactgca cggaattaca 60ggcatgggat ttccgatgct cggtacaacc gcattggctt ttatcatgcc attgtctaag 120gttgttgcct tggtggcatt accaagcctg ttaatgagct tgttggttct atgcagcaat 180aacaaaaagg gtttttggca agagattgtt tattatttaa aaacctataa attgcttgct 240atcggcagcg tcgttggcag cattttgggg gtgaagttgc ttttgatact tccagtgtct 300tggctgcttt tactgatggc aatcattaca ttgtattatt ctgtcaatgg tattttaaat 360gtatgtgcaa aagcaaaaaa tattcaagta gttgccaata ataagaatat ggttcttttt 420gggtttttgg caggcatcat cggcggttca accaatgcca tgtctcccat attgttaata 480tttttgctta gcgaaacaga aaataaaaat cgtatcgtaa aatcaagcaa tctatgctat 540cttttggcga aaattgttca aatatatatg ctaagagacc agtattggtt attaaataag 600agtgaatacg gtttaatatt tttactgtcc gtattgtctg ttattggatt gtatgttgga 660attcggttaa ggactaagat tagcccaaat ttttttaaaa tgttaatttt tattgtttta 720ttggtattgg ctctgaaaat cgggcattcg ggtttaatca aactttaa 768 76 255 PRT Neisseria meningitidis 76Met Gln Glu Ile Met Gln Ser Ile Val Phe Val Ala Ala Ala Ile Leu 1 5 10 15His Gly Ile Thr Gly Met Gly Phe Pro Met Leu Gly Thr Thr Ala Leu 20 25 30Ala Phe Ile Met Pro Leu Ser Lys Val Val Ala Leu Val Ala Leu Pro 35 40 45Ser Leu Leu Met Ser Leu Leu Val Leu Cys Ser Asn Asn Lys Lys Gly 50 55 60Phe Trp Gln Glu Ile Val Tyr Tyr Leu Lys Thr Tyr Lys Leu Leu Ala 65 70 75 80Ile Gly Ser Val Val Gly Ser Ile Leu Gly Val Lys Leu Leu Leu Ile 85 90 95Leu Pro Val Ser Trp Leu Leu Leu Leu Met Ala Ile Ile Thr Leu Tyr 100 105 110Tyr Ser Val Asn Gly Ile Leu Asn Val Cys Ala Lys Ala Lys Asn Ile 115 120 125Gln Val Val Ala Asn Asn Lys Asn Met Val Leu Phe Gly Phe Leu Ala 130 135 140Gly Ile Ile Gly Gly Ser Thr Asn Ala Met Ser Pro Ile Leu Leu Ile145 150 155 160Phe Leu Leu Ser Glu Thr Glu Asn Lys Asn Arg Ile Val Lys Ser Ser 165 170 175Asn Leu Cys Tyr Leu Leu Ala Lys Ile Val Gln Ile Tyr Met Leu Arg 180 185 190Asp Gln Tyr Trp Leu Leu Asn Lys Ser Glu Tyr Gly Leu Ile Phe Leu 195 200 205Leu Ser Val Leu Ser Val Ile Gly Leu Tyr Val Gly Ile Arg Leu Arg 210 215 220Thr Lys Ile Ser Pro Asn Phe Phe Lys Met Leu Ile Phe Ile Val Leu225 230 235 240Leu Val Leu Ala Leu Lys Ile Gly His Ser Gly Leu Ile Lys Leu 245 250 255 77 768 DNA Neisseria meningitidis 77atgcaagaaa taatgcaatc tatcgttttt gttgctgccg caatactgca cggaattaca 60ggcatgggat ttccgatgct cggtacaacc gcattggctt ttatcatgcc attgtctaag 120gttgttgcct tggtggcatt accaagcctg ttaatgagct tgttggttct atgcagcaat 180aacaaaaagg gtttttggca agagattgtt tattatttaa aaacctataa attgcttgct 240atcggcagcg tcgttggcag cattttgggg gtgaagttgc ttttgatact tccagtgtct 300tggctgcttt tactgatggc aatcattaca ttgtattatt ctgtcaatgg tattttaaat 360gtatgtgcaa aagcaaaaaa tattcaagta gttgccaata ataagaatat ggttcttttt 420gggtttttgg caggcatcat cggcggttca accaatgcca tgtctcccat attgttaata 480tttttgctta gcgaaacaga gaataaaaat cgtatcgcaa aatcaagcaa tctatgctat 540cttttggcaa aaattgttca aatatatatg ctaagagacc agtattggtt attaaataag 600agtgaatacg gtttaatatt tttactgtcc gtattgtctg ttattggatt gtatgttgga 660attcggttaa ggactaagat tagcccaaat ttttttaaaa tgttaatttt tattgtttta 720ttggtattgg ctctgaaaat cgggtattca ggtttaatca aactttaa 768 78 255 PRT Neisseria meningitidis 78Met Gln Glu Ile Met Gln Ser Ile Val Phe Val Ala Ala Ala Ile Leu 1 5 10 15His Gly Ile Thr Gly Met Gly Phe Pro Met Leu Gly Thr Thr Ala Leu 20 25 30Ala Phe Ile Met Pro Leu Ser Lys Val Val Ala Leu Val Ala Leu Pro 35 40 45Ser Leu Leu Met Ser Leu Leu Val Leu Cys Ser Asn Asn Lys Lys Gly 50 55 60Phe Trp Gln Glu Ile Val Tyr Tyr Leu Lys Thr Tyr Lys Leu Leu Ala 65 70 75 80Ile Gly Ser Val Val Gly Ser Ile Leu Gly Val Lys Leu Leu Leu Ile 85 90 95Leu Pro Val Ser Trp Leu Leu Leu Leu Met Ala Ile Ile Thr Leu Tyr 100 105 110Tyr Ser Val Asn Gly Ile Leu Asn Val Cys Ala Lys Ala Lys Asn Ile 115 120 125Gln Val Val Ala Asn Asn Lys Asn Met Val Leu Phe Gly Phe Leu Ala 130 135 140Gly Ile Ile Gly Gly Ser Thr Asn Ala Met Ser Pro Ile Leu Leu Ile145 150 155 160Phe Leu Leu Ser Glu Thr Glu Asn Lys Asn Arg Ile Ala Lys Ser Ser 165 170 175Asn Leu Cys Tyr Leu Leu Ala Lys Ile Val Gln Ile Tyr Met Leu Arg 180 185 190Asp Gln Tyr Trp Leu Leu Asn Lys Ser Glu Tyr Gly Leu Ile Phe Leu 195 200 205Leu Ser Val Leu Ser Val Ile Gly Leu Tyr Val Gly Ile Arg Leu Arg 210 215 220Thr Lys Ile Ser Pro Asn Phe Phe Lys Met Leu Ile Phe Ile Val Leu225 230 235 240Leu Val Leu Ala Leu Lys Ile Gly Tyr Ser Gly Leu Ile Lys Leu 245 250 255 79 516 DNA Neisseria meningitidis 79atgagacata tgaaaataca aaattattta ctagtattta tagttttaca tatagccttg 60atagtaatta atatagtgtt tggttatttt gtttttctat ttgatttttt tgcgtttttg 120ttttttgcaa acgtctttct tgctgtaaat ttattatttt tagaaaaaaa cataaaaaac 180aaattattgt ttttattgcc gatttctatt attatatgga tggtaattca tattagtatg 240ataaatataa aattttataa atttgagcat caaataaagg aacaaaatat atcctcgatt 300actggggtga taaaaccaca tgatagttat aattatgttt atgactcaaa tggatatgct 360aaattaaaag ataatcatag atatggtagg gtaattagag aaacacctta tattgatgta 420gttgcatctg atgttaaaaa taaatccata agattaagct tggtttgtgg tattcattca 480tatgctccat gtgccaattt tataaaattt gtcagg 516 80 172 PRT Neisseria meningitidis 80Met Arg His Met Lys Ile Gln Asn Tyr Leu Leu Val Phe Ile Val Leu 1 5 10 15His Ile Ala Leu Ile Val Ile Asn Ile Val Phe Gly Tyr Phe Val Phe 20 25 30Leu Phe Asp Phe Phe Ala Phe Leu Phe Phe Ala Asn Val Phe Leu Ala 35 40 45Val Asn Leu Leu Phe Leu Glu Lys Asn Ile Lys Asn Lys Leu Leu Phe 50 55 60Leu Leu Pro Ile Ser Ile Ile Ile Trp Met Val Ile His Ile Ser Met 65 70 75 80Ile Asn Ile Lys Phe Tyr Lys Phe Glu His Gln Ile Lys Glu Gln Asn 85 90 95Ile Ser Ser Ile Thr Gly Val Ile Lys Pro His Asp Ser Tyr Asn Tyr 100 105 110Val Tyr Asp Ser Asn Gly Tyr Ala Lys Leu Lys Asp Asn His Arg Tyr 115 120 125Gly Arg Val Ile Arg Glu Thr Pro Tyr Ile Asp Val Val Ala Ser Asp 130 135 140Val Lys Asn Lys Ser Ile Arg Leu Ser Leu Val Cys Gly Ile His Ser145 150 155 160Tyr Ala Pro Cys Ala Asn Phe Ile Lys Phe Val Arg 165 170 81 729 DNA Neisseria meningitidis 81atgagacata tgaaaaataa aaattattta ctagtattta tagttttaca tatagccttg 60atagtaatta atatagtgtt tggttatttt gtttttctat ttgatttttt tgcgtttttg 120ttttttgcaa acgtctttct tgctgtaaat ttattatttt tagaaaaaaa cataaaaaac 180aaattattgt ttttattgcc gatttctatt attatatgga tggtaattca tattagtatg 240ataaatataa aattttataa atttgagcat caaataaagg aacaaaatat atcctcgatt 300actggggtga taaaaccaca tgatagttat aattatgttt atgactcaaa tggatatgct 360aaattaaaag ataatcatag atatggtagg gtaattagag aaacacctta tattgatgta 420gttgcatctg atgttaaaaa taaatccata agattaagct tggtttgtgg tattcattca 480tatgctccat gtgccaattt tataaaattt gcaaaaaaac ctgttaaaat ttatttttat 540aatcaacctc aaggagattt tatagataat gtaatatttg aaattaatga tggaaacaaa 600agtttgtact tgttagataa gtataaaaca ttttttctta ttgaaaacag tgtttgtatc 660gtattaatta ttttatattt aaaatttaat ttgcttttat ataggactta cttcaatgag 720ttggaatag 729 82 242 PRT Neisseria meningitidis 82Met Arg His Met Lys Asn Lys Asn Tyr Leu Leu Val Phe Ile Val Leu 1 5 10 15His Ile Ala Leu Ile Val Ile Asn Ile Val Phe Gly Tyr Phe Val Phe 20 25 30Leu Phe Asp Phe Phe Ala Phe Leu Phe Phe Ala Asn Val Phe Leu Ala 35 40 45Val Asn Leu Leu Phe Leu Glu Lys Asn Ile Lys Asn Lys Leu Leu Phe 50 55 60Leu Leu Pro Ile Ser Ile Ile Ile Trp Met Val Ile His Ile Ser Met 65 70 75 80Ile Asn Ile Lys Phe Tyr Lys Phe Glu His Gln Ile Lys Glu Gln Asn 85 90 95Ile Ser Ser Ile Thr Gly Val Ile Lys Pro His Asp Ser Tyr Asn Tyr 100 105 110Val Tyr Asp Ser Asn Gly Tyr Ala Lys Leu Lys Asp Asn His Arg Tyr 115 120 125Gly Arg Val Ile Arg Glu Thr Pro Tyr Ile Asp Val Val Ala Ser Asp 130 135 140Val Lys Asn Lys Ser Ile Arg Leu Ser Leu Val Cys Gly Ile His Ser145 150 155 160Tyr Ala Pro Cys Ala Asn Phe Ile Lys Phe Ala Lys Lys Pro Val Lys 165 170 175Ile Tyr Phe Tyr Asn Gln Pro Gln Gly Asp Phe Ile Asp Asn Val Ile 180 185 190Phe Glu Ile Asn Asp Gly Asn Lys Ser Leu Tyr Leu Leu Asp Lys Tyr 195 200 205Lys Thr Phe Phe Leu Ile Glu Asn Ser Val Cys Ile Val Leu Ile Ile 210 215 220Leu Tyr Leu Lys Phe Asn Leu Leu Leu Tyr Arg Thr Tyr Phe Asn Glu225 230 235 240Leu Glu 83 729 DNA Neisseria meningitidis 83atgagacata tgaaaaataa aaattattta ctagtattta tagttttaca tataaccttg 60atagtaatta atatagtgtt tggttatttt gtttttctat ttgatttttt tgcgtttttg 120ttttttgcaa acgtctttct tgctgtaaat ttattatttt tagaaaaaaa cataaaaaac 180aaattattgt ttttattgcc gatttctatt attatatgga tggtaattca tattagtatg 240ataaatataa aattttataa atttgagcat caaataaagg aacaaaatat atcctcgatt 300actggggtga taaaaccaca tgatagttat aattatgttt atgactcaaa tggatatgct 360aaattaaaag ataatcatag atatggtagg gtaattagag aaacacctta tattgatgta 420gttgcatctg atgttaaaaa taaatccata agattaagct tggtttgtgg tattcattca 480tatgctccat gtgccaattt tataaaattt gcaaaaaaac ctgttaaaat ttatttttat 540aatcaacctc aaggagattt tatagataat gtaatatttg aaattaatga tggaaaaaaa 600agtttgtact tgttagataa gtataaaaca ttttttctta ttgaaaacag tgtttgtatc 660gtattaatta ttttatattt aaaatttaat ttgcttttat ataggactta cttcaatgag 720ttggaatag 729 84 242 PRT Neisseria meningitidis 84Met Arg His Met Lys Asn Lys Asn Tyr Leu Leu Val Phe Ile Val Leu 1 5 10 15His Ile Thr Leu Ile Val Ile Asn Ile Val Phe Gly Tyr Phe Val Phe 20 25 30Leu Phe Asp Phe Phe Ala Phe Leu Phe Phe Ala Asn Val Phe Leu Ala 35 40 45Val Asn Leu Leu Phe Leu Glu Lys Asn Ile Lys Asn Lys Leu Leu Phe 50 55 60Leu Leu Pro Ile Ser Ile Ile Ile Trp Met Val Ile His Ile Ser Met 65 70 75 80Ile Asn Ile Lys Phe Tyr Lys Phe Glu His Gln Ile Lys Glu Gln Asn 85 90 95Ile Ser Ser Ile Thr Gly Val Ile Lys Pro His Asp Ser Tyr Asn Tyr 100 105 110Val Tyr Asp Ser Asn Gly Tyr Ala Lys Leu Lys Asp Asn His Arg Tyr 115 120 125Gly Arg Val Ile Arg Glu Thr Pro Tyr Ile Asp Val Val Ala Ser Asp 130 135 140Val Lys Asn Lys Ser Ile Arg Leu Ser Leu Val Cys Gly Ile His Ser145 150 155 160Tyr Ala Pro Cys Ala Asn Phe Ile Lys Phe Ala Lys Lys Pro Val Lys 165 170 175Ile Tyr Phe Tyr Asn Gln Pro Gln Gly Asp Phe Ile Asp Asn Val Ile 180 185 190Phe Glu Ile Asn Asp Gly Lys Lys Ser Leu Tyr Leu Leu Asp Lys Tyr 195 200 205Lys Thr Phe Phe Leu Ile Glu Asn Ser Val Cys Ile Val Leu Ile Ile 210 215 220Leu Tyr Leu Lys Phe Asn Leu Leu Leu Tyr Arg Thr Tyr Phe Asn Glu225 230 235 240Leu Glu 85 552 DNA Neisseria meningitidis 85acccccaaca gcgtgaccgt cttgccgtct ttcggcggat tcgggcgtac cggcgcgacc 60atcaatgcag caggcggggt cggcatgact gccttttcga caaccttaat ttccgtagcc 120gagggcgcgg ttgtagagct gcaggccgtg agagccaaag ccgtcaatgc aaccgccgct 180tgcattttta cggtcttgag taaggacatt ttcgatttcc tttttatttt ccgttttcag 240acggctgact tccgcctgta ttttcgccaa agccatgccg acagcgtgcg ccttgacttc 300atatttaaaa gcttccgcgc gtgccagttc cagttcgcgc gcatagtttt gagccgacaa 360cagcagggct tgcgccttgt cgcgctccat cttgtcgatg accgcctgca gcttcgcaaa 420tgccgacttg tagccttgat ggtgcgacac agccaagccc gtgccgacaa gcgcgataat 480ggcaatcggt tgccagtaat tcgccagcag tttcacgaga ttcattctcg acctcctgac 540gcttcacgct ga 552 86 183 PRT Neisseria meningitidis 86Thr Pro Asn Ser Val Thr Val Leu Pro Ser Phe Gly Gly Phe Gly Arg 1 5 10 15Thr Gly Ala Thr Ile Asn Ala Ala Gly Gly Val Gly Met Thr Ala Phe 20 25 30Ser Thr Thr Leu Ile Ser Val Ala Glu Gly Ala Val Val Glu Leu Gln 35 40 45Ala Val Arg Ala Lys Ala Val Asn Ala Thr Ala Ala Cys Ile Phe Thr 50 55 60Val Leu Ser Lys Asp Ile Phe Asp Phe Leu Phe Ile Phe Arg Phe Gln 65 70 75 80Thr Ala Asp Phe Arg Leu Tyr Phe Arg Gln Ser His Ala Asp Ser Val 85 90 95Arg Leu Asp Phe Ile Phe Lys Ser Phe Arg Ala Cys Gln Phe Gln Phe 100 105 110Ala Arg Ile Val Leu Ser Arg Gln Gln Gln Gly Leu Arg Leu Val Ala 115 120 125Leu His Leu Val Asp Asp Arg Leu Gln Leu Arg Lys Cys Arg Leu Val 130 135 140Ala Leu Met Val Arg His Ser Gln Ala Arg Ala Asp Lys Arg Asp Asn145 150 155 160Gly Asn Arg Leu Pro Val Ile Arg Gln Gln Phe His Glu Ile His Ser 165 170 175Arg Pro Pro Asp Ala Ser Arg 180 87 468 DNA Neisseria meningitidis 87atgactgcct tttcgacaac cttaatttcc gtagccgagg gcgcggttgt agagctgcag 60gccgtgagag ccaaagccgt caatgcaacc gccgcttgca tttttacggt cttgagtaag 120gacattttcg atttcctttt tattttccgt tttcagacgg ctgacttccg cctgtttttt 180cgccaaagcc atgccgacag cgtgcgcctt gacttcatat tttttagctt ccgcgcgtgc 240cagttccagt tcgcgcgcat agttttgagc cgacaacagc agggcttgcg ccttgtcgcg 300ctccatcttg tcgatgaccg cctgctgctt cgcaaatgcc gacttgtagc cttgatggtg 360cgacacagcc aagcccgtgc cgacaagcgc gataatggca atcggttgcc agttattcgc 420cagcagtttc acgagattca ttctcgacct cctgacgctt cacgctga 468 88 155 PRT Neisseria meningitidis 88Met Thr Ala Phe Ser Thr Thr Leu Ile Ser Val Ala Glu Gly Ala Val 1 5 10 15Val Glu Leu Gln Ala Val Arg Ala Lys Ala Val Asn Ala Thr Ala Ala 20 25 30Cys Ile Phe Thr Val Leu Ser Lys Asp Ile Phe Asp Phe Leu Phe Ile 35 40 45Phe Arg Phe Gln Thr Ala Asp Phe Arg Leu Phe Phe Arg Gln Ser His 50 55 60Ala Asp Ser Val Arg Leu Asp Phe Ile Phe Phe Ser Phe Arg Ala Cys 65 70 75 80Gln Phe Gln Phe Ala Arg Ile Val Leu Ser Arg Gln Gln Gln Gly Leu 85 90 95Arg Leu Val Ala Leu His Leu Val Asp Asp Arg Leu Leu Leu Arg Lys 100 105 110Cys Arg Leu Val Ala Leu Met Val Arg His Ser Gln Ala Arg Ala Asp 115 120 125Lys Arg Asp Asn Gly Asn Arg Leu Pro Val Ile Arg Gln Gln Phe His 130 135 140Glu Ile His Ser Arg Pro Pro Asp Ala Ser Arg145 150 155 89 462 DNA Neisseria meningitidis 89atgaccgcct tttcgacaac cttaatttcc gtagccgagg gcgcgcttgt agagctgcaa 60gccgtgatgg ccaaagccgt caatacaacc gccgcctgca tttttacggt cttgagtaag 120gacattttcg atttcctttt tattttccgt tttcagacgg ctgacttccg cctgtttttt 180cgccaaagcc atgccgacgg cgtgcgcctt gacttcatat tttttagctt ccgcacgcgc 240ctgttccagt tcgcgggcgt agttttgagc cgacaacagc agggcttgcg ccttgtcgcg 300cttcattttc tcaatgaccg cctgctgctt cgcaaaagcc gacttgtagc cttgatggtg 360cgacaccgcc aaacccgtgc cgacaagcgc gatgatggca atcggttgcc agttattcgc 420cagcagtttc acgagattca ttctcgacct cctgacgttt ga 462 90 153 PRT Neisseria meningitidis 90Met Thr Ala Phe Ser Thr Thr Leu Ile Ser Val Ala Glu Gly Ala Leu 1 5 10 15Val Glu Leu Gln Ala Val Met Ala Lys Ala Val Asn Thr Thr Ala Ala 20 25 30Cys Ile Phe Thr Val Leu Ser Lys Asp Ile Phe Asp Phe Leu Phe Ile 35 40 45Phe Arg Phe Gln Thr Ala Asp Phe Arg Leu Phe Phe Arg Gln Ser His 50 55 60Ala Asp Gly Val Arg Leu Asp Phe Ile Phe Phe Ser Phe Arg Thr Arg 65 70 75 80Leu Phe Gln Phe Ala Gly Val Val Leu Ser Arg Gln Gln Gln Gly Leu 85 90 95Arg Leu Val Ala Leu His Phe Leu Asn Asp Arg Leu Leu Leu Arg Lys 100 105 110Ser Arg Leu Val Ala Leu Met Val Arg His Arg Gln Thr Arg Ala Asp 115 120 125Lys Arg Asp Asp Gly Asn Arg Leu Pro Val Ile Arg Gln Gln Phe His 130 135 140Glu Ile His Ser Arg Pro Pro Asp Val145 150 91 592 PRT Neisseria meningitidis 91Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Ser Val Gln 50 55 60Arg Ser Val Val Gly Ser Ile Gln Ala Ser Met Glu Gly Ser Gly Glu 65 70 75 80Leu Glu Thr Ile Ser Leu Ser Met Thr Asn Asp Ser Lys Glu Phe Val 85 90 95Asp Pro Tyr Ile Val Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile 100 105 110Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn Val Glu Thr Glu Lys Leu 130 135 140Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Ile Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly 180 185 190Ser Ser Ala Ser His Val Asp Ala Gly Asn Gln Ser Thr His Tyr Thr 195 200 205Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 210 215 220Gly Val Lys Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Ser Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val465 470 475 480Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn 485 490 495Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp 500 505 510Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr 515 520 525Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile 530 535 540Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser545 550 555 560Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly 565 570 575Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 92 594 PRT Neisseria meningitidis 92Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Asp Asp Asp Leu Tyr Leu Glu Pro Val Gln Arg 50 55 60Thr Ala Val Val Leu Ser Phe Arg Ser Asp Lys Glu Gly Thr Gly Glu 65 70 75 80Lys Glu Gly Thr Glu Asp Ser Asn Trp Ala Val Tyr Phe Asp Glu Lys 85 90 95Arg Val Leu Lys Ala Gly Ala Ile Thr Leu Lys Ala Gly Asp Asn Leu 100 105 110Lys Ile Lys Gln Asn Thr Asn Glu Asn Thr Asn Asp Ser Ser Phe Thr 115 120 125Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Glu Thr Glu 130 135 140Lys Leu Ser Phe Gly Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp145 150 155 160Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp 165 170 175Pro Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu 180 185 190Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp 195 200 205Asp Glu Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly 210 215 220Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val225 230 235 240Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr 245 250 255Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr 260 265 270Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly 275 280 285Lys Leu Val Thr Gly Lys Gly Lys Asp Glu Asn Gly Ser Ser Thr Asp 290 295 300Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn305 310 315 320Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly 325 330 335Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe 340 345 350Ala Ser Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly 355 360 365Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val 370 375 380Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala385 390 395 400Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly 405 410 415Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile 420 425 430Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln 435 440 445Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser 450 455 460Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Thr Asn Lys465 470 475 480Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val 485 490 495Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg 500 505 510Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile 515 520 525Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met 530 535 540Ala Ile Gly Gly Asp Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly545 550 555 560Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala 565 570 575Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr 580 585 590Gln Trp 93 594 PRT Neisseria meningitidis 93Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Thr Thr Asp Asp Asp Asp Leu Tyr Leu Glu Pro Val Gln Arg 50 55 60Thr Ala Pro Val Leu Ser Phe His Ala Asp Ser Glu Gly Thr Gly Glu 65 70 75 80Lys Glu Val Thr Glu Asp Ser Asn Trp Gly Val Tyr Phe Asp Lys Lys 85 90 95Gly Val Leu Thr Ala Gly Thr Ile Thr Leu Lys Ala Gly Asp Asn Leu 100 105 110Lys Ile Lys Gln Asn Thr Asp Glu Asn Thr Asn Ala Ser Ser Phe Thr 115 120 125Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Glu Thr Glu 130 135 140Lys Leu Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Thr Ser Asp145 150 155 160Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp 165 170 175Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu 180 185 190Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp 195 200 205Asp Glu Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly 210 215 220Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val225 230 235 240Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr 245 250 255Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr 260 265 270Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly 275 280 285Lys Leu Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp 290 295 300Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn305 310 315 320Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly 325 330 335Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Lys Val Thr Phe 340 345 350Ala Ser Gly Asn Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly 355 360 365Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val 370 375 380Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala385 390 395 400Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly 405 410 415Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile 420 425 430Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln 435 440 445Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser 450 455 460Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys465 470 475 480Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val 485 490 495Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn His 500 505 510Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile 515 520 525Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met 530 535 540Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly545 550 555 560Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala 565 570 575Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr 580 585 590Gln Trp 94 594 PRT Neisseria meningitidis 94Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Asp Asp Asp Leu Tyr Leu Glu Pro Val Gln Arg 50 55 60Thr Ala Val Val Leu Ser Phe Arg Ser Asp Lys Glu Gly Thr Gly Glu 65 70 75 80Lys Glu Gly Thr Glu Asp Ser Asn Trp Ala Val Tyr Phe Asp Glu Lys 85 90 95Arg Val Leu Lys Ala Gly Ala Ile Thr Leu Lys Ala Gly Asp Asn Leu 100 105 110Lys Ile Lys Gln Asn Thr Asn Glu Asn Thr Asn Asp Ser Ser Phe Thr 115 120 125Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Glu Thr Glu 130 135 140Lys Leu Ser Phe Gly Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp145 150 155 160Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp 165 170 175Pro Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu 180 185 190Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp 195 200 205Asp Glu Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly 210 215 220Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val225 230 235 240Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr 245 250 255Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr 260 265 270Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly 275 280 285Lys Leu Val Thr Gly Lys Gly Lys Asp Glu Asn Gly Ser Ser Thr Asp 290 295 300Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn305 310 315 320Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly 325 330 335Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe 340 345 350Ala Ser Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly 355 360 365Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val 370 375 380Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala385 390 395 400Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly 405 410 415Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile 420 425 430Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln 435 440 445Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser 450 455 460Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Thr Asn Lys465 470 475 480Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val 485 490 495Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg 500 505 510Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile 515 520 525Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met 530 535 540Ala Ile Gly Gly Asp Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly545 550 555 560Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala 565 570 575Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr 580 585 590Gln Trp 95 591 PRT Neisseria meningitidis 95Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Glu Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Ala Asn Asn Glu Glu Gln Glu Glu Asp Leu Tyr Leu Asp Pro 50 55 60Val Gln Arg Thr Val Ala Val Leu Ile Val Asn Ser Asp Lys Glu Gly 65 70 75 80Thr Gly Glu Lys Glu Lys Val Glu Glu Asn Ser Asp Trp Ala Val Tyr 85 90 95Phe Asn Glu Lys Gly Val Leu Thr Ala Arg Glu Ile Thr Leu Lys Ala 100 105 110Gly Asp Asn Leu Lys Ile Lys Gln Asn Gly Thr Asn Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr Glu Lys Leu 130 135 140Ser Phe Ser Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Gly Asp Ala Leu Asn Val Gly Ser Lys Lys Asp Asn Lys Pro Val Arg465 470 475 480Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val 485 490 495Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn 500 505 510Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala 515 520 525Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly 530 535 540Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser545 550 555 560Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly Asn 565 570 575Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 96 591 PRT Neisseria meningitidis 96Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Ala Asn Asn Glu Glu Gln Glu Glu Asp Leu Tyr Leu Asp Pro 50 55 60Val Gln Arg Thr Val Ala Val Leu Ile Val Asn Ser Asp Lys Glu Gly 65 70 75 80Thr Gly Glu Lys Glu Lys Val Glu Glu Asn Ser Asp Trp Ala Val Tyr 85 90 95Phe Asn Glu Lys Gly Val Leu Thr Ala Arg Glu Ile Thr Leu Lys Ala 100 105 110Gly Asp Asn Leu Lys Ile Lys Gln Asn Gly Thr Asn Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr Glu Lys Leu 130 135 140Ser Phe Ser Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Gly Asp Ala Leu Asn Val Gly Ser Lys Lys Asp Asn Lys Pro Val Arg465 470 475 480Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val 485 490 495Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn 500 505 510Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala 515 520 525Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly 530 535 540Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser545 550 555 560Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly Asn 565 570 575Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 97 595 PRT Neisseria meningitidis 97Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Glu Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Thr Asp Glu Asp Asp Glu Leu Glu Pro Val Val 50 55 60Arg Ser Ala Leu Val Leu Gln Phe Met Ile Asp Lys Glu Gly Asn Gly 65 70 75 80Glu Ile Glu Ser Thr Gly Asp Ile Gly Trp Ser Ile Tyr Tyr Asp Asp 85 90 95His Asn Thr Leu His Gly Ala Thr Val Thr Leu Lys Ala Gly Asp Asn 100 105 110Leu Lys Ile Lys Gln Asn Thr Asp Glu Asn Thr Asn Ala Ser Ser Phe 115 120 125Thr Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr 130 135 140Glu Glu Leu Ser Phe Gly Ala Asn Gly Asn Lys Val Asn Ile Thr Ser145 150 155 160Asp Thr Lys Gly Leu Asn Phe Ala Lys Lys Thr Ala Gly Thr Asn Gly 165 170 175Asp Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr 180 185 190Leu Ala Gly Ser Ser Ala Ser His Val Asp Ala Gly Asn Gln Ser Thr 195 200 205His Tyr Thr Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp 210 215 220Asn Ile Lys Gly Val Lys Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn225 230 235 240Val Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp 245 250 255Thr Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Arg 260 265 270Thr Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp 275 280 285Gly Lys Leu Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr 290 295 300Asp Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val305 310 315 320Asn Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr 325 330 335Gly Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr 340 345 350Phe Ala Ser Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln 355 360 365Gly Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp Ala Leu Asn 370 375 380Val Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val385 390 395 400Ala Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys 405 410 415Gly Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu 420 425 430Ile Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro 435 440 445Gln Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu 450 455 460Ser Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn465 470 475 480Lys Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp 485 490 495Val Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn 500 505 510His Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala 515 520 525Ile Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met 530 535 540Met Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile545 550 555 560Gly Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr 565 570 575Ala Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly 580 585 590Tyr Gln Trp 595 98 592 PRT Neisseria meningitidis 98Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Ser Val Gln 50 55 60Arg Ser Val Val Gly Ser Ile Gln Ala Ser Met Glu Gly Ser Gly Glu 65 70 75 80Leu Glu Thr Ile Ser Leu Ser Met Thr Asn Asp Ser Lys Glu Phe Val 85 90 95Asp Pro Tyr Ile Val Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile 100 105 110Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn Val Glu Thr Glu Lys Leu 130 135 140Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Ile Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly 180 185 190Ser Ser Ala Ser His Val Asp Ala Gly Asn Gln Ser Thr His Tyr Thr 195 200 205Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 210 215 220Gly Val Lys Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Ser Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val465 470 475 480Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn 485 490 495Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp 500 505 510Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr 515 520 525Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile 530 535 540Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser545 550 555 560Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly 565 570 575Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 99 594 PRT Neisseria meningitidis 99Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Thr Thr Asp Asp Asp Asp Leu Tyr Leu Glu Pro Val Gln Arg 50 55 60Thr Ala Pro Val Leu Ser Phe His Ala Asp Ser Glu Gly Thr Gly Glu 65 70 75 80Lys Glu Val Thr Glu Asp Ser Asn Trp Gly Val Tyr Phe Asp Lys Lys 85 90 95Gly Val Leu Thr Ala Gly Thr Ile Thr Leu Lys Ala Gly Asp Asn Leu 100 105 110Lys Ile Lys Gln Asn Thr Asp Glu Asn Thr Asn Ala Ser Ser Phe Thr 115 120 125Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Glu Thr Glu 130 135 140Lys Leu Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Thr Ser Asp145 150 155 160Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp 165 170 175Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu 180 185 190Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp 195 200 205Asp Glu Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly 210 215 220Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val225 230 235 240Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr 245 250 255Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr 260 265 270Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly 275 280 285Lys Leu Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp 290 295 300Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn305 310 315 320Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly 325 330 335Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Lys Val Thr Phe 340 345 350Ala Ser Gly Asn Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly 355 360 365Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val 370 375 380Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala385 390 395 400Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly 405 410 415Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile 420 425 430Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln 435 440 445Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser 450 455 460Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys465 470 475 480Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val 485 490 495Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn His 500 505 510Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile 515 520 525Ala Thr Ala Ser Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met 530 535 540Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly545 550 555 560Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala 565 570 575Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr 580 585 590Gln Trp 100 599 PRT Neisseria meningitidis 100Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Pro Val Val 50 55 60Arg Ser Ala Leu Val Leu Gln Phe Met Ile Asp Lys Glu Gly Asn Gly 65 70 75 80Glu Asn Glu Ser Thr Gly Asn Ile Gly Trp Ser Ile Tyr Tyr Asp Asn 85 90 95His Asn Thr Leu His Gly Ala Thr Val Thr Leu Lys Ala Gly Asp Asn 100 105 110Leu Lys Ile Lys Gln Asn Thr Asn Lys Asn Thr Asn Glu Asn Thr Asn 115 120 125Asp Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr 130 135 140Ser Val Glu Thr Glu Lys Leu Ser Phe Gly Ala Asn Gly Asn Lys Val145 150 155 160Asn Ile Thr Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala 165 170 175Gly Thr Asn Gly Asp Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr 180 185 190Leu Thr Asp Thr Leu Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn 195 200 205Asp Asn Val Thr Asp Asp Lys Lys Lys Arg Ala Ala Ser Val Lys Asp 210 215 220Val Leu Asn Ala Gly Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr225 230 235 240Ala Ser Asp Asn Val Asp Phe Val His Thr Tyr Asp Thr Val Glu Phe 245 250 255Leu Ser Ala Asp Thr Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp 260 265 270Asn Gly Lys Arg Thr Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile 275 280 285Lys Glu Lys Asp Gly Lys Leu Val Thr Gly Lys Gly Lys Gly Glu Asn 290 295 300Gly Ser Ser Thr Asp Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val305 310 315 320Ile Asp Ala Val Asn Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala 325 330 335Asn Gly Gln Thr Gly Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly 340 345 350Thr Asn Val Thr Phe Ala Ser Gly Lys Gly Thr Thr Ala Thr Val Ser 355 360 365Lys Asp Asp Gln Gly Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly 370 375 380Asp Ala Leu Asn Val Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp385 390 395 400Ser Lys Ala Val Ala Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val 405 410 415Ser Pro Ser Lys Gly Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly 420 425 430Asn Asn Ile Glu Ile Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr 435 440 445Ser Met Thr Pro Gln Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp 450 455 460Ala Pro Thr Leu Ser Val Asp Asp Lys Gly Ala Leu Asn Val Gly Ser465 470 475 480Lys Asp Ala Asn Lys Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val 485 490 495Lys Glu Gly Asp Val Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln 500 505 510Asn Leu Asn Asn Arg Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly 515 520 525Ile Ala Gln Ala Ile Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu Pro 530 535 540Gly Lys Ser Met Met Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu Ala545 550 555 560Gly Tyr Ala Ile Gly Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile 565 570 575Ile Lys Gly Thr Ala Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser 580 585 590Ala Ser Val Gly Tyr Gln Trp 595 101 598 PRT Neisseria meningitidis 101Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Asp Asp Asp Leu Tyr Leu Glu Pro Val Gln Arg 50 55 60Thr Ala Val Val Leu Ser Phe Arg Ser Asp Lys Glu Gly Thr Gly Glu 65 70 75 80Lys Glu Gly Thr Glu Asp Ser Asn Trp Ala Val Tyr Phe Asp Glu Lys 85 90 95Arg Val Leu Lys Ala Gly Ala Ile Thr Leu Lys Ala Gly Asp Asn Leu 100 105 110Lys Ile Lys Gln Asn Thr Asn Glu Asn Thr Asn Glu Asn Thr Asn Asp 115 120 125Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser 130 135 140Val Glu Thr Glu Lys Leu Ser Phe Gly Ala Asn Gly Asn Lys Val Asn145 150 155 160Ile Thr Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly 165 170 175Thr Asn Gly Asp Pro Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu 180 185 190Thr Asp Thr Leu Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn Asp 195 200 205Asn Val Thr Asp Asp Glu Lys Lys Arg Ala Ala Ser Val Lys Asp Val 210 215 220Leu Asn Ala Gly Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr Ala225 230 235 240Ser Asp Asn Val Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu 245 250 255Ser Ala Asp Thr Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn 260 265 270Gly Lys Lys Thr Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys 275 280 285Glu Lys Asp Gly Lys Leu Val Thr Gly Lys Gly Lys Asp Glu Asn Gly 290 295 300Ser Ser Thr Asp Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile305 310 315 320Asp Ala Val Asn Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn 325 330 335Gly Gln Thr Gly Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr 340 345 350Lys Val Thr Phe Ala Ser Gly Asn Gly Thr Thr Ala Thr Val Ser Lys 355 360 365Asp Asp Gln Gly Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp 370 375 380Ala Leu Asn Val Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser385 390 395 400Lys Ala Val Ala Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser 405 410 415Pro Ser Lys Gly Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn 420 425 430Asn Ile Glu Ile Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser 435 440 445Met Thr Pro Gln Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala 450 455 460Pro Thr Leu Ser Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys465 470 475 480Asp Ala Asn Lys Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys 485 490 495Glu Gly Asp Val Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn 500 505 510Leu Asn Asn Arg Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile 515 520 525Ala Gln Ala Ile Ala Thr Ala Gly Leu Ala Gln Ala Tyr Leu Pro Gly 530 535 540Lys Ser Met Met Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly545 550 555 560Tyr Ala Ile Gly Tyr Ser Ser Ile Ser Asp Thr Gly Asn Trp Val Ile 565 570 575Lys Gly Thr Ala Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala 580 585 590Ser Val Gly Tyr Gln Trp 595 102 594 PRT Neisseria meningitidis 102Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Thr Thr Asp Asp Asp Asp Leu Tyr Leu Glu Pro Val Gln Arg 50 55 60Thr Ala Pro Val Leu Ser Phe His Ala Asp Ser Glu Gly Thr Gly Glu 65 70 75 80Lys Glu Val Thr Glu Asp Ser Asn Trp Gly Val Tyr Phe Asp Lys Lys 85 90 95Gly Val Leu Thr Ala Gly Thr Ile Thr Leu Lys Ala Gly Asp Asn Leu 100 105 110Lys Ile Lys Gln Asn Thr Asp Glu Asn Thr Asn Ala Ser Ser Phe Thr 115 120 125Tyr Ser Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Glu Thr Glu 130 135 140Lys Leu Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Thr Ser Asp145 150 155 160Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp 165 170 175Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu 180 185 190Leu Asn Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp 195 200 205Asp Glu Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly 210 215 220Trp Asn Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val225 230 235 240Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr 245 250 255Lys Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr 260 265 270Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly 275 280 285Lys Leu Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp 290 295 300Glu Gly Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn305 310 315 320Lys Ala Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly 325 330 335Gln Ala Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Lys Val Thr Phe 340 345 350Ala Ser Gly Asn Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly 355 360 365Asn Ile Thr Val Lys Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val 370 375 380Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala385 390 395 400Gly Ser Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly 405 410 415Lys Met Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile 420 425 430Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln 435 440 445Phe Ser Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser 450 455 460Val Asp Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys465 470 475 480Pro Val Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val 485 490 495Thr Asn Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn His 500 505 510Ile Asp Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile 515 520 525Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met 530 535 540Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly545 550 555 560Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala 565 570 575Ser Gly Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr 580 585 590Gln Trp 103 591 PRT Neisseria meningitidis 103Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Ala Asn Asn Glu Glu Gln Glu Glu Asp Leu Tyr Leu Asp Pro 50 55 60Val Gln Arg Thr Val Ala Val Leu Ile Val Asn Ser Asp Lys Glu Gly 65 70 75 80Thr Gly Glu Lys Glu Lys Val Glu Glu Asn Ser Asp Trp Ala Val Tyr 85 90 95Phe Asn Glu Lys Gly Val Leu Thr Ala Arg Glu Ile Thr Leu Lys Ala 100 105 110Gly Asp Asn Leu Lys Ile Lys Gln Asn Gly Thr Asn Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr Glu Lys Leu 130 135 140Ser Phe Ser Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn His 370 375 380Leu Gln Asn Ser Gly Trp Asp Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Gly Asp Ala Leu Asn Val Gly Ser Lys Lys Asp Asn Lys Pro Val Arg465 470 475 480Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val 485 490 495Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn 500 505 510Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala 515 520 525Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly 530 535 540Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser545 550 555 560Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly Asn 565 570 575Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 104 591 PRT Neisseria meningitidis 104Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Ala Asn Asn Glu Glu Gln Glu Glu Asp Leu Tyr Leu Asp Pro 50 55 60Val Gln Arg Thr Val Ala Val Leu Ile Val Asn Ser Asp Lys Glu Gly 65 70 75 80Thr Gly Glu Lys Glu Lys Val Glu Glu Asn Ser Asp Trp Ala Val Tyr 85 90 95Phe Asn Glu Lys Gly Val Leu Thr Ala Arg Glu Ile Thr Leu Lys Ala 100 105 110Gly Asp Asn Leu Lys Ile Lys Gln Asn Gly Thr Asn Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr Glu Lys Leu 130 135 140Ser Phe Ser Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Gly Asp Ala Leu Asn Val Gly Ser Lys Lys Asp Asn Lys Pro Val Arg465 470 475 480Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val 485 490 495Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn 500 505 510Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala 515 520 525Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly 530 535 540Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser545 550 555 560Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly Asn 565 570 575Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 105 591 PRT Neisseria meningitidis 105Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Ser Ala Asn Asn Glu Glu Gln Glu Glu Asp Leu Tyr Leu Asp Pro 50 55 60Val Gln Arg Thr Val Ala Val Leu Ile Val Asn Ser Asp Lys Glu Gly 65 70 75 80Thr Gly Glu Lys Glu Lys Val Glu Glu Asn Ser Asp Trp Ala Val Tyr 85 90 95Phe Asn Glu Lys Gly Val Leu Thr Ala Arg Glu Ile Thr Leu Lys Ala 100 105 110Gly Asp Asn Leu Lys Ile Lys Gln Asn Gly Thr Asn Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Asp Leu Thr Ser Val Gly Thr Glu Lys Leu 130 135 140Ser Phe Ser Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Asp Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Gly Asp Ala Leu Asn Val Gly Ser Lys Lys Asp Asn Lys Pro Val Arg465 470 475 480Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val 485 490 495Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn 500 505 510Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala 515 520 525Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly 530 535 540Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser545 550 555 560Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly Asn 565 570 575Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 106 592 PRT Neisseria meningitidis 106Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Ser Val Gln 50 55 60Arg Ser Val Val Gly Ser Ile Gln Ala Ser Met Glu Gly Ser Gly Glu 65 70 75 80Leu Glu Thr Ile Ser Leu Ser Met Thr Asn Asp Ser Lys Glu Phe Val 85 90 95Asp Pro Tyr Ile Val Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile 100 105 110Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn Val Glu Thr Glu Lys Leu 130 135 140Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Ile Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly 180 185 190Ser Ser Ala Ser His Val Asp Ala Gly Asn Gln Ser Thr His Tyr Thr 195 200 205Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 210 215 220Gly Val Lys Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Ser Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val465 470 475 480Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn 485 490 495Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp 500 505 510Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr 515 520 525Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile 530 535 540Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser545 550 555 560Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly 565 570 575Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 107 592 PRT Neisseria meningitidis 107Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Ser Val Gln 50 55 60Arg Ser Val Val Gly Ser Ile Gln Ala Ser Met Glu Gly Ser Gly Glu 65 70 75 80Leu Glu Thr Ile Ser Leu Ser Met Thr Asn Asp Ser Lys Glu Phe Val 85 90 95Asp Pro Tyr Ile Val Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile 100 105 110Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn Val Glu Thr Glu Lys Leu 130 135 140Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Ile Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly 180 185 190Ser Ser Ala Ser His Val Asp Ala Gly Asn Gln Ser Thr His Tyr Thr 195 200 205Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 210 215 220Gly Val Lys Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Ser Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Ala Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Asp Glu Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val465 470 475 480Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn 485 490 495Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp 500 505 510Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr 515 520 525Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile 530 535 540Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser545 550 555 560Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly 565 570 575Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 108 589 PRT Neisseria meningitidis 108Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Ser Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Thr Asp Glu Asp Glu Glu Leu Glu Ser Val Val 50 55 60Arg Ser Ala Leu Val Leu Gln Phe Met Ile Asp Lys Glu Gly Asn Gly 65 70 75 80Glu Ile Glu Ser Thr Gly Asp Ile Gly Trp Ser Ile Tyr Tyr Asp Asp 85 90 95His Asn Thr Leu His Gly Ala Thr Val Thr Leu Lys Ala Gly Asp Asn 100 105 110Leu Lys Ile Lys Gln Ser Gly Lys Asp Phe Thr Tyr Ser Leu Lys Lys 115 120 125Glu Leu Lys Asp Leu Thr Ser Val Glu Thr Glu Lys Leu Ser Phe Gly 130 135 140Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys Gly Leu Asn145 150 155 160Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Pro Thr Val His Leu 165 170 175Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly Ser Ser Ala 180 185 190Ser His Val Asp Ala Gly Asn Gln Ser Thr His Tyr Thr Arg Ala Ala 195 200 205Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys Gly Val Lys 210 215 220Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe Val Arg Thr225 230 235 240Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr Thr Thr Val 245 250 255Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val Lys Ile Gly 260 265 270Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu Val Thr Gly 275 280 285Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly Glu Gly Leu 290 295 300Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala Gly Trp Arg305 310 315 320Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala Asp Lys Phe 325 330 335Glu Thr Val Thr Ser Gly Thr Lys Val Thr Phe Ala Ser Gly Asn Gly 340 345 350Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile Thr Val Lys 355 360 365Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln Leu Gln Asn 370 375 380Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser Ser Gly Lys385 390 395 400Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met Asp Glu Thr 405 410 415Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg Asn Gly Lys 420 425 430Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser Ser Val Ser 435 440 445Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp Asp Glu Gly 450 455 460Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val Arg Ile Thr465 470 475 480Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val Ala Gln 485 490 495Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn Val Asp 500 505 510Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala Gly Leu 515 520 525Ala Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly Gly Gly 530 535 540Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser Ile Ser545 550 555 560Asp Thr Gly Asn Trp Val Ile Lys Gly Thr Ala Ser Gly Asn Ser Arg 565 570 575Gly His Phe Gly Thr Ser Ala Ser Val Gly Tyr Gln Trp 580 585 109 589 PRT Neisseria meningitidis 109Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Ser Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Thr Asp Glu Asp Glu Glu Leu Glu Ser Val Val 50 55 60Arg Ser Ala Leu Val Leu Gln Phe Met Ile Asp Lys Glu Gly Asn Gly 65 70 75 80Glu Ile Glu Ser Thr Gly Asp Ile Gly Trp Ser Ile Tyr Tyr Asp Asp 85 90 95His Asn Thr Leu His Gly Ala Thr Val Thr Leu Lys Ala Gly Asp Asn 100 105 110Leu Lys Ile Lys Gln Ser Gly Lys Asp Phe Thr Tyr Ser Leu Lys Lys 115 120 125Glu Leu Lys Asp Leu Thr Ser Val Glu Thr Glu Lys Leu Ser Phe Gly 130 135 140Ala Asn Gly Asn Lys Val Asn Ile Thr Ser Asp Thr Lys Gly Leu Asn145 150 155 160Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Pro Thr Val His Leu 165 170 175Asn Gly Ile Gly Ser Thr Leu Thr Asp Thr Leu Ala Gly Ser Ser Ala 180 185 190Ser His Val Asp Ala Gly Asn Gln Ser Thr His Tyr Thr Arg Ala Ala 195 200 205Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys Gly Val Lys 210 215 220Thr Gly Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe Val Arg Thr225 230 235 240Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr Thr Thr Val 245 250 255Asn Val Glu Ser Lys Asp Asn Gly Lys Arg Thr Glu Val Lys Ile Gly 260 265 270Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu Val Thr Gly 275 280 285Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly Glu Gly Leu 290 295 300Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala Gly Trp Arg305 310 315 320Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala Asp Lys Phe 325 330 335Glu Thr Val Thr Ser Gly Thr Lys Val Thr Phe Ala Ser Gly Asn Gly 340 345 350Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile Thr Val Lys 355 360 365Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln Leu Gln Asn 370 375 380Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser Ser Gly Lys385 390 395 400Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met Asp Glu Thr 405 410 415Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg Asn Gly Lys 420 425 430Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser Ser Val Ser 435 440 445Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp Asp Glu Gly 450 455 460Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val Arg Ile Thr465 470 475 480Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn Val Ala Gln 485 490 495Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp Asn Val Asp 500 505 510Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr Ala Gly Leu 515 520 525Ala Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile Gly Gly Gly 530 535 540Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser Ser Ile Ser545 550 555 560Asp Thr Gly Asn Trp Val Ile Lys Gly Thr Ala Ser Gly Asn Ser Arg 565 570 575Gly His Phe Gly Thr Ser Ala Ser Val Gly Tyr Gln Trp 580 585 110 592 PRT Neisseria meningitidis 110Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ser Ala Leu Asn Ala Trp 1 5 10 15Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu Glu Leu Glu Ser Val Gln 50 55 60Arg Ser Val Val Gly Ser Ile Gln Ala Ser Met Glu Gly Ser Gly Glu 65 70 75 80Leu Glu Thr Ile Ser Leu Ser Met Thr Asn Asp Ser Lys Glu Phe Val 85 90 95Asp Pro Tyr Ile Val Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile 100 105 110Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser 115 120 125Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn Val Glu Thr Glu Lys Leu 130 135 140Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile Ile Ser Asp Thr Lys145 150 155 160Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly Thr Asn Gly Asp Thr Thr 165 170 175Val His Leu Asn Gly Ile Gly Ser Thr Leu Thr Asp Met Leu Leu Asn 180 185 190Thr Gly Ala Thr Thr Asn Val Thr Asn Asp Asn Val Thr Asp Asp Glu 195 200 205Lys Lys Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn 210 215 220Ile Lys Gly Val Lys Pro Gly Thr Thr Ala Ser Asp Asn Val Asp Phe225 230 235 240Val Arg Thr Tyr Asp Thr Val Glu Phe Leu Ser Ala Asp Thr Lys Thr 245 250 255Thr Thr Val Asn Val Glu Ser Lys Asp Asn Gly Lys Lys Thr Glu Val 260 265 270Lys Ile Gly Ala Lys Thr Ser Val Ile Lys Glu Lys Asp Gly Lys Leu 275 280 285Val Thr Gly Lys Gly Lys Gly Glu Asn Gly Ser Ser Thr Asp Glu Gly 290 295 300Glu Gly Leu Val Thr Ala Lys Glu Val Ile Asp Ala Val Asn Lys Ala305 310 315 320Gly Trp Arg Met Lys Thr Thr Thr Ala Asn Gly Gln Thr Gly Gln Ala 325 330 335Asp Lys Phe Glu Thr Val Thr Ser Gly Thr Asn Val Thr Phe Ala Ser 340 345 350Gly Lys Gly Thr Thr Ala Thr Val Ser Lys Asp Asp Gln Gly Asn Ile 355 360 365Thr Val Met Tyr Asp Val Asn Val Gly Asp Ala Leu Asn Val Asn Gln 370 375 380Leu Gln Asn Ser Gly Trp Asn Leu Asp Ser Lys Ala Val Ala Gly Ser385 390 395 400Ser Gly Lys Val Ile Ser Gly Asn Val Ser Pro Ser Lys Gly Lys Met 405 410 415Asp Glu Thr Val Asn Ile Asn Ala Gly Asn Asn Ile Glu Ile Thr Arg 420 425 430Asn Gly Lys Asn Ile Asp Ile Ala Thr Ser Met Thr Pro Gln Phe Ser 435 440 445Ser Val Ser Leu Gly Ala Gly Ala Asp Ala Pro Thr Leu Ser Val Asp 450 455 460Asp Lys Gly Ala Leu Asn Val Gly Ser Lys Asp Ala Asn Lys Pro Val465 470 475 480Arg Ile Thr Asn Val Ala Pro Gly Val Lys Glu Gly Asp Val Thr Asn 485 490 495Val Ala Gln Leu Lys Gly Val Ala Gln Asn Leu Asn Asn Arg Ile Asp 500 505 510Asn Val Asp Gly Asn Ala Arg Ala Gly Ile Ala Gln Ala Ile Ala Thr 515 520 525Ala Gly Leu Val Gln Ala Tyr Leu Pro Gly Lys Ser Met Met Ala Ile 530 535 540Gly Gly Gly Thr Tyr Arg Gly Glu Ala Gly Tyr Ala Ile Gly Tyr Ser545 550 555 560Ser Ile Ser Asp Gly Gly Asn Trp Ile Ile Lys Gly Thr Ala Ser Gly 565 570 575Asn Ser Arg Gly His Phe Gly Ala Ser Ala Ser Val Gly Tyr Gln Trp 580 585 590 111 600 PRT Neisseria meningitidis 111Met Asn Lys Ile Tyr Arg Ile Ile Trp Asn Ile Ala Leu Asn Ala Trp 1 5 10 15Val Val Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 20 25 30Thr Val Ala Thr Ala Val Leu Ala Thr Leu Leu Ser Ala Thr Val Gln 35 40 45Ala Asn Ala Thr Asp Glu Glu Asp Asn Glu Asp Leu Glu Pro Val Val 50 55 60Arg Thr Ala Pro Val Leu Ser Phe His Ser Asp Lys Glu Gly Thr Gly 65 70 75 80Glu Lys Glu Glu Val Gly Ala Ser Ser Asn Leu Thr Val Tyr Phe Asp 85 90 95Lys Asn Arg Val Leu Lys Ala Gly Thr Ile Thr Leu Lys Ala Gly Asp 100 105 110Asn Leu Lys Ile Lys Gln Asn Thr Asn Glu Asn Thr Asn Glu Asn Thr 115 120 125Asn Ala Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu Thr Gly Leu 130 135 140Ile Asn Val Glu Thr Glu Lys Leu Ser Phe Gly Ala Asn Gly Lys Lys145 150 155 160Val Asn Ile Ile Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr 165 170 175Ala Gly Thr Asn Gly Asp Pro Thr Val His Leu Asn Gly Ile Gly Ser 180 185 190Thr Leu Thr Asp Thr Leu Ala Gly Ser Ser Ala Ser His Val Asp Ala 195 200 205Gly Asn Gln Ser Thr His Tyr Thr Arg Ala Ala Ser Ile Lys Asp Val 210 215 220Leu Asn Ala Gly Trp Asn Ile Lys Gly Val Lys Thr Gly Ser Thr Thr225 230 235 240Gly Gln Ser Glu Asn Val Asp Phe Val Arg Thr Tyr Asp Thr Val Glu 245 250 255Phe Leu Ser Ala Asp Thr Lys Thr Thr Thr Val Asn Val Glu Ser Lys 260 265 270Asp Asn Gly Lys Arg Thr Glu Val Lys Ile Gly Ala Lys Thr Ser Val 275 280 285Ile Lys Glu Lys Asp Gly Lys Leu Val Thr Gly Lys Gly Lys Gly Glu 290 295 300Asn Gly Ser Ser Thr Asp Glu Gly Glu Gly Leu Val Thr Ala Lys Glu305 310 315 320Val Ile Asp Ala Val Asn Lys Ala Gly Trp Arg Met Lys Thr Thr Thr 325 330 335Ala Asn Gly Gln Thr Gly Gln Ala Asp Lys Phe Glu Thr Val Thr Ser 340 345 350Gly Thr Lys Val Thr Phe Ala Ser Gly Asn Gly Thr Thr Ala Thr Val 355 360 365Ser Lys Asp Asp Gln Gly Asn Ile Thr Val Lys Tyr Asp Val Asn Val 370 375 380Gly Asp Ala Leu Asn Val Asn Gln Leu Gln Asn Ser Gly Trp Asn Leu385 390 395 400Asp Ser Lys Ala Val Ala Gly Ser Ser Gly Lys Val Ile Ser Gly Asn 405 410 415Val Ser Pro Ser Lys Gly Lys Met Asp Glu Thr Val Asn Ile Asn Ala 420 425 430Gly Asn Asn Ile Glu Ile Thr Arg Asn Gly Lys Asn Ile Asp Ile Ala 435 440 445Thr Ser Met Thr Pro Gln Phe Ser Ser Val Ser Leu Gly Ala Gly Ala 450 455 460Asp Ala Pro Thr Leu Ser Val Asp Asp Glu Gly Ala Leu Asn Val Gly465 470 475 480Ser Lys Asp Ala Asn Lys Pro Val Arg Ile Thr Asn Val Ala Pro Gly 485 490 495Val Lys Glu Gly Asp Val Thr Asn Val Ala Gln Leu Lys Gly Val Ala 500 505 510Gln Asn Leu Asn Asn Arg Ile Asp Asn Val Asp Gly Asn Ala Arg Ala 515 520 525Gly Ile Ala Gln Ala Ile Ala Thr Ala Gly Leu Val Gln Ala Tyr Leu 530 535 540Pro Gly Lys Ser Met Met Ala Ile Gly Gly Gly Thr Tyr Arg Gly Glu545 550 555 560Ala Gly Tyr Ala Ile Gly Tyr Ser Ser Ile Ser Asp Gly Gly Asn Trp 565 570 575Ile Ile Lys Gly Thr Ala Ser Gly Asn Ser Arg Gly His Phe Gly Ala 580 585 590Ser Ala Ser Val Gly Tyr Gln Trp 595 600 112 32 DNA Neisseria meningitidis 112cgcggatccc atatgtcgcc gcaaaattcc ga 32 113 28 DNA Neisseria meningitidis 113cccgctcgag ttttgccgcg ttaaaagc 28 114 30 DNA Neisseria meningitidis 114cgcggatccc atatgaccgt gaagaccgcc 30 115 28 DNA Neisseria meningitidis 115cccgctcgag ccactgataa ccgacaga 28 116 34 DNA Neisseria meningitidis 116cgcggatccc atatgtattt gaaacagctc caag 34 117 26 DNA Neisseria meningitidis 117cccgctcgag ttctgggtga atgtta 26 118 29 DNA Neisseria meningitidis 118gcggatccca tatgggcacg gacaacccc 29 119 26 DNA Neisseria meningitidis 119cccgctcgag acgtggggaa cagtct 26 120 34 DNA Neisseria meningitidis 120gcggatccca tatgaaaaat attcaagtag ttgc 34 121 27 DNA Neisseria meningitidis 121cccgctcgag aagtttgatt aaacccg 27 122 31 DNA Neisseria meningitidis 122cgcggatccc atatgtgcca accgcaatcc g 31 123 27 DNA Neisseria meningitidis 123cccgctcgag tttttccagc tccggca 27 124 33 DNA Neisseria meningitidis 124gcggatccca tatggttatc ggaatattac tcg 33 125 25 DNA Neisseria meningitidis 125cccgctcgag ggctgcagaa gctgg 25 126 32 DNA Neisseria meningitidis 126cgcggatccc atatgcggac gtggttggtt tt 32 127 30 DNA Neisseria meningitidis 127cccgctcgag atatcttccg tttttttcac 30 128 36 DNA Neisseria meningitidis 128cgcggatccg ctagcgtaaa tttattattt ttagaa 36 129 28 DNA Neisseria meningitidis 129cccgctcgag ttccaactca ttgaagta 28 130 35 DNA Neisseria meningitidis 130cgcggatccc atatgaataa aggtttacat cgcat 35 131 26 DNA Neisseria meningitidis 131cccgctcgag aatcgctgca ccggct 26 132 30 DNA Neisseria meningitidis 132cgcggatccc atatgactgc cttttcgaca 30 133 26 DNA Neisseria meningitidis 133cccgctcgag gcgtgaagcg tcagga 26 134 15 DNA Artificial Sequence Description of Artificial Sequence BamHI - NdeI 134cgcggatccc atatg 15 135 15 DNA Artificial Sequence Description of Artificial Sequence BamHI - NheI 135cgcggatccg ctagc 15 136 17 DNA Artificial Sequence Description of Artificial Sequence EcoRI - NheI 136ccggaattct agctagc 17 137 10 DNA Artificial Sequence Description of Artificial Sequence XhoI 137cccgctcgag 10 138 291 PRT Artificial Sequence Description of Artificial Sequence ORF40a 138Ser Ala Leu Asn Ala Xaa Val Ala Val Ser Glu Leu Thr Arg Asn His 1 5 10 15Thr Lys Arg Ala Ser Ala Thr Val Lys Thr Ala Val Leu Ala Thr Leu 20 25 30Leu Phe Ala Thr Val Gln Ala Asn Ala Thr Asp Glu Asp Glu Glu Glu 35 40 45Glu Leu Glu Ser Val Gln Arg Ser Val Val Gly Ser Ile Gln Ala Ser 50 55 60Met Glu Gly Ser Gly Glu Leu Glu Thr Ile Ser Leu Ser Met Thr Asn 65 70 75 80Asp Ser Lys Glu Phe Val Asp Pro Tyr Ile Val Val Thr Leu Lys Ala 85 90 95Gly Asp Asn Leu Lys Ile Lys Gln Asn Thr Asn Glu Asn Thr Asn Ala 100 105 110Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu Thr Gly Leu Ile Asn 115 120 125Val Xaa Thr Glu Lys Leu Ser Phe Gly Ala Asn Gly Lys Lys Val Asn 130 135 140Ile Ile Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr Ala Gly145 150 155 160Thr Asn Gly Asp Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu 165 170 175Thr Asp Thr Leu Ala Gly Ser Ser Ala Ser His Val Asp Ala Gly Asn 180 185 190Xaa Ser Thr His Tyr Thr Arg Ala Ala Ser Ile Lys Asp Val Leu Asn 195 200 205Ala Gly Trp Asn Ile Lys Gly Val Lys Xaa Gly Ser Thr Thr Gly Gln 210 215 220Ser Glu Asn Val Asp Phe Val Arg Thr Tyr Asp Thr Val Glu Phe Leu225 230 235 240Ser Ala Asp Thr Xaa Thr Thr Thr Val Asn Val Glu Ser Lys Asp Asn 245 250 255Gly Lys Arg Thr Glu Val Lys Ile Gly Ala Lys Thr Ser Val Ile Lys 260 265 270Glu Lys Asp Gly Lys Leu Val Thr Gly Lys Gly Lys Gly Glu Asn Gly 275 280 285Ser Ser Thr 290 139 240 PRT Artificial Sequence Description of Artificial Sequence Hsf 139Thr Leu Leu Phe Ala Thr Val Gln Ala Asn Ala Thr Asp Glu Asp Glu 1 5 10 15Glu Leu Asp Pro Val Val Arg Thr Ala Pro Val Leu Ser Phe His Ser 20 25 30Asp Lys Glu Gly Thr Gly Glu Lys Glu Val Thr Glu Asn Ser Asn Trp 35 40 45Gly Ile Tyr Phe Asp Asn Lys Gly Val Leu Lys Ala Gly Ala Ile Thr 50 55 60Leu Lys Ala Gly Asp Asn Leu Lys Ile Lys Gln Asn Thr Asp Glu Ser 65 70 75 80Thr Asn Ala Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu Thr Asp 85 90 95Leu Thr Ser Val Ala Thr Glu Lys Leu Ser Phe Gly Ala Asn Gly Asp 100 105 110Lys Val Asp Ile Thr Ser Asp Ala Asn Gly Leu Lys Leu Ala Lys Thr 115 120 125Gly Asn Gly Asn Val His Leu Asn Gly Leu Asp Ser Thr Leu Pro Asp 130 135 140Ala Val Thr Asn Thr Gly Val Leu Ser Ser Ser Ser Phe Thr Pro Asn145 150 155 160Asp Val Glu Lys Thr Arg Ala Ala Thr Val Lys Asp Val Leu Asn Ala 165 170 175Gly Trp Asn Ile Lys Gly Ala Lys Thr Ala Gly Gly Asn Val Glu Ser 180 185 190Val Asp Leu Val Ser Ala Tyr Asn Asn Val Glu Phe Ile Thr Gly Asp 195 200 205Lys Asn Thr Leu Asp Val Val Leu Thr Ala Lys Glu Asn Gly Lys Thr 210 215 220Thr Glu Val Lys Phe Thr Pro Lys Thr Ser Val Ile Lys Glu Lys Asp225 230 235 240 140 251 PRT Artificial Sequence Description of Artificial Sequence overlap identity 140Thr Leu Leu Phe Ala Thr Val Gln Ala Xaa Ala Xaa Xaa Glu Xaa Xaa 1 5 10 15Glu Xaa Xaa Xaa Xaa Leu Asp Pro Val Xaa Arg Thr Xaa Xaa Val Leu 20 25 30Xaa Xaa Xaa Ser Asp Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45Xaa Asn Ser Xaa Trp Xaa Xaa Tyr Phe Xaa Xaa Lys Gly Val Leu Xaa 50 55 60Ala Xaa Xaa Ile Thr Xaa Lys Ala Gly Asp Asn Leu Lys Ile Lys Gln 65 70 75 80Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Thr Tyr Ser Leu Lys 85 90 95Lys Asp Leu Thr Asp Leu Thr Ser Val Xaa Thr Glu Lys Leu Ser Phe 100 105 110Xaa Ala Asn Gly Xaa Lys Val Xaa Ile Thr Ser Asp Xaa Xaa Gly Leu 115 120 125Xaa Xaa Ala Lys Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Val His 130 135 140Leu Asn Gly Xaa Xaa Ser Thr Leu Xaa Asp Xaa Xaa Xaa Asn Thr Gly145 150 155 160Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Glu Lys Xaa 165 170 175Arg Ala Ala Xaa Val Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 180 185 190Gly Xaa Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Val Asp Xaa Val Xaa 195 200 205Xaa Tyr Xaa Xaa Val Glu Phe Xaa Xaa Xaa Asp Xaa Xaa Thr Xaa Xaa 210 215 220Val Xaa Xaa Xaa Xaa Lys Xaa Asn Gly Lys Xaa Thr Glu Val Lys Xaa225 230 235 240Xaa Xaa Lys Thr Ser Val Ile Lys Glu Lys Asp 245 250 141 36 PRT Artificial Sequence Description of Artificial Sequence ORF40a 141Val Ala Val Ser Glu Leu Thr Arg Asn His Thr Lys Arg Ala Ser Ala 1 5 10 15Thr Val Lys Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 20 25 30Ala Asn Ala Thr 35 142 36 PRT Artificial Sequence Description of Artificial Sequence homology 142Val Xaa Val Ser Glu Leu Thr Arg Xaa His Thr Lys Arg Ala Ser Ala 1 5 10 15Thr Val Xaa Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 20 25 30Ala Asn Ala Thr 35 143 36 PRT Artificial Sequence Description of Artificial Sequence Hsf 143Val Val Val Ser Glu Leu Thr Arg Thr His Thr Lys Arg Ala Ser Ala 1 5 10 15Thr Val Glu Thr Ala Val Leu Ala Thr Leu Leu Phe Ala Thr Val Gln 20 25 30Ala Asn Ala Thr 35 144 38 PRT Artificial Sequence Description of Artificial Sequence Orf40a 144Val Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile Lys Gln Asn Thr Asn 1 5 10 15Glu Asn Thr Asn Ala Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu 20 25 30Thr Gly Leu Ile Asn Val 35 145 38 PRT Artificial Sequence Description of Artificial Sequence homology 145Xaa Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile Lys Gln Asn Thr Xaa 1 5 10 15Glu Xaa Thr Asn Ala Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu 20 25 30Thr Xaa Leu Xaa Xaa Val 35 146 38 PRT Artificial Sequence Description of Artificial Sequence Hsf 146Ile Thr Leu Lys Ala Gly Asp Asn Leu Lys Ile Lys Gln Asn Thr Asp 1 5 10 15Glu Ser Thr Asn Ala Ser Ser Phe Thr Tyr Ser Leu Lys Lys Asp Leu 20 25 30Thr Asp Leu Thr Ser Val 35 147 29 PRT Artificial Sequence Description of Artificial Sequence Orf40a 147Val Thr Glu Lys Leu Ser Phe Gly Ala Asn Gly Lys Lys Val Asn Ile 1 5 10 15Ile Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Glu Thr 20 25 148 29 PRT Artificial Sequence Description of Artificial Sequence homology 148Val Xaa Xaa Lys Leu Ser Xaa Gly Xaa Asn Gly Xaa Lys Val Asn Ile 1 5 10 15Xaa Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Xaa Xaa 20 25 149 29 PRT Artificial Sequence Description of Artificial Sequence Hsf 149Val Ser Asp Lys Leu Ser Leu Gly Thr Asn Gly Asn Lys Val Asn Ile 1 5 10 15Thr Ser Asp Thr Lys Gly Leu Asn Phe Ala Lys Asp Ser 20 25 150 32 PRT Artificial Sequence Description of Artificial Sequence ORF40a 150Thr Asn Gly Asp Thr Thr Val His Leu Asn Gly Ile Gly Ser Thr Leu 1 5 10 15Thr Asp Thr Leu Ala Gly Ser Ser Ala Ser His Val Asp Ala Gly Asn 20 25 30 151 32 PRT Artificial Sequence Description of Artificial Sequence homology 151Thr Xaa Xaa Asp Xaa Xaa Xaa His Leu Asn Gly Ile Xaa Ser Thr Leu 1 5 10 15Thr Asp Thr Leu Xaa Xaa Ser Xaa Ala Xaa Xaa Xaa Xaa Xaa Gly Asn 20 25 30 152 32 PRT Artificial Sequence Description of Artificial Sequence Hsf 152Thr Gly Asp Asp Ala Asn Ile His Leu Asn Gly Ile Ala Ser Thr Leu 1 5 10 15Thr Asp Thr Leu Leu Asn Ser Gly Ala Thr Thr Asn Leu Gly Gly Asn 20 25 30 153 19 PRT Artificial Sequence Description of Artificial Sequence ORF40a 153Arg Ala Ala Ser Ile Lys Asp Val Leu Asn Ala Gly Trp Asn Ile Lys 1 5 10 15Gly Val Lys 154 19 PRT Artificial Sequence Description of Artificial Sequence homology 154Arg Ala Ala Ser Xaa Lys Asp Val Leu Asn Ala Gly Trp Asn Xaa Xaa 1 5 10 15Gly Val Lys 155 19 PRT Artificial Sequence Description of Artificial Sequence Hsf 155Arg Ala Ala Ser Val Lys Asp Val Leu Asn Ala Gly Trp Asn Val Arg 1 5 10 15Gly Val Lys 156 28 PRT Artificial Sequence Description of Artificial Sequence ORF40a 156Ser Thr Thr Gly Gln Ser Glu Asn Val Asp Phe Val Arg Thr Tyr Asp 1 5 10 15Thr Val Glu Phe Leu Ser Ala Asp Thr Thr Thr Thr 20 25 157 28 PRT Artificial Sequence Description of Artificial Sequence homology 157Ser Xaa Xaa Xaa Gln Xaa Glu Asn Xaa Asp Phe Val Xaa Thr Tyr Asp 1 5 10 15Thr Val Xaa Phe Xaa Ser Xaa Asp Xaa Xaa Thr Thr 20 25 158 28 PRT Artificial Sequence Description of Artificial Sequence Hsf 158Ser Ala Asn Asn Gln Val Glu Asn Ile Asp Phe Val Ala Thr Tyr Asp 1 5 10 15Thr Val Asp Phe Val Ser Gly Asp Lys Asp Thr Thr 20 25 159 240 PRT Artificial Sequence Description of Artificial Sequence ORF38a 159Met Leu Arg Leu Thr Ala Leu Ala Val Cys Thr Ala Leu Ala Leu Gly 1 5 10 15Ala Cys Ser Pro Gln Asn Ser Asp Ser Ala Pro Gln Ala Lys Glu Gln 20 25 30Ala Val Ser Ala Ala Gln Ser Glu Gly Val Ser Val Thr Val Lys Thr 35 40 45Ala Arg Gly Asp Val Gln Ile Pro Gln Asn Pro Glu Arg Ile Ala Val 50 55 60Tyr Asp Leu Gly Met Leu Asp Thr Leu Ser Lys Leu Gly Val Lys Thr 65 70 75 80Gly Leu Ser Val Asp Lys Asn Arg Leu Pro Tyr Leu Glu Glu Tyr Phe 85 90 95Lys Thr Thr Lys Pro Ala Gly Thr Leu Phe Glu Pro Asp Tyr Glu Thr 100 105 110Leu Asn Ala Tyr Lys Pro Gln Leu Ile Ile Ile Gly Ser Arg Ala Ala 115 120 125Lys Ala Phe Asp Lys Leu Asn Glu Ile Ala Pro Thr Ile Glu Met Thr 130 135 140Ala Asp Thr Ala Asn Leu Lys Glu Ser Ala Lys Glu Arg Ile Asp Ala145 150 155 160Leu Ala Gln Ile Phe Gly Lys Lys Ala Glu Ala Asp Lys Leu Lys Ala 165 170 175Glu Ile Asp Ala Ser Phe Glu Ala Ala Lys Thr Ala Ala Gln Gly Lys 180 185 190Gly Lys Gly Leu Val Ile Leu Val Asn Gly Gly Lys Met Ser Ala Phe 195 200 205Gly Pro Ser Ser Arg Leu Gly Gly Trp Leu His Lys Asp Ile Gly Val 210 215 220Pro Ala Val Asp Glu Ala Ile Lys Glu Gly Ser His Gly Gln Pro Ile225 230 235 240 160 94 PRT Artificial Sequence Description of Artificial Sequence ORF38 160Glu Gly Ala Ser Val Thr Val Lys Thr Ala Arg Gly Asp Val Gln Ile 1 5 10 15Pro Gln Asn Pro Glu Arg Ile Ala Val Tyr Asp Leu Gly Met Leu Asp 20 25 30Thr Leu Ser Lys Leu Gly Val Lys Thr Gly Leu Ser Val Asp Lys Asn 35 40 45Arg Leu Pro Tyr Leu Glu Glu Tyr Phe Lys Thr Thr Lys Pro Ala Gly 50 55 60Thr Leu Phe Glu Pro Asp Tyr Glu Thr Leu Asn Ala Tyr Lys Pro Gln 65 70 75 80Leu Ile Ile Ile Gly Ser Arg Ala Ala Lys Ala Phe Asp Lys 85 90 161 96 PRT Artificial Sequence Description of Artificial Sequence overlap identity 161Glu Gly Xaa Ser Xaa Xaa Val Lys Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa 1 5 10 15Pro Xaa Asn Pro Xaa Xaa Xaa Xaa Xaa Xaa Asp Leu Gly Xaa Leu Asp 20 25 30Thr Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Val Xaa Xaa 35 40 45Xaa Xaa Leu Pro Xaa Xaa Xaa Xaa Xaa Phe Lys Xaa Xaa Xaa Xaa Xaa 50 55 60Gly Xaa Xaa Xaa Xaa Xaa Asp Xaa Glu Xaa Xaa Asn Ala Xaa Lys Pro 65 70 75 80Xaa Leu Ile Ile Ile Xaa Xaa Arg Xaa Xaa Lys Xaa Xaa Asp Lys Leu 85 90 95 162 96 PRT Artificial Sequence Description of Artificial Sequence Lipo 162Glu Gly Asp Ser Phe Leu Val Lys Asp Ser Leu Gly Glu Asn Lys Thr 1 5 10 15Pro Lys Asn Pro Ser Lys Val Val Ile Leu Asp Leu Gly Ile Leu Asp 20 25 30Thr Phe Asp Ala Leu Lys Leu Asn Asp Lys Val Ala Gly Val Pro Ala 35 40 45Lys Asn Leu Pro Lys Tyr Leu Gln Gln Phe Lys Asn Lys Pro Ser Val 50 55 60Gly Gly Val Gln Gln Val Asp Phe Glu Ala Ile Asn Ala Leu Lys Pro 65 70 75 80Asp Leu Ile Ile Ile Ser Gly Arg Gln Ser Lys Phe Tyr Asp Lys Leu 85 90 95 163 91 PRT Artificial Sequence Description of Artificial Sequence ORF44 163Thr Val Ser Tyr Val Cys Gln Gln Gly Lys Lys Val Lys Val Thr Tyr 1 5 10 15Gly Phe Asn Lys Gln Gly Leu Thr Thr Tyr Ala Ser Ala Val Ile Asn 20 25 30Gly Lys Arg Val Gln Met Pro Val Asn Leu Asp Lys Ser Asp Asn Val 35 40 45Glu Thr Phe Tyr Gly Lys Glu Gly Gly Tyr Val Leu Gly Thr Gly Val 50 55 60Met Asp Gly Lys Ser Tyr Arg Lys Gln Pro Ile Met Ile Thr Ala Pro 65 70 75 80Asp Asn Gln Ile Val Phe Lys Asp Cys Ser Pro 85 90 164 91 PRT Artificial Sequence Description of Artificial Sequence overlap identity 164Xaa Val Xaa Tyr Val Cys Gln Gln Gly Xaa Xaa Xaa Xaa Val Xaa Tyr 1 5 10 15Xaa Phe Asn Xaa Xaa Gly Xaa Xaa Thr Xaa Ala Xaa Xaa Xaa Xaa Asn 20 25 30Xaa Xaa Xaa Xaa Xaa Xaa Pro Xaa Asn Leu Xaa Xaa Ser Asp Asn Val 35 40 45Xaa Thr Xaa Xaa Xaa Xaa Xaa Xaa Gly Tyr Xaa Leu Xaa Thr Xaa Xaa 50 55 60Met Asp Xaa Xaa Xaa Tyr Arg Xaa Gln Xaa Ile Xaa Xaa Xaa Ala Pro 65 70 75 80Xaa Xaa Gln Xaa Xaa Xaa Lys Asp Cys Ser Pro 85 90 165 90 PRT Artificial Sequence Description of Artificial Sequence LecA 165Ser Val Ala Tyr Val Cys Gln Gln Gly Arg Arg Leu Asn Val Asn Tyr 1 5 10 15Arg Phe Asn Ser Ala Gly Val Pro Thr Ser Ala Glu Leu Arg Val Asn 20 25 30Asn Arg Asn Leu Arg Leu Pro Tyr Asn Leu Ser Ala Ser Asp Asn Val 35 40 45Asp Thr Val Phe Ser Ala Asn Gly Tyr Arg Leu Thr Thr Asn Ala Met 50 55 60Asp Ser Ala Asn Tyr Arg Ser Gln Asp Ile Ile Val Ser Ala Pro Asn 65 70 75 80Gly Gln Met Leu Tyr Lys Asp Cys Ser Pro 85 90 166 240 PRT Artificial Sequence Description of Artificial Sequence ORF49a 166Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val Arg Val Val Ala 1 5 10 15Gln Xaa Ala Ala Thr Arg Ser Gly Trp Asp Thr Val Leu Glu Gly Thr 20 25 30Glu Phe Lys Thr Thr Leu Ala Gly Ala Asp Ile Gln Ala Gly Val Xaa 35 40 45Glu Lys Ala Arg Val Asp Ala Lys Ile Ile Leu Lys Gly Ile Val Asn 50 55 60Arg Ile Gln Ser Glu Glu Lys Leu Glu Thr Asn Ser Thr Val Trp Gln 65 70 75 80Lys Gln Ala Gly Arg Gly Ser Thr Ile Glu Thr Leu Lys Leu Pro Ser 85 90 95Phe Glu Ser Pro Thr Pro Pro Lys Leu Ser Ala Pro Gly Gly Tyr Ile 100 105 110Val Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile Glu Lys Leu Ser 115 120 125Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln Val Ala Lys Asn 130 135 140Ile Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Arg Trp Asp Tyr Lys145 150 155 160Gln Glu Gly Leu Thr Glu Ala Gly Ala Ala Ile Ile Ala Leu Ala Val 165 170 175Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val Leu Gly Leu Asn 180 185 190Gly Ala Xaa Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser 195 200 205Gln Ala Ser Val Ser Phe Ile Asn Asn Lys Gly Asp Val Gly Lys Thr 210 215 220Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Val Val Ala225 230 235 240 167 540 PRT Artificial Sequence Description of Artificial Sequence ORF49a 167Xaa Gln Leu Leu Ala Glu Glu Gly Ile His Lys His Glu Leu Asp Val 1 5 10 15Gln Lys Ser Arg Arg Phe Ile Gly Ile Lys Val Gly Xaa Ser Asn Tyr 20 25 30Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val Arg Val Val Ala 35 40 45Gln Xaa Ala Ala Thr Arg Ser Gly Trp Asp Thr Val Leu Glu Gly Thr 50 55 60Glu Phe Lys Thr Thr Leu Ala Gly Ala Asp Ile Gln Ala Gly Val Xaa 65 70 75 80Glu Lys Ala Arg Val Asp Ala Lys Ile Ile Leu Lys Gly Ile Val Asn 85 90 95Arg Ile Gln Ser Glu Glu Lys Leu Glu Thr Asn Ser Thr Val Trp Gln 100 105 110Lys Gln Ala Gly Arg Gly Ser Thr Ile Glu Thr Leu Lys Leu Pro Ser 115 120 125Phe Glu Ser Pro Thr Pro Pro Lys Leu Ser Ala Pro Gly Gly Tyr Ile 130 135 140Val Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile Glu Lys Leu Ser145 150 155 160Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln Val Ala Lys Asn 165 170 175Ile Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Arg Trp Asp Tyr Lys 180 185 190Gln Glu Gly Leu Thr Glu Ala Gly Ala Ala Ile Ile Ala Leu Ala Val 195 200 205Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val Leu Gly Leu Asn 210 215 220Gly Ala Xaa Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser225 230 235 240Gln Ala Ser Val Ser Phe Ile Asn Asn Lys Gly Asp Val Gly Lys Thr 245 250 255Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Val Val Ala 260 265 270Ala Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala Ser Ala Leu Xaa 275 280 285Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr Val Asn Leu Ala 290 295 300Asn Ala Gly Ser Ala Ala Leu Ile Asn Thr Ala Val Asn Gly Gly Ser305 310 315 320Leu Lys Asp Xaa Leu Glu Ala Asn Ile Leu Ala Ala Leu Val Asn Thr 325 330 335Ala His Gly Glu Ala Ala Ser Lys Ile Lys Gln Leu Asp Gln His Tyr 340 345 350Ile Val His Lys Ile Ala His Ala Ile Ala Gly Cys Ala Ala Ala Ala 355 360 365Ala Asn Lys Gly Lys Cys Gln Asp Gly Ala Ile Gly Ala Ala Val Gly 370 375 380Glu Ile Val Gly Glu Ala Leu Thr Asn Gly Lys Asn Pro Asp Thr Leu385 390 395 400Thr Ala Lys Glu Arg Glu Gln Ile Leu Ala Tyr Ser Lys Leu Val Ala 405 410 415Gly Thr Val Ser Gly Val Val Gly Gly Asp Val Asn Ala Ala Ala Asn 420 425 430Ala Ala Glu Val Ala Val Lys Asn Asn Gln Leu Ser Asp Xaa Glu Gly 435 440 445Arg Glu Phe Asp Asn Glu Met Thr Ala Cys Ala Lys Gln Asn Xaa Pro 450 455 460Gln Leu Cys Arg Lys Asn Thr Val Lys Lys Tyr Gln Asn Val Ala Asp465 470 475 480Lys Arg Leu Ala Ala Ser Ile Ala Ile Cys Thr Asp Ile Ser Arg Ser 485 490 495Thr Glu Cys Arg Thr Ile Arg Lys Gln His Leu Ile Asp Ser Arg Ser 500 505 510Leu His Ser Ser Trp Glu Ala Gly Leu Ile Gly Lys Asp Asp Glu Trp 515 520 525Tyr Lys Leu Phe Ser Lys Ser Tyr Thr Gln Ala Asp 530 535 540 168 540 PRT Artificial Sequence Description of Artificial Sequence ORF49-1 168Met Gln Leu Leu Ala Ala Glu Gly Ile His Gln His Gln Leu Asn Val 1 5 10 15Gln Lys Ser Thr Arg Phe Ile Gly Ile Lys Val Gly Lys Ser Asn Tyr 20 25 30Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val Arg Val Ile Ala 35 40 45Gln Thr Ala Lys Thr Arg Ser Gly Trp Asp Thr Val Leu Glu Gly Thr 50 55 60Glu Phe Lys Thr Thr Leu Ser Gly Ala Asp Ile Gln Ala Gly Val Gly 65 70 75 80Glu Lys Ala Arg Ala Asp Ala Lys Ile Ile Leu Lys Gly Ile Val Asn 85 90 95Arg Ile Gln Thr Glu Glu Lys Leu Glu Ser Asn Ser Thr Val Trp Gln 100 105 110Lys Gln Ala Gly Ser Gly Ser Thr Val Glu Thr Leu Lys Leu Pro Ser 115 120 125Phe Glu Gly Pro Ala Leu Pro Lys Leu Thr Ala Pro Gly Gly Tyr Ile 130 135 140Ala Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile Glu Lys Leu Ala145 150 155 160Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln Thr Val Lys Asp 165 170 175Val Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Lys Trp Asp Tyr Lys 180 185 190Gln Glu Gly Leu Thr Gly Ala Gly Ala Ala Ile Ile Ala Leu Ala Val 195 200 205Thr Val Val Thr Ser Gly Ala Gly Thr Gly Ala Val Leu Gly Leu Asn 210 215 220Gly Ala Ala Ala Ala Ala Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser225 230 235 240Gln Ala Ser Val Ser Phe Ile Asn Asn Lys Gly Asn Ile Gly Asn Thr 245 250 255Leu Lys Glu Leu Gly Arg Ser Ser Thr Val Lys Asn Leu Met Val Ala 260 265 270Val Ala Thr Ala Gly Val Ala Asp Lys Ile Gly Ala Ser Ala Leu Asn 275 280 285Asn Val Ser Asp Lys Gln Trp Ile Asn Asn Leu Thr Val Asn Leu Ala 290 295 300Asn Ala Gly Ser Ala Ala Leu Ile Asn Thr Ala Val Asn Gly Gly Ser305 310 315 320Leu Lys Asp Asn Leu Glu Ala Asn Ile Leu Ala Ala Leu Val Asn Thr 325 330 335Ala His Gly Glu Ala Ala Ser Lys Ile Lys Gln Leu Asp Gln His Tyr 340 345 350Ile Ala His Lys Ile Ala His Ala Ile Ala Gly Cys Ala Ala Ala Ala 355 360 365Ala Asn Lys Gly Lys Cys Gln Asp Gly Ala Ile Gly Ala Ala Val Gly 370 375 380Glu Ile Leu Gly Glu Thr Leu Leu Asp Gly Arg Asp Pro Gly Ser Leu385 390 395 400Asn Val Lys Asp Arg Ala Lys Ile Ile Ala Lys Ala Lys Leu Ala Ala 405 410 415Gly Ala Val Ala Ala Leu Ser Lys Gly Asp Val Ser Thr Ala Ala Asn 420 425 430Ala Ala Ala Val Ala Val Glu Asn Asn Ser Leu Asn Asp Ile Gln Asp 435 440 445Arg Leu Leu Ser Gly Asn Tyr Ala Leu Cys Met Ser Ala Gly Gly Ala 450 455 460Glu Ser Phe Cys Glu Ser Tyr Arg Pro Leu Gly Leu Pro His Phe Val465 470 475 480Ser Val Ser Gly Glu Met Lys Leu Pro Asn Lys Phe Gly Asn Arg Met 485 490 495Val Asn Gly Lys Leu Ile Ile Asn Thr Arg Asn Gly Asn Val Tyr Phe 500 505 510Ser Val Gly Lys Ile Trp Ser Thr Val Lys Ser Thr Lys Ser Asn Ile 515 520 525Ser Gly Val Ser Val Gly Trp Val Leu Asn Val Ser 530 535 540 169 210 PRT Artificial Sequence Description of Artificial Sequence ORF39 169Lys Phe Asp Phe Thr Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Arg 1 5 10 15Leu Phe Phe Glu Val Leu Val Val Ser Val Val Leu Gln Leu Phe Ala 20 25 30Leu Ile Thr Pro Leu Phe Phe Gln Val Val Met Asp Lys Val Leu Val 35 40 45His Arg Gly Phe Ser Thr Leu Asp Val Val Ser Val Ala Leu Leu Val 50 55 60Val Ser Leu Phe Glu Ile Val Leu Gly Gly Leu Arg Thr Tyr Leu Phe 65 70 75 80Ala His Thr Thr Ser Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe 85 90 95Arg His Leu Leu Ser Leu Pro Leu Ser Tyr Phe Glu His Arg Arg Val 100 105 110Gly Asp Thr Val Ala Arg Val Arg Glu Leu Glu Gln Ile Arg Asn Phe 115 120 125Leu Thr Gly Gln Ala Leu Thr Ser Val Leu Asp Leu Ala Phe Ser Phe 130 135 140Ile Phe Leu Ala Val Met Trp Tyr Tyr Ser Ser Thr Leu Thr Trp Val145 150 155 160Val Leu Ala Ser Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165 170 175Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 180 185 190Xaa Ile Cys Ala Asn Arg Thr Val Leu Ile Ile Ala His Arg Leu Ser 195 200 205Thr Val 210 170 240 PRT Artificial Sequence Description of Artificial Sequence ORF39a 170Ala Val Leu Ser Phe Ala Glu Phe Ser Asn Arg Tyr Ser Gly Lys Leu 1 5 10 15Ile Leu Val Ala Ser Arg Ala Ser Val Leu Gly Ser Leu Ala Lys Phe 20 25 30Asp Phe Thr Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Arg Leu Phe 35 40 45Phe Glu Val Leu Val Val Ser Val Val Leu Gln Leu Phe Ala Leu Ile 50 55 60Thr Pro Leu Phe Phe Gln Val Val Met Asp Lys Val Leu Val His Arg 65 70 75 80Gly Phe Ser Thr Leu Asp Val Val Ser Val Ala Leu Leu Val Val Ser 85 90 95Leu Phe Glu Ile Val Leu Gly Gly Leu Arg Thr Tyr Leu Phe Ala His 100 105 110Thr Thr Ser Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe Arg His 115 120 125Leu Leu Ser Leu Pro Leu Ser Tyr Phe Glu His Arg Arg Val Gly Asp 130 135 140Thr Val Ala Arg Val Arg Glu Leu Glu Gln Ile Arg Asn Phe Leu Thr145 150 155 160Gly Gln Ala Leu Thr Ser Val Leu Asp Leu Ala Phe Ser Phe Ile Phe 165 170 175Leu Ala Val Met Trp Tyr Tyr Ser Ser Thr Leu Thr Trp Val Val Leu 180 185 190Ala Ser Leu Pro Ala Tyr Ala Phe Trp Ser Ala Phe Ile Ser Pro Ile 195 200 205Leu Arg Thr Arg Leu Asn Asp Lys Phe Ala Arg Asn Ala Asp Asn Gln 210 215 220Ser Phe Leu Val Glu Ser Ile Thr Ala Val Gly Thr Val Lys Ala Met225 230 235 240 171 690 PRT Artificial Sequence Description of Artificial Sequence ORF39a 171Tyr His Gly Ile Ala Ala Asn Pro Ala Asp Ile Gln His Glu Phe Cys 1 5 10 15Thr Ser Ala Gln Ser Asp Leu Asn Glu Thr Gln Trp Xaa Xaa Xaa Xaa 20 25 30Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Val Val Arg Gln Pro Ile Lys Arg 35 40 45Leu Ala Met Ala Thr Leu Pro Ala Leu Val Trp Cys Asp Asp Gly Asn 50 55 60His Phe Ile Leu Ala Lys Thr Asp Gly Gly Gly Glu His Ala Gln Tyr 65 70 75 80Leu Ile Gln Asp Leu Thr Thr Asn Lys Ser Ala Val Leu Ser Phe Ala 85 90 95Glu Phe Ser Asn Arg Tyr Ser Gly Lys Leu Ile Leu Val Ala Ser Arg 100 105 110Ala Ser Val Leu Gly Ser Leu Ala Lys Phe Asp Phe Thr Trp Phe Ile 115 120 125Pro Ala Val Ile Lys Tyr Arg Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 130 135 140Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Thr Pro Leu Phe Phe Gln145 150 155 160Val Val Met Asp Lys Val Leu Val His Arg Gly Phe Xaa Xaa Xaa Xaa 165 170 175Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Glu Ile Val Leu 180 185 190Gly Gly Leu Arg Thr Tyr Leu Phe Ala His Thr Thr Ser Arg Ile Asp 195 200 205Val Glu Leu Gly Ala Arg Leu Phe Arg His Leu Leu Ser Leu Pro Leu 210 215 220Ser Tyr Phe Glu His Arg Arg Val Gly Asp Thr Val Ala Arg Val Arg225 230 235 240Glu Leu Glu Gln Ile Arg Asn Phe Leu Thr Gly Gln Ala Leu Thr Ser 245 250 255Val Leu Asp Leu Ala Phe Ser Phe Ile Phe Leu Ala Val Met Trp Tyr 260 265 270Tyr Ser Ser Thr Leu Thr Trp Val Val Leu Ala Ser Leu Pro Ala Tyr 275 280 285Ala Phe Trp Ser Ala Phe Ile Ser Pro Ile Leu Arg Thr Arg Leu Asn 290 295 300Asp Lys Phe Ala Arg Asn Ala Asp Asn Gln Ser Phe Leu Val Glu Ser305 310 315 320Ile Thr Ala Val Gly Thr Val Lys Ala Met Ala Val Glu Pro Gln Met 325 330 335Thr Gln Arg Trp Asp Asn Gln Leu Ala Ala Tyr Val Ala Ser Gly Phe 340 345 350Arg Val Thr Lys Leu Ala Val Val Gly Gln Gln Gly Val Gln Leu Ile 355 360 365Gln Lys Leu Val Thr Val Ala Thr Leu Trp Ile Gly Ala Arg Leu Val 370 375 380Ile Glu Ser Lys Leu Thr Val Gly Gln Leu Ile Ala Phe Asn Met Leu385 390 395 400Ser Gly Gln Val Ala Ala Pro Val Ile Arg Leu Ala Gln Leu Trp Gln 405 410 415Asp Phe Gln Gln Val Gly Ile Ser Val Ala Arg Leu Gly Asp Ile Leu 420 425 430Asn Ala Pro Thr Glu Asn Ala Ser Ser His Leu Ala Leu Pro Asp Ile 435 440 445Arg Gly Glu Ile Thr Phe Glu His Val Asp Phe Arg Tyr Lys Ala Asp 450 455 460Gly Arg Leu Ile Leu Gln Asp Leu Asn Leu Arg Ile Arg Ala Gly Glu465 470 475 480Val Leu Gly Ile Val Gly Arg Ser Gly Ser Gly Lys Ser Thr Leu Thr 485 490 495Lys Leu Val Gln Arg Leu Tyr Val Pro Ala Gln Gly Arg Val Leu Val 500 505 510Asp Gly Asn Asp Leu Ala Leu Ala Ala Pro Ala Trp Leu Arg Arg Gln 515 520 525Val Gly Val Val Leu Gln Glu Asn Val Leu Leu Asn Arg Ser Ile Arg 530 535 540Asp Asn Ile Ala Leu Thr Asp Thr Gly Met Pro Leu Glu Arg Ile Ile545 550 555 560Glu Ala Ala Lys Leu Ala Gly Ala His Glu Phe Ile Met Glu Leu Pro 565 570 575Glu Gly Tyr Gly Thr Val Val Gly Glu Gln Gly Ala Gly Leu Ser Gly 580 585 590Gly Gln Arg Gln Arg Ile Ala Ile Ala Arg Ala Leu Ile Thr Asn Pro 595 600 605Arg Ile Leu Ile Phe Asp Glu Ala Thr Ser Ala Leu Asp Tyr Glu Ser 610 615 620Glu Arg Ala Ile Met Gln Asn Met Gln Ala Ile Cys Ala Asn Arg Thr625 630 635 640Val Leu Ile Ile Ala His Arg Leu Ser Thr Val Lys Thr Ala His Arg 645 650 655Ile Ile Ala Met Asp Lys Gly Arg Ile Val Glu Ala Gly Thr Gln Gln 660 665 670Glu Leu Leu Ala Lys Pro Asn Gly Tyr Tyr Arg Tyr Leu Tyr Asp Leu 675 680 685Gln Asn 690 172 690 PRT Artificial Sequence Description of Artificial Sequence homology 172Tyr His Xaa Ile Ala Xaa Asn Pro Xaa Xaa Xaa Xaa His Xaa Phe Xaa 1 5 10 15Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Thr Xaa Trp Xaa Xaa Xaa Xaa 20 25 30Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Val Xaa Xaa Xaa Ile Xaa Arg 35 40 45Leu Ala Xaa Xaa Xaa Leu Pro Ala Leu Val Trp Xaa Xaa Asp Gly Xaa 50 55 60His Phe Ile Leu Xaa Lys Xaa Asp Xaa Xaa Xaa Glu Xaa Xaa Xaa Tyr 65 70 75 80Leu Ile Xaa Asp Leu Xaa Thr Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Ala 85 90 95Glu Phe Xaa Xaa Xaa Tyr Xaa Gly Lys Leu Ile Leu Val Ala Ser Arg 100 105 110Ala Ser Xaa Xaa Gly Xaa Leu Ala Lys Phe Asp Phe Thr Trp Phe Ile 115 120 125Pro Ala Val Ile Lys Tyr Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 130 135 140Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Thr Pro Leu Phe Phe Gln145 150 155 160Val Val Met Asp Lys Val Leu Val His Arg Gly Phe Xaa Xaa Xaa Xaa 165 170 175Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Glu Ile Val Leu 180 185 190Xaa Gly Leu Arg Thr Tyr Xaa Phe Ala His Xaa Thr Ser Arg Ile Asp 195 200 205Val Glu Leu Gly Ala Arg Leu Phe Arg His Leu Leu Xaa Leu Pro Xaa 210 215 220Ser Tyr Phe Glu Xaa Arg Arg Val Gly Asp Thr Val Ala Arg Val Arg225 230 235 240Glu Leu Xaa Gln Ile Arg Asn Phe Leu Thr Gly Gln Ala Leu Thr Ser 245 250 255Val Leu Asp Leu Xaa Phe Ser Phe Ile Phe Xaa Ala Val Met Trp Tyr 260 265 270Tyr Ser Xaa Xaa Leu Thr Xaa Val Xaa Leu Xaa Ser Leu Pro Xaa Tyr 275 280 285Xaa Xaa Trp Ser Xaa Phe Ile Ser Pro Ile Leu Arg Xaa Arg Leu Xaa 290 295 300Xaa Lys Phe Ala Arg Xaa Ala Asp Asn Gln Ser Phe Leu Val Glu Ser305 310 315 320Xaa Thr Ala Xaa Xaa Thr Xaa Lys Ala Xaa Ala Val Xaa Pro Gln Met 325 330 335Thr Xaa Xaa Trp Asp Xaa Gln Leu Ala Xaa Tyr Val Xaa Xaa Gly Phe 340 345 350Arg Val Thr Xaa Leu Ala Xaa Xaa Gly Gln Gln Gly Val Gln Xaa Ile 355 360 365Gln Lys Xaa Val Xaa Val Xaa Thr Leu Trp Xaa Gly Ala Xaa Leu Val 370 375 380Ile Xaa Xaa Xaa Leu Xaa Xaa Gly Gln Leu Ile Ala Phe Asn Met Leu385 390 395 400Ser Gly Gln Val Xaa Ala Pro Val Ile Arg Leu Ala Gln Leu Trp Gln 405 410 415Asp Phe Gln Gln Val Gly Ile Ser Val Xaa Arg Leu Gly Asp Xaa Leu 420 425 430Asn Xaa Pro Thr Glu Xaa Xaa Xaa Xaa Xaa Leu Ala Leu Pro Xaa Ile 435 440 445Xaa Gly Xaa Ile Thr Phe Xaa Xaa Xaa Xaa Phe Arg Tyr Lys Xaa Asp 450 455 460Xaa Xaa Xaa Ile Leu Xaa Asp Xaa Asn Leu Xaa Ile Xaa Xaa Gly Glu465 470 475 480Val Xaa Gly Ile Val Gly Arg Ser Gly Ser Gly Lys Ser Thr Leu Thr 485 490 495Lys Leu Xaa Gln Arg Xaa Tyr Xaa Pro Xaa Xaa Gly Xaa Val Leu Xaa 500 505 510Asp Gly Xaa Asp Leu Ala Leu Ala Xaa Pro Xaa Trp Leu Arg Arg Gln 515 520 525Val Gly Val Val Leu Gln Xaa Asn Val Leu Leu Asn Arg Ser Ile Arg 530 535 540Asp Asn Ile Ala Leu Xaa Asp Xaa Gly Met Pro Xaa Glu Xaa Ile Xaa545 550 555 560Xaa Ala Ala Lys Leu Ala Gly Ala His Glu Phe Ile Xaa Glu Leu Xaa 565 570 575Glu Gly Tyr Xaa Thr Xaa Val Gly Glu Gln Gly Ala Gly Leu Ser Gly 580 585 590Gly Gln Arg Gln Arg Ile Ala Ile Ala Arg Ala Leu Xaa Xaa Asn Pro 595 600 605Xaa Ile Leu Ile Phe Asp Glu Ala Thr Ser Ala Leu Asp Tyr Glu Ser 610 615 620Glu Xaa Xaa Ile Met Xaa Asn Met Xaa Xaa Ile Cys Xaa Xaa Arg Thr625 630 635 640Val Xaa Ile Ile Ala His Arg Leu Ser Thr Val Lys Xaa Ala Xaa Arg 645 650 655Ile Ile Xaa Met Xaa Lys Gly Xaa Ile Val Glu Xaa Gly Xaa Xaa Xaa 660 665 670Glu Leu Leu Ala Xaa Pro Asn Gly Xaa Tyr Xaa Tyr Leu Xaa Xaa Leu 675 680 685Gln Xaa 690 173 687 PRT Artificial Sequence Description of Artificial Sequence cytolysin from A. pleuropneumoniae 173Tyr His Asn Ile Ala Val Asn Pro Glu Glu Leu Lys His Lys Phe Asp 1 5 10 15Leu Glu Gly Lys Gly Leu Asp Leu Thr Ala Trp Leu Leu Ala Ala Lys 20 25 30Ser Leu Glu Leu Lys Ala Lys Gln Val Lys Lys Ala Ile Asp Arg Leu 35 40 45Ala Phe Ile Ala Leu Pro Ala Leu Val Trp Arg Glu Asp Gly Lys His 50 55 60Phe Ile Leu Thr Lys Ile Asp Asn Glu Ala Lys Lys Tyr Leu Ile Phe 65 70 75 80Asp Leu Glu Thr His Asn Pro Arg Ile Leu Glu Gln Ala Glu Phe Glu 85 90 95Ser Leu Tyr Gln Gly Lys Leu Ile Leu Val Ala Ser Arg Ala Ser Ile 100 105 110Val Gly Lys Leu Ala Lys Phe Asp Phe Thr Trp Phe Ile Pro Ala Val 115 120 125Ile Lys Tyr Arg Lys Ile Phe Ile Glu Thr Leu Ile Val Ser Ile Phe 130 135 140Leu Gln Ile Phe Ala Leu Ile Thr Pro Leu Phe Phe Gln Val Val Met145 150 155 160Asp Lys Val Leu Val His Arg Gly Phe Ser Thr Leu Asn Val Ile Thr 165 170 175Val Ala Leu Ala Ile Val Val Leu Phe Glu Ile Val Leu Asn Gly Leu 180 185 190Arg Thr Tyr Ile Phe Ala His Ser Thr Ser Arg Ile Asp Val Glu Leu 195 200 205Gly Ala Arg Leu Phe Arg His Leu Leu Ala Leu Pro Ile Ser Tyr Phe 210 215 220Glu Asn Arg Arg Val Gly Asp Thr Val Ala Arg Val Arg Glu Leu Asp225 230 235 240Gln Ile Arg Asn Phe Leu Thr Gly Gln Ala Leu Thr Ser Val Leu Asp 245 250 255Leu Met Phe Ser Phe Ile Phe Phe Ala Val Met Trp Tyr Tyr Ser Pro 260 265 270Lys Leu Thr Leu Val Ile Leu Gly Ser Leu Pro Phe Tyr Met Gly Trp 275 280 285Ser Ile Phe Ile Ser Pro Ile Leu Arg Arg Arg Leu Asp Glu Lys Phe 290 295 300Ala Arg Gly Ala Asp Asn Gln Ser Phe Leu Val Glu Ser Val Thr Ala305 310 315 320Ile Asn Thr Ile Lys Ala Leu Ala Val Thr Pro Gln Met Thr Asn Thr 325 330 335Trp Asp Lys Gln Leu Ala Ser Tyr Val Ser Ala Gly Phe Arg Val Thr 340 345 350Thr Leu Ala Thr Ile Gly Gln Gln Gly Val Gln Phe Ile Gln Lys Val 355 360 365Val Met Val Ile Thr Leu Trp Leu Gly Ala His Leu Val Ile Ser Gly 370 375 380Asp Leu Ser Ile Gly Gln Leu Ile Ala Phe Asn Met Leu Ser Gly Gln385 390 395 400Val Ile Ala Pro Val Ile Arg Leu Ala Gln Leu Trp Gln Asp Phe Gln 405 410 415Gln Val Gly Ile Ser Val Thr Arg Leu Gly Asp Val Leu Asn Ser Pro 420 425 430Thr Glu Ser Tyr Gln Gly Lys Leu Ala Leu Pro Glu Ile Lys Gly Asp 435 440 445Ile Thr Phe Arg Asn Ile Arg Phe Arg Tyr Lys Pro Asp Ala Pro Val 450 455 460Ile Leu Asn Asp Val Asn Leu Ser Ile Gln Gln Gly Glu Val Ile Gly465 470 475 480Ile Val Gly Arg Ser Gly Ser Gly Lys Ser Thr Leu Thr Lys Leu Ile 485 490 495Gln Arg Phe Tyr Ile Pro Glu Asn Gly Gln Val Leu Ile Asp Gly His 500 505 510Asp Leu Ala Leu Ala Asp Pro Asn Trp Leu Arg Arg Gln Val Gly Val 515 520 525Val Leu Gln Asp Asn Val Leu Leu Asn Arg Ser Ile Arg Asp Asn Ile 530 535 540Ala Leu Ala Asp Pro Gly Met Pro Met Glu Lys Ile Val His Ala Ala545 550 555 560Lys Leu Ala Gly Ala His Glu Phe Ile Ser Glu Leu Arg Glu Gly Tyr 565 570 575Asn Thr Ile Val Gly Glu Gln Gly Ala Gly Leu Ser Gly Gly Gln Arg 580 585 590Gln Arg Ile Ala Ile Ala Arg Ala Leu Val Asn Asn Pro Lys Ile Leu 595 600 605Ile Phe Asp Glu Ala Thr Ser Ala Leu Asp Tyr Glu Ser Glu His Ile 610 615 620Ile Met Arg Asn Met His Gln Ile Cys Lys Gly Arg Thr Val Ile Ile625 630 635 640Ile Ala His Arg Leu Ser Thr Val Lys Asn Ala Asp Arg Ile Ile Val 645 650 655Met Glu Lys Gly Gln Ile Val Glu Gln Gly Lys His Lys Glu Leu Leu 660 665 670Ala Asp Pro Asn Gly Leu Tyr His Tyr Leu His Gln Leu Gln Ser 675 680 685 174 222 PRT Artificial Sequence Description of Artificial Sequence ORF39 174Lys Phe Asp Phe Thr Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Arg 1 5 10 15Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30Xaa Ile Thr Pro Leu Phe Phe Gln Val Val Met Asp Lys Val Leu Val 35 40 45His Arg Gly Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Phe Glu Ile Val Leu Gly Gly Leu Arg Thr Tyr Leu Phe 65 70 75 80Ala His Thr Thr Ser Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe 85 90 95Arg His Leu Leu Ser Leu Pro Leu Ser Tyr Phe Glu His Arg Arg Val 100 105 110Gly Asp Thr Val Ala Arg Val Arg Glu Leu Glu Gln Ile Arg Asn Phe 115 120 125Leu Thr Gly Gln Ala Leu Thr Ser Val Leu Asp Leu Ala Phe Ser Phe 130 135 140Ile Phe Leu Ala Val Met Trp Tyr Tyr Ser Ser Thr Leu Thr Trp Val145 150 155 160Val Leu Ala Ser Leu Ile Cys Ile Cys Ala Asn Arg Thr Val Leu Ile 165 170 175Ile Ala His Arg Leu Ser Thr Val Lys Thr Ala His Arg Ile Ile Ala 180 185 190Met Asp Lys Gly Arg Ile Val Glu Ala Gly Thr Gln Gln Glu Leu Leu 195 200 205Ala Asn Xaa Asn Gly Tyr Tyr Arg Tyr Leu Tyr Asp Leu Gln 210 215 220 175 222 PRT Artificial Sequence Description of Artificial Sequence overlap identity 175Lys Phe Asp Phe Thr Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Xaa 1 5 10 15Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30Xaa Ile Thr Pro Leu Phe Phe Gln Val Val Met Asp Lys Val Leu Val 35 40 45His Arg Gly Phe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Phe Glu Ile Xaa Leu Gly Gly Leu Arg Thr Tyr Xaa Phe 65 70 75 80Ala His Xaa Thr Ser Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe 85 90 95Arg His Leu Leu Xaa Leu Pro Xaa Ser Tyr Phe Glu Xaa Arg Arg Val 100 105 110Gly Asp Thr Val Ala Arg Val Arg Glu Leu Xaa Gln Ile Arg Asn Phe 115 120 125Leu Thr Gly Gln Ala Leu Thr Ser Xaa Leu Asp Leu Xaa Phe Ser Phe 130 135 140Ile Phe Xaa Ala Val Met Trp Tyr Tyr Ser Xaa Xaa Leu Thr Xaa Val145 150 155 160Val Leu Xaa Ser Leu Xaa Cys Ile Cys Xaa Asn Arg Thr Val Leu Ile 165 170 175Ile Ala His Arg Leu Ser Thr Val Lys Xaa Ala Xaa Arg Ile Ile Xaa 180 185 190Met Asp Lys Gly Xaa Ile Xaa Glu Xaa Gly Xaa Xaa Gln Glu Leu Leu 195 200 205Xaa Xaa Xaa Xaa Gly Xaa Tyr Xaa Tyr Leu Xaa Xaa Leu Gln 210 215 220 176 222 PRT Artificial Sequence Description of Artificial Sequence HlyB 176Lys Phe Asp Phe Thr Trp Phe Ile Pro Ala Val Ile Lys Tyr Arg Lys 1 5 10 15Ile Phe Ile Glu Thr Leu Ile Val Ser Ile Phe Leu Gln Ile Phe Ala 20 25 30Leu Ile Thr Pro Leu Phe Phe Gln Val Val Met Asp Lys Val Leu Val 35 40 45His Arg Gly Phe Ser Thr Leu Asn Val Ile Thr Val Ala Leu Ala Ile 50 55 60Val Val Leu Phe Glu Ile Ile Leu Gly Gly Leu Arg Thr Tyr Val Phe 65 70 75 80Ala His Ser Thr Ser Arg Ile Asp Val Glu Leu Gly Ala Arg Leu Phe 85 90 95Arg His Leu Leu Ala Leu Pro Ile Ser Tyr Phe Glu Ala Arg Arg Val 100 105 110Gly Asp Thr Val Ala Arg Val Arg Glu Leu Asp Gln Ile Arg Asn Phe 115 120 125Leu Thr Gly Gln Ala Leu Thr Ser Ile Leu Asp Leu Leu Phe Ser Phe 130 135 140Ile Phe Phe Ala Val Met Trp Tyr Tyr Ser Pro Lys Leu Thr Leu Val145 150 155 160Val Leu Gly Ser Leu Pro Cys Ile Cys Gln Asn Arg Thr Val Leu Ile 165 170 175Ile Ala His Arg Leu Ser Thr Val Lys Asn Ala Asp Arg Ile Ile Val 180 185 190Met Asp Lys Gly Glu Ile Ile Glu Gln Gly Lys His Gln Glu Leu Leu 195 200 205Lys Asp Glu Lys Gly Leu Tyr Ser Tyr Leu His Gln Leu Gln 210 215 220 177 240 PRT Artificial Sequence Description of Artificial Sequence ORF112a 177Met Asn Leu Ile Ser Arg Tyr Ile Ile Arg Gln Met Ala Val Met Ala 1 5 10 15Val Tyr Ala Leu Leu Ala Phe Leu Ala Leu Tyr Ser Phe Phe Glu Ile 20 25 30Leu Tyr Glu Thr Gly Asn Leu Gly Lys Gly Ser Tyr Gly Ile Trp Glu 35 40 45Met Xaa Gly Tyr Thr Ala Leu Lys Met Xaa Ala Arg Ala Tyr Glu Leu 50 55 60Met Pro Leu Ala Val Leu Ile Gly Gly Leu Val Ser Xaa Ser Gln Leu 65 70 75 80Ala Ala Gly Ser Glu Leu Xaa Val Ile Lys Ala Ser Gly Met Ser Thr 85 90 95Lys Lys Leu Leu Leu Ile Leu Ser Gln Phe Gly Phe Ile Phe Ala Ile 100 105 110Ala Thr Val Ala Leu Gly Glu Trp Val Ala Pro Thr Leu Ser Gln Lys 115 120 125Ala Glu Asn Ile Lys Ala Ala Ala Ile Asn Gly Lys Ile Ser Thr Gly 130 135 140Asn Thr Gly Leu Trp Leu Lys Glu Lys Asn Ser Ile Ile Asn Val Arg145 150 155 160Glu Met Leu Pro Asp His Thr Leu Leu Gly Ile Lys Ile Trp Ala Arg 165 170 175Asn Asp Lys Asn Glu Leu Ala Glu Ala Val Glu Ala Asp Ser Ala Val 180 185 190Leu Asn Ser Asp Gly Ser Trp Gln Leu Lys Asn Ile Arg Arg Ser Thr 195 200 205Leu Gly Glu Asp Lys Val Glu Val Ser Ile Ala Ala Glu Glu Xaa Trp 210 215 220Pro Ile Ser Val Lys Arg Asn Leu Met Asp Val Leu Leu Val Lys Pro225 230 235 240 178 360 PRT Artificial Sequence Description of Artificial Sequence ORF114a 178Met Asn Lys Gly Leu His Arg Ile Ile Phe Ser Lys Lys His Ser Thr 1 5 10 15Met Val Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln 20 25 30Ala Gly Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Leu Cys 35 40 45Gly Lys Leu Lys Thr Thr Leu Lys Thr Leu Val Cys Ser Leu Val Ser 50 55 60Leu Ser Met Xaa Xaa Xaa Xaa Xaa Xaa Gln Ile Thr Thr Asp Lys Ser 65 70 75 80Ala Pro Lys Asn Xaa Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala 85 90 95Pro Leu Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn 100 105 110Arg Tyr Thr Gln Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn 115 120 125Asp Arg Asn Asn Asn Pro Phe Leu Val Lys Gly Ser Ala Gln Leu Ile 130 135 140Leu Asn Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr145 150 155 160Val Gly Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile 165 170 175Thr Val Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr 180 185 190Ile Gly Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp 195 200 205Val Arg Gln Gly Thr Leu Thr Val Gly Ala Ala Gly Trp Asn Asp Lys 210 215 220Gly Gly Ala Asp Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln225 230 235 240Gly Lys Leu Gln Gly Lys Asn Leu Ala Val Ser Thr Gly Pro Gln Lys 245 250 255Val Asp Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr 260 265 270Lys Pro Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala 275 280 285Asp Ser Ile Thr Leu Ile Ala Xaa Glu Lys Gly Val Gly Val Lys Asn 290 295 300Ala Gly Thr Leu Glu Ala Ala Lys Gln Leu Ile Val Thr Ser Ser Gly305 310 315 320Arg Ile Glu Asn Ser Gly Arg Ile Ala Thr Thr Ala Asp Gly Thr Glu 325 330 335Ala Ser Pro Thr Tyr Leu Xaa Ile Glu Thr Thr Glu Lys Gly Ala Xaa 340 345 350Gly Thr Phe Ile Ser Asn Gly Gly 355 360 179 1574 PRT Artificial Sequence Description of Artificial Sequence ORF114-1 179Met Asn Lys Gly Leu His Arg Ile Ile Phe Ser Lys Lys His Ser Thr 1 5 10 15Met Val Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln 20 25 30Ala Gly Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Leu Cys 35 40 45Gly Lys Leu Lys Thr Thr Leu Lys Thr Leu Val Cys Ser Leu Val Ser 50 55 60Leu Ser Met Val Leu Pro Ala His Ala Gln Ile Thr Thr Asp Lys Ser 65 70 75 80Ala Pro Lys Asn Gln Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala 85 90 95Pro Leu Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn 100 105 110Arg Tyr Thr Gln Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn 115 120 125Asp Arg Asn Asn Asn Pro Phe Val Val Lys Gly Ser Ala Gln Leu Ile 130 135 140Leu Asn Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr145 150 155 160Val Gly Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile 165 170 175Thr Val Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr 180 185 190Thr Gly Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp 195 200 205Val Arg Gln Gly Thr Leu Thr Val Gly Ala Ala Gly Trp Asn Asp Lys 210 215 220Gly Gly Ala Asp Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln225 230 235 240Gly Lys Leu Gln Gly Lys Asn Leu Ala Val Ser Thr Gly Pro Gln Lys 245 250 255Val Asp Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr 260 265 270Lys Pro Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala 275 280 285Asp Ser Ile Thr Leu Ile Ala Asn Glu Lys Gly Val Gly Val Lys Asn 290 295 300Ala Gly Thr Leu Glu Ala Ala Lys Gln Leu Ile Val Thr Ser Ser Gly305 310 315 320Arg Ile Glu Asn Ser Gly Arg Ile Ala Thr Thr Ala Asp Gly Thr Glu 325 330 335Ala Ser Pro Thr Tyr Leu Ser Ile Glu Thr Thr Glu Lys Gly Ala Ala 340 345 350Gly Thr Phe Ile Ser Asn Gly Gly Arg Ile Glu Ser Lys Gly Leu Leu 355 360 365Val Ile Glu Thr Gly Glu Asp Ile Ser Leu Arg Asn Gly Ala Val Val 370 375 380Gln Asn Asn Gly Ser Arg Pro Ala Thr Thr Val Leu Asn Ala Gly His385 390 395 400Asn Leu Val Ile Glu Ser Lys Thr Asn Val Asn Asn Ala Lys Gly Pro 405 410 415Ala Thr Leu Ser Ala Asp Gly Arg Thr Val Ile Lys Glu Ala Ser Ile 420 425 430Gln Thr Gly Thr Thr Val Tyr Ser Ser Ser Lys Gly Asn Ala Glu Leu 435 440 445Gly Asn Asn Thr Arg Ile Thr Gly Ala Asp Val Thr Val Leu Ser Asn 450 455 460Gly Thr Ile Ser Ser Ser Ala Val Ile Asp Ala Lys Asp Thr Ala His465 470 475 480Ile Glu Ala Gly Lys Pro Leu Ser Leu Glu Ala Ser Thr Val Thr Ser 485 490 495Asp Ile Arg Leu Asn Gly Gly Ser Ile Lys Gly Gly Lys Gln Leu Ala 500 505 510Leu Leu Ala Asp Asp Asn Ile Thr Ala Lys Thr Thr Asn Leu Asn Thr 515 520 525Pro Gly Asn Leu Tyr Val His Thr Gly Lys Asp Leu Asn Leu Asn Val 530 535 540Asp Lys Asp Leu Ser Ala Ala Ser Ile His Leu Lys Ser Asp Asn Ala545 550 555 560Ala His Ile Thr Gly Thr Ser Lys Thr Leu Thr Ala Ser Lys Asp Met 565 570 575Gly Val Glu Ala Gly Ser Leu Asn Val Thr Asn Thr Asn Leu Arg Thr 580 585 590Asn Ser Gly Asn Leu His Ile Gln Ala Ala Lys Gly Asn Ile Gln Leu 595 600 605Arg Asn Thr Lys Leu Asn Ala Ala Lys Ala Leu Glu Thr Thr Ala Leu 610 615 620Gln Gly Asn Ile Val Ser Asp Gly Leu His Ala Val Ser Ala Asp Gly625 630 635 640His Val Ser Leu Leu Ala Asn Gly Asn Ala Asp Phe Thr Gly His Asn 645 650 655Thr Leu Thr Ala Lys Ala Asp Val Asn Ala Gly Ser Val Gly Lys Gly 660 665 670Arg Leu Lys Ala Asp Asn Thr Asn Ile Thr Ser Ser Ser Gly Asp Ile 675 680 685Thr Leu Val Ala Gly Asn Gly Ile Gln Leu Gly Asp Gly Lys Gln Arg 690 695 700Asn Ser Ile Asn Gly Lys His Ile Ser Ile Lys Asn Asn Gly Gly Asn705 710 715 720Ala Asp Leu Lys Asn Leu Asn Val His Ala Lys Ser Gly Ala Leu Asn 725 730 735Ile His Ser Asp Arg Ala Leu Ser Ile Glu Asn Thr Lys Leu Glu Ser 740 745 750Thr His Asn Thr His Leu Asn Ala Gln His Glu Arg Val Thr Leu Asn 755 760 765Gln Val Asp Ala Tyr Ala His Arg His Leu Ser Ile Thr Gly Ser Gln 770 775 780Ile Trp Gln Asn Asp Lys Leu Pro Ser Ala Asn Lys Leu Val Ala Asn785 790 795 800Gly Val Leu Ala Leu Asn Ala Arg Tyr Ser Gln Ile Ala Asp Asn Thr 805 810 815Thr Leu Arg Ala Gly Ala Ile Asn Leu Thr Ala Gly Thr Ala Leu Val 820 825 830Lys Arg Gly Asn Ile Asn Trp Ser Thr Val Ser Thr Lys Thr Leu Glu 835 840 845Asp Asn Ala Glu Leu Lys Pro Leu Ala Gly Arg Leu Asn Ile Glu Ala 850 855 860Gly Ser Gly Thr Leu Thr Ile Glu Pro Ala Asn Arg Ile Ser Ala His865 870 875 880Thr Asp Leu Ser Ile Lys Thr Gly Gly Lys Leu Leu Leu Ser Ala Lys 885 890 895Gly Gly Asn Ala Gly Ala Pro Ser Ala Gln Val Ser Ser Leu Glu Ala 900 905 910Lys Gly Asn Ile Arg Leu Val Thr Gly Glu Thr Asp Leu Arg Gly Ser 915 920 925Lys Ile Thr Ala Gly Lys Asn Leu Val Val Ala Thr Thr Lys Gly Lys 930 935 940Leu Asn Ile Glu Ala Val Asn Asn Ser Phe Ser Asn Tyr Phe Pro Thr945 950 955 960Gln Lys Ala Ala Glu Leu Asn Gln Lys Ser Lys Glu Leu Glu Gln Gln 965 970 975Ile Ala Gln Leu Lys Lys Ser Ser Pro Lys Ser Lys Leu Ile Pro Thr 980 985 990Leu Gln Glu Glu Arg Asp Arg Leu Ala Phe Tyr Ile Gln Ala Ile Asn 995 1000 1005Lys Glu Val Lys Gly Lys Lys Pro Lys Gly Lys Glu Tyr Leu Gln Ala 1010 1015 1020Lys Leu Ser Ala Gln Asn Ile Asp Leu Ile Ser Ala Gln Gly Ile Glu1025 1030 1035 1040Ile Ser Gly Ser Asp Ile Thr Ala Ser Lys Lys Leu Asn Leu His Ala 1045 1050 1055Ala Gly Val Leu Pro Lys Ala Ala Asp Ser Glu Ala Ala Ala Ile Leu 1060 1065 1070Ile Asp Gly Ile Thr Asp Gln Tyr Glu Ile Gly Lys Pro Thr Tyr Lys 1075 1080 1085Ser His Tyr Asp Lys Ala Ala Leu Asn Lys Pro Ser Arg Leu Thr Gly 1090 1095 1100Arg Thr Gly Val Ser Ile His Ala Ala Ala Ala Leu Asp Asp Ala Arg1105 1110 1115 1120Ile Ile Ile Gly Ala Ser Glu Ile Lys Ala Pro Ser Gly Ser Ile Asp 1125 1130 1135Ile Lys Ala His Ser Asp Ile Val Leu Glu Ala Gly Gln Asn Asp Ala 1140 1145 1150Tyr Thr Phe Leu Lys Thr Lys Gly Lys Ser Gly Lys Ile Ile Arg Lys 1155 1160 1165Thr Lys Phe Thr Ser Thr Arg Asp His Leu Ile Met Pro Ala Pro Val 1170 1175 1180Glu Leu Thr Ala Asn Gly Ile Thr Leu Gln Ala Gly Gly Asn Ile Glu1185 1190 1195 1200Ala Asn Thr Thr Arg Phe Asn Ala Pro Ala Gly Lys Val Thr Leu Val 1205 1210 1215Ala Gly Glu Glu Leu Gln Leu Leu Ala Glu Glu Gly Ile His Lys His 1220 1225 1230Glu Leu Asp Val Gln Lys Ser Arg Arg Phe Ile Gly Ile Lys Val Gly 1235 1240 1245Lys Ser Asn Tyr Ser Lys Asn Glu Leu Asn Glu Thr Lys Leu Pro Val 1250 1255 1260Arg Val Val Ala Gln Thr Ala Ala Thr Arg Ser Gly Trp Asp Thr Val1265 1270 1275 1280Leu Glu Gly Thr Glu Phe Lys Thr Thr Leu Ala Gly Ala Asp Ile Gln 1285 1290 1295Ala Gly Val Gly Glu Lys Ala Arg Ala Asp Ala Lys Ile Ile Leu Lys 1300 1305 1310Gly Ile Val Asn Arg Ile Gln Ser Glu Glu Lys Leu Glu Thr Asn Ser 1315 1320 1325Thr Val Trp Gln Lys Gln Ala Gly Arg Gly Ser Thr Ile Glu Thr Leu 1330 1335 1340Lys Leu Pro Ser Phe Glu Ser Pro Thr Pro Pro Lys Leu Thr Ala Pro1345 1350 1355 1360Gly Gly Tyr Ile Val Asp Ile Pro Lys Gly Asn Leu Lys Thr Glu Ile 1365 1370 1375Glu Lys Leu Ala Lys Gln Pro Glu Tyr Ala Tyr Leu Lys Gln Leu Gln 1380 1385 1390Val Ala Lys Asn Val Asn Trp Asn Gln Val Gln Leu Ala Tyr Asp Lys 1395 1400 1405Trp Asp Tyr Lys Gln Glu Gly Leu Thr Arg Ala Gly Ala Ala Ile Val 1410 1415 1420Thr Ile Ile Val Thr Ala Leu Thr Tyr Gly Tyr Gly Ala Thr Ala Ala1425 1430 1435 1440Gly Gly Val Ala Ala Ser Gly Ser Ser Thr Ala Ala Ala Ala Gly Thr 1445 1450 1455Ala Ala Thr Thr Thr Ala Ala Ala Thr Thr Val Ser Thr Ala Thr Ala 1460 1465 1470Met Gln Thr Ala Ala Leu Ala Ser Leu Tyr Ser Gln Ala Ala Val Ser 1475 1480 1485Ile Ile Asn Asn Lys Gly Asp Val Gly Lys Ala Leu Lys Asp Leu Gly 1490 1495 1500Thr Ser Asp Thr Val Lys Gln Ile Val Thr Ser Ala Leu Thr Ala Gly1505 1510 1515 1520Ala Leu Asn Gln Met Gly Ala Asp Ile Ala Gln Leu Asn Ser Lys Val 1525 1530 1535Arg Thr Glu Leu Phe Ser Ser Thr Gly Asn Gln Thr Ile Ala Asn Leu 1540 1545 1550Gly Gly Arg Leu Ala Thr Asn Leu Ser Asn Ala Gly Ile Ser Ala Gly 1555 1560 1565Ile Asn Thr Ala Val Asn 1570 180 281 PRT Artificial Sequence Description of Artificial Sequence ORF114 180Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln Ala Gly 1 5 10 15Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Xaa Xaa Xaa Xaa 20 25 30Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45Xaa Xaa Xaa Pro Ala His Ala Gln Ile Thr Thr Asp Lys Ser Ala Pro 50 55 60Lys Asn Gln Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala Pro Leu 65 70 75 80Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn Arg Xaa 85 90 95Tyr Ala Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn Asp Arg 100 105 110Asn Asn Asn Pro Phe Val Val Lys Gly Ser Ala Gln Leu Ile Leu Asn 115 120 125Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr Val Gly 130 135 140Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile Thr Val145 150 155 160Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr Thr Gly 165 170 175Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp Val Val 180 185 190Lys Ala His Trp Thr Val Xaa Ala Ala Gly Trp Asn Asp Lys Gly Gly 195 200 205Ala Xaa Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln Gly Lys 210 215 220Xaa Xaa Gly Lys Xaa Leu Ala Val Ser Thr Gly Pro Gln Lys Val Asp225 230 235 240Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr Lys Pro 245 250 255Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala Asp Ser 260 265 270Ile Thr Leu Ile Ala Asn Glu Lys Gly 275 280 181 302 PRT Artificial Sequence Description of Artificial Sequence overlap identity 181Ala Val Ala Glu Xaa Xaa Xaa Xaa Xaa Gly Lys Xaa Xaa Gln Xaa Xaa 1 5 10 15Xaa Xaa Xaa Ser Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser Xaa Xaa 20 25 30Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45Xaa Xaa Xaa Xaa Xaa Xaa Xaa Pro Ala Xaa Ala Xaa Xaa Ile Xaa Xaa 50 55 60Asp Lys Ser Ala Pro Lys Asn Gln Gln Xaa Val Ile Leu Xaa Thr Xaa 65 70 75 80Xaa Gly Xaa Pro Xaa Val Asn Ile Gln Thr Pro Xaa Xaa Xaa Gly Xaa 85 90 95Ser Xaa Asn Arg Xaa Xaa Xaa Phe Asp Val Asp Xaa Lys Gly Xaa Xaa 100 105 110Leu Asn Asn Xaa Arg Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 115 120 125Xaa Xaa Asn Pro Xaa Xaa Xaa Xaa Gly Xaa Ala Xaa Xaa Ile Xaa Asn 130 135 140Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser Xaa Leu Asn Gly Xaa Xaa Xaa Val145 150 155 160Gly Gly Xaa Xaa Ala Xaa Val Xaa Xaa Ala Asn Pro Xaa Gly Ile Xaa 165 170 175Val Asn Gly Gly Gly Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Leu Thr Xaa 180 185 190Gly Xaa Pro Xaa Xaa Xaa Xaa Xaa Gly Xaa Leu Thr Gly Phe Asp Val 195 200 205Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Asp Xaa Xaa 210 215 220Xaa Ala Xaa Tyr Thr Xaa Xaa Leu Xaa Arg Ala Xaa Xaa Xaa Xaa Xaa225 230 235 240Xaa Xaa Xaa Gly Lys Xaa Xaa Xaa Val Xaa Xaa Gly Xaa Xaa Lys Xaa 245 250 255Asp Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Xaa Xaa 260 265 270Xaa Xaa Xaa Xaa Pro Thr Xaa Ala Xaa Asp Thr Ala Xaa Leu Gly Gly 275 280 285Met Tyr Ala Asp Xaa Ile Thr Leu Ile Xaa Xaa Xaa Xaa Gly 290 295 300 182 300 PRT Artificial Sequence Description of Artificial Sequence pspA 182Ala Val Ala Glu Asn Val His Arg Asp Gly Lys Ser Met Gln Asp Ser 1 5 10 15Glu Ala Ala Ser Val Arg Val Thr Gly Ala Ala Ser Val Ser Ser Ala 20 25 30Arg Ala Ala Phe Gly Phe Arg Met Ala Ala Phe Ser Val Met Leu Ala 35 40 45Leu Gly Val Ala Ala Phe Ser Pro Ala Pro Ala Ser Gly Ile Ile Ala 50 55 60Asp Lys Ser Ala Pro Lys Asn Gln Gln Ala Val Ile Leu Gln Thr Ala 65 70 75 80Asn Gly Leu Pro Gln Val Asn Ile Gln Thr Pro Ser Ser Gln Gly Val 85 90 95Ser Val Asn Arg Phe Lys Gln Phe Asp Val Asp Glu Lys Gly Val Ile 100 105 110Leu Asn Asn Ser Arg Ser Asn Thr Gln Thr Gln Leu Gly Gly Trp Ile 115 120 125Gln Gly Asn Pro His Leu Ala Arg Gly Glu Ala Arg Val Ile Val Asn 130 135 140Gln Ile Asp Ser Ser Asn Pro Ser Leu Leu Asn Gly Tyr Ile Glu Val145 150 155 160Gly Gly Lys Arg Ala Glu Val Val Val Ala Asn Pro Ser Gly Ile Arg 165 170 175Val Asn Gly Gly Gly Leu Ile Asn Ala Ala Ser Val Thr Leu Thr Ser 180 185 190Gly Val Pro Val Leu Asn Asn Gly Asn Leu Thr Gly Phe Asp Val Ser 195 200 205Ser Gly Lys Val Val Ile Gly Gly Lys Gly Leu Asp Thr Ser Asp Ala 210 215 220Asp Tyr Thr Arg Ile Leu Ser Arg Ala Ala Glu Ile Asn Ala Gly Val225 230 235 240Trp Gly Lys Asp Val Lys Val Val Ser Gly Lys Asn Lys Leu Asp Phe 245 250 255Asp Gly Ser Leu Ala Lys Thr Ala Ser Ala Pro Ser Ser Ser Asp Ser 260 265 270Val Thr Pro Thr Val Ala Ile Asp Thr Ala Thr Leu Gly Gly Met Tyr 275 280 285Ala Asp Lys Ile Thr Leu Ile Ser Thr Asp Asn Gly 290 295 300 183 628 PRT Artificial Sequence Description of Artificial Sequence ORF114a 183Met Asn Lys Gly Leu His Arg Ile Ile Phe Ser Lys Lys His Ser Thr 1 5 10 15Met Val Ala Val Ala Glu Thr Ala Asn Ser Gln Gly Lys Gly Lys Gln 20 25 30Ala Gly Ser Ser Val Ser Val Ser Leu Lys Thr Ser Gly Asp Xaa Xaa 35 40 45Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gln Ile Thr Thr Asp Lys Ser 65 70 75 80Ala Pro Lys Asn Xaa Gln Val Val Ile Leu Lys Thr Asn Thr Gly Ala 85 90 95Pro Leu Val Asn Ile Gln Thr Pro Asn Gly Arg Gly Leu Ser His Asn 100 105 110Arg Tyr Thr Gln Phe Asp Val Asp Asn Lys Gly Ala Val Leu Asn Asn 115 120 125Asp Arg Asn Asn Asn Pro Phe Leu Val Lys Gly Ser Ala Gln Leu Ile 130 135 140Leu Asn Glu Val Arg Gly Thr Ala Ser Lys Leu Asn Gly Ile Val Thr145 150 155 160Val Gly Gly Gln Lys Ala Asp Val Ile Ile Ala Asn Pro Asn Gly Ile 165 170 175Thr Val Asn Gly Gly Gly Phe Lys Asn Val Gly Arg Gly Ile Leu Thr 180 185 190Ile Gly Ala Pro Gln Ile Gly Lys Asp Gly Ala Leu Thr Gly Phe Asp 195 200 205Val Arg Gln Gly Thr Leu Thr Val Gly Ala Ala Gly Trp Asn Asp Lys 210 215 220Gly Gly Ala Asp Tyr Thr Gly Val Leu Ala Arg Ala Val Ala Leu Gln225 230 235 240Gly Lys Leu Gln Gly Lys Asn Leu Ala Val Ser Thr Gly Pro Gln Lys 245 250 255Val Asp Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala Gly Thr 260 265 270Lys Pro Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met Tyr Ala 275 280 285Asp Ser Ile Thr Leu Ile Ala Xaa Glu Lys Gly Val Gly Val Lys Asn 290 295 300Ala Gly Thr Leu Glu Ala Ala Lys Gln Leu Ile Val Thr Ser Ser Gly305 310 315 320Arg Ile Glu Asn Ser Gly Arg Ile Ala Thr Thr Ala Asp Gly Thr Glu 325 330 335Ala Ser Pro Thr Tyr Leu Xaa Ile Glu Thr Thr Glu Lys Gly Ala Xaa 340 345 350Gly Thr Phe Ile Ser Asn Gly Gly Arg Ile Glu Ser Lys Gly Leu Leu 355 360 365Val Ile Glu Thr Gly Glu Asp Ile Xaa Leu Arg Asn Gly Ala Val Val 370 375 380Gln Asn Asn Gly Ser Arg Pro Ala Thr Thr Val Leu Asn Ala Gly His385 390 395 400Asn Leu Val Ile Glu Ser Lys Thr Asn Val Asn Asn Ala Lys Gly Ser 405 410 415Xaa Asn Leu Ser Ala Gly Gly Arg Thr Thr Ile Asn Asp Ala Thr Ile 420 425 430Gln Ala Gly Ser Ser Val Tyr Ser Ser Thr Lys Gly Asp Thr Xaa Leu 435 440 445Gly Glu Asn Thr Arg Ile Ile Ala Glu Asn Val Thr Val Leu Ser Asn 450 455 460Gly Ser Ile Gly Ser Ala Ala Val Ile Glu Ala Lys Asp Thr Ala His465 470 475 480Ile Glu Ser Gly Lys Pro Leu Ser Leu Glu Thr Ser Thr Val Ala Ser 485 490 495Asn Ile Arg Leu Asn Asn Gly Asn Ile Lys Gly Gly Lys Gln Leu Ala 500 505 510Leu Leu Ala Asp Asp Asn Ile Thr Ala Lys Thr Thr Asn Leu Asn Thr 515 520 525Pro Gly Asn Leu Tyr Val His Thr Gly Lys Asp Leu Asn Leu Asn Val 530 535 540Asp Lys Asp Leu Ser Ala Ala Ser Ile His Leu Lys Ser Asp Asn Ala545 550 555 560Ala His Ile Thr Gly Thr Ser Lys Thr Leu Thr Ala Ser Lys Asp Met 565 570 575Gly Val Glu Ala Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 580 585 590Xaa Ser Gly Asn Leu His Ile Gln Ala Ala Lys Gly Asn Ile Gln Leu 595 600 605Arg Asn Thr Lys Leu Asn Ala Ala Lys Ala Leu Glu Thr Thr Ala Leu 610 615 620Gln Gly Asn Ile625 184 663 PRT Artificial Sequence Description of Artificial Sequence homology 184Met Asn Lys Xaa Xaa Xaa Xaa Xaa Ile Phe Xaa Lys Lys Xaa Ser Xaa 1 5 10 15Met Xaa Ala Val Ala Glu Xaa Xaa Xaa Xaa Xaa Gly Lys Xaa Xaa Gln 20 25 30Xaa Xaa Xaa Xaa Xaa Ser Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 45Ser Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile 65 70 75 80Xaa Xaa Asp Lys Ser Ala Pro Lys Asn Xaa Gln Xaa Val Ile Leu Xaa 85 90 95Thr Xaa Xaa Gly Xaa Pro Xaa Val Asn Ile Gln Thr Pro Xaa Xaa Xaa 100 105 110Gly Xaa Ser Xaa Asn Arg Xaa Xaa Gln Phe Asp Val Asp Xaa Lys Gly 115 120 125Xaa Xaa Leu Asn Asn Xaa Arg Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa 130 135 140Xaa Xaa Xaa Xaa Asn Pro Xaa Leu Xaa Xaa Gly Xaa Ala Xaa Xaa Ile145 150 155 160Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser Xaa Leu Asn Gly Xaa Xaa 165 170 175Xaa Val Gly Gly Xaa Xaa Ala Xaa Val Xaa Xaa Ala Asn Pro Xaa Gly 180 185 190Ile Xaa Val Asn Gly Gly Gly Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Leu 195 200 205Thr Xaa Gly Xaa Pro Xaa Xaa Xaa Xaa Xaa Gly Xaa Leu Thr Gly Phe 210 215 220Asp Val Xaa Xaa Gly Xaa Xaa Xaa Xaa Gly Xaa Xaa Gly Xaa Xaa Asp225 230 235 240Xaa Xaa Xaa Ala Asp Tyr Thr Xaa Xaa Leu Xaa Arg Ala Xaa Xaa Xaa 245 250 255Xaa Xaa Xaa Xaa Xaa Gly Lys Xaa Xaa Xaa Val Xaa Xaa Gly Xaa Xaa 260 265 270Lys Xaa Asp Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa 275 280 285Xaa Xaa Xaa Xaa Xaa Xaa Pro Thr Xaa Ala Xaa Asp Thr Ala Xaa Leu 290 295 300Gly Gly Met Tyr Ala Asp Xaa Ile Thr Leu Ile Xaa Xaa Xaa Xaa Gly305 310 315 320Xaa Xaa Xaa Xaa Asn Xaa Gly Xaa Xaa Xaa Ala Ala Xaa Xaa Xaa Xaa 325 330 335Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Asn Ser Gly Xaa Ile Xaa Xaa 340 345 350Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr 355 360 365Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Ile 370 375 380Xaa Ser Xaa Xaa Xaa Xaa Val Xaa Xaa Xaa Xaa Xaa Xaa Ile Xaa Xaa385 390 395 400Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Gly Ser Xaa Xaa Xaa Xaa Xaa 405 410 415Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Ser Xaa Xaa Xaa Xaa 420 425 430Asn Asn Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser Xaa Xaa 435 440 445Xaa Xaa Xaa Xaa Xaa Asn Asp Xaa Xaa Xaa Xaa Ala Xaa Xaa Xaa Val 450 455 460Xaa Ser Xaa Xaa Xaa Xaa Asp Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa465 470 475 480Xaa Xaa Xaa Xaa Xaa Thr Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa 485 490 495Xaa Xaa Ile Xaa Ala Xaa Asp Thr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 500 505 510Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser Xaa Xaa Arg Xaa Xaa Xaa 515 520 525Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa 530 535 540Ile Thr Xaa Xaa Xaa Xaa Xaa Ala Lys Xaa Xaa Asn Xaa Xaa Thr Xaa545 550 555 560Gly Xaa Xaa Tyr Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp 565 570 575Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Ala 580 585 590Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Xaa Xaa Xaa 595 600 605Xaa Xaa Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 610 615 620Ser Gly Xaa Leu His Ile Xaa Xaa Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa625 630 635 640Gln Xaa Xaa Asn Thr Xaa Leu Xaa Asn Xaa Xaa Xaa Ala Xaa Glu Xaa 645 650 655Xaa Xaa Xaa Xaa Gly Asn Ile 660 185 622 PRT Artificial Sequence Description of Artificial Sequence pspA 185Met Asn Lys Arg Cys Tyr Lys Val Ile Phe Asn Lys Lys Arg Ser Cys 1 5 10 15Met Met Ala Val Ala Glu Asn Val His Arg Asp Gly Lys Ser Met Gln 20 25 30Asp Ser Glu Ala Ala Ser Val Arg Val Thr Gly Ala Ala Ser Val Ser 35 40 45Ser Ala Arg Ala Ala Phe Gly Phe Arg Met Ala Ala Phe Ser Val Met 50 55 60Leu Ala Leu Gly Val Ala Ala Phe Ser Pro Ala Pro Ala Ser Gly Ile 65 70 75 80Ile Ala Asp Lys Ser Ala Pro Lys Asn Gln Gln Ala Val Ile Leu Gln 85 90 95Thr Ala Asn Gly Leu Pro Gln Val Asn Ile Gln Thr Pro Ser Ser Gln 100 105 110Gly Val Ser Val Asn Arg Phe Lys Gln Phe Asp Val Asp Glu Lys Gly 115 120 125Val Ile Leu Asn Asn Ser Arg Ser Asn Thr Gln Thr Gln Leu Gly Gly 130 135 140Trp Ile Gln Gly Asn Pro His Leu Ala Arg Gly Glu Ala Arg Val Ile145 150 155 160Val Asn Gln Ile Asp Ser Ser Asn Pro Ser Leu Leu Asn Gly Tyr Ile 165 170 175Glu Val Gly Gly Lys Arg Ala Glu Val Val Val Ala Asn Pro Ser Gly 180 185 190Ile Arg Val Asn Gly Gly Gly Leu Ile Asn Ala Ala Ser Val Thr Leu 195 200 205Thr Ser Gly Val Pro Val Leu Asn Asn Gly Asn Leu Thr Gly Phe Asp 210 215 220Val Ser Ser Gly Lys Val Val Ile Gly Gly Lys Gly Leu Asp Thr Ser225 230 235 240Asp Ala Asp Tyr Thr Arg Ile Leu Ser Arg Ala Ala Glu Ile Asn Ala 245 250 255Gly Val Trp Gly Lys Asp Val Lys Val Val Ser Gly Lys Asn Lys Leu 260 265 270Asp Phe Asp Gly Ser Leu Ala Lys Thr Ala Ser Ala Pro Ser Ser Ser 275 280 285Asp Ser Val Thr Pro Thr Val Ala Ile Asp Thr Ala Thr Leu Gly Gly 290 295 300Met Tyr Ala Asp Lys Ile Thr Leu Ile Ser Thr Asp Asn Gly Ala Val305 310 315 320Ile Arg Asn Lys Gly Arg Ile Phe Ala Ala Thr Gly Gly Val Thr Leu 325 330 335Ser Ala Asp Gly Lys Leu Ser Asn Ser Gly Ser Ile Asp Ala Ala Glu 340 345 350Ile Thr Ile Ser Ala Gln Thr Val Asp Asn Arg Gln Gly Phe Ile Arg 355 360 365Ser Gly Lys Gly Ser Val Leu Lys Val Ser Asp Gly Ile Asn Asn Gln 370 375 380Ala Gly Leu Ile Gly Ser Ala Gly Leu Leu Asp Ile Arg Asp Thr Gly385 390 395 400Lys Ser Ser Leu His Ile Asn Asn Thr Asp Gly Thr Ile Ile Ala Gly 405 410 415Lys Asp Val Ser Leu Gln Ala Lys Ser Leu Asp Asn Asp Gly Ile Leu 420 425 430Thr Ala Ala Arg Asp Val Ser Val Ser Leu His Asp Asp Phe Ala Gly 435 440 445Lys Arg Asp Ile Glu Ala Gly Arg Thr Leu Thr Phe Ser Thr Gln Gly 450 455 460Arg Leu Lys Asn Thr Arg Ile Ile Gln Ala Gly Asp Thr Val Ser Leu465 470 475 480Thr Ala Ala Gln Ile Asp Asn Thr Val Ser Gly Lys Ile Gln Ser Gly 485 490 495Asn Arg Thr Gly Leu Asn Gly Lys Asn Gly Ile Thr Asn Arg Gly Leu 500 505 510Ile Asn Ser Asn Gly Ile Thr Leu Leu Gln Thr Glu Ala Lys Ser Asp 515 520 525Asn Ala Gly Thr Gly Arg Ile Tyr Gly Ser Arg Val Ala Val Glu Ala 530 535 540Asp Thr Leu Leu Asn Arg Glu Glu Thr Val Asn Gly Glu Thr Lys Ala545 550 555 560Ala Val Ile Ala Ala Arg Glu Arg Leu Asp Ile Gly Ala Arg Glu Ile 565 570 575Glu Asn Arg Glu Ala Ala Leu Leu Ser Ser Ser Gly Asp Leu His Ile 580 585 590Gly Ser Ala Leu Asn Gly Ser Arg Gln Val Gln Gly Ala Asn Thr Ser 595 600 605Leu His Asn Arg Ser Ala Ala Ile Glu Ser Ser Gly Asn Ile 610 615 620 186 400 PRT Artificial Sequence Description of Artificial Sequence ORF114a 186Leu Gln Gly Lys Leu Gln Gly Lys Asn Leu Ala Val Ser Thr Gly Pro 1 5 10 15Gln Lys Val Asp Tyr Ala Ser Gly Glu Ile Ser Ala Gly Thr Ala Ala 20 25 30Gly Thr Lys Pro Thr Ile Ala Leu Asp Thr Ala Ala Leu Gly Gly Met 35 40 45Tyr Ala Asp Ser Ile Thr Leu Ile Ala Xaa Glu Lys Gly Val Gly Val 50 55 60Lys Asn Ala Gly Thr Leu Glu Ala Ala Lys Gln Leu Ile Val Thr Ser 65 70 75 80Ser Gly Arg Ile Glu Asn Ser Gly Arg Ile Ala Thr Thr Ala Asp Gly 85 90 95Thr Glu Ala Ser Pro Thr Tyr Leu Xaa Ile Glu Thr Thr Glu Lys Gly 100 105 110Ala Xaa Gly Thr Phe Ile Ser Asn Gly Gly Arg Ile Glu Ser Lys Gly 115 120 125Leu Leu Val Ile Glu Thr Gly Glu Asp Ile Xaa Leu Arg Asn Gly Ala 130 135 140Val Val Gln Asn Asn Gly Ser Arg Pro Ala Thr Thr Val Leu Asn Ala145 150 155 160Gly His Asn Leu Val Ile Glu Ser Lys Thr Asn Val Asn Asn Ala Lys 165 170 175Gly Ser Xaa Asn Leu Ser Ala Gly Gly Arg Thr Thr Ile Asn Asp Ala 180 185 190Thr Ile Gln Ala Gly Ser Ser Val Tyr Ser Ser Thr Lys Gly Asp Thr 195 200 205Xaa Leu Gly Glu Asn Thr Arg Ile Ile Ala Glu Asn Val Thr Val Leu 210 215 220Ser Asn Gly Ser Ile Gly Ser Ala Ala Val Ile Glu Ala Lys Asp Thr225 230 235 240Ala His Ile Glu Ser Gly Lys Pro Leu Ser Leu Glu Thr Ser Thr Val 245 250 255Ala Ser Asn Ile Arg Leu Asn Asn Gly Asn Ile Lys Gly Gly Lys Gln 260 265 270Leu Ala Leu Leu Ala Asp Asp Asn Ile Thr Ala Lys Thr Thr Asn Leu 275 280 285Asn Thr Pro Gly Asn Leu Tyr Val His Thr Gly Lys Asp Leu Asn Leu 290 295 300Asn Val Asp Lys Asp Leu Ser Ala Ala Ser Ile His Leu Lys Ser Asp305 310 315 320Asn Ala Ala His Ile Thr Gly Thr Ser Lys Thr Leu Thr Ala Ser Lys 325 330 335Asp Met Gly Val Glu Ala Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 340 345 350Xaa Xaa Xaa Ser Gly Asn Leu His Ile Gln Ala Ala Lys Gly Asn Ile 355 360 365Gln Leu Arg Asn Thr Lys Leu Asn Ala Ala Lys Ala Leu Glu Thr Thr 370 375 380Ala Leu Gln Gly Asn Ile Val Ser Asp Gly Leu His Ala Val Ser Ala385 390 395 400 187 432 PRT Artificial Sequence Description of Artificial Sequence homology 187Leu Gln Gly Xaa Leu Gln Gly Lys Asn Xaa Xaa Xaa Xaa Xaa Gly Xaa 1 5 10 15Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Ile Xaa Ala Xaa Xaa Ala Xaa 20 25 30Xaa Xaa Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Xaa Xaa 35 40 45Xaa Xaa Xaa Ser Xaa Thr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Val 50 55 60Xaa Asn Xaa Gly Xaa Xaa Xaa Ala Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa 65 70 75 80Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Xaa Xaa Thr Ala 85 90 95Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Glu Xaa Thr Xaa 100 105 110Xaa Xaa Xaa Xaa Gly Xaa Thr Xaa Xaa Xaa Xaa Gly Gly Xaa Ile Xaa 115 120 125Ser Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Xaa Xaa Xaa 130 135 140Xaa Xaa Xaa Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr Xaa145 150 155 160Xaa Xaa Xaa Gly Xaa Asn Leu Xaa Xaa Xaa Xaa Lys Xaa Xaa Xaa Xaa 165 170 175Xaa Xaa Xaa Xaa Xaa Val Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Leu Xaa 180 185 190Ala Gly Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa Xaa Xaa Ala Gly Xaa 195 200 205Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Xaa Xaa Xaa Gly 210 215 220Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa225 230 235 240Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ile Xaa Xaa Xaa 245 250 255Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 260 265 270Thr Xaa Xaa Ser Xaa Xaa Xaa Xaa Asn Asn Xaa Xaa Xaa Lys Xaa Xaa 275 280 285Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Asn Xaa Xaa Xaa Lys Xaa Xaa 290 295 300Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Asp305 310 315 320Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Ser Xaa Xaa Xaa Xaa Xaa Xaa 325 330 335Xaa Ser Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa Thr 340 345 350Xaa Thr Xaa Xaa Thr Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Gly Xaa Xaa 355 360 365Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 370 375 380Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa385 390 395 400Xaa Xaa Xaa Asn Thr Xaa Xaa Xaa Ala Xaa Xaa Ala Xaa Xaa Xaa Xaa 405 410 415Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Ala 420 425 430 188 402 PRT Artificial Sequence Description of Artificial Sequence pspA 188Leu Gln Gly Asp Leu Gln Gly Lys Asn Ile Phe Ala Ala Ala Gly Ser 1 5 10 15Asp Ile Thr Asn Thr Gly Ser Ile Gly Ala Glu Asn Ala Leu Leu Leu 20 25 30Lys Ala Ser Asn Asn Ile Glu Ser Arg Ser Glu Thr Arg Ser Asn Gln 35 40 45Asn Glu Gln Gly Ser Val Arg Asn Ile Gly Arg Val Ala Gly Ile Tyr 50 55 60Leu Thr Gly Arg Gln Asn Gly Ser Val Leu Leu Asp Ala Gly Asn Asn 65 70 75 80Ile Val Leu Thr Ala Ser Glu Leu Thr Asn Gln Ser Glu Asp Gly Gln 85 90 95Thr Val Leu Asn Ala Gly Gly Asp Ile Arg Ser Asp Thr Thr Gly Ile 100 105 110Ser Arg Asn Gln Asn Thr Ile Phe Asp Ser Asp Asn Tyr Val Ile Arg 115 120 125Lys Glu Gln Asn Glu Val Gly Ser Thr Ile Arg Thr Arg Gly Asn Leu 130 135 140Ser Leu Asn Ala Lys Gly Asp Ile Arg Ile Arg Ala Ala Glu Val Gly145 150 155 160Ser Glu Gln Gly Arg Leu Lys Leu Ala Ala Gly Arg Asp Ile Lys Val 165 170 175Glu Ala Gly Lys Ala His Thr Glu Thr Glu Asp Ala Leu Lys Tyr Thr 180 185 190Gly Arg Ser Gly Gly Gly Ile Lys Gln Lys Met Thr Arg His Leu Lys 195 200 205Asn Gln Asn Gly Gln Ala Val Ser Gly Thr Leu Asp Gly Lys Glu Ile 210 215 220Ile Leu Val Ser Gly Arg Asp Ile Thr Val Thr Gly Ser Asn Ile Ile225 230 235 240Ala Asp Asn His Thr Ile Leu Ser Ala Lys Asn Asn Ile Val Leu Lys 245 250 255Ala Ala Glu Thr Arg Ser Arg Ser Ala Glu Met Asn Lys Lys Glu Lys 260 265 270Ser Gly Leu Met Gly Ser Gly Gly Ile Gly Phe Thr Ala Gly Ser Lys 275 280 285Lys Asp Thr Gln Thr Asn Arg Ser Glu Thr Val Ser His Thr Glu Ser 290 295 300Val Val Gly Ser Leu Asn Gly Asn Thr Leu Ile Ser Ala Gly Lys His305 310 315 320Tyr Thr Gln Thr Gly Ser Thr Ile Ser Ser Pro Gln Gly Asp Val Gly 325 330 335Ile Ser Ser Gly Lys Ile Ser Ile Asp Ala Ala Gln Asn Arg Tyr Ser 340 345 350Gln Glu Ser Lys Gln Val Tyr Glu Gln Lys Gly Val Thr Val Ala Ile 355 360 365Ser Val Pro Val Val Asn Thr Val Met Gly Ala Val Asp Ala Val Lys 370 375 380Ala Val Gln Thr Val Gly Lys Ser Lys Asn Ser Arg Val Asn Ala Met385 390 395 400Ala Ala 189 497 PRT Artificial Sequence Description of Artificial Sequence ORF116 189Glu Ala Val Gly Ser Asn Ile Gly Gly Gly Lys Met Ile Val Ala Ala 1 5 10 15Gly Gln Asp Ile Asn Val Arg Gly Xaa Ser Leu Ile Ser Asp Lys Gly 20 25 30Ile Val Leu Lys Ala Gly His Asp Ile Asp Ile Ser Thr Ala His Asn 35 40 45Arg Tyr Thr Gly Asn Glu Tyr His Glu Ser Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Arg Lys Xaa Xaa Xaa 65 70 75 80Xaa Xaa Xaa Arg Thr Asn Ile Val His Thr Gly Ser Ile Ile Gly Ser 85 90 95Leu Asn Gly Asp Thr Val Thr Val Ala Gly Asn Arg Tyr Arg Gln Thr 100 105 110Gly Ser Thr Val Ser Ser Pro Glu Gly Arg Asn Thr Val Thr Ala Lys 115 120 125Xaa Ile Asp Val Glu Phe Ala Asn Asn Arg Tyr Ala Thr Asp Tyr Ala 130 135 140His Thr Gln Glu Gln Lys Gly Leu Thr Val Ala Leu Asn Val Pro Xaa145 150 155 160Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Lys 165 170 175Ser Lys Asn Lys Arg Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp 180 185 190Gln Ser Tyr Gln Ala Thr Gln Gln Met Gln Gln Phe Ala Pro Ser Ser 195 200 205Ser Ala Gly Gln Gly Gln Asn Tyr Asn Gln Ser Pro Ser Ile Ser Val 210 215 220Ser Ile Xaa Tyr Gly Glu Gln Lys Ser Arg Asn Glu Gln Lys Arg His225 230 235 240Tyr Thr Glu Ala Ala Ala Ser Gln Ile Ile Gly Lys Gly Gln Thr Thr 245 250 255Leu Ala Ala Thr Gly Ser Gly Glu Gln Ser Asn Ile Asn Ile Thr Gly 260 265 270Ser Asp Val Ile Gly His Ala Gly Thr Xaa Leu Ile Ala Asp Asn His 275 280 285Ile Arg Leu Gln Ser Ala Lys Gln Asp Gly Ser Glu Gln Ser Lys Asn 290 295 300Lys Ser Ser Gly Trp Asn Ala Gly Val Arg Xaa Lys Ile Gly Asn Gly305 310 315 320Ile Arg Phe Gly Ile Thr Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 325 330 335Xaa Xaa Xaa Ser Thr Thr His Arg His Thr His Val Gly Ser Thr Thr 340 345 350Gly Lys Thr Thr Ile Arg Ser Gly Gly Asp Thr Thr Leu Lys Gly Val 355 360 365Gln Leu Ile Gly Lys Gly Ile Gln Ala Asp Thr Arg Asn Leu His Ile 370 375 380Glu Ser Val Gln Asp Thr Glu Thr Tyr Gln Ser Lys Gln Gln Asn Gly385 390 395 400Asn Val Gln Val Thr Val Gly Tyr Gly Phe Ser Ala Ser Gly Ser Tyr 405 410 415Arg Gln Ser Lys Val Lys Ala Asp His Ala Ser Val Thr Gly Gln Ser 420 425 430Gly Ile Tyr Ala Gly Glu Asp Gly Tyr Gln Ile Lys Val Arg Asp Asn 435 440 445Thr Asp Leu Lys Gly Gly Ile Ile Thr Ser Ser Gln Ser Ala Glu Asp 450 455 460Lys Gly Lys Asn Leu Phe Gln Thr Ala Thr Leu Thr Ala Ser Asp Ile465 470 475 480Gln Asn His Ser Arg Tyr Glu Gly Arg Ser Phe Gly Ile Gly Gly Ser 485 490 495Phe 190 502 PRT Artificial Sequence Description of Artificial Sequence overlap identity 190Xaa Ala Val Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Ile Xaa Xaa Xaa 1 5 10 15Gly Xaa Asp Ile Xaa Val Xaa Gly Xaa Xaa Xaa Ile Xaa Asp Xaa Xaa 20 25 30Xaa Xaa Leu Xaa Ala Xaa Xaa Xaa Ile Xaa Xaa Xaa Xaa Ala Xaa Xaa 35 40 45Arg Xaa Xaa Xaa Xaa Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 50 55 60Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Xaa Xaa Xaa 65 70 75 80Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Thr Xaa Ser Xaa Xaa Gly Ser 85 90 95Leu Asn Gly Xaa Thr Xaa Xaa Xaa Ala Gly Xaa Xaa Tyr Xaa Gln Thr 100 105 110Gly Ser Thr Xaa Ser Ser Pro Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa 115 120 125Xaa Ile Xaa Xaa Xaa Xaa Ala Xaa Asn Arg Tyr Xaa Xaa Xaa Xaa Xaa 130 135 140Xaa Xaa Xaa Glu Gln Lys Gly Xaa Thr Val Ala Xaa Xaa Val Pro Xaa145 150 155 160Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165 170 175Xaa Gly Lys Ser Lys Asn Xaa Arg Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa 180 185 190Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ala 195 200 205Xaa Xaa Pro Xaa Xaa Xaa Ala Gly Gln Gly Xaa Xaa Xaa Xaa Xaa Xaa 210 215 220Xaa Xaa Ile Ser Val Ser Xaa Xaa Tyr Gly Glu Gln Lys Xaa Xaa Xaa225 230 235 240Glu Xaa Xaa Xaa Xaa Xaa Thr Xaa Xaa Xaa Xaa Xaa Xaa Ile Xaa Gly 245 250 255Xaa Gly Xaa Xaa Xaa Leu Xaa Ala Xaa Gly Xaa Gly Xaa Xaa Ser Xaa 260 265 270Ile Xaa Ile Thr Gly Ser Asp Val Xaa Gly Xaa Xaa Gly Thr Xaa Leu 275 280 285Xaa Ala Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Gln Xaa Xaa Xaa 290 295 300Glu Xaa Ser Xaa Asn Lys Ser Xaa Gly Xaa Asn Ala Gly Val Xaa Xaa305 310 315 320Xaa Ile Xaa Xaa Gly Ile Xaa Phe Gly Xaa Thr Ala Xaa Xaa Xaa Xaa 325 330 335Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr Xaa Xaa Arg Xaa Xaa His 340 345 350Xaa Gly Ser Xaa Xaa Xaa Xaa Thr Xaa Ile Xaa Ser Gly Gly Asp Thr 355 360 365Xaa Xaa Lys Gly Xaa Gln Leu Xaa Gly Lys Gly Xaa Xaa Xaa Xaa Xaa 370 375 380Xaa Xaa Leu His Ile Glu Ser Xaa Gln Asp Thr Xaa Xaa Xaa Xaa Xaa385 390 395 400Lys Gln Xaa Asn Xaa Xaa Xaa Gln Val Thr Val Gly Tyr Gly Phe Ser 405 410 415Xaa Xaa Gly Ser Tyr Xaa Xaa Ser Lys Xaa Xaa Xaa Asp Xaa Ala Ser 420 425 430Val Xaa Xaa Gln Ser Gly Ile Xaa Ala Gly Xaa Asp Gly Tyr Xaa Ile 435 440 445Xaa Val Xaa Xaa Xaa Thr Xaa Leu Xaa Gly Xaa Xaa Xaa Xaa Ser Xaa 450 455 460Xaa Xaa Xaa Xaa Asp Lys Xaa Lys Asn Leu Xaa Xaa Thr Xaa Xaa Xaa465 470 475 480Xaa Xaa Xaa Asp Ile Gln Asn His Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 485 490 495Gly Xaa Xaa Gly Xaa Phe 500 191 491 PRT Artificial Sequence Description of Artificial Sequence pspA 191Gln Ala Val Ser Gly Thr Leu Asp Gly Lys Glu Ile Ile Leu Val Ser 1 5 10 15Gly Arg Asp Ile Thr Val Thr Gly Ser Asn Ile Ile Ala Asp Asn His 20 25 30Thr Ile Leu Ser Ala Lys Asn Asn Ile Val Leu Lys Ala Ala Glu Thr 35 40 45Arg Ser Arg Ser Ala Glu Met Asn Lys Lys Glu Lys Ser Gly Leu Met 50 55 60Gly Ser Gly Gly Ile Gly Phe Thr Ala Gly Ser Lys Lys Asp Thr Gln 65 70 75 80Thr Asn Arg Ser Glu Thr Val Ser His Thr Glu Ser Val Val Gly Ser 85 90 95Leu Asn Gly Asn Thr Leu Ile Ser Ala Gly Lys His Tyr Thr Gln Thr 100 105 110Gly Ser Thr Ile Ser Ser Pro Gln Gly Asp Val Gly Ile Ser Ser Gly 115 120 125Lys Ile Ser Ile Asp Ala Ala Gln Asn Arg Tyr Ser Gln Glu Ser Lys 130 135 140Gln Val Tyr Glu Gln Lys Gly Val Thr Val Ala Ile Ser Val Pro Val145 150 155 160Val Asn Thr Val Met Gly Ala Val Asp Ala Val Lys Ala Val Gln Thr 165 170 175Val Gly Lys Ser Lys Asn Ser Arg Val Asn Ala Met Ala Ala Ala Asn 180 185 190Ala Leu Asn Lys Gly Val Asp Ser Gly Val Ala Leu Tyr Asn Ala Ala 195 200 205Arg Asn Pro Lys Lys Ala Ala Gly Gln Gly Ile Ser Val Ser Val Thr 210 215 220Tyr Gly Glu Gln Lys Asn Thr Ser Glu Ser Arg Ile Lys Gly Thr Gln225 230 235 240Val Gln Glu Gly Lys Ile Thr Gly Gly Gly Lys Val Ser Leu Thr Ala 245 250 255Ser Gly Ala Gly Lys Asp Ser Arg Ile Thr Ile Thr Gly Ser Asp Val 260 265 270Tyr Gly Gly Lys Gly Thr Arg Leu Lys Ala Glu Asn Ala Val Gln Ile 275 280 285Glu Ala Ala Arg Gln Thr His Gln Glu Arg Ser Glu Asn Lys Ser Ala 290 295 300Gly Phe Asn Ala Gly Val Ala Ile Ala Ile Asn Lys Gly Ile Ser Phe305 310 315 320Gly Phe Thr Ala Gly Ala Asn Tyr Gly Lys Gly Tyr Gly Asn Gly Asp 325 330 335Glu Thr Ala Tyr Arg Asn Ser His Ile Gly Ser Lys Asp Ser Gln Thr 340 345 350Ala Ile Glu Ser Gly Gly Asp Thr Val Ile Lys Gly Gly Gln Leu Lys 355 360 365Gly Lys Gly Val Gly Val Thr Ala Glu Ser Leu His Ile Glu Ser Leu 370 375 380Gln Asp Thr Ala Val Phe Lys Gly Lys Gln Glu Asn Val Ser Ala Gln385 390 395 400Val Thr Val Gly Tyr Gly Phe Ser Val Gly Gly Ser Tyr Asn Arg Ser 405 410 415Lys Ser Ser Ser Asp Tyr Ala Ser Val Asn Glu Gln Ser Gly Ile Phe 420 425 430Ala Gly Gly Asp Gly Tyr Arg Ile Arg Val Asn Gly Lys Thr Gly Leu 435 440 445Val Gly Ala Ala Val Val Ser Asp Ala Asp Lys Ser Lys Asn Leu Leu 450 455 460Lys Thr Ser Glu Ile Trp His Lys Asp Ile Gln Asn His Ala Ser Ala465 470 475 480Ala Ala Ser Ala Leu Gly Leu Ser Gly Gly Phe 485 490 192 310 PRT Artificial Sequence Description of Artificial Sequence ORF41 192Tyr Arg Arg His Leu Leu Cys Lys Tyr Ile Tyr Arg Phe Pro Ile Tyr 1 5 10 15Cys Pro Xaa Ala Cys Val Ala Glu Asp Thr Pro Tyr Ala Cys Tyr Leu 20 25 30Xaa Gln Leu Gln Val Thr Lys Asp Val Asn Trp Asn Gln Val Xaa Leu 35 40 45Ala Tyr Asp Lys Trp Asp Tyr Lys Gln Glu Gly Leu Thr Gly Ala Gly 50 55 60Ala Ala Ile Ile Ala Leu Ala Val Thr Val Val Thr Ala Gly Ala Gly 65 70 75 80Ala Gly Ala Ala Leu Gly Leu Asn Gly Ala Ala Ala Ala Ala Thr Asp 85 90 95Ala Ala Phe Ala Ser Leu Ala Ser Gln Ala Ser Val Ser Leu Ile Asn 100 105 110Asn Lys Gly Asn Ile Gly Asn Thr Leu Lys Glu Leu Gly Arg Ser Ser 115 120 125Thr Val Lys Asn Leu Met Val Ala Val Ala Thr Ala Gly Val Ala Asp 130 135 140Lys Ile Gly Ala Ser Ala Leu Asn Asn Val Ser Asp Lys Gln Trp Ile145 150 155 160Asn Asn Leu Thr Val Asn Leu Ala Asn Ala Gly Ser Ala Ala Leu Ile 165 170 175Asn Thr Ala Val Asn Gly Gly Ser Leu Lys Asp Asn Leu Glu Ala Asn 180 185 190Ile Leu Ala Ala Leu Val Asn Thr Ala His Gly Glu Ala Ala Ser Lys 195 200 205Ile Lys Gln Leu Asp Gln His Tyr Ile Thr His Lys Ile Ala His Ala 210 215 220Ile Ala Gly Cys Ala Ala Ala Ala Ala Asn Lys Gly Lys Cys Gln Asp225 230 235 240Gly Ala Ile Gly Ala Ala Val Gly Glu Ile Val Gly Glu Ala Leu Thr 245 250 255Asn Gly Lys Asn Pro Asp Thr Leu Thr Ala Lys Glu Arg Glu Gln Ile 260 265 270Leu Ala Tyr Ser Lys Leu Val Ala Gly Thr Val Ser Gly Val Val Gly 275 280 285Gly Asp Val Asn Ala Ala Ala Asn Ala Ala Glu Val Ala Val Lys Asn 290 295 300Asn Gln Leu Ser Asp Lys305 310 193 330 PRT Artificial Sequence Description of Artificial Sequence ORF41a 193Tyr Leu Lys Gln Leu Gln Val Ala Lys Asn Ile Asn Trp Asn Gln Val 1 5 10 15Gln Leu Ala Tyr Asp Arg Trp Asp Tyr Lys Gln Glu Gly Leu Thr Glu 20 25 30Ala Gly Ala Ala Ile Ile Ala Leu Ala Val Thr Val Val Thr Ser Gly 35 40 45Ala Gly Thr Gly Ala Val Leu Gly Leu Asn Gly Ala Xaa Ala Ala Ala 50 55 60Thr Asp Ala Ala Phe Ala Ser Leu Ala Ser Gln Ala Ser Val Ser Phe 65 70 75 80Ile Asn Asn Lys Gly Asp Val Gly Lys Thr Leu Lys Glu Leu Gly Arg 85 90 95Ser Ser Thr Val Lys Asn Leu Val Val Ala Ala Ala Thr Ala Gly Val 100 105 110Ala Asp Lys Ile Gly Ala Ser Ala Leu Xaa Asn Val Ser Asp Lys Gln 115 120 125Trp Ile Asn Asn Leu Thr Val Asn Leu Ala Asn Ala Gly Ser Ala Ala 130 135 140Leu Ile Asn Thr Ala Val Asn Gly Gly Ser Leu Lys Asp Xaa Leu Glu145 150 155 160Ala Asn Ile Leu Ala Ala Leu Val Asn Thr Ala His Gly Glu Ala Ala 165 170 175Ser Lys Ile Lys Gln Leu Asp Gln His Tyr Ile Val His Lys Ile Ala 180 185 190His Ala Ile Ala Gly Cys Ala Ala Ala Ala Ala Asn Lys Gly Lys Cys 195 200 205Gln Asp Gly Ala Ile Gly Ala Ala Val Gly Glu Ile Val Gly Glu Ala 210 215 220Leu Thr Asn Gly Lys Asn Pro Asp Thr Leu Thr Ala Lys Glu Arg Glu225 230 235 240Gln Ile Leu Ala Tyr Ser Lys Leu Val Ala Gly Thr Val Ser Gly Val 245 250 255Val Gly Gly Asp Val Asn Ala Ala Ala Asn Ala Ala Glu Val Ala Val 260 265 270Lys Asn Asn Gln Leu Ser Asp Xaa Glu Gly Arg Glu Phe Asp Asn Glu 275 280 285Met Thr Ala Cys Ala Lys Gln Asn Xaa Pro Gln Leu Cys Arg Lys Asn 290 295 300Thr Val Lys Lys Tyr Gln Asn Val Ala Asp Lys Arg Leu Ala Ala Ser305 310 315 320Ile Ala Ile Cys Thr Asp Ile Ser Arg Ser 325 330 194 180 PRT Artificial Sequence Description of Artificial Sequence ORF51a 194Tyr Lys Leu Leu Ala Ile Gly Ser Val Val Gly Ser Ile Leu Gly Val 1 5 10 15Lys Leu Leu Leu Ile Leu Pro Val Ser Trp Leu Leu Leu Leu Met Ala 20 25 30Ile Ile Thr Leu Tyr Tyr Ser Val Asn Gly Ile Leu Asn Val Cys Ala 35 40 45Lys Ala Lys Asn Ile Gln Val Val Ala Asn Asn Lys Asn Met Val Leu 50 55 60Phe Gly Phe Leu Ala Gly Ile Ile Gly Gly Ser Thr Asn Ala Met Ser 65 70 75 80Pro Ile Leu Leu Ile Phe Leu Leu Ser Glu Thr Glu Asn Lys Asn Arg 85 90 95Ile Ala Lys Ser Ser Asn Leu Cys Tyr Leu Leu Ala Lys Ile Val Gln 100 105 110Ile Tyr Met Leu Arg Asp Gln Tyr Trp Leu Leu Asn Lys Ser Glu Tyr 115 120 125Gly Leu Ile Phe Leu Leu Ser Val Leu Ser Val Ile Gly Leu Tyr Val 130 135 140Gly Ile Arg Leu Arg Thr Lys Ile Ser Pro Asn Phe Phe Lys Met Leu145 150 155 160Ile Phe Ile Val Leu Leu Val Leu Ala Leu Lys Ile Gly Tyr Ser Gly 165 170 175Leu Ile Lys Leu 180 195 180 PRT Artificial Sequence Description of Artificial Sequence ORF82a 195Met Arg His Met Lys Asn Lys Asn Tyr Leu Leu Val Phe Ile Val Leu 1 5 10 15His Ile Thr Leu Ile Val Ile Asn Ile Val Phe Gly Tyr Phe Val Phe 20 25 30Leu Phe Asp Phe Phe Ala Phe Leu Phe Phe Ala Asn Val Phe Leu Ala 35 40 45Val Asn Leu Leu Phe Leu Glu Lys Asn Ile Lys Asn Lys Leu Leu Phe 50 55 60Leu Leu Pro Ile Ser Ile Ile Ile Trp Met Val Ile His Ile Ser Met 65 70 75 80Ile Asn Ile Lys Phe Tyr Lys Phe Glu His Gln Ile Lys Glu Gln Asn 85 90 95Ile Ser Ser Ile Thr Gly Val Ile Lys Pro His Asp Ser Tyr Asn Tyr 100 105 110Val Tyr Asp Ser Asn Gly Tyr Ala Lys Leu Lys Asp Asn His Arg Tyr 115 120 125Gly Arg Val Ile Arg Glu Thr Pro Tyr Ile Asp Val Val Ala Ser Asp 130 135 140Val Lys Asn Lys Ser Ile Arg Leu Ser Leu Val Cys Gly Ile His Ser145 150 155 160Tyr Ala Pro Cys Ala Asn Phe Ile Lys Phe Ala Lys Lys Pro Val Lys 165 170 175Ile Tyr Phe Tyr 180

Claims

1. An isolated or recombinant protein comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 4, and 6.

2. An isolated or recombinant protein comprising an immunogenic fragment having 60% or greater sequence identity to the contiguous sequence of amino acids depicted in SEQ ID NOS: 2, 4, or 6.

3. A composition comprising a protein of claim 1 or 2.

4. A diagnostic composition comprising a protein of claim 1 or 2.

5. A pharmaceutical composition according to claim 3.

6. An isolated or recombinant protein of claim 2, where in the protein has 80% or greater sequence identity to the contiguous sequence of amino acids depicted in SEQ ID NOS: 2, 4, or 6.

7. An isolated or recombinant protein of claim 2, wherein the protein has 90% or greater sequence identity to the contiguous sequence of amino acids depicted in SEQ ID NOS: 2, 4, or 6.

8. An isolated or recombinant protein of claim 2, wherein the protein has 95% or greater sequence identity to the contiguous sequence of amino acids depicted in SEQ ID NOS: 2, 4, or 6.

9. A pharmaceutical composition according to claim 4.