Fusion proteins for tuberculosis vaccines

Abstract

The present invention relates to fusion proteins based on Mycobacterium tuberculosis antigenic polypeptides for use in preventing, inhibiting, or treating infections and / or diseases caused by species of the tuberculosis complex. Specifically, the present invention relates to fusion proteins comprising antigens that do not elicit an immune response to BCG and / or ESAT-6 repeats. The fusion protein may comprise a combination of early and late antigens. In addition, the present invention relates to vaccines, immunogenic compositions, and pharmaceutical compositions comprising the fusion protein.

Description

Technical Field

[0001] The present invention relates to fusion proteins based on Mycobacterium tuberculosis antigenic polypeptides for use in preventing, inhibiting, or treating infections and / or diseases caused by species of the tuberculosis complex. Specifically, the present invention relates to fusion proteins comprising antigens that do not elicit an immune response against BCG and / or ESAT-6 repeats. The fusion protein may comprise a combination of early and late antigens. Furthermore, the present invention relates to vaccines, immunogenic compositions, and pharmaceutical compositions comprising the fusion protein. Background Art

[0002] According to the World Health Organization (WHO), human tuberculosis, caused by Mycobacterium tuberculosis (M. tuberculosis), is a serious global health problem, killing millions of people each year. New tuberculosis cases declined in the 1960s and 1970s, but this positive trend has been disrupted, in part due to the advent of HIV / AIDS and the emergence of multidrug-resistant strains of M. tuberculosis.

[0003] The only vaccine currently available for clinical use is BCG (Bacillus Calmette-Guérin), the efficacy of which remains controversial. BCG is a live attenuated vaccine that is widely used in infants in most areas where tuberculosis is endemic. BCG often induces high levels of acquired drug resistance in animal models of tuberculosis, and in children it protects against disseminated forms of tuberculosis, such as meningitis and miliary tuberculosis. When given to young children, it can prevent tuberculosis for many years, but then its efficacy declines. Comparisons of various controlled trials have shown that the protective efficacy of BCG in adults varies greatly, ranging from no protection to 80% protection. This makes the development of new, improved vaccines against Mycobacterium tuberculosis an urgent matter, and the WHO has made it a high priority.

[0004] Currently, several new tuberculosis (TB) vaccines are in clinical trials. Because a robust cellular immune response is generally accepted to prevent TB infection and disease, most current clinical TB vaccine candidates aim to induce classic TH1 cytokines, such as IFN-γ or TNF-α, from CD4+ or CD8+ T cells without addressing the question of improving T cell quality, which plays an important role in protective immunity. Current TB vaccine candidates include various vaccine platforms, such as inactivated whole cells or whole cell extracts, viral vector-based vaccines, live recombinant BCG or attenuated BCG vaccines, and DNA vaccines. However, to date, none of these vaccine platforms have produced convincing clinical results.

[0005] Alternative strategies for delivering effective tuberculosis vaccines revolve around subunit vaccines based on fusion proteins of Mycobacterium tuberculosis antigens. This subunit approach has been proposed to offer numerous advantages, such as improved safety and stability, as well as the ability to augment prior BCG vaccination. Efforts have been made to combine fusion proteins of multiple Mycobacterium tuberculosis antigens with suitable adjuvants to induce robust cellular immune responses.

[0006] WO2010006607A2 demonstrated that a simple fusion consisting of only two Mycobacterium tuberculosis antigens can function as a tuberculosis vaccine when administered with the adjuvant IC31. WO2006136162A2, WO2014063704A2, and WO2015161853A1 all disclose fusion proteins for combating tuberculosis infection and disease, in which multiple Mycobacterium tuberculosis antigens are combined to provide subunit vaccines capable of inducing a strong immune response. However, a common feature of these fusion proteins is that no corresponding subunit vaccine currently exists as a commercially available anti-tuberculosis vaccine.

[0007] Therefore, improved tuberculosis vaccines with the potential to produce clinically acceptable vaccines would be advantageous. In particular, fusion proteins that are able to induce strong, high-quality immune responses and thus have enhanced protective capabilities would be advantageous. Summary of the Invention

[0008] Provided herein are fusion proteins that are conceptually different from previously developed fusion proteins for preventing, inhibiting, or treating tuberculosis infection and / or disease. The fusion proteins described herein are based on antigens that do not elicit an immune response to BCG (referred to herein as "BCG - The fusion proteins provided herein generate an improved immune response compared to existing tuberculosis vaccines.

[0009] Therefore, one object of the present invention relates to providing a method comprising BCG - Fusion proteins of antigens and / or ESAT-6 repeats that induce strong cellular immune responses while improving the quality of T cells.

[0010] Another object of the present invention relates to providing a fusion protein comprising a combination of early and late tuberculosis antigens, which can both accelerate the immune response after Mtb infection and induce T cells with the potential to specifically recognize bacteria in the late chronic stage of infection.

[0011] Another object of the present invention is to provide fusion proteins that can be effectively used as stand-alone vaccines or in combination with BCG in vaccines or immunogenic compositions to prevent, inhibit or treat tuberculosis infection and / or disease.

[0012] Thus, one aspect of the present invention relates to a fusion protein comprising at least five antigens derived from Mycobacterium tuberculosis.

[0013] One embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises early and late antigens.

[0014] Another embodiment of the present invention relates to the fusion protein as described herein, wherein the antigen is absent from BCG, is not secreted, or has low expression.

[0015] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises at least two ESAT-6 repeats, such as at least three ESAT-6 repeats, such as at least four ESAT-6 repeats, such as at least five ESAT-6 repeats.

[0016] Another aspect of the present invention relates to a vaccine or immunogenic composition comprising a fusion protein as described herein.

[0017] Another aspect of the present invention is to provide a fusion protein as described herein or a vaccine or immunogenic composition as described herein for use in vaccinating or immunizing a subject against infection and / or disease caused by virulent mycobacteria.

[0018] Another aspect of the present invention is to provide a kit comprising:

[0019] i) a fusion protein as described herein or a vaccine or immunogenic composition as described herein,

[0020] ii) BCG, and

[0021] iii) Optionally, instructions for use.

[0022] Another aspect of the present invention is to provide a nucleic acid sequence comprising a sequence encoding the fusion protein as described herein.

[0023] Another aspect of the present invention is to provide a recombinant expression vector comprising a nucleotide sequence as described herein operably linked to one or more control sequences suitable for directing the production of a fusion protein in a suitable host.

[0024] Another aspect of the present invention is to provide a recombinant host cell comprising the expression vector described herein.

[0025] BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 shows immune recognition of selected antigens. Mycobacterium bovis (M. bovis) BCG Danish does not induce T cell responses against this antigen (BCG -(A) Mycobacterium bovis BCG Danish immunization, (B) Mycobacterium tuberculosis Erdman infection, or (C) immune responses after immunization with each antigen.

[0027] Figure 2 Show selected BCG-containing - Schematic diagram of the fusion protein of the antigen.

[0028] Figure 3 Schematic diagrams showing additional selected fusion proteins described herein.

[0029] Figure 4 shows the immunokinetic recognition of ESAT-6 and MPT70 during Mtb infection. (A) Antigen-specific interferon-γ release or (B) fold increase in the frequency of antigen-specific CD4 T cells was measured in the lungs 3, 12, and 20 weeks after Mtb Erdman infection.

[0030] Figure 5 shows that vaccines containing late-expressing antigens induce long-term protection. Lung CFUs from Mtb-infected animals immunized with H104 or H105 or injected with saline at (A) 4 and (B) 18 weeks after aerosol Mtb infection. H105 contains antigens MPT64, MPT70, and MPT83.

[0031] Figure 6 shows that co-administration of BCG and H107 / CAF01 increases specific H107 immune responses in two vaccination models. In (A) the standard prophylaxis model and (B) the BCG revaccination model, Mycobacterium bovis BCG Danish was administered 12 months before the H107 vaccine, and the frequency of H107-specific CD4 T cells in the spleen was measured three weeks after H107 / CAF01 immunization.

[0032] Figure 7 shows that BCG co-administration affects the differentiation of fusion protein-specific CD4 T cells, depending on whether the fusion protein contains BCG - Antigen (H107) or BCG + Antigen (H65). (A) Summary of T cell differentiation based on cytokine expression by single cells, adapted from Seder et al. (2008). (B) Cytokine expression profile of CD4 T cells located in the spleen based on single cell expression of the cytokines IFN-γ, TNF-α, and / or IL-2. (C) Frequency of CD4 T cells specific for the KLRG1 fusion protein.

[0033] Figure 8 shows that when BCG - Co-administration of a fusion protein of the antigen (H107) with BCG enhances protection against Mtb infection. Lung CFU at 4 weeks (A) and 18 weeks (B) after Mtb Erdman infection.

[0034] Figure 9 Co-vaccination with M. bovis BCG and H104-H107 fusion protein formulated in CAF01 was shown to induce significant protection against Mtb infection. Lung CFU at week 4 after Mtb Erdman infection are shown.

[0035] Figure 10 shows that adding free ESAT-6 protein to the vaccine formulation did not increase the immune response elicited by the vaccine nor reduce the number of mycobacteria after Mtb infection. The number of ESAT-6-specific CD4 T cells in the vaccinated group was measured (A) three weeks after vaccination and (B) three weeks after Mtb infection. (C) The number of mycobacteria in the lungs was measured six weeks after infection.

[0036] Figure 11 shows that repeating ESAT-6 in the fusion protein leads to an increased immune response against ESAT-6 and provides better protection in a prophylactic vaccination model. (A) Antigen-specific responses were measured in the spleen after immunization with H64 (one copy of ESAT-6) and H76 (five copies of ESAT-6) three weeks after immunization. (B) Mycobacterium counts were measured in the lungs six weeks after Mtb infection.

[0037] Figure 12 shows that repeating ESAT-6 in the fusion molecule improves the recruitment of ESAT-6-specific T cells to the site of infection and improves the protective efficacy of the vaccine in a post-exposure vaccination model. Animals infected with Mtb Erdman were treated with antibiotics. Two weeks before treatment termination, immunization with H83 / CAF01 (one copy of ESAT-6) and H84 / CAF01 (four copies of ESAT-6) was initiated. (A) Number of ESAT-6-specific CD4 T cells in the lungs two weeks after immunization. (B) Number of mycobacteria in the lungs 22 and 35 weeks after infection (6 and 19 weeks after the last immunization).

[0038] Figure 13 shows that repeating ESAT-6 in the fusion protein increases the immune response and improves the protective efficacy of the vaccine. Specific ESAT-6 responses in spleens isolated from (A) 129sc-vaccinated mice and (B) CB6F1 mice. (C) ESAT-6-specific immune responses in spleens from BCG- and subunit-co-immunized mice and protective efficacy in the lungs 4 weeks after infection.

[0039] Figure 14 The H107 fusion protein formulated in CAF01 adjuvant was shown to be a better vaccine than H56 formulated in CAF01. Protective efficacy (Δlog 10 CFU 生理盐水 -Δlog 10 CFU 免疫 ).

[0040] Figure 15 It was shown that three weeks after the first immunization (one week after the second dose of H107), an increased TB10.4 response was observed in the BCG+H107 group compared with BCG alone, which means that co-administration of H107+BCG increased the BCG-specific immune response and thus H107 played a role as a BCG adjuvant.

[0041] Figure 16 Schematic diagram showing the composition of H107 and H107e fusion proteins. Figure 16 A shows SDS-PAGE and western blots of H107 and H107e fusion protein expression from E. coli. H107e increased protein expression compared to H107.

[0042] Figure 17

[0043] After co-immunization with BCG, CD4 T cells expressing cytokines ( Figure 17 A, left) and IFNγ release detected by ELISA ( Figure 17 A, right) Immunogenicity was measured using both H107 and H107e. Figure 17 B shows that H107e induces immune responses to the same respective antigens as H107. Figure 17 C shows that after infection, H107e confers similar or better protection than BCG, and co-vaccination of BCG+H107e results in a significant increase in protection compared with BCG and H107e alone. Figure 17 D shows that in BCG-memory mice, H107e (BCG-) vaccination resulted in less differentiated (better quality) CD4 T cells compared with H65 (BCG+), as measured by functional differentiation score, FDS, and the proportion of IL-17-producing CD4 T cells.

[0044] The present invention will be described in more detail below. DETAILED DESCRIPTION

[0045] definition

[0046] Before discussing the present invention in further detail, the following terms and conventions are first defined:

[0047] antigen

[0048] In this context, the term "antigen" refers to a molecule capable of inducing an immune response, such as an immunogenic polypeptide. The immune response generated by an antigen can be B cell-driven (antibody-mediated immune response) and / or T cell-driven (cellular immune response).

[0049] The antigens described herein are derived from Mycobacterium tuberculosis (M. tuberculosis or Mtb).

[0050] Early and late antigens

[0051] The course of Mycobacterium tuberculosis infection essentially progresses through three phases: (i) an acute phase, (ii) a latent / chronic phase, and possibly (iii) a reactivation phase. The gene expression pattern of Mycobacterium tuberculosis changes during these phases.

[0052] Thus, in this context, the term "early antigen" refers to an antigen that is primarily expressed in the acute phase, whereas "late antigen" refers to an antigen that is primarily expressed in the latent / chronic phase.

[0053] Fusion protein

[0054] As used herein, the term "fusion protein" refers to a polypeptide comprising two or more antigens from Mycobacterium tuberculosis or its analogs in random order. These antigens can be fused together with or without amino acid linkers of varying lengths and sequences.

[0055] Fusion proteins can be produced by operably linking two or more heterologous nucleic acid sequences encoding the amino acid sequences of an antigen of interest. To avoid protein aggregation during downstream production, all cysteines in the fusion protein can be replaced with any amino acid, but serine is a preferred alternative due to its high structural similarity to cysteine.

[0056] Fusion proteins or antigens may contain appropriate purification tags (or affinity tags) to allow purification from crude biological sources (e.g., recombinant expression systems). Purification tags include, but are not limited to, His tags, chitin binding protein (CBP), maltose binding protein (MBP), and glutathione-S-transferase (GST).

[0057] Herein, the term "fusion protein" is used interchangeably with the term "subunit vaccine".

[0058] peptides

[0059] As used herein, the term "polypeptide" refers to a polymer composed of amino acid residues linked by peptide bonds, related naturally occurring structural variants, and / or synthetic non-naturally occurring analogs thereof. Conventional notation is used herein to describe polypeptide sequences: the left-hand end of a polypeptide sequence is the amino terminus (N-terminus); the right-hand end of a polypeptide sequence is the carboxyl terminus (C-terminus).

[0060] Polypeptides can be chemically modified by glycosylation, lipidation, prosthetic groups, or by the inclusion of additional amino acids, such as purification tags (e.g., His-tags) or signal peptides. Purification tags are used to obtain highly purified protein preparations. If used at the N-terminus, a His-tag can contain a methionine as the first amino acid, followed by 6-8 histidines, and if used at the C-terminus, 6-8 histidines followed by a stop codon. When used at the N-terminus, the methionine start codon in the gene encoding the polypeptide fusion can be deleted to avoid incorrect translation initiation sites.

[0061] Each polypeptide is encoded by a specific nucleic acid sequence. It should be understood that such sequences include analogs and variants thereof, wherein such nucleic acid sequences have been modified by substitution, insertion, addition, or deletion of one or more nucleic acids. Substitutions are preferably conservative substitutions in codon usage that do not result in any changes in the amino acid sequence, but may be introduced to enhance protein expression.

[0062] Polypeptides can be produced recombinantly or synthetically, for example, using an automated polypeptide synthesizer.

[0063] Mycobacterium tuberculosis

[0064] As used herein, the term "Mycobacterium tuberculosis" refers to the pathogenic species Mycobacterium tuberculosis of the family Mycobacteriaceae, which can be the causative agent of tuberculosis infection and disease. Mycobacterium tuberculosis may be abbreviated herein as Mtb.

[0065] As described herein, Mycobacterium tuberculosis infection progresses through three phases. As used herein, the term "infection" refers to any of these three phases, i.e., vaccination or immunization against disease caused by virulent mycobacteria may include an acute phase, a latent / chronic phase, and a reactivation phase.

[0066] Bacillus Calmette-Guérin (BCG)

[0067] As used herein, the term "Bacille Calmette-Guérin (BCG)" refers to a live, attenuated strain of tuberculosis bacteria derived from Mycobacterium bovis. BCG is currently the only commercially available vaccine against tuberculosis infection and disease.

[0068] BCG was developed approximately 100 years ago at the Pasteur Institute by attenuating Mycobacterium bovis. During this process, the strain lost a key gene segment encoding virulence-associated antigens, such as ESAT-6. This original mutation is known as RD1. Over the following 30–40 years, BCG vaccine was distributed to various laboratories and production facilities worldwide. This resulted in various vaccine substrains, often named after the location of the production laboratory (BCG Danish, Prague, Tokyo, etc.). Because many of these strains were originally propagated in continuous culture, numerous deletions from the original genome have been observed, with varying distributions among different substrains. A total of at least 12 major alterations / deletions (RD1-11 and SigK mutations) have been reported in analyzed BCG vaccine strains. Some of these deletions contain immunologically important antigens with vaccine potential, while others are immunologically silent.

[0069] The initial RD1 deletion is an example of a region containing immunodominant antigens that are very important for vaccines. This article discloses antigens that are strongly recognized during natural infection: RD1, RD2, and antigens whose expression is regulated by SigK. The coverage of antigens from all three regions provides enough antigens to construct a powerful polyprotein-based vaccine, and also provides antigens expressed at different stages of infection.

[0070] For the purposes of this article, BCG strains are divided into two main categories: early BCG strains and late BCG strains. Early and late BCG strains should not be confused with early and late antigens.

[0071] Early BCG strains lack the RD1 region. Early BCG strains include BCG Russia, BCG Japan, BCG Moreau, BCG Sweden, and BCG Birkhaug.

[0072] Late BCG strains lack the RD1 and RD2 regions and have mutations in sigK. Late BCG strains include BCGTice, BCG Frappier, BCG Pasteur, BCG Denmark, BCG Glaxo, BCG Prague, BCG China, and the genetically modified BCG strain VPM1002.

[0073] Late and early BCG strains do not share the same genotype and phenotype. Consequently, some antigens encoded by the RD2 region have been deleted in late BCG strains, while others are poorly expressed or not secreted. Consequently, vaccination with these late BCG strains does not induce an immune response to these deleted, poorly expressed, or non-secreted antigens.

[0074] BCG+ antigen

[0075] In this article, the term "BCG + "Antigen" refers to an antigen that is expressed by a given BCG strain and that elicits an immune response (measured by IFN-γ release in cultures of stimulated cells) following vaccination with a given BCG strain. + The fusion proteins of the antigens, when administered after an initial vaccination with an already administered BCG strain, can be used to enhance the immune response previously induced by the already administered BCG strain.

[0076] In this article, BCG + Antigens may be defined based on a related BCG strain, selected from BCG Danish, BCG Pasteur and BCG Prague and / or any derivative of these strains (e.g. VPM1002). + Antigens include, but are not limited to: Rv1886c (Ag85b), Rv3804c (Ag85a), Rv0288 (TB10.4), Rv0287 (EsxG), Rv3478 (PPE60), Rv0475 (HBHA), Rv3890c (EsxC), Rv3891c (EsxD), Rv1284 (CanA), Rv3019c (EsxR), Rv3020c (EsxS), Rv3017c (EsxQ), Rv2031c (HspX), Rv0983 (PepD), Rv1196 (PPE18), Rv2608 (PPE42), Rv3619 (EsxV) and Rv3620 (EsxW), and variants thereof.

[0077] BCG - antigen

[0078] In this article, the term "BCG - An "antigen" is an antigen that does not elicit an immune response (measured by IFN-γ release) after vaccination with the supplied BCG strain because it is either deleted, not secreted, or has low expression levels. - The fusion protein of the antigen, when administered after initial vaccination with an already administered BCG strain, does not enhance the immune response previously induced by the administered BCG strain. - Antigens can be defined as functionally negative.

[0079] In this article, BCG -Antigens may be defined according to related BCG strains, selected from BCG Danish, BCG Pasteur and BCG Prague and / or any derivatives of these strains (eg VPM 1002). Thus, BCG antigens may be defined as functional negatives related to these BCG strains.

[0080] Therefore, BCG - Antigens include, but are not limited to, Rv3875 (ESAT-6), Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC), Rv3616c (espA), Rv1980c (MPT64), Rv2875 (MPT70), Rv2873 (MPT83), and variants thereof.

[0081] BCG - Antigens can be used to elicit a complementary immune response, ie, an immune response directed against an Mtb antigen different from the antigen used in the initial BCG vaccination.

[0082] Immunogenic epitopes

[0083] An immunogenic epitope of a polypeptide is a portion of a polypeptide that elicits an immune response in an animal or human and / or in a biological sample as determined by any of the bioassays described herein. An immunogenic epitope of a polypeptide can be a T cell epitope or a B cell epitope. An immunogenic epitope can be associated with one or several relatively small portions of a polypeptide, they can be dispersed throughout the polypeptide sequence or located in a specific portion of the polypeptide. For some polypeptides, it has even been demonstrated that the epitopes are dispersed throughout the polypeptide, covering the entire sequence (Ravn, 1999).

[0084] To identify relevant T cell epitopes that are recognized during an immune response, a "brute force" approach can be used: because T cell epitopes are linear, deletion mutants of a polypeptide, if systematically constructed, will reveal which regions of the polypeptide are essential for immune recognition, for example, by subjecting these deletion mutants to an IFN-γ assay, such as described herein. Another approach utilizes overlapping peptides to detect MHC class II epitopes, preferably synthetic, of, for example, 20 amino acid residues from the polypeptide. These peptides can be tested in a bioassay (such as the IFN-γ assay described herein), and some will produce a positive reaction (and therefore be immunogenic), providing evidence for the presence of a T cell epitope in the peptide. To detect MHC class I epitopes, peptides that will bind can be predicted (Stryhn, 1996), then synthesized and tested in a relevant bioassay, such as the IFN-γ assay described herein. The peptides are preferably, for example, 8 to 11 amino acid residues from the polypeptide in length.

[0085] Although it has been shown that the minimum length of a T cell epitope is at least 6 amino acids, it is normal for such epitopes to be composed of longer amino acid fragments. Therefore, it is preferred that the length of the polypeptide fragments of the present invention has at least 7 amino acid residues, such as at least 8, at least 9, at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 22, at least 24 and at least 30 amino acid residues. Therefore, in an important embodiment of the present invention, it is preferred that the length of the polypeptide fragment is at most 50 amino acid residues, such as at most 40, 35, 30, 25 and 20 amino acid residues. It is expected that peptides with a length of 10 to 30 amino acid residues will prove to be the most effective MHC class II epitopes, and therefore the particularly preferred length of the polypeptide fragments used in the method of the present invention is 18, such as 15, 14, 13, 12 and even 11 amino acid residues. It is expected that peptides with a length between 7 and 12 amino acid residues will prove to be the most effective MHC class I epitopes, and therefore the particularly preferred length of the polypeptide fragments used in the method of the present invention is 11, such as 10, 9, 8 and even 7 amino acid residues.

[0086] An immunogenic portion of a polypeptide (a fragment comprising an immunogenic epitope), including the immunogenic epitope, may be recognized by a large portion (high frequency) or a small portion (low frequency) of a genetically heterogeneous population. In addition, some immunogenic portions induce a high immune response (dominant), while other immunogenic portions induce a lower but still significant response (subdominant). High frequency < low frequency may be associated with the immunogenic portion being bound by a wide distribution of MHC molecules (HLA types) or even multiple MHC molecules (Sinigaglia, 1988; Kilgus, 1991). Fragments comprising immunogenic epitopes from the polypeptide may exist as overlapping peptides of at least 10 amino acids in length, thereby spanning several epitopes.

[0087] Variants

[0088] As used herein, the term "variant" refers to an antigen, polypeptide, or fusion protein described herein that is "substantially homologous" and / or retains at least substantial immunogenicity of the referenced antigen, polypeptide, or fusion protein.

[0089] As used herein, the term "substantially homologous" refers to an antigen, polypeptide, or fusion protein having an amino acid sequence that has at least 80% sequence identity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity to the antigen, polypeptide, or fusion protein to which it is referenced. As used herein, the term "substantially immunogenic" refers to an antigen, polypeptide, or fusion protein that retains at least 80% immunogenicity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% immunogenicity compared to the antigen, polypeptide, or fusion protein to which it is referenced.

[0090] Thus, variants of an antigen, polypeptide, or fusion protein as described herein may have the same immunogenicity, or at least the same immunogenicity, as the antigen, polypeptide, or fusion protein to which it is referenced. Similarly, an antigen, polypeptide, or fusion protein having a defined sequence identity to an antigen, polypeptide, or fusion protein as described herein may have the same immunogenicity, or at least the same immunogenicity, as the antigen, polypeptide, or fusion protein to which it is referenced.

[0091] Antigenic repeats

[0092] As used herein, the term "antigenic repeat" refers to an antigen in which more than one copy of the antigen is present in the fusion protein. The different copies of the antigenic repeat can be positioned in the fusion protein (i) consecutively, (ii) alternating with antigens different from the antigenic repeat, or (iii) separated by antigens different from the antigenic repeat. The antigenic repeat can include at least two antigenic repeats, such as at least three antigenic repeats, such as at least four antigenic repeats, such as at least five antigenic repeats. Thus, a fusion protein having, for example, at least three antigenic repeats includes at least three copies of the antigen.

[0093] In this context, preferred are fusion proteins comprising antigenic repeats of ESAT-6 (ie ESAT-6 repeats).

[0094] linker molecules

[0095] As used herein, the term "linker molecule" or "linker" refers to a peptide sequence that appears between the antigens in a fusion protein. Linkers are typically composed of flexible residues such as glycine and serine, allowing the adjacent domains of the fusion protein to move freely relative to each other and fold independently and correctly during secretion / manufacturing. If necessary, longer linkers can be used to ensure that the two adjacent domains of the fusion protein do not interfere with each other sterically.

[0096] It should be understood that the linker at the genetic level is composed of nucleic acids. Therefore, the nucleic acid encoding the fusion protein described herein may include a linker represented by a nucleic acid sequence.

[0097] Co-inoculation

[0098] As used herein, the term "co-vaccination" refers to the simultaneous administration of two different vaccines and / or immunogenic compositions, for example, the simultaneous administration of a subunit vaccine (or fusion protein) as described herein and a BCG vaccine, or vaccination with BCG followed by subunit vaccination within the period of time during which BCG is expected to persist in the host.

[0099] The terms "co-vaccination," "co-immunization," and "co-administration" are used interchangeably herein.

[0100] Vaccines and immunogenic compositions

[0101] As used herein, the terms "vaccine" and "immunogenic composition" refer to a composition comprising at least one antigen that provides active acquired immunity to tuberculosis infection or disease. A "vaccine" or "immunogenic composition" may preferably include a fusion protein as described herein that provides active acquired immunity to tuberculosis infection or disease. Thus, following challenge with virulent mycobacteria, the vaccines or immunogenic compositions described herein can reduce bacterial loads in target organs, prolong survival, and / or reduce weight loss in animals compared to unvaccinated animals.

[0102] Vaccines or immunogenic compositions may include an immunologically and pharmaceutically acceptable carrier or vehicle. Suitable carriers include, but are not limited to, polymers to which polypeptides are bound via hydrophobic, non-covalent interactions, such as plastics, e.g., polystyrene; polymers to which polypeptides are covalently bound, such as polysaccharides; or polypeptides, such as bovine serum albumin, ovalbumin, or keyhole limpet hemocyanin. Suitable vehicles include, but are not limited to, diluents and suspending agents.

[0103] Additionally, the vaccine or immunogenic composition preferably includes one or more adjuvants.

[0104] adjuvant

[0105] As used herein, the term "adjuvant" refers to a compound or mixture that enhances the immune response to an antigen. Adjuvants can act as tissue depots that slowly release antigens and as lymphatic system activators, which nonspecifically enhance the immune response. Typically, without an adjuvant, a primary challenge with an antigen alone will fail to elicit either a humoral or cellular immune response.

[0106] Adjuvants include, but are not limited to, neutral adjuvant preparations, anionic adjuvant preparations, cationic adjuvant preparations (e.g., "CAF01," "CAF04," "CAF09," and "CAF10"), cationic liposomes (e.g., dimethyldioctadecyl ammonium bromide (DDA)), Quil A, QS21, poly I:C, aluminum hydroxide, Freund's incomplete adjuvant, IFN-γ, IL-2, IL-12, monophosphoryl lipid A (MPL), trehalose dimycolate (TDM), trehalose dibehenate (TDB), muramyl dipeptide (MDP), monobranched acylglycerol (MMG), CpG, and "IC31," or a combination thereof.

[0107] expression vector

[0108] In this article, the term "expression vector" refers to a DNA molecule used as a medium that recombinant genetic material is transferred to a host cell. The four main types of expression vectors are plasmids, bacteriophages and other viruses, cosmids and artificial chromosomes. The expression vector itself is typically a DNA sequence, which consists of an insert (heterogeneous nucleic acid sequence, transgene) and a larger sequence that serves as the "skeleton" of the expression vector. The purpose of the expression vector is to transfer genetic information to the host, typically to separate, propagate or express the insert in the target cell. The expression vector is particularly suitable for expressing heterologous sequences in the target cell and typically has a promoter sequence that drives the expression of the heterologous sequence.

[0109] operably connected

[0110] As used herein, the term "operably linked" refers to the connection of elements to form part of a functional unit (e.g., a gene or open reading frame). Thus, by operably linking a promoter to a nucleic acid sequence encoding a polypeptide, the two elements become part of a functional unit, the gene. The connection of an expression control sequence (promoter) to a nucleic acid sequence makes it possible for the promoter to direct transcription of the nucleic acid sequence. By operably linking two heterologous nucleic acid sequences encoding polypeptides, these sequences become part of a functional unit, the open reading frame; the open reading frame encodes a fusion protein comprising an amino acid sequence encoded by the heterologous nucleic acid sequence. By operably linking two amino acid sequences, these sequences become part of the same functional unit, the polypeptide. Operatively linking two heterologous amino acid sequences produces a hybrid (fusion) polypeptide.

[0111] mammal

[0112] As used herein, the term "mammal" refers to any animal belonging to the class Mammalia, including but not limited to rodents, primates, ungulates, and carnivores. Ungulates include but are not limited to cattle, horses, pigs, sheep, goats, and camels.

[0113] In this context, the mammal is preferably a human or a domestic animal.

[0114] Sequence identity

[0115] As used herein, the term "sequence identity" refers to sequence identity between genes or proteins at the nucleotide, base or amino acid level, respectively. Specifically, if the transcript of the DNA sequence can be transcribed into the same RNA sequence, then the DNA and RNA sequences are considered to be identical.

[0116] Thus, as used herein, "sequence identity" is a measure of identity between proteins at the amino acid level and a measure of identity between nucleic acids at the nucleotide level. When sequences are aligned, protein sequence identity can be determined by comparing the amino acid sequence at a given position in each sequence. Similarly, nucleic acid sequence identity can be determined by comparing the nucleotide sequence at a given position in each sequence when sequences are aligned.

[0117] To determine the percent identity of two amino acid sequences or two nucleic acids, the sequences are aligned for optimal comparison purposes (e.g., a gap can be introduced into the first amino acid sequence or nucleic acid sequence for optimal alignment with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules at that position are identical. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = # number of identical positions / # total number of positions (e.g., overlapping positions) × 100). In one embodiment, the two sequences are the same length.

[0118] In another embodiment, the two sequences have different lengths and gaps are considered different positions. One can manually align the sequences and count the number of identical amino acids. Alternatively, aligning two sequences to determine percent identity can be accomplished using a mathematical algorithm. This algorithm is incorporated into the NBLAST and XBLAST programs of (Altschul et al., 1990). BLAST nucleotide searches can be performed with the NBLAST program, score = 100, word length = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules of the present invention. BLAST protein searches can be performed with the XBLAST program, score = 50, word length = 3, to obtain amino acid sequences homologous to the protein molecules of the present invention.

[0119] In order to obtain sequence alignments that introduce gaps for comparison purposes, Gapped BLAST can be used. Alternatively, PSI-Blast can be used to perform an iterative search to detect the close or distant relationship between molecules. When using BLAST, XBLAST, and Gapped BLAST programs, the default parameters of each program can be used. See http: / / www.ncbi.nlm.nih.gov. Alternatively, sequence identity can be calculated after sequence alignment, for example, using the BLAST program in the EMBL database (www.ncbi.nlm.gov / cgi-bin / BLAST). Typically, the default settings for, for example, "scoring matrix" and "gap penalty" can be used for comparison. In the context of the present invention, the BLASTN and PSI BLAST default settings may be advantageous.

[0120] The percent identity between two sequences can be determined using techniques similar to those described above, with or without gaps. When calculating percent identity, only exact matches are counted. Thus, the sequences of the present invention to which one embodiment of the present invention relates have a certain degree of sequence variation.

[0121] Fusion proteins for tuberculosis vaccines

[0122] An estimated one-quarter of the world's population is infected with tuberculosis (TB), a disease that kills more than one million people annually. This makes TB the deadliest infectious disease worldwide. Despite this, no effective vaccine has been developed to replace and enhance the controversial and unreliable BCG vaccine developed a century ago. Strategies based on different platforms, such as inactivated whole cells or whole-cell extracts, viral vector-based vaccines, live recombinant BCG, attenuated BCG vaccines, or DNA vaccines, have largely failed to deliver encouraging results and hold the promise of an enhanced TB vaccine.

[0123] Another approach is to design fusion proteins, including antigens selected for the production of tuberculosis subunit vaccines. This type of tuberculosis vaccine is believed to have a series of advantages over other vaccine strategies, including improved vaccine safety and stability. By carefully designing fusion proteins, strong cellular immune responses of high-quality T cells can be induced. Therefore, the present invention provides fusion proteins that can be effectively used in vaccines or immunogenic compositions for the prevention, suppression or treatment of tuberculosis infection and / or disease.

[0124] Therefore, one aspect of the present invention relates to a fusion protein comprising at least two antigens derived from Mycobacterium tuberculosis.

[0125] The genome of Mycobacterium tuberculosis contains approximately 4000 genes, many of which encode proteins that are not suitable as antigens in subunit vaccines against tuberculosis infection or disease because they do not induce an immune response of sufficient strength or quality. Therefore, for immunogenicity, the antigens of the fusion proteins described herein are carefully selected, including but not limited to Rv3875 (ESAT-6), Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC), Rv3616c (espA), Rv1980c (MPT64), Rv2875 (MPT70), Rv2873 (MPT83), Rv1886c (Ag85b), Rv3804c (Ag85a), Rv0288 (TB10.4), Rv0287 (EsxG), Rv3478 (PPE60), Rv0475 (HBHA), Rv 3890c(EsxC), Rv3891c(EsxD), Rv1284(CanA), Rv3019c(EsxR), Rv3020c(EsxS), Rv3017c(EsxQ), Rv2031c(HspX), Rv0983(PepD), Rv1196(PP) E18), Rv2608(PPE42), Rv3619(EsxV), Rv3620(EsxW), Rv2660c, Rv3614(EspD), Rv3865(EspF), Rv3849(EspR), Rv3872(PE35) and Rv3881(EspB).

[0126] Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is selected from the group consisting of: SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70), SEQ ID NO: 8 (MPT83), SEQ ID NO: 10 (Ag85b), SEQ ID NO: 11 (Ag85a), SEQ ID NO: 12 (TB10.4), SEQ ID NO: 13 (EsxG), SEQ ID NO: 14 (PPE60), SEQ ID NO: 15 (HBHA), SEQ ID NO: 16 (EsxC), SEQ ID NO: 17 (EsxD), SEQ ID NO: 18 (CanA), SEQ ID NO: 19 (EsxR), SEQ ID NO: 20 (EsxS), SEQ ID NO: 21 (EsxQ), SEQ ID NO: 22 (EsxQ), SEQ ID NO: 23 (EsxQ), SEQ ID NO: 24 (EsxQ), SEQ ID NO: 25 (EsxQ), SEQ ID NO: 26 (EsxQ), SEQ ID NO: 27 (EsxQ), SEQ ID NO: 28 (EsxQ), SEQ ID NO: 29 (EsxR), SEQ ID NO: 30 (EsxS), SEQ ID NO: 31 (EsxQ), SEQ ID NO: 32 (EsxQ), SEQ ID NO: 33 (EsxQ), SEQ ID NO: 34 (EsxQ), SEQ ID NO: 35 (EsxQ), SEQ ID NO: 36 (Esx NO:22 (HspX), SEQ ID NO:23 (PepD), SEQ ID NO:24 (PPE18), SEQ ID NO:25 (PPE42), SEQ ID NO:26 (EsxV), SEQ ID NO:27 (EsxW), SEQ ID NO:28 (Rv2660c), SEQ ID NO:29 (Rv3614), SEQ ID NO:30 (Rv3865), SEQ ID NO:31 (Rv3849), SEQ ID NO:32 (Rv3872) and SEQ ID NO:33 (Rv3881), and variants thereof.

[0127] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises at least three antigens, such as at least four antigens, such as at least five antigens, such as at least six antigens, such as at least seven antigens, such as at least eight antigens.

[0128] The course of Mycobacterium tuberculosis infection generally progresses through three phases. During the acute phase, the bacteria proliferate in organs until an immune response mounts. Specifically sensitized CD4 T lymphocytes mediate control of the infection, with interferon gamma (IFN-γ) appearing to be the most important mediator. The bacterial load begins to decline, and a latent / chronic phase is established, in which the bacterial load remains stable at a low level. During this phase, Mycobacterium tuberculosis transitions from actively proliferating to dormancy, becoming essentially non-replicating and remaining within granulomas. In some cases, the infection enters a reactivation phase, in which dormant bacteria begin replicating again.

[0129] The transition from initial infection to latency in Mycobacterium tuberculosis is accompanied by changes in gene expression. The antigenic specificity of the immune response may also shift, as bacteria modulate gene expression during the transition from active replication to dormancy. The full nature of the immune response governing latent / chronic infection and the factors that lead to reactivation are not yet fully elucidated. However, some evidence suggests a shift in the primary cell type responsible. While CD4 T cells are essential and sufficient to control infection during the acute phase, studies suggest that CD8 T cell responses are more important during the latent / chronic phase.

[0130] Several new TB vaccines are currently in clinical trials. However, they are primarily classical prophylactic vaccines based on a limited number of antigens expressed early in the infection phase. However, as a direct result of these expression dynamics, the pattern of epitopes presented to T cells can change radically over time. Therefore, without being bound by theory, it is contemplated herein that a fusion protein comprising two antigens that are highly expressed in the early and late stages of infection, respectively, could be used in a potent vaccine that could (i) accelerate the immune response following Mtb infection and (ii) induce T cells that may specifically recognize the bacterium in the late chronic phase. Benefits of this fusion protein design also include improved epitope coverage and the ability to target both acute and latent / chronic infections.

[0131] Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises an early and a late antigen.

[0132] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the early antigen is selected from the group consisting of: Rv3875 (ESAT-6), Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC), Rv3616c (espA), Rv1886c (Ag85b), Rv3804c (Ag85a), Rv0288 (TB10.4), Rv0287 (EsxG), Rv3478 (PPE60), Rv0475 (HBHA), Rv3890c (EsxC), Rv 3891c (EsxD), Rv1284 (CanA), Rv3019c (EsxR), Rv3020c (EsxS), Rv3017c (EsxQ), Rv0983 (PepD), Rv1196 (PPE18), Rv2608 (PPE42), Rv3619 (EsxV), Rv3620 (EsxW), Rv3614 (EspD), Rv3865 (EspF), Rv3849 (EspR), Rv3872 (PE35) and Rv3881 (EspB), and their variants.

[0133] Another embodiment of the present invention relates to the fusion protein as described herein, wherein the late antigen is selected from the group consisting of Rv2875 (MPT70), Rv2873 (MPT83), Rv2031c (HspX) and Rv2660c, and variants thereof.

[0134] In countries plagued by tuberculosis, most of the population is vaccinated with BCG, a live, attenuated strain of tuberculosis bacteria derived from Mycobacterium bovis. A common strategy for enhancing protection against tuberculosis is to boost the immune response generated in response to the initial BCG vaccination. This can be achieved by administering tuberculosis antigens that are expressed by BCG and that already elicit an immune response after the initial BCG vaccination. Such antigens are referred to herein as BCG. + Antigens can be administered as part of a subunit vaccine with the goal of boosting the immune response induced by the initial BCG vaccination. However, mycobacterial infections, including BCG vaccination and Mtb infection, induce T cells of poor quality, and they have proven extremely difficult to reprogram after boosting with subunit vaccines, with disappointing results.

[0135] This article discloses a method that is conceptually different from the above-mentioned BCG + Fusion protein variants for boosting vaccines. Surprisingly, it has been found that tuberculosis antigens that are functionally negative for BCG (i.e., antigens that do not elicit an immune response following BCG vaccination) can be used in fusion proteins to provide significantly enhanced protection against Mycobacterium tuberculosis. Such antigens are referred to herein as BCG - The antigens can also be used in fusion proteins as part of a stand-alone vaccine or in combination with BCG to elicit a complementary immune response directed against a different BCG antigen than the antigen of the initial BCG vaccination.

[0136] Without being bound by theory, it is expected that at least part of the increased protection is due to the quality of the immune response induced by the fusion proteins disclosed herein. BCG vaccination is primarily a mycobacterial infection that, due to chronic antigenic stimulation, induces short-lived T cells that can only penetrate lung tissue and fight infection to a very poor extent. This footprint or immunological legacy left by the initial immune response to BCG vaccination is subsequently difficult to change. The fusion proteins disclosed herein are based on BCG. - Variant forms of the antigen that are not restricted by this immunological heritage and can therefore bypass T cell quality barriers; based on BCG + Traditional subunit vaccines of antigens are often limited by this obstacle.

[0137] Thus, the variants of the fusion proteins described herein are based on antigens that do not elicit an immune response against BCG, namely BCG - Antigens. The fusion proteins provided herein generate an improved immune response compared to existing tuberculosis vaccines.

[0138] Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein at least one antigen does not elicit an immune response against BCG. Another embodiment of the present invention relates to a fusion protein as described herein, wherein at least one antigen is functionally negative for BCG. Another embodiment of the present invention relates to a fusion protein as described herein, wherein at least one antigen is BCG. - antigen.

[0139] It is important to note that there are many different strains of BCG, and the genotypes and phenotypes of different strains vary. Therefore, to determine whether the antigen is BCG + or BCG - Antigens, herein with reference to late BCG strains, preferably BCG Danish, BCG Pasteur, BCG Prague or VPM1002. Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein BCG is selected from a late BCG strain, preferably BCG Danish, BCG Pasteur or BCG Prague. Another embodiment of the present invention relates to a fusion protein as described herein, wherein at least one antigen does not elicit an immune response against a late BCG strain. Another embodiment of the present invention relates to a fusion protein as described herein, wherein at least one antigen does not elicit an immune response against a BCG strain selected from BCG Danish, BCG Pasteur and BCG Prague.

[0140] Late BCG strains may also have the following characteristics: (i) some antigens encoded by the deleted RD2 region are missing; (ii) other antigens are poorly expressed or not secreted. Therefore, for these missing, poorly expressed or non-secreted antigens, no immune response will be induced after vaccination with late BCG strains. Therefore, one embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is missing, not secreted or has low expression from BCG. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is missing, not secreted or has low expression in late BCG strains, preferably the late BCG strains are selected from BCG Danish, BCG Pasteur and BCG Prague.

[0141] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is selected from the group consisting of an antigen located within RD1, an antigen located within RD2, an antigen whose expression is regulated by SigK, and combinations thereof.

[0142] Although the genome of Mycobacterium tuberculosis contains approximately 4000 genes, it has a large overlap of approximately 98% with the genome of BCG. - The number of antigens is limited to approximately 100 antigens against which BCG does not elicit a T cell response. BCG is identified herein as suitable for inclusion in a fusion protein for use in a vaccine against tuberculosis infection and / or disease. - Antigen. Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is selected from the group consisting of: Rv3875 (ESAT-6), Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC), Rv3616c (espA), Rv1980c (MPT64), Rv2875 (MPT70) and Rv2873 (MPT83), and variants thereof. It should be understood that variants of the antigens, polypeptides or fusion proteins described herein may have the same immunogenicity, i.e., they are equally effective in inducing an immune response, or at least have the same immunogenicity as the antigen, polypeptide or fusion protein to which they are based. Yet another embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is selected from the group consisting of:

[0143] a) an amino acid sequence selected from the group consisting of SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70) and SEQ ID NO: 8 (MPT83), and variants or immunogenic epitopes thereof, or

[0144] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0145] It has been found that some antigen fragments are advantageous. Therefore, one embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises an amino acid sequence selected from the group consisting of:

[0146] a) SEQ ID NO: 34 (H107b), SEQ ID NO: 35 (H107c) or SEQ ID NO: 91 (H107e), or a variant or immunogenic epitope thereof, or

[0147] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0148] A preferred embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises:

[0149] a) SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC) and SEQ ID NO: 5 (espA), or variants or immunogenic epitopes thereof, or

[0150] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0151] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises an amino acid sequence selected from the group consisting of:

[0152] a) SEQ ID NO: 36 (H106) or SEQ ID NO: 38 (H104), or a variant or immunogenic epitope thereof, or

[0153] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0154] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises:

[0155] a) SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70) and SEQ ID NO: 8 (MPT83), or variants or immunogenic epitopes thereof, or

[0156] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0157] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises an amino acid sequence selected from the group consisting of:

[0158] a) SEQ ID NO: 37 (H105) or a variant or immunogenic epitope thereof, or

[0159] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0160] Another embodiment of the present invention relates to a fusion protein as described herein, wherein the amino acid sequence of b) has at least 90% sequence identity, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity with any of the amino acid sequences of a). It should be understood that an antigen, polypeptide or fusion protein having a defined sequence identity with an antigen, polypeptide or fusion protein described herein may have the same immunogenicity, i.e., they are equally effective in inducing an immune response, or at least have the same immunogenicity as the antigen, polypeptide or fusion protein to which it refers.

[0161] As described herein, vaccination with both early and late antigens can be expected to produce an improved immune response. - Therefore, a preferred embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises an early antigen and a late antigen, neither of which elicits an immune response against BCG (i.e., BCG - Antigen). Another embodiment of the present invention relates to a fusion protein as described herein, wherein the late antigen that does not elicit an immune response to BCG is Rv2875 (MPT70) and / or Rv2873 (MPT83), and variants thereof. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the early antigen that does not elicit an immune response to BCG is selected from Rv3875 (ESAT-6), Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC) and Rv3616c (espA), and variants thereof.

[0162] Some variants of the fusion proteins disclosed herein are limited to containing only BCG- Antigen. Therefore, one embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises only antigens that do not elicit an immune response against BCG (i.e. BCG - antigen).

[0163] The fusion protein described herein can also be improved by changing the content of the 6kDa early secretory antigen target (ESAT-6). ESAT-6 is an important secretory protein and a potent T cell antigen of Mycobacterium tuberculosis. The ESAT-6 protein is constitutively expressed and secreted in large quantities by the bacteria and can be recognized after infection in humans and animals. Therefore, ESAT-6 is considered to be an important antigen and can be advantageously included in subunit vaccines for tuberculosis infection and disease. However, since the natural ESAT-6 molecule is a small protein with only 95 amino acids, the degree of immune response generated against ESAT-6 in humans and animals is relatively low.

[0164] Disclosed herein are variants of fusion proteins comprising multiple ESAT-6 repeats, whereby ESAT-6 relatively represents a larger portion of the fusion protein. Without being bound by theory, it is contemplated herein that increasing the relative amount of ESAT-6 antigen in the fusion protein mitigates the challenge of native ESAT-6 having low immunogenicity. For fusion proteins comprising more than one copy of ESAT-6, each occurrence of the antigen is referred to as an ESAT-6 repeat, e.g., a fusion protein comprising four copies of the ESAT-6 antigen can be said to comprise four ESAT-6 repeats. Therefore, one embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises at least two ESAT-6 repeats, e.g., at least three ESAT-6 repeats, e.g., at least four ESAT-6 repeats, e.g., at least five ESAT-6 repeats. A preferred embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises at least four ESAT-6 repeats. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises four ESAT-6 repeats. Another embodiment of the present invention relates to a fusion protein as described herein, wherein each ESAT-6 repeat is represented by SEQ ID NO: 1 or an amino acid sequence having at least 80% sequence identity (e.g., at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity) to SEQ ID NO: 1.

[0165] In principle, the ESAT-6 repeats can be arranged in any order in the fusion protein, for example, at the C-terminus, N-terminus, consecutively, alternately, or spaced apart. One embodiment of the present invention relates to a fusion protein as described herein, wherein the ESAT-6 repeats are separated by at least one antigen other than ESAT-6. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the ESAT-6 repeats are positioned alternately with an antigen other than ESAT-6.

[0166] Antigens suitable for use in fusion proteins containing ESAT-6 repeats are preferably highly immunogenic and are recognized in both humans and animals after tuberculosis infection. Therefore, antigens suitable for use in fusion proteins with ESAT-6 repeats include, but are not limited to: Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC), Rv3616c (espA), Rv1980c (MPT64), Rv2875 (MPT70), Rv2873 (MPT83), Rv1886c (Ag85b), Rv3804c (Ag85a), Rv0288 (TB10.4), Rv0287 (EsxG), Rv3478 (PPE60), Rv0475 (HBHA), Rv3890c (Es xC), Rv3891c(EsxD), Rv1284(CanA), Rv3019c(EsxR), Rv3020c(EsxS), Rv3017c(EsxQ), Rv2031c(HspX), Rv0983(PepD), Rv1196(PPE18) , Rv2608(PPE42), Rv3619(EsxV), Rv3620(EsxW), Rv2660c, Rv3614(EspD), Rv3865(EspF), Rv3849(EspR), Rv3872(PE35) and Rv3881(EspB).

[0167] Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen is selected from the group consisting of SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70), SEQ ID NO: 8 (MPT83), SEQ ID NO: 10 (Ag85b), SEQ ID NO: 11 (Ag85a), SEQ ID NO: 12 (TB10.4), SEQ ID NO: 13 (EsxG), SEQ ID NO: 14 (PPE60), SEQ ID NO: 15 (HBHA), SEQ ID NO: 16 (EsxC), SEQ ID NO: 17 (EsxD), SEQ ID NO: 18 (CanA), SEQ ID NO: 19 (EsxR), SEQ ID NO: 20 (EsxS), SEQ ID NO: 21 (EsxQ), SEQ ID NO: 22 (HspX), SEQ ID NO: 23 (HspX), SEQ ID NO: 24 (HspX), SEQ ID NO: 25 (HspX), SEQ ID NO: 26 (HspX), SEQ ID NO: 27 (HspX), SEQ ID NO: 28 (HspX), SEQ ID NO: 29 (HspX), SEQ ID NO: 30 (HspX), SEQ ID NO: 31 (HspX), SEQ ID NO: 32 (HspX), SEQ ID NO: 33 (HspX), SEQ ID NO: 34 (HspX), SEQ ID NO: 35 (HspX), SEQ ID NO: 36 (HspX), SEQ ID NO: 37 (HspX), SEQ ID NO: 38 (HspX), SEQ ID NO: 3 ID NO: 23 (PepD), SEQ ID NO: 24 (PPE18), SEQ ID NO: 25 (PPE42), SEQ ID NO: 26 (EsxV), SEQ ID NO: 27 (EsxW), SEQ ID NO: 28 (Rv2660c), SEQ ID NO: 29 (Rv3614), SEQ ID NO: 30 (Rv3865), SEQ ID NO: 31 (Rv3849), SEQ ID NO: 32 (Rv3872) and SEQ ID NO: 33 (Rv3881), and variants thereof, and wherein the fusion protein comprises at least two ESAT-6 repeats, such as at least three ESAT-6 repeats, such as at least four ESAT-6 repeats, such as at least five ESAT-6 repeats.

[0168] The fusion proteins described herein also include those containing both early and late antigens or BCG -A fusion protein comprising an antigen and an ESAT-6 repeat. Thus, a preferred embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises both early and late antigens, and at least two ESAT-6 repeats, such as at least three ESAT-6 repeats, such as at least four ESAT-6 repeats, such as at least five ESAT-6 repeats. Another preferred embodiment of the present invention relates to a fusion protein as described herein, wherein at least one antigen does not elicit an immune response against BCG, and wherein the fusion protein comprises at least two ESAT-6 repeats, such as at least three ESAT-6 repeats, such as at least four ESAT-6 repeats, such as at least five ESAT-6 repeats.

[0169] One embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises:

[0170] a) SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70) and SEQ ID NO: 8 (MPT83), or variants or immunogenic epitopes thereof, or

[0171] b) an amino acid sequence that has at least 80% sequence identity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity to any of the amino acid sequences of a), and

[0172] The fusion protein comprises four ESAT-6 repeats alternately positioned with an antigen other than ESAT-6.

[0173] Another particularly preferred embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises both early and late BCG - The antigen binds to the ESAT-6 repeats. A variant of this fusion protein is called H107 (SEQ ID NO: 9). Therefore, a preferred embodiment of the present invention relates to the fusion protein as described herein, wherein the fusion protein comprises the amino acid sequence represented by SEQ ID NO: 9. Another embodiment of the present invention relates to the fusion protein as described herein, wherein the fusion protein comprises:

[0174] a) SEQ ID NO: 9 (H107) or a variant or immunogenic epitope thereof, or

[0175] b) an amino acid sequence that has at least 80% sequence identity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity to any of the amino acid sequences of a), and

[0176] The amino acid sequence of b) has the same immunogenicity or at least the same immunogenicity as SEQ ID NO: 9.

[0177] A variant of the H107 fusion protein is referred to as H107e (SEQ ID NO: 91). Therefore, a preferred embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises the amino acid sequence represented by SEQ ID NO: 91. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises:

[0178] a) SEQ ID NO: 91 (H107e) or a variant or immunogenic epitope thereof, or

[0179] b) an amino acid sequence that has at least 80% sequence identity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity to any of the amino acid sequences of a), and

[0180] The amino acid sequence of b) has the same immunogenicity or at least the same immunogenicity as SEQ ID NO: 91.

[0181] The antigen of the fusion protein described herein may be linked via a linker, such as a peptide linker. Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein the antigen of the fusion protein is linked to a linker molecule.

[0182] In order to be able to produce fusion proteins or antigens, they may comprise appropriate purification tags to allow purification from, for example, recombinant expression systems. Thus, one embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises a purification tag. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the purification tag is selected from the group consisting of a His-tag, a chitin binding protein (CBP), a maltose binding protein (MBP) and a glutathione-S-transferase (GST). Another embodiment of the present invention relates to a fusion protein as described herein, wherein the purification tag is a His-tag.

[0183] Variants of the fusion protein may include BCG +Antigen. Therefore, one embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises at least one antigen that elicits an immune response against BCG. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the fusion protein comprises at least one BCG + Antigen. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the at least one antigen that elicits an immune response against BCG is selected from the group consisting of Rv1886c (Ag85b), Rv3804c (Ag85a), Rv0288 (TB10.4), Rv0287 (EsxG), Rv3478 (PPE60), Rv0475 (HBHA), Rv3890c (EsxC), Rv3 891c (EsxD), Rv1284 (CanA), Rv3019c (EsxR), Rv3020c (EsxS), Rv3017c (EsxQ), Rv2031c (HspX), Rv0983 (PepD), Rv1196 (PPE18), Rv2608 (PPE42), Rv3619 (EsxV) and Rv3620 (EsxW), and variants thereof. Another embodiment of the present invention relates to a fusion protein as described herein, wherein the at least one antigen that elicits an immune response against BCG is selected from the group consisting of:

[0184] a) an amino acid selected from the group consisting of SEQ ID NO: 10 (Ag85b), SEQ ID NO: 11 (Ag85a), SEQ ID NO: 12 (TB10.4), SEQ ID NO: 13 (EsxG), SEQ ID NO: 14 (PPE60), SEQ ID NO: 15 (HBHA), SEQ ID NO: 16 (EsxC), SEQ ID NO: 17 (EsxD), SEQ ID NO: 18 (CanA), SEQ ID NO: 19 (EsxR), SEQ ID NO: 20 (EsxS), SEQ ID NO: 21 (EsxQ), SEQ ID NO: 22 (HspX), SEQ ID NO: 23 (PepD), SEQ ID NO: 24 (PPE18), SEQ ID NO: 25 (PPE42), SEQ ID NO: 26 (EsxV) and SEQ ID NO: 27 (EsxW), and variants or immunogenic epitopes thereof, or

[0185] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0186] The fusion proteins disclosed herein are immunogenic and can be effectively used in vaccines or immunogenic compositions to prevent, inhibit or treat tuberculosis infection and / or disease. Such vaccines or immunogenic compositions can be used prophylactically or therapeutically. Thus, one aspect of the present invention relates to vaccines or immunogenic compositions comprising the fusion proteins described herein.

[0187] To ensure optimal performance of the vaccine or immunogenic composition, it is preferred that it comprises an immunologically and pharmaceutically acceptable carrier, vehicle, or adjuvant. Thus, one embodiment of the present invention relates to a vaccine or immunogenic composition as described herein, wherein the vaccine or immunogenic composition further comprises an immunologically and pharmaceutically acceptable carrier, vehicle, or adjuvant. Another embodiment of the present invention relates to a vaccine or immunogenic composition as described herein, wherein the vaccine or immunogenic composition further comprises one or more adjuvants. Another embodiment of the present invention relates to a vaccine or immunogenic composition as described herein, wherein the adjuvant is selected from a neutral adjuvant preparation, an anionic adjuvant preparation, a cationic adjuvant preparation, a cationic liposome (e.g., dimethyldioctadecyl ammonium bromide (DDA), Quil A, QS21, poly I:C, aluminum hydroxide, Freund's incomplete adjuvant, IFN-γ, IL-2, IL-12, monophosphoryl lipid A (MPL), trehalose dimycoate (TDM), trehalose dibehenate (TDB), muramyl dipeptide (MDP), monomycoacylglycerol (MMG), CpC and "IC31", or a combination thereof. One embodiment of the present invention relates to a vaccine or immunogenic composition as described herein, wherein the adjuvant is cationic adjuvant preparation 1 (CAF01). In another preferred embodiment, the adjuvant is cationic adjuvant preparation 10 (CAF10) comprising DDA, MMG and CpG. Other cationic adjuvant preparations that can be used as adjuvants in the vaccines or immunogenic compositions of the present invention are listed in Table 1 below:

[0188] Table 1 CAF adjuvant nomenclature

[0189]

[0190] Abbreviations: DDA: N,N-dimethyl-N,N-dioctadecyl ammonium (bromide salt). DSPC: 1,2-distearoyl-sn-glycero-3-phosphocholine. DSPE-PEG: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[carboxy(polyethylene glycol)-2000] (sodium salt). TDB: α,α-trehalose 6,6'-dibehenate. MMG: synthetic monobranched acylglycerol. Poly-IC: polyinosinic-polycytidylic acid (sodium salt). CpG: 5'-C-phospho-G-3' oligonucleotide. MPL: monophosphoryl lipid A.

[0191] The fusion proteins disclosed herein can be used as stand-alone vaccines or in combination with BCG vaccines.Thus, one embodiment of the present invention relates to a vaccine or immunogenic composition as described herein, wherein the vaccine or immunogenic composition further comprises BCG.

[0192] Another aspect of the invention relates to a fusion protein as described herein or a vaccine or immunogenic composition as described herein for use in vaccinating or immunizing a subject against infection and / or disease caused by virulent mycobacteria.

[0193] Toxicity mycobacteria can infect a wide variety of animals and, in some cases, can pose a challenge to, for example, farm animals. Thus, one embodiment of the present invention relates to a fusion protein, vaccine, or immunogenic composition for use as described herein, wherein the subject is a mammal. However, a primary purpose of providing the fusion proteins disclosed herein is to improve the potential for combating the current global challenge of tuberculosis infection and / or disease in humans. Thus, a preferred embodiment of the present invention relates to a fusion protein, vaccine, or immunogenic composition for use as described herein, wherein the mammal is a human.

[0194] Tuberculosis can be caused by different virulent mycobacteria. Therefore, one embodiment of the present invention relates to a fusion protein, vaccine or immunogenic composition for use as described herein, wherein the virulent mycobacterium is selected from the group consisting of Mycobacterium tuberculosis (M. tuberculosis), Mycobacterium bovis (M. bovis), Mycobacterium africanum (M. africanum), Mycobacterium canetti (M. canetti), and Mycobacterium microti (M. microti), preferably Mycobacterium tuberculosis (M. tuberculosis). A preferred embodiment of the present invention relates to a fusion protein, vaccine or immunogenic composition for use as described herein, wherein the virulent mycobacterium is Mycobacterium tuberculosis.

[0195] The mode of application can be varied. Any conventional method for administering a vaccine is suitable, including but not limited to oral formulations, suppositories, parenteral, and by injection, e.g., subcutaneous or intramuscular. One embodiment of the present invention relates to a fusion protein, vaccine, or immunogenic composition for use as described herein, wherein the fusion protein, vaccine, or immunogenic composition is administered by a route selected from the group consisting of oral, parenteral, subcutaneous, and intramuscular. The dosage of the vaccine will depend on the route of administration and will vary according to the age of the person to be vaccinated, and to a lesser extent, the size of the person to be vaccinated.

[0196] As described herein, the fusion protein, vaccine or immunogenic composition can be advantageously administered in combination with a BCG vaccine. Therefore, one embodiment of the present invention relates to a fusion protein, vaccine or immunogenic composition used as described herein, wherein BCG is administered before, simultaneously with or after the administration of the fusion protein, vaccine or immunogenic composition. Surprisingly, it has been found that when administered simultaneously with the fusion protein, vaccine or immunogenic composition disclosed herein, BCG can provide a strong adjuvant effect, producing a high-quality strong immune response. Therefore, a preferred embodiment of the present invention relates to a fusion protein, vaccine or immunogenic composition used as described herein, wherein the administration of BCG is carried out simultaneously with the administration of the fusion protein, vaccine or immunogenic composition. Another embodiment of the present invention relates to a fusion protein, vaccine or immunogenic composition used as described herein, wherein the subject has previously been vaccinated with a BCG vaccine.

[0197] Since the fusion protein, vaccine or immunogenic composition disclosed herein can act synergistically with the BCG vaccine, they can be conveniently provided together for ease of use. Therefore, one aspect of the present invention relates to a kit comprising:

[0198] i) a fusion protein as described herein or a vaccine or immunogenic composition as described herein,

[0199] ii) BCG, and

[0200] iii) Optionally, instructions for use.

[0201] One embodiment of the present invention relates to a kit as described herein, wherein i) and ii) are for simultaneous, separate or sequential administration.

[0202] The fusion proteins disclosed herein can be recombinantly produced by designing an expression vector construct encoding the fusion protein and introducing the expression vector into an appropriate recombinant expression system. Therefore, one aspect of the present invention relates to a nucleic acid sequence comprising a sequence encoding a fusion protein as described herein.

[0203] Another aspect of the present invention relates to a recombinant expression vector comprising a nucleotide sequence as described herein operably linked to one or more control sequences suitable for directing the production of a fusion protein in a suitable host.

[0204] Another aspect of the present invention relates to a recombinant host cell comprising an expression vector as described herein.

[0205] An expression vector suitable for producing a fusion protein disclosed herein may include a nucleic acid selected from the group consisting of SEQ ID NO: 46 (ESAT-6), SEQ ID NO: 47 (PPE68), SEQ ID NO: 48 (espI), SEQ ID NO: 49 (espC), SEQ ID NO: 50 (espA), SEQ ID NO: 51 (MPT64), SEQ ID NO: 52 (MPT70), SEQ ID NO: 53 (MPT83), SEQ ID NO: 55 (Ag85b), SEQ ID NO: 56 (Ag85a), SEQ ID NO: 57 (TB10.4), SEQ ID NO: 58 (EsxG), SEQ ID NO: 59 (PPE60), SEQ ID NO: 60 (HBHA), SEQ ID NO: 61 (EsxC), SEQ ID NO: 62 (EsxD), SEQ ID NO: 63 (CanA), SEQ ID NO: 64 (EsxR), SEQ ID NO: 65 (EsxS), SEQ ID NO: 66 (EsxS), SEQ ID NO: 67 (EsxA), SEQ ID NO: 68 (EsxB), SEQ ID NO: 69 (EsxC), SEQ ID NO: 70 (EsxD), SEQ ID NO: 71 (EsxB), SEQ ID NO: 72 (EsxC), SEQ ID NO: 73 (EsxD), SEQ ID NO: 74 (EsxR), SEQ ID NO: 75 (EsxS), SEQ ID NO: 76 (EsxB), SEQ ID NO: 77 (EsxB), SEQ ID NO: 78 (EsxC), SEQ ID NO: 79 (EsxD), SEQ ID NO: 80 (EsxB), SEQ ID NO: NO:66 (EsxQ), SEQ ID NO:67 (HspX), SEQ ID NO:68 (PepD), SEQ ID NO:69 (PPE18), SEQ ID NO:70 (PPE42), SEQ ID NO:71 (EsxV), SEQ ID NO:72 (EsxW), SEQ ID NO:73 (Rv2660c), SEQ ID NO:74 (Rv3614), SEQ ID NO:75 (Rv3865), SEQ ID NO:76 (Rv3849), SEQ ID NO:77 (Rv3872) and SEQ ID NO:78 (Rv3881), and variants thereof.

[0206] In addition, the selected fusion protein can be encoded by a nucleic acid selected from the group consisting of SEQ ID NO: 54 (H107), SEQ ID NO: 79 (H107b), SEQ ID NO: 80 (H107c), SEQ ID NO: 92 (H107e), SEQ ID NO: 81 (H106), SEQ ID NO: 82 (H105) and SEQ ID NO: 83 (H104), and variants thereof.

[0207] Table 2 summarizes the antigens and fusion proteins along with their sequence numbers and expression patterns:

[0208]

[0209]

[0210]

[0211] All patent and non-patent references cited in this application are hereby incorporated by reference in their entirety.

[0212] It should be noted that embodiments and features described in the context of one aspect of the invention also apply to other aspects of the invention. Embodiments and features of the invention are also summarized in the following items.

[0213] project

[0214] 1. A fusion protein comprising at least two antigens derived from Mycobacterium tuberculosis.

[0215] 2. The fusion protein according to item 1, wherein the fusion protein comprises at least three antigens, such as at least four antigens, such as at least five antigens, such as at least six antigens, such as at least seven antigens, such as at least eight antigens.

[0216] 3. The fusion protein according to any one of items 1 or 2, wherein the fusion protein comprises an early antigen and a late antigen.

[0217] 4. The fusion protein according to any one of the preceding items, wherein at least one antigen does not elicit an immune response against BCG.

[0218] 5. The fusion protein according to any one of the preceding items, wherein the antigen is absent from BCG, not secreted, or has low expression in BCG.

[0219] 6. The fusion protein according to any one of the preceding items, wherein the antigen is selected from:

[0220] Rv3875 (ESAT-6), Rv3873 (PPE68), Rv3876 (espI), Rv3615c (espC), Rv3616c (espA), Rv1980c (MPT64), Rv2875 (MPT70) and Rv2873 (MPT83), and variants thereof.

[0221] 7. The fusion protein according to any one of the preceding items, wherein the antigen is selected from:

[0222] a) an amino acid sequence selected from the group consisting of SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70) and SEQ ID NO: 8 (MPT83), and variants or immunogenic epitopes thereof, or

[0223] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0224] 8. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises:

[0225] a) SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC) and SEQ ID NO: 5 (espA), or variants thereof or immunogenic epitopes thereof, or

[0226] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0227] 9. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises:

[0228] a) SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70) and SEQ ID NO: 8 (MPT83), or variants or immunogenic epitopes thereof, or

[0229] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0230] 10. The fusion protein according to any one of items 7 to 9, wherein the amino acid sequence of b) has at least 90% sequence identity, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity with any of the amino acid sequences of a).

[0231] 11. The fusion protein according to any of the preceding items, wherein the fusion protein comprises at least two ESAT-6 repeats, such as at least three ESAT-6 repeats, such as at least four ESAT-6 repeats, such as at least five ESAT-6 repeats.

[0232] 12. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises at least four ESAT-6 repeats.

[0233] 13. The fusion protein according to any one of items 11 or 12, wherein the ESAT-6 repeats are separated by at least one antigen different from ESAT-6.

[0234] 14. The fusion protein according to any one of items 11 to 13, wherein the ESAT-6 repeats are positioned alternately with an antigen different from ESAT-6.

[0235] 15. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises:

[0236] a) SEQ ID NO: 1 (ESAT-6), SEQ ID NO: 2 (PPE68), SEQ ID NO: 3 (espI), SEQ ID NO: 4 (espC), SEQ ID NO: 5 (espA), SEQ ID NO: 6 (MPT64), SEQ ID NO: 7 (MPT70) and SEQ ID NO: 8 (MPT83), or variants or immunogenic epitopes thereof, or

[0237] b) an amino acid sequence that has at least 80% sequence identity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity to any of the amino acid sequences of a); and

[0238] The fusion protein comprises four ESAT-6 repeats alternately positioned with an antigen other than ESAT-6.

[0239] 19. The fusion protein according to any one of the preceding claims, wherein the fusion protein comprises an amino acid sequence selected from any one of the following:

[0240] a) SEQ ID NO: 9, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38 or SEQ ID NO: 91, or a variant or immunogenic epitope thereof; or

[0241] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0242] 20. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises the amino acid sequence represented by SEQ ID NO: 9 or SEQ ID NO: 91.

[0243] 21. The fusion protein according to any one of claims 1 to 3, wherein the fusion protein is encoded by a nucleic acid sequence selected from the group consisting of:

[0244] a) SEQ ID NO: 54 (H107), SEQ ID NO: 79 (H107b), SEQ ID NO: 80 (H107c), SEQ ID NO: 92 (H107e), SEQ ID NO: 81 (H106), SEQ ID NO: 82 (H105) and SEQ ID NO: 83 (H104), or variants or fragments thereof; or

[0245] b) a nucleic acid sequence having at least 80% sequence identity, such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity with any of the amino acid sequences of a).

[0246] 22. The fusion protein according to any one of the preceding items, wherein the antigen of the fusion protein is linked to a linker molecule.

[0247] 23. The fusion protein according to any one of the preceding items, wherein the fusion protein comprises at least one antigen that elicits an immune response against BCG.

[0248] 24. The fusion protein according to item 23, wherein the at least one antigen that triggers an immune response against BCG is selected from Rv1886c (Ag85b), Rv3804c (Ag85a), Rv0288 (TB10.4), Rv0287 (EsxG), Rv3478 (PPE60), Rv0475 (HBHA), Rv3890c (EsxC), Rv3891c (EsxD), Rv1284 (CanA), Rv3019c (EsxR), Rv3020c (EsxS), Rv3017c (EsxQ), Rv2031c (HspX), Rv0983 (PepD), Rv1196 (PPE18), Rv2608 (PPE42), Rv3619 (EsxV) and Rv3620 (EsxW), and variants thereof.

[0249] 25. The fusion protein according to any one of items 23 or 24, wherein the at least one antigen that elicits an immune response against BCG is selected from:

[0250] a) an amino acid selected from the group consisting of SEQ ID NO: 10 (Ag85b), SEQ ID NO: 11 (Ag85a), SEQ ID NO: 12 (TB10.4), SEQ ID NO: 13 (EsxG), SEQ ID NO: 14 (PPE60), SEQ ID NO: 15 (HBHA), SEQ ID NO: 16 (EsxC), SEQ ID NO: 17 (EsxD), SEQ ID NO: 18 (CanA), SEQ ID NO: 19 (EsxR), SEQ ID NO: 20 (EsxS), SEQ ID NO: 21 (EsxQ), SEQ ID NO: 22 (HspX), SEQ ID NO: 23 (PepD), SEQ ID NO: 24 (PPE18), SEQ ID NO: 25 (PPE42), SEQ ID NO: 26 (EsxV) and SEQ ID NO: 27 (EsxW), and variants or immunogenic epitopes thereof, or

[0251] b) an amino acid sequence that has at least 80% sequence identity with any of the amino acid sequences of a), such as at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity.

[0252] 26. A vaccine or immunogenic composition comprising the fusion protein of any one of the preceding items.

[0253] 27. The vaccine or immunogenic composition according to item 26, wherein the vaccine or immunogenic composition further comprises one or more adjuvants.

[0254] 28. The vaccine or immunogenic composition according to claim 27, wherein the adjuvant is selected from a neutral adjuvant preparation, an anionic adjuvant preparation, a cationic adjuvant preparation, a cationic liposome (e.g., dimethyldioctadecyl ammonium bromide (DDA), Quil A, QS21, poly I:C, aluminum hydroxide, Freund's incomplete adjuvant, IFN-γ, IL-2, IL-12, monophosphoryl lipid A (MPL), trehalose dimycoate (TDM), trehalose dibehenate (TDB), muramyl dipeptide (MDP), monobranched acylglycerol (MMG), CpG and "IC31", or a combination thereof.

[0255] 29. The vaccine or immunogenic composition according to any one of items 26 to 28, wherein the vaccine or immunogenic composition further comprises BCG.

[0256] 30. The fusion protein according to any one of items 1 to 25 or the vaccine or immunogenic composition according to any one of items 26 to 29 for use in vaccinating or immunizing a subject against infection and / or disease caused by virulent mycobacteria.

[0257] 31. The fusion protein, vaccine or immunogenic composition for use according to item 30, wherein the subject is a mammal.

[0258] 32. The fusion protein, vaccine or immunogenic composition for use according to item 31, wherein the mammal is a human.

[0259] 33. The fusion protein, vaccine or immunogenic composition for use according to any one of items 30 to 32, wherein the virulent mycobacterium is selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium canetti and Mycobacterium microti, preferably Mycobacterium tuberculosis.

[0260] 34. The fusion protein, vaccine or immunogenic composition for use according to any one of items 30 to 33, wherein BCG is administered before, simultaneously with or after the administration of the fusion protein, vaccine or immunogenic composition.

[0261] 35. The fusion protein, vaccine or immunogenic composition for use according to item 34, wherein BCG is administered simultaneously with the administration of the fusion protein, vaccine or immunogenic composition.

[0262] 36. The fusion protein, vaccine or immunogenic composition for use according to any one of items 30 to 35, wherein the subject has previously been vaccinated with BCG.

[0263] 37. A kit comprising:

[0264] i) a fusion protein according to any one of items 1 to 25 or a vaccine or immunogenic composition according to any one of items 26 to 29,

[0265] ii) BCG, and

[0266] iii) Optionally, instructions for use.

[0267] 38. The kit according to item 37, wherein i) and ii) are for simultaneous, separate or sequential administration.

[0268] 39. A nucleic acid sequence comprising a sequence encoding the fusion protein according to any one of items 1 to 25.

[0269] 40. A recombinant expression vector comprising the nucleotide sequence of item 39, which is operably linked to one or more control sequences suitable for directing the production of a fusion protein in a suitable host.

[0270] 41. A recombinant host cell comprising the expression vector according to item 40.

[0271] The invention will now be described in more detail in the following non-limiting examples.

[0272] Example

[0273] Example 1: BCG vaccination does not elicit immunity against Mycobacterium tuberculosis antigens of the H104-H107 fusion protein Epidemic Response

[0274] Materials and methods

[0275] Six- to ten-week-old female CB6F1 mice were immunized with Mycobacterium bovis (BCG) Danish, infected with 25-50 CFU of virulent Mtb Erdman via aerosol, or immunized three times with 2 μg of a single protein formulated in a total volume of 200 μL of cationic adjuvant formulation 1 (CAF01). Each protein vaccine was administered two weeks apart. Single-cell suspensions of splenocytes were obtained from each animal by passing the spleen through a 100 μm cell strainer 10 weeks after BCG immunization, 18 weeks after Mtb infection, or 2 weeks after the last single-protein immunization. The cells were washed twice in RPMI, and the number of cells obtained from each spleen was determined and adjusted to 2 × 10 6 To stimulate cells, 100 μL (2×10 5Cells were aliquoted into each well of a 96-well plate and purified recombinant single Mtb antigens (2 μg / ml) were added. The total volume per well was adjusted to 200 μL using RPMI medium, and the cells were then incubated with the respective antigens at 37°C in a CO2 incubator for 3-5 days. At this time point, the concentration of the cytokine IFN-γ in the cell supernatant was measured by sandwich ELISA.

[0276] result

[0277] A panel of potential candidate antigens was selected for design of non-BCG (BCG - ) antigen, and study whether they cause immune response after BCG vaccination. The splenocytes isolated from mice were stimulated with the specified single antigen (2 μg / ml) purified as recombinant protein (see also Example 2). After stimulation, the cytokine IFN-γ released by the cells into the culture medium was measured. The cytokine IFN-γ concentration measured in the culture medium of cells incubated without antigen ("none") established the baseline of non-stimulated cells. In animals immunized with BCG, none of the candidate antigens caused an immune response (Figure 1A). On the contrary, the TB10.4 antigen expressed by BCG caused a significant immune response (Figure 1A). In animals infected with Mtb, all antigens caused an immune response, confirming that these antigens are immunogenic and expressed during Mtb infection (Figure 1B). Similarly, in animals vaccinated with single proteins, all antigens caused an immune response, showing that these antigens are immunogenic (Figure 1C) in the CB6F1 mouse strain.

[0278] in conclusion

[0279] Immunization with Mycobacterium bovis BCG Danish did not induce a cellular immune response against any of the eight candidate vaccine antigens. In contrast, all candidate antigens elicited an immune response following either Mtb infection or single protein immunization. Thus, antigens H104-H107 do not share antigens with M. bovis BCG Danish and are specific for Mtb. Such antigens are referred to herein as BCG. - In contrast, those antigens to which BCG immunization elicits a T cell response are referred to herein as BCG + Antigens, such as TB10.4.

[0280] Example 2: Construction of fusion protein

[0281] Materials and Methods

[0282] Genes encoding single and fusion proteins were synthesized based on protein sequences translated from the published genomic sequence of Mycobacterium tuberculosis strain H37Rv. The synthesized DNA sequences were codon-optimized for expression in E. coli and cloned into the pJ411 expression vector under the control of the inducible T7 promoter and transformed into E. coli. The pJ411 vector encodes an N-terminal His tag in frame with the coding sequence for the inserted recombinant protein. For each protein, the protein was expressed in vitro when the culture reached an OD value of ∼0.5. 600 When the density of the His-tagged recombinant protein is reached, expression of the His-tagged recombinant protein is induced. The next day, the bacteria are harvested, lysed, and the recombinant protein is purified using a three-step purification process. This process involves removal of soluble E. coli proteins from the precipitated recombinant protein (inclusion bodies), immobilized metal affinity chromatography, and anion or cation exchange depending on the protein charge. After purification, the identity of the purified protein is confirmed by mass spectrometry, and purity is confirmed by scanning Coomassie-stained SDS-gels. Finally, the protein concentration is determined.

[0283] result

[0284] Based on the results of Example 1, a BCG - The selection of recombinant fusion protein is as follows ( Figure 2 and 3 ). Yields of purified fusion protein from a 6-L starter culture ranged from 0.6 mg to 20 mg. Concentrations of purified protein varied from 0.2 mg / ml to 0.7 mg / ml. Purity was 95% or greater.

[0285] in conclusion

[0286] All single and fusion proteins were produced in sufficient quantity and quality to be suitable antigens for use in vaccines and to perform all required immune and cellular stimulations described in the following examples.

[0287] Example 3: MTP70 is an antigen that is upregulated during late Mtb infection (i.e., a "late antigen")

[0288] Materials and Methods

[0289] Female CB6F1 mice were infected with 25-50 CFU of virulent Mtb Erdman aerosol. Mice were sacrificed after three, twelve, and twenty weeks for immunoassay analysis. Lungs were gently homogenized using a T-tube GentleMACS (Miltenyi Biotec) and digested with collagenase IV (Sigma-Aldrich) at 37°C for 30-60 minutes. Single cell suspensions were obtained by passing the tissue through a 100 μm cell strainer. Cells were washed twice in RPMI and then subjected to antigen stimulation for intracellular cytokine staining (ICS) and ELISA.

[0290] ICS : 1-2×10 6 Lung cells were stimulated in vitro for 1 hour in RPMI + 10% FCS containing 1 μg / ml anti-CD28 and anti-CD49d, with or without 5 μg ESAT-6 or MPT70 pepmix, and then incubated in the presence of 10 μg / ml brefeldin A (Sigma-Aldrich) at 37°C in an automated heating pad for 5 hours. After incubation, the cells were cooled to 4°C. The next day, cells were washed in FACS buffer (1× PBS containing 1% FCS) and stained for surface markers using anti-CD3, anti-CD4, and anti-CD44 at 4°C. After 15-30 minutes of surface staining, cells were washed in FACS buffer and permeabilized using the Cytofix / Cytoperm kit (BD) according to the manufacturer's instructions, followed by intracellular staining with anti-IFN-γ, anti-TNF-α, anti-IL-2, and anti-IL-17A. The cells were then washed, resuspended in FACS buffer and analyzed using an LSR FACSFortessa (BD).All flow cytometric data were analyzed using FlowJo software v.10 (TreeStar, Ashland, OR, USA).

[0291] ELISA Lung cells were stimulated with ESAT-6 or MPT70 pepmix at 37°C and 5% CO2. Three days later, secreted IFN-γ was measured by sandwich ELISA of culture supernatants.

[0292] result

[0293] Three weeks after Mtb infection, there was a significant ESAT-6 response, as measured by both IFN-γ ELISA (Figure 4A) and cytokine-secreting CD4 T cells (Figure 4B). However, at this time point, there was no immune response to MPT70. At 12 weeks after infection, there was a small but detectable immune response to MPT70, which increased at week 20. This supports ESAT-6 as an "early antigen" and MPT70 as a "late antigen," characterized by immune recognition in the late / chronic phase of infection.

[0294] in conclusion

[0295] Immune response kinetics against ESAT-6 and MPT70 support: in BCG - Among the antigens, ESAT-6 is an “early antigen”; whereas MPT70 is a “late antigen” because its expression is upregulated during late infection.

[0296] Example 4: Late antigens improve long-term protection

[0297] Materials and Methods

[0298] Each group of mice was immunized three times with 2 μg of the fusion protein in a total volume of 200 μL of cationic adjuvant formulation 1 (CAF01). Each round of vaccination was separated by two weeks. Six weeks after the third immunization, all animals were aerosol-challenged with 20-50 CFU of Mycobacterium tuberculosis strain Erdman. After four or eighteen weeks, the number of mycobacteria in each lung was determined by plating serially diluted lung homogenates.

[0299] result

[0300] To test the importance of "late antigens" for long-term protection, bacterial loads in infected animals immunized with a fusion protein containing both early and late antigens (H105) (SEQ ID NO: 37) were compared to infected animals immunized with a fusion protein containing only early expressed antigens (H104) (SEQ ID NO: 38). In the early stages of infection (4 weeks post-infection), all four fusion proteins induced significant protection, with no difference in the level of protection ( FIG. 5A ). In the chronic phase of infection (18 weeks), the addition of late antigens had an impact, as H105 provided better protection than H104, which lacks late antigens (p=0.044) ( FIG. 5B ).

[0301] in conclusion

[0302] Inclusion of late antigens in the vaccine improved long-term protection / containment (Figure 5).

[0303] Example 5: BCG - The antigen is highly effective and similar to BCG + Compared with antigen, it can "synergize" with BCG

[0304] Materials and Methods

[0305] Each group of CB6F1 mice (n=4) was co-immunized with H107 and BCG (BCG+H107). BCG was first administered subcutaneously (sc) on the left side of the base of the tail, and then 1 μg of H107 (SEQ ID NO: 9) in CAF01 was administered at the same injection site the next day. H107 was inoculated again two weeks (right side) and four weeks (left side) later. Two weeks after the last immunization, the animals were euthanized and single-cell suspensions were obtained by passing the spleen through a 100 μm cell strainer. The cells were washed twice in RPMI and then subjected to antigen stimulation for ICS as described in Example 3, except that this time, anti-KLRG1 was included in the surface staining. In the second experiment, all animals, except a small saline control group (n=4), received Mycobacterium bovis BCG Danish immunization and rested for 12 months. At this time point, BCG-immunized animals were divided into four groups (n = 4 per group) and re-immunized once with Mycobacterium bovis BCG or three times with H107 / CAF01 or saline. Three weeks after the third immunization, animals were euthanized and single cell suspensions were obtained from the spleen and stimulated with H107 as an antigen for ICS (as described in Example 3).

[0306] In separate experiments, BCG and H104-H107 (BCG - antigen), H74 (BCG - antigen, SEQ ID NO: 42) or H65 (BCG + Each group of mice was co-immunized with one of the antigens (SEQ ID NO: 43) (described above). A control group was immunized with H107 / CAF01 or BCG or injected with saline. Bacterial loads in each lung were measured 4 and 18 weeks after infection with the Mtb strain Erdman.

[0307] result

[0308] In saline and BCG immunized animals, as expected, there was no immune response to H107, as H107 does not share antigens with BCG. Immunization with H107 resulted in the development of a strong CD4 T cell response, and importantly, this response was significantly enhanced by co-administration of BCG (BCG+H107, Figure 6A). This suggests that co-administration of BCG can enhance the immune response of H107 (BCG - antigens) even though the two vaccines do not share antigens.

[0309] Most of the world's population is immunized with BCG as neonates, and BCG revaccination is gaining increasing attention due to promising results from recent clinical trials. To investigate whether BCG's co-adjuvant effect also occurs in BCG-vaccinated animals, a BCG revaccination model was employed, in which two BCG immunizations were separated by a year (Figure 6B). As expected, no responses to H107 were seen in the saline, BCG, or BCG revaccination groups. In contrast, vaccination with H107 resulted in a pronounced CD4 T cell response in animals that had been initially vaccinated with BCG; this response was significantly increased when H107 was co-administered with the secondary BCG vaccination. This suggests that BCG can also function as an adjuvant in animals that have already been vaccinated with BCG.

[0310] Vaccination with live BCG induces a highly differentiated phenotype of CD4 T cells, which are known to have poor protective capacity. In contrast, subunit vaccines induce less differentiation of CD4 T cells. Compared with less differentiated T cells (such as central memory cells), highly differentiated CD4 T cells (effector memory and effector cells) express IFN-γ, but the co-expression of TNF-γ and IL-2 gradually decreases (Figure 7A). To characterize T cell differentiation, the functional differentiation score (FDS) can be defined as IFN-γ+ cells divided by IFN-γ- cells. BCG vaccination mainly leads to IFN-γ-producing effector and effector memory CD4 T cells (and very few IFN-γ cells), resulting in a differentiation index of 13.3 (Figure 7B, top). BCG and BCG + Co-immunization with vaccine (H65) resulted in an improved cytokine profile with more IFN-γ cells and an FDS of 1.9 (Fig. 7B, middle). However, BCG and H107 (BCG - Co-vaccination with BCG resulted in a greater improvement in the cytokine profile, with IFN-γ being the dominant central memory CD4 T cell, producing an FDS of 0.5. This suggests that when the vaccine is administered together with BCG, + Compared with BCG antigen - Improved T cell quality can be obtained when the antigen is present. Highly differentiated T cells are also characterized by the expression of KLRG1, and it was demonstrated that BCG+H107 produced the lowest frequency of KLRG1-expressing CD4 T cells compared with both BCG and BCG+H65 (Figure 7C).

[0311] To compare BCG with BCG - Antigen (H107) or BCG +Protective effect of co-immunization with antigen (H65), mice immunized with MtbErdman aerosol infection. H74 was included as a reference for the previously described fusion protein (WO2015161853A1). Four weeks after infection, co-immunization with both H107 and H74 increased the protective efficacy of the Mycobacterium bovis BCG vaccine, while H65 did not. For H107, this increase was significant, not only compared with H107 or BCG alone (p<0.0001), but also compared with co-immunization of BCG with H74 (p=0.0489) and H65 (p<0.0001) (Figure 8A). It is worth noting that H107, administered as an independent vaccine, was more effective than BCG and BCG+H65, and comparable to BCG+H74. At late time points (Figure 8B), the protective efficacy of BCG alone and BCG+H65 co-vaccination was lost, but H107 / CAF01 still significantly protected against Mtb (p < 0.0001), and co-vaccination with BCG and H107 fusion protein remained the best protection group. - Fusion H104-H106, when these vaccines were co-administered with BCG at week 4, also induced similar levels of protection ( Figure 9 ).

[0312] in conclusion

[0313] Mycobacterium bovis BCG and BCG - Co-administration of subunit vaccines composed of 100 antigens and 100 mcg of 100 mcg of 100 mcg of 100 mcg of 2 ... Figure 9 H107 was more effective than BCG and BCG+H65 as a stand-alone vaccine and comparable to BCG+H74, indicating that the BCG disclosed herein is more effective when co-administered with BCG and as a stand-alone vaccine. - The antigen combination is potent.

[0314] Example 6: ESAT-6 repeats in fusion proteins increase immunogenicity

[0315] Materials and Methods

[0316] In the first experiment, groups of female CB6F1 mice were immunized three times with 5 μg H56 (SEQ ID NO: 45) in CAF01 or 5 μg H56 + 5 μg ESAT-6 in CAF01. Three weeks after the third immunization, splenocytes were isolated from two animals in each group and 2×10 5Cells / well were stimulated in vitro with ESAT-6 protein at 37°C for 6 hours. The number of CD4 T cells, which produce the cytokines IFN-γ, TNF-α, or IL-2 in response to stimulation, was determined by ICS. Six weeks after immunization, mice were infected with the Mtb strain Erdman. Three weeks later, lung cells were isolated from four animals per group and processed as described above for splenocytes to determine the number of cytokine-producing CD4 T cells. Six weeks after infection, bacterial counts were determined by plating lung homogenates from each mouse on 7H11 agar (n=8 per group) ( FIG10 ).

[0317] In the second experiment ( FIG11 ), groups of mice were immunized with either H64 / CAF01 or H76 / CAF01 containing one (H64, SEQ ID NO: 44) or five (H76, SEQ ID NO: 41) copies of the ESAT-6 molecule. Three weeks after the third immunization, splenocytes were stimulated with the single antigens present in both H64 and H76 fusion proteins, and the frequency of CD4 T cells producing IFN-γ, TNF-α, or IL-2 was determined by ICS. Stimulation with the Rv0287 protein was included as a negative control. Six weeks after infection with the Mtb strain Erdman, bacterial counts in the lungs were determined.

[0318] In a third experiment (Figure 12), the effect of ESAT-6 repetition in another molecular framework was studied using an Mtb relapse model. Six weeks after Mtb Erdman aerosol infection, mice were given isoniazid (0.1g / L) and rifabutin (0.1g / L) ad libitum in drinking water for six weeks. Starting at week 10 post-infection, mice were immunized three times with 0.5μg H83 (one ESAT-6 copy, SEQ ID NO:40) and H84 (four ESAT-6 copies, SEQ ID NO:39) or saline (control), each with a 3-week interval. Two weeks after the last immunization (week 18 of infection), the lungs were removed and immunoassayed by ICS, and at weeks 22 and 35 post-infection, the bacterial load of the lungs was assessed by plating the homogenate on 7H11 agar.

[0319] result

[0320] ESAT-6 is a low immunogenic antigen after protein vaccination, so ESAT-6 specific immunogenicity needs to be increased. First, it was studied whether adding more free ESAT-6 in combination with a fusion protein containing ESAT-6 (H56) would increase the immune response. Three weeks after vaccination, the ESAT-6 specific immune response was higher in mice that received H56 alone compared to mice that received H56+ESAT-6 (Figure 10A). This difference was more obvious in the lungs three weeks after aerosol infection. At this time point, the number of cytokine-producing CD4 T cells was significantly different (p=0.0060) (Figure 10B). Both vaccination regimens provided similar significant protection in the lungs six weeks after infection (p=0.0016 and p=0.0121 relative to unvaccinated animals). This suggests that adding more ESAT-6 protein to the vaccine did not enhance immunogenicity or protection.

[0321] As a solution to the low immunogenicity of ESAT6, several fusion proteins were designed with the ESAT-6 sequence repeated throughout the molecular construct. Comparison of the immune responses induced by H64 (containing one copy of the ESAT-6 molecule) and H76 fusion proteins (containing five copies of the ESAT-6 molecule) revealed that the number of ESAT-6-specific CD4 T cells elicited by the vaccine was three- to five-fold higher in animals immunized with H76 (Figure 11A). Six weeks after aerosol Mtb Erdman infection, we compared bacterial loads in H64- and H76-immunized animals, and the enhanced ESAT-6-specific immune response also led to enhanced protection (Figure 11B).

[0322] Similarly, increasing the ESAT-6 copy number from one (H83) to four (H84) using different molecular constructs enhanced immunogenicity and protection in an Mtb relapse model treated with partial antibiotics. Two weeks after the last immunization, the ESAT-6-specific CD4 T cell response was significantly enhanced in the H84 group compared with the H83 group (Figure 12A). After the end of partial antibiotic treatment, the bacterial load in the control group and H83-vaccinated animals increased from week 22 to week 35 (relapse) (Figure 12B). In contrast, the bacterial load in H84-vaccinated animals decreased from week 22 to week 35 (Figure 12B), indicating that the ESAT-6 duplication pattern both increased the ESAT-6-specific immune response and increased protection in the Mtb relapse model.

[0323] in conclusion

[0324] The addition of free ESAT-6 protein ("free ESAT-6" means that the ESAT-6 protein is not part of a fusion protein) as well as ESAT-6-containing fusion proteins to the vaccine formulation was unlikely to increase the ESAT-6 response elicited by the vaccine (Figure 10). In contrast, incorporating more copies of ESAT-6 into the fusion protein increased the number of ESAT-6-specific CD4 T cells elicited by the vaccine and improved protection in different animal models (Figures 11 and 12).

[0325] Example 7: Fusion protein can be used alone or in combination with BCG

[0326] Materials and Methods

[0327] In the first ( FIG13A ) and second ( FIG13B ) experiments, female 129 / Sv and CB6F1 mice were immunized three times with 1 μg of either H74 (SEQ ID NO: 42), H105 (SEQ ID NO: 37), or H107 (SEQ ID NO: 9) in CAF01. Two weeks after the third immunization, splenocytes were isolated from four animals in each group and 1–2 × 10 6 Cells / well were stimulated with ESAT-6 peptide in vitro for 6 h at 37°C, and the number of cytokine-producing CD4 T cells was measured by ICS. The level of IFN-γ secretion in the 3-day culture supernatant was measured by ELISA as described above.

[0328] In the third experiment ( FIG13C ), female CB6F1 mice were co-immunized with BCG and 1 μg of H105 or H107 in CAF01 (BCG+H105 or BCG+H107). BCG was administered by subcutaneous injection (sc) in the left side at the base of the tail, followed by H105 or H107 in CAF01 at the same injection site the next day. Subsequently, two weeks (right side) and four weeks (left side) later, the animals were immunized again with the subunit vaccine. Two weeks after the third immunization, splenocytes were isolated from four animals in each group, and 1-2×10 6 Cells / well were stimulated in vitro with ESAT-6 peptide for 6 hours at 37°C, and the number of cytokine-producing CD4 T cells was measured by ICS. IFN-γ secretion levels were measured in 3-day culture supernatants by ELISA as described. Six weeks after the last immunization, all animals were aerosol-infected with the Erdman strain of Mtb, and 4 weeks after infection, lung bacterial burdens were measured by plating organ homogenates.

[0329] result

[0330] In 129 / Sv mice, increasing the ESAT-6 copy number from one (H74 and H105) to four (H107) resulted in an enhanced ESAT-6 response (Figure 13A). This was also true in CB6F1 mice, as measured by ICS and ELISA (Figure 13B). Finally, the test was conducted to see whether this was also true when these vaccines were co-administered with BCG. In this approach, again, H107 significantly increased the ESAT-6 specific immune response compared to H105, which also resulted in an increased protective effect against Mtb aerosol infection (Figure 13C).

[0331] in conclusion

[0332] In three independent experiments (using two different mouse strains), it was demonstrated that increasing the ESAT-6 copy number from one (H74 and H105) to four (H107) increased the ESAT-6-specific immune response. This was also true when these vaccines were co-administered with BCG. This also reduced bacterial load, showing that repeating ESAT-6 in the vaccine backbone is a powerful approach to improve immunogenicity and vaccine-mediated protection. Furthermore, the data support that the fusion is effective whether used as a standalone vaccine or with BCG.

[0333] Example 8: H107 is a better stand-alone vaccine than the prior art subunit vaccine H56

[0334] Materials and Methods

[0335] Each group of mice was immunized three times with 2 μg of the fusion protein in a total volume of 200 μL of cationic adjuvant formulation 1 (CAF01). Each round of vaccination was separated by two weeks. Six weeks after the third immunization, all animals were aerosol-challenged with 20-50 CFU of Mycobacterium tuberculosis strain Erdman. Four to twelve weeks later, the number of mycobacteria in each lung was determined by plating serially diluted lung homogenates.

[0336] result

[0337] To compare the protective efficacy of H107 (SEQ ID NO: 9) relative to the most advanced subunit vaccine (H56; SEQ ID NO: 45), groups of mice were immunized with the fusion protein in CAF01 adjuvant in two (H107) and five (H56) independent experiments. Animals were challenged with the Mtb strain Erdman, and the number of bacteria in the lungs of each animal was counted ( Figure 14 Depending on the experiment, protective efficacy was measured four to twelve weeks after challenge with the Mtb strain Erdman. On average, the H107 vaccine provided 2.0 log10 of protection, corresponding to a 100-fold reduction in bacterial load. In contrast, the H56 vaccine reduced bacterial load by an average of approximately 7-fold (0.82 log10).

[0338] in conclusion

[0339] Compared with the most advanced subunit TB vaccine (H56 / CAF01), the H107 / CAF01 vaccine provides superior protection against aerosol challenge with virulent Mtb ( Figure 14 ).

[0340] Example 9: Co-administration of BCG+H107 enhances BCG-specific immune response

[0341] Materials and Methods

[0342] As described in the previous examples, each group of blank (naive) CB6F1 mice was co-immunized with BCG and three 1 μg H107 (SEQ ID NO: 9) (n = 4 per time point). Animals were euthanized at different time points after immunization, and single cell suspensions were obtained by passing the spleen through a 100 μm cell strainer. As described in the previous examples, cells were washed twice in RPMI and then antigen-stimulated for ICS. TB10.4 was used for stimulation to assess BCG-specific immune responses because this antigen is contained in BCG but not in H107.

[0343] result

[0344] Three weeks after the first immunization (one week after the second H107), an increased TB10.4 response was observed in the BCG+H107 group compared to BCG alone ( Figure 15 This difference was maintained until the end of the experiment at week 9. This suggests that co-administration of H107 with BCG increased BCG-specific immune responses, even though they do not share antigens.

[0345] in conclusion

[0346] The previous example showed that co-administration of BCG+H107 resulted in an increase in the H107 response (BCG acted as an H107 adjuvant). This example shows that co-administration of H107+BCG also increased the BCG-specific immune response, which means that H107 acted as a BCG adjuvant ( Figure 15 ). In this way, there is a true synergy between the two vaccines.

[0347] Example 10: H107e increases protein expression in E. coli compared to H107

[0348] Among other things, the yield of recombinant protein expression depends on the protein's amino acid sequence, and large-scale vaccine production requires an optimized expression process. To address this issue, a high-expression version of H107, H107e, was developed.

[0349] Materials and Methods

[0350] To optimize expression, the proline-rich sequence AA298-AA517 in the Rv3876 portion of H107 (SEQ ID NO: 9) was deleted to generate H107e (SEQ ID NO: 91). The DNA sequences corresponding to H107 (SEQ ID NO: 54) and H107e (SEQ ID NO: 92) were chemically synthesized, inserted into the pJ411 expression vector (ATUM, Menlo Park, CA, US), and transformed into Escherichia coli BL21 (DE3) strain (Agilent, DK) grown in Luria-Bertani (LB) medium. Recombinant protein expression levels were assessed by SDS-PAGE or Western blotting (using a primary antibody that recognizes the MPT70 portion of H107 / H107e) at 0, 1, and 3 hours after induction with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). Equal numbers of cells (same OD600) were loaded into the lanes on the gel. After expression, the recombinant antigen was purified from inclusion bodies and subjected to metal ion affinity chromatography using a 5 ml HisTrap HP column (GE Healthcare), followed by anion exchange using a HiTrap Q HP column (GE Healthcare). Final protein yield was estimated by bicinchoninic acid assay.

[0351] result

[0352] It is clear from the culture induction experiments that the expression of H107e in E. coli is significantly increased. At 1 and 3 hours after induction, the band corresponding to H107e is much stronger than that of H107 in both SDS-PAGE gel and Western blot ( Figure 16 A) Comparative yields of purified protein from 6 L fermentation: 2.4-7.2 mg for H107 and 15.2 mg for H107e.

[0353] in conclusion:

[0354] Compared with H107, the sequence changes in H107e resulted in a significant increase in protein expression and yield.

[0355] Example 11: H107e is as immunogenic as H107 and also acts synergistically with BCG

[0356] Materials and methods

[0357] As described in the previous example, groups of naive CB6F1 mice (n=8) were co-immunized with BCG and 1 μg of H107 (SEQ ID NO: 9) or H107e (SEQ ID NO: 91). Two weeks after the last immunization, the animals were euthanized and single-cell suspensions were obtained by passing the spleens through a 100 μm cell strainer. As described in the previous example, the cells were washed twice in RPMI and then antigen-stimulated for ICS and IFN-γ ELISA.

[0358] In the second experiment, groups of naive CB6F1 mice (n=6) were co-immunized with BCG and H107e as described above, and controls were immunized with H107e / CAF01 or BCG or injected with saline. Eight weeks after infection with Mtb strain Erdman, bacterial loads in each lung were measured.

[0359] In a third experiment, groups of BCG-remembered CB6F1 mice (n=6) were immunized three times with 2 μg of H107e or H65 subcutaneously at the base of the tail. Two weeks after the last immunization, the animals were euthanized and single-cell suspensions were obtained by passing inguinal lymph nodes through a 100 μm cell strainer. The cells were washed twice in RPMI and then antigen-stimulated for ICS.

[0360] result

[0361] After co-immunization with BCG, H107e and H107 were confirmed to have the same immunogenicity, which was demonstrated by the expression of cytokines by CD4 T cells ( Figure 17 A, left) and IFN-γ release measured by ELISA ( Figure 17 A, right). It was also confirmed that H107e induced immune responses to the same antigens as H107 ( Figure 17 B). Notably, deletion of the Rv3876 portion of H107e resulted in a slight decrease in the Rv3876-specific immune response, but an increase in the immune response against MPT70 and MPT83 ( Figure 17 B). After infection, H107e provided similar or better protection than BCG, and co-vaccination of BCG + H107e resulted in a significant increase in protection compared to BCG and H107e alone ( Figure 17 C), as previously observed with H107 in Example 9. In BCG memory mice, H107e (BCG-) vaccination resulted in less differentiated (better quality) CD4 T cells compared to H65 (BCG+), as measured by the functional differentiation score (FDS) ( Figure 17D). In addition, as Th17 cells have gained increasing attention over the past decade and are thought to have protective properties during Mtb infection, IL-17-expressing CD4 T cells were measured by flow cytometry. H107e induced a much higher proportion of Th17 cells compared to H65, suggesting that BCG- vaccines produce an overall broader immune response in BCG-primed individuals compared to traditional BCG+ vaccines ( Figure 17 D).

[0362] in conclusion

[0363] H107e has similar immunogenicity to H107 and induces significant protection after Mtb challenge. Like H107, H107e acts synergistically with BCG, and the level of protection achieved with co-vaccination of BCG+H107e is significantly higher than that achieved with either H107e or BCG alone. In BCG-primed animals, H107e (BCG-) vaccination induced less T cell differentiation (as with H107) and increased the proportion of Th17 cells, as measured by CD4 T cells expressing IL-17, compared to H65 (BCG+).

[0364] References

[0365] ·WO2010006607A2

[0366] ·WO2006136162A2

[0367] ·WO2014063704A2

[0368] ·WO2015161853A1

[0369] ·Sederetal(2008),Nat.Rev.Immunol.8,247-258 Sequence Listing <110> Steins Serum Institute <120> Fusion proteins for tuberculosis vaccines <130> 68271PC01 <150> EP19180280 <151> 2019-06-14 <160> 92 <170> BiSSAP 1.3.6 <210> 1 <211> 95 <212> PRT <213> Mycobacterium tuberculosis <400> 1 Met Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 1 5 10 15 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 20 25 30 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 35 40 45 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu 50 55 60 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 65 70 75 80 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala 85 90 95 <210> 2 <211> 368 <212> PRT <213> Mycobacterium tuberculosis <400> 2 Met Leu Trp His Ala Met Pro Pro Glu Leu Asn Thr Ala Arg Leu Met 1 5 10 15 Ala Gly Ala Gly Pro Ala Pro Met Leu Ala Ala Ala Ala Gly Trp Gln 20 25 30 Thr Leu Ser Ala Ala Leu Asp Ala Gln Ala Val Glu Leu Thr Ala Arg 35 40 45 Leu Asn Ser Leu Gly Glu Ala Trp Thr Gly Gly Gly Ser Asp Lys Ala 50 55 60 Leu Ala Ala Ala Thr Pro Met Val Val Trp Leu Gln Thr Ala Ser Thr 65 70 75 80 Gln Ala Lys Thr Arg Ala Met Gln Ala Thr Ala Gln Ala Ala Ala Tyr 85 90 95 Thr Gln Ala Met Ala Thr Thr Pro Ser Leu Pro Glu Ile Ala Ala Asn 100 105 110 His Ile Thr Gln Ala Val Leu Thr Ala Thr Asn Phe Phe Gly Ile Asn 115 120 125 Thr Ile Pro Ile Ala Leu Thr Glu Met Asp Tyr Phe Ile Arg Met Trp 130 135 140 Asn Gln Ala Ala Leu Ala Met Glu Val Tyr Gln Ala Glu Thr Ala Val 145 150 155 160 Asn Thr Leu Phe Glu Lys Leu Glu Pro Met Ala Ser Ile Leu Asp Pro 165 170 175 Gly Ala Ser Gln Ser Thr Thr Asn Pro Ile Phe Gly Met Pro Ser Pro 180 185 190 Gly Ser Ser Thr Pro Val Gly Gln Leu Pro Pro Ala Ala Thr Gln Thr 195 200 205 Leu Gly Gln Leu Gly Glu Met Ser Gly Pro Met Gln Gln Leu Thr Gln 210 215 220 Pro Leu Gln Gln Val Thr Ser Leu Phe Ser Gln Val Gly Gly Thr Gly 225 230 235 240 Gly Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met Gly Leu Leu Gly 245 250 255 Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly Ser Gly Pro Ser 260 265 270 Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro Gly Ala Gly Gly 275 280 285 Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile Glu Lys Pro Val 290 295 300 Ala Pro Ser Val Met Pro Ala Ala Ala Ala Gly Ser Ser Ala Thr Gly 305 310 315 320 Gly Ala Ala Pro Val Gly Ala Gly Ala Met Gly Gln Gly Ala Gln Ser 325 330 335 Gly Gly Ser Thr Arg Pro Gly Leu Val Ala Pro Ala Pro Leu Ala Gln 340 345 350 Glu Arg Glu Glu Asp Asp Glu Asp Asp Trp Asp Glu Glu Asp Asp Trp 355 360 365 <210> 3 <211> 666 <212> PRT <213> Combined <400> 3 Met Ala Ala Asp Tyr Asp Lys Leu Phe Arg Pro His Glu Gly Met Glu 1 5 10 15 Ala Pro Asp Asp Met Ala Ala Gln Pro Phe Phe Asp Pro Ser Ala Ser 20 25 30 Phe Pro Pro Ala Pro Ala Ser Ala Asn Leu Pro Lys Pro Asn Gly Gln 35 40 45 Thr Pro Pro Pro Thr Ser Asp Asp Leu Ser Glu Arg Phe Val Ser Ala 50 55 60 Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr Pro Met 65 70 75 80 Pro Ile Ala Ala Gly Glu Pro Ser Pro Glu Pro Ala Ala Ser Lys 85 90 95 Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala Pro Pro 100 105 110 Lys Pro Pro Thr Pro Met Pro Ile Ala Gly Pro Glu Pro Ala Pro 115 120 125 For Lys For Thr For Met For Ile Ala Gly For Ala For Thr 130 135 140 Pro Thr Glu Ser Gln Leu Ala Pro Pro Arg Pro Pro Thr Pro Gln Thr 145 150 155 160 For Thr Gly Ala For Gln Gln For Glu Ser For Ala For His Val For 165 170 175 Ser His Gly Pro His Gln Pro Arg Arg Thr Ala Pro Ala Pro Pro Trp 180 185 190 Ala Lys Met Pro Ile Gly Glu Pro Pro Pro Ala Pro Ser Arg Pro Ser 195 200 205 Ala Ser Pro Ala Glu Pro Pro Thr Arg Pro Ala Pro Gln His Ser Arg 210 215 220 Arg Ala Arg Arg Gly His Arg Tyr Arg Thr Asp Thr Glu Arg Asn Val 225 230 235 240 Gly Lys Val Ala Thr Gly Pro Ser Ile Gln Ala Arg Leu Arg Ala Glu 245 250 255 Glu Ala Ser Gly Ala Gln Leu Ala Pro Gly Thr Glu Pro Ser Pro Ala 260 265 270 Pro Leu Gly Gln Pro Arg Ser Tyr Leu Ala Pro Pro Thr Arg Pro Ala 275 280 285 Pro Thr Glu Pro Pro Pro Ser Pro Ser Pro Gln Arg Asn Ser Gly Arg 290 295 300 Arg Ala Glu Arg Arg Val His Pro Asp Leu Ala Ala Gln His Ala Ala 305 310 315 320 Ala Gln Pro Asp Ser Ile Thr Ala Ala Thr Thr Gly Gly Arg Arg Arg 325 330 335 Lys Arg Ala Ala Pro Asp Leu Asp Ala Thr Gln Lys Ser Leu Arg Pro 340 345 350 Ala Ala Lys Gly Pro Lys Val Lys Lys Val Lys Pro Gln Lys Pro Lys 355 360 365 Ala Thr Lys Pro Pro Lys Val Val Ser Gln Arg Gly Trp Arg His Trp 370 375 380 Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser Pro Asp Glu Lys 385 390 395 400 Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn Pro Arg Gly Ser 405 410 415 Tyr Gln Ile Ala Val Val Gly Leu Lys Gly Gly Ala Gly Lys Thr Thr 420 425 430 Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val Arg Ala Asp Arg 435 440 445 Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn Leu Ala Asp Arg 450 455 460 Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val Leu Ala Glu Lys 465 470 475 480 Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr Ser Val Asn Ala 485 490 495 Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser Ser Ala Gln Arg 500 505 510 Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp Pro Ala Ser Arg 515 520 525 Phe Tyr Asn Leu Val Leu Ala Asp Cys Gly Ala Gly Phe Phe Asp Pro 530 535 540 Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val Val Val Val Ala 545 550 555 560 Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val Ala Leu Asp Trp 565 570 575 Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg Ala Cys Val Val 580 585 590 Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala Val Lys Asp Leu 595 600 605 Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg Val Val Val Met 610 615 620 Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile Ser Leu Asp Leu 625 630 635 640 Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu Ala Ala Ala Leu 645 650 655 Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg 660 665 <210> 4 <211> 103 <212> PRT <213> Combined <400> 4 Met Thr Glu Asn Leu Thr Val Gln Pro Glu Arg Leu Gly Val Leu Ala 1 5 10 15 Ser His His Asp Asn Ala Ala Val Asp Ala Ser Ser Gly Val Glu Ala 20 25 30 Ala Ala Gly Leu Gly Glu Ser Val Ala Ile Thr His Gly Pro Tyr Cys 35 40 45 Ser Gln Phe Asn Asp Thr Leu Asn Val Tyr Leu Thr Ala His Asn Ala 50 55 60 Leu Gly Ser Ser Leu His Thr Ala Gly Val Asp Leu Ala Lys Ser Leu 65 70 75 80 Arg Ile Ala Ala Lys Ile Tyr Ser Glu Ala Asp Glu Ala Trp Arg Lys 85 90 95 Ala Ile Asp Gly Leu Phe Thr 100 <210> 5 <211> 392 <212> PRT <213> Combined <400> 5 Met Ser Arg Ala Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly 1 5 10 15 Leu Tyr Asp Leu Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu 20 25 30 Tyr Ser Ser Leu Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala 35 40 45 Ala Phe Pro Gly Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala 50 55 60 Gly Lys Asn Arg Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu 65 70 75 80 Asp Arg Gln Leu Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln 85 90 95 Thr Thr Arg Asp Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val 100 105 110 Arg Pro Val Ala Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala 115 120 125 Leu Ser Ala Ala Phe Gln Ala Pro Phe Cys Ala Gly Ala Met Ala Val 130 135 140 Val Gly Gly Ala Leu Ala Tyr Leu Val Val Lys Thr Leu Ile Asn Ala 145 150 155 160 Thr Gln Leu Leu Lys Leu Leu Ala Lys Leu Ala Glu Leu Val Ala Ala 165 170 175 Ala Ile Ala Asp Ile Ile Ser Asp Val Ala Asp Ile Ile Lys Gly Thr 180 185 190 Leu Gly Glu Val Trp Glu Phe Ile Thr Asn Ala Leu Asn Gly Leu Lys 195 200 205 Glu Leu Trp Asp Lys Leu Thr Gly Trp Val Thr Gly Leu Phe Ser Arg 210 215 220 Gly Trp Ser Asn Leu Glu Ser Phe Phe Ala Gly Val Pro Gly Leu Thr 225 230 235 240 Gly Ala Thr Ser Gly Leu Ser Gln Val Thr Gly Leu Phe Gly Ala Ala 245 250 255 Gly Leu Ser Ala Ser Ser Gly Leu Ala His Ala Asp Ser Leu Ala Ser 260 265 270 Ser Ala Ser Leu Pro Ala Leu Ala Gly Ile Gly Gly Gly Ser Gly Phe 275 280 285 Gly Gly Leu Pro Ser Leu Ala Gln Val His Ala Ala Ser Thr Arg Gln 290 295 300 Ala Leu Arg Pro Arg Ala Asp Gly Pro Val Gly Ala Ala Ala Glu Gln 305 310 315 320 Val Gly Gly Gln Ser Gln Leu Val Ser Ala Gln Gly Ser Gln Gly Met 325 330 335 Gly Gly Pro Val Gly Met Gly Gly Met His Pro Ser Ser Gly Ala Ser 340 345 350 Lys Gly Thr Thr Thr Lys Lys Tyr Ser Glu Gly Ala Ala Ala Gly Thr 355 360 365 Glu Asp Ala Glu Arg Ala Pro Val Glu Ala Asp Ala Gly Gly Gly Gln 370 375 380 Lys Val Leu Val Arg Asn Val Val 385 390 <210> 6 <211> 228 <212> PRT[[ID=4*]] <213> Binding Mycobacterium <400> 6 Val Arg Ile Lys Ile Phe Met Leu Val Thr Ala Val Val Leu Leu Cys 1 5 10 15 Cys Ser Gly Val Ala Thr Ala Ala Pro Lys Thr Tyr Cys Glu Glu Leu 20 25 30 Lys Gly Thr Asp Thr Gly Gln Ala Cys Gln Ile Gln Met Ser Asp Pro 35 40 45 Ala Tyr Asn Ile Asn Ile Ser Leu Pro Ser Tyr Tyr Pro Asp Gln Lys 50 55 60 Ser Leu Glu Asn Tyr Ile Ala Gln Thr Arg Asp Lys Phe Leu Ser Ala 65 70 75 80 Ala Thr Ser Ser Thr Pro Arg Glu Ala Pro Tyr Glu Leu Asn Ile Thr 85 90 95 Ser Ala Thr Tyr Gln Ser Ala Ile Pro Pro Arg Gly Thr Gln Ala Val 100 105 110 Val Leu Lys Val Tyr Gln Asn Ala Gly Gly Thr His Pro Thr Thr Thr 115 120 125 Tyr Lys Ala Phe Asp Trp Asp Gln Ala Tyr Arg Lys Pro Ile Thr Tyr 130 135 140 Asp Thr Leu Trp Gln Ala Asp Thr Asp Pro Leu Pro Val Val Phe Pro 145 150 155 160 Ile Val Gln Gly Glu Leu Ser Lys Gln Thr Gly Gln Gln Val Ser Ile 165 170 175 Ala Pro Asn Ala Gly Leu Asp Pro Val Asn Tyr Gln Asn Phe Ala Val 180 185 190 Thr Asn Asp Gly Val Ile Phe Phe Phe Asn Pro Gly Glu Leu Leu Pro 195 200 205 Glu Ala Ala Gly Pro Thr Gln Val Leu Val Pro Arg Ser Ala Ile Asp 210 215 220 Ser Met Leu Ala 225 <210> 7 <211> 193 <212> PRT <213> Combined <400> 7 Met Lys Val Lys Asn Thr Ile Ala Ala Thr Ser Phe Ala Ala Ala Gly 1 5 10 15 Leu Ala Ala Leu Ala Val Ala Val Ser Pro Pro Ala Ala Ala Gly Asp 20 25 30 Leu Val Gly Pro Gly Cys Ala Glu Tyr Ala Ala Ala Asn Pro Thr Gly 35 40 45 Pro Ala Ser Val Gln Gly Met Ser Gln Asp Pro Val Ala Val Ala Ala 50 55 60 Ser Asn Asn Pro Glu Leu Thr Thr Leu Thr Ala Ala Leu Ser Gly Gln 65 70 75 80 Leu Asn Pro Gln Val Asn Leu Val Asp Thr Leu Asn Ser Gly Gln Tyr 85 90 95 Thr Val Phe Ala Pro Thr Asn Ala Ala Phe Ser Lys Leu Pro Ala Ser 100 105 110 Thr Ile Asp Glu Leu Lys Thr Asn Ser Ser Leu Leu Thr Ser Ile Leu 115 120 125 Thr Tyr His Val Val Ala Gly Gln Thr Ser Pro Ala Asn Val Val Gly 130 135 140 Thr Arg Gln Thr Leu Gln Gly Ala Ser Val Thr Val Thr Gly Gln Gly 145 150 155 160 Asn Ser Leu Lys Val Gly Asn Ala Asp Val Val Cys Gly Gly Val Ser 165 170 175 Thr Ala Asn Ala Thr Val Tyr Met Ile Asp Ser Val Leu Met Pro Pro 180 185 190 Ala <210> 8 <211> 220 <212> PRT <213> Combined <400> 8 Met Ile Asn Val Gln Ala Lys Pro Ala Ala Ala Ala Ser Leu Ala Ala 1 5 10 15 Ile Ala Ile Ala Phe Leu Ala Gly Cys Ser Ser Thr Lys Pro Val Ser 20 25 30 Gln Asp Thr Ser Pro Lys Pro Ala Thr Ser Pro Ala Ala Pro Val Thr 35 40 45 Thr Ala Ala Met Ala Asp Pro Ala Ala Asp Leu Ile Gly Arg Gly Cys 50 55 60 Ala Gln Tyr Ala Ala Gln Asn Pro Thr Gly Pro Gly Ser Val Ala Gly 65 70 75 80 Met Ala Gln Asp Pro Val Ala Thr Ala Ala Ser Asn Asn Pro Met Leu 85 90 95 Ser Thr Leu Thr Ser Ala Leu Ser Gly Lys Leu Asn Pro Asp Val Asn 100 105 110 Leu Val Asp Thr Leu Asn Gly Gly Glu Tyr Thr Val Phe Ala Pro Thr 115 120 125 Asn Ala Ala Phe Asp Lys Leu Pro Ala Ala Thr Ile Asp Gln Leu Lys 130 135 140 Thr Asp Ala Lys Leu Leu Ser Ser Ile Leu Thr Tyr His Val Ile Ala 145 150 155 160 Gly Gln Ala Ser Pro Ser Arg Ile Asp Gly Thr His Gln Thr Leu Gln 165 170 175 Gly Ala Asp Leu Thr Val Ile Gly Ala Arg Asp Asp Leu Met Val Asn 180 185 190 Asn Ala Gly Leu Val Cys Gly Gly Val His Thr Ala Asn Ala Thr Val 195 200 205 Tyr Met Ile Asp Thr Val Leu Met Pro Pro Ala Gln 210 215 220 <210> 9 <211> 2153<000!198><212> PRT <213> binds to Mycobacterium <400> 9 Met Gln Ala Glu Thr Ala Val Asn Thr Leu Phe Glu Lys Leu Glu Pro 1 5 10 1 !5 Met Ala Ser Ile Leu Asp Pro Gly Ala Ser Gln Ser Thr Thr Asn Pro 20 25 30 Ile Phe Gly Met Pro Ser Pro Gly Ser Ser Thr Pro Val Gly Gln Leu 35 40 45 Pro Pro Ala Ala Thr Gln Thr Leu Gly Gln Leu Gly Glu Met Ser Gly 50 55 60 Pro Met Gly Gly Ser Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met 65 70 75 80 Gly Leu Leu Gly Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly 85 90 95 It should be noted that there seems to be an error in the original text where "1!5" is likely a typo and is corrected to "15" in the translation. Also, "<000!198>" is likely a typo and should be " ".Ser Gly Pro Ser Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro 100 105 110 Gly Ala Gly Gly Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile 115 120 125 Glu Lys Pro Val Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu 130 135 140 Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu 145 150 155 160 Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly 165 170 175 Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala 180 185 190 Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile 195 200 205 Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly 210 215 220 Met Phe Ala Ala Ala Asp Tyr Asp Lys Leu Phe Arg Pro His Glu Gly 225 230 235 240 Met Glu Ala Pro Asp Asp Met Ala Ala Gln Pro Phe Phe Asp Pro Ser 245 250 255 Ala Ser Phe Pro Pro Ala Pro Ala Ser Ala Asn Leu Pro Lys Pro Asn 260 265 270 Gly Gln Thr Pro Pro Pro Thr Ser Asp Asp Leu Ser Glu Arg Phe Val 275 280 285 Ser Ala Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr 290 295 300 Pro Met Pro Ile Ala Ala Gly Glu Pro Pro Ser Pro Glu Pro Ala Ala Ala 305 310 315 320 Ser Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala 325 330 335 Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro 340 345 350 Ala Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Ala 355 360 365 Pro Thr Pro Thr Glu Ser Gln Leu Ala Pro Pro Arg Pro Pro Thr Pro 370 375 380 Gln Thr Pro Thr Gly Ala Pro Gln Gln Pro Glu Ser Pro Ala Pro His 385 390 395 400 Val Pro Ser His Gly Pro His Gln Pro Arg Arg Thr Ala Pro Ala Pro 405 410 415 Pro Trp Ala Lys Met Pro Ile Gly Glu Pro Pro Pro Ala Pro Ser Arg 420 425 430 Pro Ser Ala Ser Pro Ala Glu Pro Pro Thr Arg Pro Ala Pro Gln His 435 440 445 Ser Arg Arg Ala Arg Arg Gly His Arg Tyr Arg Thr Asp Thr Glu Arg 450 455 460 Asn Val Gly Lys Val Ala Thr Gly Pro Ser Ile Gln Ala Arg Leu Arg 465 470 475 480 Ala Glu Glu Ala Ser Gly Ala Gln Leu Ala Pro Gly Thr Glu Pro Ser 485 490 495 Pro Ala Pro Leu Gly Gln Pro Arg Ser Tyr Leu Ala Pro Pro Thr Arg 500 505 510 Pro Ala Pro Thr Glu Pro Pro Pro Ser Pro Ser Pro Gln Arg Asn Ser 515 520 525 Gly Arg Arg Ala Glu Arg Arg Val His Pro Asp Leu Ala Ala Gln His 530 535 540 Ala Ala Ala Gln Pro Asp Ser Ile Thr Ala Ala Thr Thr Gly Gly Arg 545 550 555 560 Arg Arg Lys Arg Ala Ala Pro Asp Leu Asp Ala Thr Gln Lys Ser Leu 565 570 575 Arg Pro Ala Ala Lys Gly Pro Lys Val Lys Lys Val Lys Pro Gln Lys 580 585 590 Pro Lys Ala Thr Lys Pro Pro Lys Val Val Ser Gln Arg Gly Trp Arg 595 600 605 His Trp Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser Pro Asp 610 615 620 Glu Lys Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn Pro Arg 625 630 635 640 Gly Ser Tyr Gln Ile Ala Val Val Gly Leu Gln Gly Gly Ala Gly Lys 645 650 655 Thr Thr Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val Arg Ala 660 665 670 Asp Arg Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn Leu Ala 675 680 685 Asp Arg Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val Leu Ala 690 695 700 Glu Lys Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr Ser Val 705 710 715 720 Asn Ala Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser Ser Ala 725 730 735 Gln Arg Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp Pro Ala 740 745 750 Ser Arg Phe Tyr Asn Leu Val Leu Ala Asp Ser Gly Ala Gly Phe Phe 755 760 765 Asp Pro Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val Val Val 770 775 780 Val Ala Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val Ala Leu 785 790 795 800 Asp Trp Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg Ala Ser 805 810 815 Val Val Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala Val Lys 820 825 830 Asp Leu Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg Val Val 835 840 845 Val Met Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile Ser Leu 850 855 860 Asp Leu Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu Ala Ala 865 870 875 880 Ala Leu Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg Thr Glu Gln Gln 885 890 895 Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn 900 905 910 Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr 915 920 925 Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly 930 935 940 Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu 945 950 955 960 Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser 965 970 975 Thr Glu Gly Asn Val Thr Gly Met Phe Ala Thr Glu Asn Leu Thr Val 980 985 990 Gln Pro Glu Arg Leu Gly Val Leu Ala Ser His His Asp Asn Ala Ala 995 1000 1005 Val Asp Ala Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly Glu Ser 1010 1015 1020 Val Ala Ile Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp Thr Thr 1025 1030 1035 1040 Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile 1045 1050 1055 Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln 1060 1065 1070 Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala 1075 1080 1085 Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn 1090 1095 1100 Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala 1105 1110 1115 1120 Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ser Arg Ala 1125 1130 1135 Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly Leu Tyr Asp Leu 1140 1145 1150 Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu Tyr Ser Ser Leu 1155 1160 1165 Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala Ala Phe Pro Gly 1170 1175 1180 Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala Gly Lys Asn Arg 1185 1190 1195 1200 Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu Asp Arg Gln Leu 1205 1210 1215 Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln Thr Thr Arg Asp 1220 1225 1230 Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val Arg Pro Val Ala 1235 1240 1245 Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala Leu Ser Ala Lys 1250 1255 1260 Thr Leu Ile Asn Ala Thr Gln Leu Leu Lys Leu Leu Ala Lys Leu Ala 1265 1270 1275 1280 Glu Leu Val Ala Ala Ala Ile Ala Asp Ile Ile Ser Asp Val Ala Asp 1285 1290 1295 Ile Ile Lys Gly Thr Leu Gly Glu Val Trp Glu Phe Ile Thr Asn Ala 1300 1305 1310 Leu Asn Gly Leu Lys Glu Leu Trp Asp Lys Leu Thr Gly Trp Val Thr 1315 1320 1325 Gly Leu Phe Ser Arg Gly Trp Ser Asn Leu Glu Ser Phe Phe Ala Gly 1330 1335 1340 Val Pro Gly Leu Thr Gly Ala Thr Ser Gly Leu Ser Gln Val Thr Gly 1345 1350 1355 1360 Leu Phe Gly Ala Ala Gly Leu Ser Ala Ser Ser Gly Leu Ala His Ala 1365 1370 1375 Asp Ser Leu Ala Ser Ser Ala Ser Leu Pro Ala Leu Ala Gly Ile Gly 1380 1385 1390 Gly Gly Ser Gly Phe Gly Gly Leu Pro Ser Leu Ala Gln Val His Ala 1395 1400 1405 Ala Ser Thr Arg Gln Ala Leu Arg Pro Arg Ala Asp Gly Pro Val Gly 1410 1415 1420 Ala Ala Ala Glu Gln Val Gly Gly Gln Ser Gln Leu Val Ser Ala Gln 1425 1430 1435 1440 Gly Ser Gln Gly Met Gly Gly Pro Val Gly Met Gly Gly Met His Pro 1445 1450 1455 Ser Ser Gly Ala Ser Lys Gly Thr Thr Thr Lys Lys Tyr Ser Glu Gly 1460 1465 1470 Ala Ala Ala Gly Thr Glu Asp Ala Glu Arg Ala Pro Val Glu Ala Asp 1475 1480 1485 Ala Gly Gly Gly Gln Lys Val Leu Val Arg Asn Val Val Thr Glu Gln 1490 1495 1500 Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly 1505 1510 1515 1520 Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu 1525 1530 1535 Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln 1540 1545 1550 Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala 1555 1560 1565 Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala 1570 1575 1580 Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ala Pro Lys Thr Tyr 1585 1590 1595 1600 Ser Glu Glu Leu Lys Gly Thr Asp Thr Gly Gln Ala Ser Gln Ile Gln 1605 1610 1615 Met Ser Asp Pro Ala Tyr Asn Ile Asn Ile Ser Leu Pro Ser Tyr Tyr 1620 1625 1630 Pro Asp Gln Lys Ser Leu Glu Asn Tyr Ile Ala Gln Thr Arg Asp Lys 1635 1640 1645 Phe Leu Ser Ala Ala Thr Ser Ser Thr Pro Arg Glu Ala Pro Tyr Glu 1650 1655 1660 Leu Asn Ile Thr Ser Ala Thr Tyr Gln Ser Ala Ile Pro Pro Arg Gly 1665 1670 1675 1680 Thr Gln Ala Val Val Leu Lys Val Tyr Gln Asn Ala Gly Gly Thr His 1685 1690 1695 Pro Thr Thr Thr Tyr Lys Ala Phe Asp Trp Asp Gln Ala Tyr Arg Lys 1700 1705 1710 Pro Ile Thr Tyr Asp Thr Leu Trp Gln Ala Asp Thr Asp Pro Leu Pro 1715 1720 1725 Val Val Phe Pro Ile Val Gln Gly Glu Leu Ser Lys Gln Thr Gly Gln 1730 1735 1740 Gln Val Ser Ile Ala Pro Asn Ala Gly Leu Asp Pro Val Asn Tyr Gln 1745 1750 1755 1760 Asn Phe Ala Val Thr Asn Asp Gly Val Ile Phe Phe Phe Asn Pro Gly 1765 1770 1775 Glu Leu Leu Pro Glu Ala Ala Gly Pro Thr Gln Val Leu Val Pro Arg 1780 1785 1790 Ser Ala Ile Asp Ser Met Leu Ala Gly Asp Leu Val Gly Pro Gly Ser 1795 1800 1805 Ala Glu Tyr Ala Ala Ala Asn Pro Thr Gly Pro Ala Ser Val Gln Gly 1810 1815 1820 Met Ser Gln Asp Pro Val Ala Val Ala Ala Ser Asn Asn Pro Glu Leu 1825 1830 1835 1840 Thr Thr Leu Thr Ala Ala Leu Ser Gly Gln Leu Asn Pro Gln Val Asn 1845 1850 1855 Leu Val Asp Thr Leu Asn Ser Gly Gln Tyr Thr Val Phe Ala Pro Thr 1860 1865 1870 Asn Ala Ala Phe Ser Lys Leu Pro Ala Ser Thr Ile Asp Glu Leu Lys 1875 1880 1885 Thr Asn Ser Ser Leu Leu Thr Ser Ile Leu Thr Tyr His Val Val Ala 1890 1895 1900 Gly Gln Thr Ser Pro Ala Asn Val Val Gly Thr Arg Gln Thr Leu Gln 1905 1910 1915 1920 Gly Ala Ser Val Thr Val Thr Gly Gln Gly Asn Ser Leu Lys Val Gly 1925 1930 1935 Asn Ala Asp Val Val Ser Gly Gly Val Ser Thr Ala Asn Ala Thr Val 1940 1945 1950 Tyr Met Ile Asp Ser Val Leu Met Pro Pro Ala Val Ser Gln Asp Thr 1955 1960 1965 Ser Pro Lys Pro Ala Thr Ser Pro Ala Ala Pro Val Thr Thr Ala Ala 1970 1975 1980 Met Ala Asp Pro Ala Ala Asp Leu Ile Gly Arg Gly Ser Ala Gln Tyr 1985 1990 1995 2000 Ala Ala Gln Asn Pro Thr Gly Pro Gly Ser Val Ala Gly Met Ala Gln 2005 2010 2015 Asp Pro Val Ala Thr Ala Ala Ser Asn Asn Pro Met Leu Ser Thr Leu 2020 2025 2030 Thr Ser Ala Leu Ser Gly Lys Leu Asn Pro Asp Val Asn Leu Val Asp 2035 2040 2045 Thr Leu Asn Gly Gly Glu Tyr Thr Val Phe Ala Pro Thr Asn Ala Ala 2050 2055 2060 Phe Asp Lys Leu Pro Ala Ala Thr Ile Asp Gln Leu Lys Thr Asp Ala 2065 2070 2075 2080 Lys Leu Leu Ser Ser Ile Leu Thr Tyr His Val Ile Ala Gly Gln Ala 2085 2090 2095 Ser Pro Ser Arg Ile Asp Gly Thr His Gln Thr Leu Gln Gly Ala Asp 2100 2105 2110 Leu Thr Val Ile Gly Ala Arg Asp Asp Leu Met Val Asn Asn Ala Gly 2115 2120 2125 Leu Val Ser Gly Gly Val His Thr Ala Asn Ala Thr Val Tyr Met Ile 2130 2135 2140 Asp Thr Val Leu Met Pro Pro Ala Gln 2145 2150 <210> 10 <211> 325 <212> PRT <213> Combined <400> 10 Met Thr Asp Val Ser Arg Lys Ile Arg Ala Trp Gly Arg Arg Leu Met 1 5 10 15 Ile Gly Thr Ala Ala Ala Val Val Leu Pro Gly Leu Val Gly Leu Ala 20 25 30 Gly Gly Ala Ala Thr Ala Gly Ala Phe Ser Arg Pro Gly Leu Pro Val 35 40 45 Glu Tyr Leu Gln Val Pro Ser Pro Ser Met Gly Arg Asp Ile Lys Val 50 55 60 Gln Phe Gln Ser Gly Gly Asn Asn Ser Pro Ala Val Tyr Leu Leu Asp 65 70 75 80 Gly Leu Arg Ala Gln Asp Asp Tyr Asn Gly Trp Asp Ile Asn Thr Pro 85 90 95 Ala Phe Glu Trp Tyr Tyr Gln Ser Gly Leu Ser Ile Val Met Pro Val 100 105 110 Gly Gly Gln Ser Ser Phe Tyr Ser Asp Trp Tyr Ser Pro Ala Cys Gly 115 120 125 Lys Ala Gly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu Thr Ser Glu 130 135 140 Leu Pro Gln Trp Leu Ser Ala Asn Arg Ala Val Lys Pro Thr Gly Ser 145 150 155 160 Ala Ala Ile Gly Leu Ser Met Ala Gly Ser Ser Ala Met Ile Leu Ala 165 170 175 Ala Tyr His Pro Gln Gln Phe Ile Tyr Ala Gly Ser Leu Ser Ala Leu 180 185 190 Leu Asp Pro Ser Gln Gly Met Gly Pro Ser Leu Ile Gly Leu Ala Met 195 200 205 Gly Asp Ala Gly Gly Tyr Lys Ala Ala Asp Met Trp Gly Pro Ser Ser 210 215 220 Asp Pro Ala Trp Glu Arg Asn Asp Pro Thr Gln Gln Ile Pro Lys Leu 225 230 235 240 Val Ala Asn Asn Thr Arg Leu Trp Val Tyr Cys Gly Asn Gly Thr Pro 245 250 255 Asn Glu Leu Gly Gly Ala Asn Ile Pro Ala Glu Phe Leu Glu Asn Phe 260 265 270 Val Arg Ser Ser Asn Leu Lys Phe Gln Asp Ala Tyr Asn Ala Ala Gly 275 280 285 Gly His Asn Ala Val Phe Asn Phe Pro Pro Asn Gly Thr His Ser Trp 290 295 300 Glu Tyr Trp Gly Ala Gln Leu Asn Ala Met Lys Gly Asp Leu Gln Ser 305 310 315 320[[ID=_{20}]] Ser Leu Gly Ala Gly 325 <210> 11 <211> 338 <212> PRT <213> Binding to Mycobacterium <400> 11 Met Gln Leu Val Asp Arg Val Arg Gly Ala Val Thr Gly Met Ser Arg 1 5 10 15 Arg Leu Val Val Gly Ala Val Gly Ala Ala Leu Val Ser Gly Leu Val 20 25 30 Gly Ala Val Gly Gly Thr Ala Thr Ala Gly Ala Phe Ser Arg Pro Gly 35 40 45 Leu Pro Val Glu Tyr Leu Gln Val Pro Ser Pro Ser Met Gly Arg Asp 50 55 60 Ile Lys Val Gln Phe Gln Ser Gly Gly Ala Asn Ser Pro Ala Leu Tyr 65 70 75 80 Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Phe Ser Gly Trp Asp Ile 85 90 95 Asn Thr Pro Ala Phe Glu Trp Tyr Asp Gln Ser Gly Leu Ser Val Val 100 105 110 Met Pro Val Gly Gly Gln Ser Ser Phe Tyr Ser Asp Trp Tyr Gln Pro 115 120 125 Ala Cys Gly Lys Ala Gly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu 130 135 140 Thr Ser Glu Leu Pro Gly Trp Leu Gln Ala Asn Arg His Val Lys Pro 145 150 155 160 Thr Gly Ser Ala Val Val Gly Leu Ser Met Ala Ala Ser Ser Ala Leu 165 170 175 Thr Leu Ala Ile Tyr His Pro Gln Gln Phe Val Tyr Ala Gly Ala Met 180 185 190 Ser Gly Leu Leu Asp Pro Ser Gln Ala Met Gly Pro Thr Leu Ile Gly 195 200 205 Leu Ala Met Gly Asp Ala Gly Gly Tyr Lys Ala Ser Asp Met Trp Gly 210 215 220 Pro Lys Glu Asp Pro Ala Trp Gln Arg Asn Asp Pro Leu Leu Asn Val 225 230 235 240 Gly Lys Leu Ile Ala Asn Asn Thr Arg Val Trp Val Tyr Cys Gly Asn 245 250 255 Gly Lys Pro Ser Asp Leu Gly Gly Asn Asn Leu Pro Ala Lys Phe Leu 260 265 270 Glu Gly Phe Val Arg Thr Ser Asn Ile Lys Phe Gln Asp Ala Tyr Asn 275 280 285 Ala Gly Gly Gly His Asn Gly Val Phe Asp Phe Pro Asp Ser Gly Thr 290 295 300 His Ser Trp Glu Tyr Trp Gly Ala Gln Leu Asn Ala Met Lys Pro Asp 305 310 315 320 Leu Gln Arg Ala Leu Gly Ala Thr Pro Asn Thr Gly Pro Ala Pro Gln 325 330 335 Gly Ala <210> 12 <211> 96 <212> PRT <213> Binding to Mycobacterium <400> 12 Met Ser Gln Ile Met Tyr Asn Tyr Pro Ala Met Leu Gly His Ala Gly 1 5 10 15 Asp Met Ala Gly Tyr Ala Gly Thr Leu Gln Ser Leu Gly Ala Glu Ile 20 25 30 Ala Val Glu Gln Ala Ala Leu Gln Ser Ala Trp Gln Gly Asp Thr Gly 35 40 45 Ile Thr Tyr Gln Ala Trp Gln Ala Gln Trp Asn Gln Ala Met Glu Asp 50 55 60 Leu Val Arg Ala Tyr His Ala Met Ser Ser Thr His Glu Ala Asn Thr 65 70 75 80 Met Ala Met Met Ala Arg Asp Thr Ala Glu Ala Ala Lys Trp Gly Gly 85 90 95 <210> 13 <211> 97 <212> PRT <213> Combined <400> 13 Met Ser Leu Leu Asp Ala His Ile Pro Gln Leu Val Ala Ser Gln Ser 1 5 10 15 Ala Phe Ala Ala Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala 20 25 30 Glu Gln Ala Ala Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ser 35 40 45 Ala Ala Phe Gln Ala Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys 50 55 60 Val Asn Thr Leu Leu Asp Val Ala Gln Ala Asn Leu Gly Glu Ala Ala 65 70 75 80 Gly Thr Tyr Val Ala Ala Asp Ala Ala Ala Ala Ser Thr Tyr Thr Gly 85 90 95 Phew <210> 14 <211> 393 <212> PRT <213> Combined <400> 14 Val Val Asp Phe Gly Ala Leu Pro Pro Glu Ile Asn Ser Ala Arg Met 1 5 10 15 Tyr Ala Gly Pro Gly Ser Ala Ser Leu Val Ala Ala Ala Lys Met Trp 20 25 30 Asp Ser Val Ala Ser Asp Leu Phe Ser Ala Ala Ser Ala Phe Gln Ser 35 40 45 Val Val Trp Gly Leu Thr Val Gly Ser Trp Ile Gly Ser Ser Ala Gly 50 55 60 Leu Met Ala Ala Ala Ala Ser Pro Tyr Val Ala Trp Met Ser Val Thr 65 70 75 80 Ala Gly Gln Ala Gln Leu Thr Ala Ala Gln Val Arg Val Ala Ala Ala 85 90 95 Ala Tyr Glu Thr Ala Tyr Arg Leu Thr Val Pro Pro Pro Val Ile Ala 100 105 110 Glu Asn Arg Thr Glu Leu Met Thr Leu Thr Ala Thr Asn Leu Leu Gly 115 120 125 Gln Asn Thr Pro Ala Ile Glu Ala Asn Gln Ala Ala Tyr Ser Gln Met 130 135 140 Trp Gly Gln Asp Ala Glu Ala Met Tyr Gly Tyr Ala Ala Thr Ala Ala 145 150 155 160 Thr Ala Thr Glu Ala Leu Leu Pro Phe Glu Asp Ala Pro Leu Ile Thr 165 170 175 Asn Pro Gly Gly Leu Leu Glu Gln Ala Val Ala Val Glu Glu Ala Ile 180 185 190 Asp Thr Ala Ala Ala Asn Gln Leu Met Asn Asn Val Pro Gln Ala Leu 195 200 205 Gln Gln Leu Ala Gln Pro Ala Gln Gly Val Val Pro Ser Ser Lys Leu 210 215 220 Gly Gly Leu Trp Thr Ala Val Ser Pro His Leu Ser Pro Leu Ser Asn 225 230 235 240 Val Ser Ser Ile Ala Asn Asn His Met Ser Met Met Gly Thr Gly Val 245 250 255 Ser Met Thr Asn Thr Leu His Ser Met Leu Lys Gly Leu Ala Pro Ala 260 265 270 Ala Ala Gln Ala Val Glu Thr Ala Ala Glu Asn Gly Val Trp Ala Met 275 280 285 Ser Ser Leu Gly Ser Gln Leu Gly Ser Ser Leu Gly Ser Ser Gly Leu 290 295 300 Gly Ala Gly Val Ala Ala Asn Leu Gly Arg Ala Ala Ser Val Gly Ser 305 310 315 320 Leu Ser Val Pro Pro Ala Trp Ala Ala Ala Asn Gln Ala Val Thr Pro 325 330 335 Ala Ala Arg Ala Leu Pro Leu Thr Ser Leu Thr Ser Ala Ala Gln Thr 340 345 350 Ala Pro Gly His Met Leu Gly Gly Leu Pro Leu Gly His Ser Val Asn 355 360 365 Ala Gly Ser Gly Ile Asn Asn Ala Leu Arg Val Pro Ala Arg Ala Tyr 370 375 380 Ala Ile Pro Arg Thr Pro Ala Ala Gly 385 390 <210> 15 <211> 199 <212> PRT <213> Combined <400> 15 Met Ala Glu Asn Ser Asn Ile Asp Asp Ile Lys Ala Pro Leu Leu Ala 1 5 10 15 Ala Leu Gly Ala Ala Asp Leu Ala Leu Ala Thr Val Asn Glu Leu Ile 20 25 30 Thr Asn Leu Arg Glu Arg Ala Glu Glu Thr Arg Thr Asp Thr Arg Ser 35 40 45 Arg Val Glu Glu Ser Arg Ala Arg Leu Thr Lys Leu Gln Glu Asp Leu 50 55 60 Pro Glu Gln Leu Thr Glu Leu Arg Glu Lys Phe Thr Ala Glu Glu Leu 65 70 75 80 Arg Lys Ala Ala Glu Gly Tyr Leu Glu Ala Ala Thr Ser Arg Tyr Asn 85 90 95 Glu Leu Val Glu Arg Gly Glu Ala Ala Leu Glu Arg Leu Arg Ser Gln 100 105 110 Gln Ser Phe Glu Glu Val Ser Ala Arg Ala Glu Gly Tyr Val Asp Gln 115 120 125 Ala Val Glu Leu Thr Gln Glu Ala Leu Gly Thr Val Ala Ser Gln Thr 130 135 140 Arg Ala Val Gly Glu Arg Ala Ala Lys Leu Val Gly Ile Glu Leu Pro 145 150 155 160 Lys Lys Ala Ala Pro Ala Lys Lys Ala Ala Pro Ala Lys Lys Ala Ala 165 170 175 Pro Ala Lys Lys Ala Ala Ala Lys Lys Ala Pro Ala Lys Lys Ala Ala 180 185 190 Ala Lys Lys Val Thr Gln Lys 195 <210> 16 <211> 95 <212> PRT <213> Binding to Mycobacterium <400> 16 Met Ser Asp Gln Ile Thr Tyr Asn Pro Gly Ala Val Ser Asp Phe Ala 1 5 10 15 Ser Asp Val Gly Ser Arg Ala Gly Gln Leu His Met Ile Tyr Glu Asp 20 25 30 Thr Ala Ser Lys Thr Asn Ala Leu Gln Glu Phe Phe Ala Gly His Gly 35 40 45 Ala Gln Gly Phe Phe Asp Ala Gln Ala Gln Met Leu Ser Gly Leu Gln<s 50 55 60 Gly Leu Ile Glu Thr Val Gly Gln His Gly Thr Thr Thr Gly His Val 65 70 75 80 Leu Asp Asn Ala Ile Gly Thr Asp Gln Ala Ile Ala Gly Leu Phe 85 90 95 <210> 17 <211> 107 <212> PRT <213> Mycobacterium conjugatum <400> 17 Val Ala Asp Thr Ile Gln Val Thr Pro Gln Met Leu Arg Ser Thr Ala 1 5 10 15 Asn Asp Ile Gln Ala Asn Met Glu Gln Ala Met Gly Ile Ala Lys Gly 20 25 30 Tyr Leu Ala Asn Gln Glu Asn Val Met Asn Pro Ala Thr Trp Ser Gly 35 40 45 Thr Gly Val Val Ala Ser His Met Thr Ala Thr Glu Ile Thr Asn Glu 50 55 60 Leu Asn Lys Val Leu Thr Gly Gly Thr Arg Leu Ala Glu Gly Leu Val 65 70 75 80 Gln Ala Ala Ala Leu Met Glu Gly His Glu Ala Asp Ser Gln Thr Ala 85 90 95 Phe Gln Ala Leu Phe Gly Ala Ser His Gly Ser 100 105 <210> 18 <211> 163 <212> PRT <213> Mycobacterium conjugatum <400> 18 Val Thr Val Thr Asp Asp Tyr Leu Ala Asn Asn Val Asp Tyr Ala Ser 1 5 10 15 Gly Phe Lys Gly Pro Leu Pro Met Pro Pro Ser Lys His Ile Ala Ile 20 25 30 Val Ala Cys Met Asp Ala Arg Leu Asp Val Tyr Arg Met Leu Gly Ile 35 40 45 Lys Glu Gly Glu Ala His Val Ile Arg Asn Ala Gly Cys Val Val Thr 50 55 60 Asp Asp Val Ile Arg Ser Leu Ala Ile Ser Gln Arg Leu Leu Gly Thr 65 70 75 80 Arg Glu Ile Ile Leu Leu His His Thr Asp Cys Gly Met Leu Thr Phe 85 90 95 ​​​​​​​​​​​​​​​​​​​​​​​​​​​​Met Ser Gln Ile Met Tyr Asn Tyr Pro Ala Met Met Ala His Ala Gly 1 5 10 15 Asp Met Ala Gly Tyr Ala Gly Thr Leu Gln Ser Leu Gly Ala Asp Ile 20 25 30 Ala Ser Glu Gln Ala Val Leu Ser Ser Ala Trp Gln Gly Asp Thr Gly 35 40 45 Ile Thr Tyr Gln Gly Trp Gln Thr Gln Trp Asn Gln Ala Leu Glu Asp 50 55 60 Leu Val Arg Ala Tyr Gln Ser Met Ser Gly Thr His Glu Ser Asn Thr 65 70 75 80 Met Ala Met Leu Ala Arg Asp Gly Ala Glu Ala Ala Lys Trp Gly Gly 85 90 95 <210> 20 <211> 97 <212> PRT <213> Mycobacterium-binding <400> 20 Met Ser Leu Leu Asp Ala His Ile Pro Gln Leu Ile Ala Ser His Thr 1 5 10 15 Ala Phe Ala Ala Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala 20 25 30 Glu Gln Gln Ala Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ala 35 40 45 Ala Ala Phe Gln Gly Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys 50 55 60 Val Asn Thr Leu Leu Asp Ile Ala Gln Ala Asn Leu Gly Glu Ala Ala 65 70 75 80 Gly Thr Tyr Val Ala Ala Asp Ala Ala Ala Ala Ser Ser Tyr Thr Gly 85 90 95 Phew <210> 21 <211> 120 <212> PRT <213> Combined <400> 21 Val Ser Gln Ser Met Tyr Ser Tyr Pro Ala Met Thr Ala Asn Val Gly 1 5 10 15 Asp Met Ala Gly Tyr Thr Gly Thr Thr Gln Ser Leu Gly Ala Asp Ile 20 25 30 Ala Ser Glu Arg Thr Ala Pro Ser Arg Ala Cys Gln Gly Asp Leu Gly 35 40 45 Met Ser His Gln Asp Trp Gln Ala Gln Trp Asn Gln Ala Met Glu Ala 50 55 60 Leu Ala Arg Ala Tyr Arg Arg Cys Arg Arg Ala Leu Arg Gln Ile Gly 65 70 75 80 Val Leu Glu Arg Pro Val Gly Asp Ser Ser Asp Cys Gly Thr Ile Arg 85 90 95 Val Gly Ser Phe Arg Gly Arg Trp Leu Asp Pro Arg His Ala Gly Pro 100 105 110 Ala Thr Ala Ala Asp Ala Gly Asp 115 120 <210> 22 <211> 144 <212> PRT <213> Binding Mycobacterium <400> 22 Met Ala Thr Thr Leu Pro Val Gln Arg His Pro Arg Ser Leu Phe Pro 1 5 10 15 Glu Phe Ser Glu Leu Phe Ala Ala Phe Pro Ser Phe Ala Gly Leu Arg 20 25 30 Pro Thr Phe Asp Thr Arg Leu Met Arg Leu Glu Asp Glu Met Lys Glu 35 40 45 Gly Arg Tyr Glu Val Arg Ala Glu Leu Pro Gly Val Asp Pro Asp Lys 50 55 60 Asp Val Asp Ile Met Val Arg Asp Gly Gln Leu Thr Ile Lys Ala Glu 65 70 75 ⑧ Arg Thr Glu Gln Lys Asp Phe Asp Gly Arg Ser Glu Phe Ala Tyr Gly [[ID=④]]85 90 95 Ser Phe Val Arg Thr Val Ser Leu Pro Val Gly Ala Asp Glu Asp Asp 100 105 110 It should be noted that there seems to be an error in "⑧" in line and "④" in line in the original text. They are likely incorrect characters and should be corrected for a more accurate translation.Ile Lys Ala Thr Tyr Asp Lys Gly Ile Leu Thr Val Ser Val Ala Val 115 120 125 Ser Glu Gly Lys Pro Thr Glu Lys His Ile Gln Ile Arg Ser Thr Asn 130 135 140 <210> 23 <211> 464 <212> PRT <213> Combined <400> 23 Met Ala Lys Leu Ala Arg Val Val Gly Leu Val Gln Glu Glu Gln Pro 1 5 10 15 Ser Asp Met Thr Asn His Pro Arg Tyr Ser Pro Pro Pro Gln Gln Pro 20 25 30 Gly Thr Pro Gly Tyr Ala Gln Gly Gln Gln Gln Thr Tyr Ser Gln Gln 35 40 45 Phe Asp Trp Arg Tyr Pro Pro Ser Pro Pro Pro Gln Pro Thr Gln Tyr 50 55 60 Arg Gln Pro Tyr Glu Ala Leu Gly Gly Thr Arg Pro Gly Leu Ile Pro 65 70 75 80 Gly Val Ile Pro Thr Met Thr Pro Pro Pro Gly Met Val Arg Gln Arg 85 90 95 Pro Arg Ala Gly Met Leu Ala Ile Gly Ala Val Thr Ile Ala Val Val 100 105 110 Ser Ala Gly Ile Gly Gly Ala Ala Ala Ser Leu Val Gly Phe Asn Arg 115 120 125 Ala Pro Ala Gly Pro Ser Gly Gly Pro Val Ala Ala Ser Ala Ala Pro 130 135 140 Ser Ile Pro Ala Ala Asn Met Pro Pro Gly Ser Val Glu Gln Val Ala 145 150 155 160 Ala Lys Val Val Pro Ser Val Val Met Leu Glu Thr Asp Leu Gly Arg 165 170 175 Gln Ser Glu Glu Gly Ser Gly Ile Ile Leu Ser Ala Glu Gly Leu Ile 180 185 190 Leu Thr Asn Asn His Val Ile Ala Ala Ala Ala Lys Pro Pro Leu Gly 195 200 205 Ser Pro Pro Pro Lys Thr Thr Val Thr Phe Ser Asp Gly Arg Thr Ala 210 215 220 Pro Phe Thr Val Val Gly Ala Asp Pro Thr Ser Asp Ile Ala Val Val 225 230 235 240 Arg Val Gln Gly Val Ser Gly Leu Thr Pro Ile Ser Leu Gly Ser Ser 245 250 255 Ser Asp Leu Arg Val Gly Gln Pro Val Leu Ala Ile Gly Ser Pro Leu 260 265 270 Gly Leu Glu Gly Thr Val Thr Thr Gly Ile Val Ser Ala Leu Asn Arg 275 280 285 Pro Val Ser Thr Thr Gly Glu Ala Gly Asn Gln Asn Thr Val Leu Asp 290 295 300 Ala Ile Gln Thr Asp Ala Ala Ile Asn Pro Gly Asn Ser Gly Gly Ala 305 310 315 320 Leu Val Asn Met Asn Ala Gln Leu Val Gly Val Asn Ser Ala Ile Ala 325 330 335 Thr Leu Gly Ala Asp Ser Ala Asp Ala Gln Ser Gly Ser Ile Gly Leu 340 345 350 Gly Phe Ala Ile Pro Val Asp Gln Ala Lys Arg Ile Ala Asp Glu Leu 355 360 365 Ile Ser Thr Gly Lys Ala Ser His Ala Ser Leu Gly Val Gln Val Thr 370 375 380 Asn Asp Lys Asp Thr Leu Gly Ala Lys Ile Val Glu Val Val Ala Gly 385 390 395 400 Gly Ala Ala Ala Asn Ala Gly Val Pro Lys Gly Val Val Val Thr Lys 405 410 415 Val Asp Asp Arg Pro Ile Asn Ser Ala Asp Ala Leu Val Ala Ala Val 420 425 430 Arg Ser Lys Ala Pro Gly Ala Thr Val Ala Leu Thr Phe Gln Asp Pro 435 440 445 Ser Gly Gly Ser Arg Thr Val Gln Val Thr Leu Gly Lys Ala Glu Gln 450 455 460 <210> 24 <211> 391 <212> PRT <213> Combined <400> 24 Met Val Asp Phe Gly Ala Leu Pro Pro Glu Ile Asn Ser Ala Arg Met 1 5 10 15 Tyr Ala Gly Pro Gly Ser Ala Ser Leu Val Ala Ala Ala Gln Met Trp 20 25 30 Asp Ser Val Ala Ser Asp Leu Phe Ser Ala Ala Ser Ala Phe Gln Ser 35 40 45 Val Val Trp Gly Leu Thr Val Gly Ser Trp Ile Gly Ser Ser Ala Gly 50 55 60 Leu Met Val Ala Ala Ala Ser Pro Tyr Val Ala Trp Met Ser Val Thr 65 70 75 80 Ala Gly Gln Ala Glu Leu Thr Ala Ala Gln Val Arg Val Ala Ala Ala 85 90 95 Ala Tyr Glu Thr Ala Tyr Gly Leu Thr Val Pro Pro Pro Val Ile Ala 100 105 110 Glu Asn Arg Ala Glu Leu Met Ile Leu Ile Ala Thr Asn Leu Leu Gly 115 120 125 Gln Asn Thr Pro Ala Ile Ala Val Asn Glu Ala Glu Tyr Gly Glu Met 130 135 140 Trp Ala Gln Asp Ala Ala Ala Met Phe Gly Tyr Ala Ala Ala Thr Ala 145 150 155 160 Thr Ala Thr Ala Thr Leu Leu Pro Phe Glu Glu Ala Pro Glu Met Thr 165 170 175 Ser Ala Gly Gly Leu Leu Glu Gln Ala Ala Ala Val Glu Glu Ala Ser 180 185 190 Asp Thr Ala Ala Ala Asn Gln Leu Met Asn Asn Val Pro Gln Ala Leu 195 200 205 Gln Gln Leu Ala Gln Pro Thr Gln Gly Thr Thr Pro Ser Ser Lys Leu 210 215 220 Gly Gly Leu Trp Lys Thr Val Ser Pro His Arg Ser Pro Ile Ser Asn 225 230 235 240 Met Val Ser Met Ala Asn Asn His Met Ser Met Thr Asn Ser Gly Val 245 250 255 Ser Met Thr Asn Thr Leu Ser Ser Met Leu Lys Gly Phe Ala Pro Ala 260 265 270 Ala Ala Ala Gln Ala Val Gln Thr Ala Ala Gln Asn Gly Val Arg Ala 275 280 285 Met Ser Ser Leu Gly Ser Ser Leu Gly Ser Ser Gly Leu Gly Gly Gly 290 295 300 Val Ala Ala Asn Leu Gly Arg Ala Ala Ser Val Gly Ser Leu Ser Val 305 310 315 320 Pro Gln Ala Trp Ala Ala Ala Asn Gln Ala Val Thr Pro Ala Ala Arg 325 330 335 Ala Leu Pro Leu Thr Ser Leu Thr Ser Ala Ala Glu Arg Gly Pro Gly 340 345 350 Gln Met Leu Gly Gly Leu Pro Val Gly Gln Met Gly Ala Arg Ala Gly 355 360 365 Gly Gly Leu Ser Gly Val Leu Arg Val Pro Pro Arg Pro Tyr Val Met 370 375 380 Pro His Ser Pro Ala Ala Gly 385 390 <210> 25 <211> 580 <212> PRT <213> Combined <400> 25 Met Asn Phe Ala Val Leu Pro Pro Glu Val Asn Ser Ala Arg Ile Phe 1 5 10 15 Path Gly Path Gly Leu Gly Pro Met Leu Path Path Path Ser Path Trp Asp 20 25 30 Gly Leu Ala Glu Glu Leu His Ala Ala Ala Gly Ser Phe Ala Ser Val 35 40 45 Thr Thr Gly Leu Ala Gly Asp Ala Trp His Gly Pro Ala Ser Leu Ala 50 55 60 Met Thr Arg Ala Ala Ser Pro Tyr Val Gly Trp Leu Asn Thr Ala Ala 65 70 75 80 Gly Gln Ala Ala Gln Ala Ala Gly Gln Ala Arg Leu Ala Ala Ser Ala 85 90 95 Phe Glu Ala Thr Leu Ala Ala Thr Val Ser Pro Ala Met Val Ala Ala 100 105 110 Asn Arg Thr Arg Leu Ala Ser Leu Val Ala Ala Asn Leu Leu Gly Gln 115 120 125 Asn Ala Pro Ala Ile Ala Ala Glu Ala Glu Tyr Glu Gln Ile Trp 130 135 140 Ala Gln Asp Val Ala Ala Met Phe Gly Tyr His Ser Ala Ala Ser Ala 145 150 155 160 Val Ala Thr Gln Leu Ala Pro Ile Gln Glu Gly Leu Gln Gln Gln Leu 165 170 175 Gln Asn Val Leu Ala Gln Leu Ala Ser Gly Asn Leu Gly Ser Gly Asn 180 185 190 Val Gly Val Gly Asn Ile Gly Asn Asp Asn Ile Gly Asn Ala Asn Ile 195 200 205 Gly Phe Gly Asn Arg Gly Asp Ala Asn Ile Gly Ile Gly Asn Ile Gly 210 215 220 Asp Arg Asn Leu Gly Ile Gly Asn Thr Gly Asn Trp Asn Ile Gly Ile 225 230 235 240 Gly Ile Thr Gly Asn Gly Gln Ile Gly Phe Gly Lys Pro Ala Asn Pro 245 250 255 Asp Val Leu Val Val Gly Asn Gly Gly Pro Gly Val Thr Ala Leu Val 260 265 270 Met Gly Gly Thr Asp Ser Leu Leu Pro Leu Pro Asn Ile Pro Leu Leu 275 280 285 Glu Tyr Ala Ala Arg Phe Ile Thr Pro Val His Pro Gly Tyr Thr Ala 290 295 300 Thr Phe Leu Glu Thr Pro Ser Gln Phe Phe Pro Phe Thr Gly Leu Asn 305 310 315 320 Ser Leu Thr Tyr Asp Val Ser Val Ala Gln Gly Val Thr Asn Leu His 325 330 335 Thr Ala Ile Met Ala Gln Leu Ala Ala Gly Asn Glu Val Val Val Phe 340 345 350 Gly Thr Ser Gln Ser Ala Thr Ile Ala Thr Phe Glu Met Arg Tyr Leu 355 360 365 Gln Ser Leu Pro Ala His Leu Arg Pro Gly Leu Asp Glu Leu Ser Phe 370 375 380 Thr Leu Thr Gly Asn Pro Asn Arg Pro Asp Gly Gly Ile Leu Thr Arg 385 390 395 400 Phe Gly Phe Ser Ile Pro Gln Leu Gly Phe Thr Leu Ser Gly Ala Thr 405 410 415 Pro Ala Asp Ala Tyr Pro Thr Val Asp Tyr Ala Phe Gln Tyr Asp Gly 420 425 430 Val Asn Asp Phe Pro Lys Tyr Pro Leu Asn Val Phe Ala Thr Ala Asn 435 440 445 Ala Ile Ala Gly Ile Leu Phe Leu His Ser Gly Leu Ile Ala Leu Pro 450 455 460 Pro Asp Leu Ala Ser Gly Val Val Gln Pro Val Ser Ser Pro Asp Val 465 470 475 480 Leu Thr Thr Tyr Ile Leu Leu Pro Ser Gln Asp Leu Pro Leu Leu Val 485 490 495 Pro Leu Arg Ala Ile Pro Leu Leu Gly Asn Pro Leu Ala Asp Leu Ile 500 505 510 Gln Pro Asp Leu Arg Val Leu Val Glu Leu Gly Tyr Asp Arg Thr Ala 515 520 525 His Gln Asp Val Pro Ser Pro Phe Gly Leu Phe Pro Asp Val Asp Trp 530 535 540 Ala Glu Val Ala Ala Asp Leu Gln Gln Gly Ala Val Gln Gly Val Asn 545 550 555 560 Asp Ala Leu Ser Gly Leu Gly Leu Pro Pro Pro Trp Gln Pro Ala Leu 565 570 575 Pro Arg Leu Phe 580 <210> 26 <211> 94 <212> PRT <213> Mycobacterium tuberculosis <400> 26 Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met 1 5 10 15 Ile Arg Ala Gln Ala Gly Ser Leu Glu Ala Glu His Gln Ala Ile Ile 20 25 30 Ser Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 35 40 45 Ala Cys Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 50 55 60 Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn 65 70 75 80 Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 85 90 <210> 27 <211> 98 <212> PRT <213> Mycobacterium tuberculosis <400> 27 Met Thr Ser Arg Phe Met Thr Asp Pro His Ala Met Arg Asp Met Ala 1 5 10 15 Gly Arg Phe Glu Val His Ala Gln Thr Val Glu Asp Glu Ala Arg Arg 20 25 30 Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala Gly Trp Ser Gly Met 35 40 45 Ala Glu Ala Thr Ser Leu Asp Thr Met Thr Gln Met Asn Gln Ala Phe 50 55 60 Arg Asn Ile Val Asn Met Leu His Gly Val Arg Asp Gly Leu Val Arg 65 70 75 80 Asp Ala Asn Asn Tyr Glu Gln Gln Glu Gln Ala Ser Gln Gln Ile Leu 85 90 95 Ser Ser <210> 28 <211> 75 <212> PRT <213> Mycobacterium tuberculosis <400> 28 Val Ile Ala Gly Val Asp Gln Ala Leu Ala Ala Thr Gly Gln Ala Ser 1 5 10 15 Gln Arg Ala Ala Gly Ala Ser Gly Gly Val Thr Val Gly Val Gly Val 20 25 30 Gly Thr Glu Gln Arg Asn Leu Ser Val Val Ala Pro Ser Gln Phe Thr 35 40 45 Phe Ser Ser Arg Ser Pro Asp Phe Val Asp Glu Thr Ala Gly Gln Ser 50 55 60 Trp Cys Ala Ile Leu Gly Leu Asn Gln Phe His 65 70 75 <210> 29 <211> 184 <212> PRT <213> Mycobacterium tuberculosis <400> 29 Val Asp Leu Pro Gly Asn Asp Phe Asp Ser Asn Asp Phe Asp Ala Val 1 5 10 15 Asp Leu Trp Gly Ala Asp Gly Ala Glu Gly Trp Thr Ala Asp Pro Ile 20 25 30 Ile Gly Val Gly Ser Ala Ala Thr Pro Asp Thr Gly Pro Asp Leu Asp 35 40 45 Asn Ala His Gly Gln Ala Glu Thr Asp Thr Glu Gln Glu Ile Ala Leu 50 55 60 Phe Thr Val Thr Asn Pro Pro Arg Thr Val Ser Val Ser Thr Leu Met 65 70 75 80 Asp Gly Arg Ile Asp His Val Glu Leu Ser Ala Arg Val Ala Trp Met 85 90 95 Ser Glu Ser Gln Leu Ala Ser Glu Ile Leu Val Ile Ala Asp Leu Ala 100 105 110 Arg Gln Lys Ala Gln Ser Ala Gln Tyr Ala Phe Ile Leu Asp Arg Met 115 120 125 Ser Gln Gln Val Asp Ala Asp Glu His Arg Val Ala Leu Leu Arg Lys 130 135 140 Thr Val Gly Glu Thr Trp Gly Leu Pro Ser Pro Glu Glu Ala Ala Ala 145 150 155 160 Ala Glu Ala Glu Val Phe Ala Thr Arg Tyr Ser Asp Asp Cys Pro Ala 165 170 175 Pro Asp Asp Glu Ser Asp Pro Trp 180 <210> 30 <211> 103 <212> PRT <213> Mycobacterium tuberculosis <400> 30 Met Thr Gly Phe Leu Gly Val Val Pro Ser Phe Leu Lys Val Leu Ala 1 5 10 15 Gly Met His Asn Glu Ile Val Gly Asp Ile Lys Arg Ala Thr Asp Thr 20 25 30 Val Ala Gly Ile Ser Gly Arg Val Gln Leu Thr His Gly Ser Phe Thr 35 40 45 Ser Lys Phe Asn Asp Thr Leu Gln Glu Phe Glu Thr Thr Arg Ser Ser 50 55 60 Thr Gly Thr Gly Leu Gln Gly Val Thr Ser Gly Leu Ala Asn Asn Leu 65 70 75 80 Leu Ala Ala Ala Gly Ala Tyr Leu Lys Ala Asp Asp Gly Leu Ala Gly 85 90 95 Val Ile Asp Lys Ile Phe Gly 100 <210> 31 <211> 132 <212> PRT <213> Mycobacterium tuberculosis <400> 31 Met Ser Thr Thr Phe Ala Ala Arg Leu Asn Arg Leu Phe Asp Thr Val 1 5 10 15 Tyr Pro Pro Gly Arg Gly Pro His Thr Ser Ala Glu Val Ile Ala Ala 20 25 30 Leu Lys Ala Glu Gly Ile Thr Met Ser Ala Pro Tyr Leu Ser Gln Leu 35 40 45 Arg Ser Gly Asn Arg Thr Asn Pro Ser Gly Ala Thr Met Ala Ala Leu 50 55 60 Ala Asn Phe Phe Arg Ile Lys Ala Ala Tyr Phe Thr Asp Asp Glu Tyr 65 70 75 80 Tyr Glu Lys Leu Asp Lys Glu Leu Gln Trp Leu Cys Thr Met Arg Asp 85 90 95 Asp Gly Val Arg Arg Ile Ala Gln Arg Ala His Gly Leu Pro Ser Ala 100 105 110 Ala Gln Gln Lys Val Leu Asp Arg Ile Asp Glu Leu Arg Arg Ala Glu 115 120 125 Gly Ile Asp Ala 130 <210> 32 <211> 99 <212> PRT <213> Mycobacterium tuberculosis <400> 32 Met Glu Lys Met Ser His Asp Pro Ile Ala Ala Asp Ile Gly Thr Gln 1 5 10 15<00​​​​Thr Ser Val Thr Gly Leu Val Pro Ala Gly Ala Asp Glu Val Ser Ala 35 40 45 Gln Ala Ala Thr Ala Phe Thr Ser Glu Gly Ile Gln Leu Leu Ala Ser 50 55 60 Asn Ala Ser Ala Gln Asp Gln Leu His Arg Ala Gly Glu Ala Val Gln 65 70 75 80 Asp Val Ala Arg Thr Tyr Ser Gln Ile Asp Asp Gly Ala Ala Gly Val 85 90 95 Phe Ala Glu <210> 33 <211> 460 <212> PRT <213> Tuberculosis <400> 33 Met Thr Gln Ser Gln Thr Val Thr Val Asp Gln Gln Glu Ile Leu Asn 1 5 10 15 Arg Ala Asn Glu Val Glu Ala Pro Met Ala Asp Pro Pro Thr Asp Val 20 25 30 Pro Ile Thr Pro Cys Glu Leu Thr Ala Ala Lys Asn Ala Ala Gln Gln 35 40 45 Leu Val Leu Ser Ala Asp Asn Met Arg Glu Tyr Leu Ala Ala Gly Ala 50 55 60 Lys Glu Arg Gln Arg Leu Ala Thr Ser Leu Arg Asn Ala Ala Lys Ala 65 70 75 80 Tyr Gly Glu Val Asp Glu Glu Ala Ala Thr Ala Leu Asp Asn Asp Gly 85 90 95 Glu Gly Thr Val Gln Ala Glu Ser Ala Gly Ala Val Gly Gly Asp Ser 100 105 110 Ser Ala Glu Leu Thr Asp Thr Pro Arg Val Ala Thr Ala Gly Glu Pro 115 120 125 Asn Phe Met Asp Leu Lys Glu Ala Ala Arg Lys Leu Glu Thr Gly Asp 130 135 140 Gln Gly Ala Ser Leu Ala His Phe Ala Asp Gly Trp Asn Thr Phe Asn 145 150 155 160 Leu Thr Leu Gln Gly Asp Val Lys Arg Phe Arg Gly Phe Asp Asn Trp 165 170 175 Glu Gly Asp Ala Ala Thr Ala Cys Glu Ala Ser Leu Asp Gln Gln Arg 180 185 190 Gln Trp Ile Leu His Met Ala Lys Leu Ser Ala Ala Met Ala Lys Gln 195 200 205 Ala Gln Tyr Val Ala Gln Leu His Val Trp Ala Arg Arg Glu His Pro 210 215 220 Thr Tyr Glu Asp Ile Val Gly Leu Glu Arg Leu Tyr Ala Glu Asn Pro 225 230 235 240 Ser Ala Arg Asp Gln Ile Leu Pro Val Tyr Ala Glu Tyr Gln Gln Arg 245 250 255 Ser Glu Lys Val Leu Thr Glu Tyr Asn Asn Lys Ala Ala Leu Glu Pro 260 265 270 Val Asn Pro Pro Lys Pro Pro Pro Ala Ile Lys Ile Asp Pro Pro Pro 275 280 285 Pro Pro Gln Glu Gln Gly Leu Ile Pro Gly Phe Leu Met Pro Pro Ser 290 295 300 Asp Gly Ser Gly Val Thr Pro Gly Thr Gly Met Pro Ala Ala Pro Met 305 310 315 320 Val Pro Pro Thr Gly Ser Pro Gly Gly Gly Leu Pro Ala Asp Thr Ala 325 330 335 Ala Gln Leu Thr Ser Ala Gly Arg Glu Ala Ala Ala Leu Ser Gly Asp 340 345 350 Val Ala Val Lys Ala Ala Ser Leu Gly Gly Gly Gly Gly Gly Gly Val 355 360 365 Pro Ser Ala Pro Leu Gly Ser Ala Ile Gly Gly Ala Glu Ser Val Arg 370 375 380 Pro Ala Gly Ala Gly Asp Ile Ala Gly Leu Gly Gln Gly Arg Ala Gly 385 390 395 400 Gly Gly Ala Ala Leu Gly Gly Gly Gly Met Gly Met Pro Met Gly Ala 405 410 415 Ala His Gln Gly Gln Gly Gly Ala Lys Ser Lys Gly Ser Gln Gln Glu 420 425 430 Asp Glu Ala Leu Tyr Thr Glu Asp Arg Ala Trp Thr Glu Ala Val Ile 435 440 445 Gly Asn Arg Arg Arg Gln Asp Ser Lys Glu Ser Lys 450 455 460 <210> 34 <211> 2100 <212> PRT <213> Mycobacterium tuberculosis <400> 34 Met Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 1 5 10 15 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 20 25 30 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 35 40 45 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu[[ID=4%]] 50 55 60 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 65 70 75 80 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Gln 85 90 95 Ala Glu Thr Ala Val Asn Thr Leu Phe Glu Lys Leu Glu Pro Met Ala 100 105 110 Ser Ile Leu Asp Pro Gly Ala Ser Gln Ser Thr Thr Asn Pro Ile Phe 115 120 125 Gly Met Pro Ser Pro Gly Ser Ser Thr Pro Val Gly Gln Leu Pro Pro 130 135 140 Ala Ala Thr Gln Thr Leu Gly Gln Leu Gly Glu Met Ser Gly Pro Met 145 150 155 160 Gly Gly Ser Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met Gly Leu 165 170 175 Leu Gly Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly Ser Gly 180 185 190 Pro Ser Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro Gly Ala 195 200 205 Gly Gly Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile Glu Lys 210 215 220 Pro Val Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala 225 230 235 240 Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp 245 250 255 Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser 260 265 270 Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala 275 280 285 Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu 290 295 300 Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe 305 310 315 320 Ala Ala Ala Asp Tyr Asp Lys Leu Phe Arg Pro His Glu Gly Met Glu 325 330 335 Ala Pro Asp Asp Met Ala Ala Gln Pro Phe Phe Asp Pro Ser Ala Ser 340 345 350 Phe Pro Pro Ala Pro Ala Ser Ala Asn Leu Pro Lys Pro Asn Gly Gln 355 360 365 Thr Pro Pro Pro Thr Ser Asp Asp Leu Ser Glu Arg Phe Val Ser Ala 370 375 380 Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr Pro Met 385 390 395 400 Pro Ile Ala Ala Gly Glu Pro Pro Ser Pro Glu Pro Ala Ala Ser Lys 405 410 415 Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala Pro Pro 420 425 430 Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala Pro 435 440 445 For Lys For Thr For Met For Ile Ala Gly For Ala For Thr 450 455 460 Pro Thr Glu Ser Gln Leu Ala Pro Pro Arg Pro Pro Thr Pro Gln Thr 465 470 475 480 For Thr Gly Ala For Gln Gln For Glu Ser For Ala For His Val For 485 490 495 Ser His Gly Pro His Gln Pro Arg Arg Thr Ala Pro Ala Pro Pro Trp 500 505 510 Ala Lys Met Pro Ile Gly Glu Pro Pro Pro Ala Pro Ser Arg Pro Ser 515 520 525 Ala Ser Pro Ala Glu Pro Pro Thr Arg Pro Ala Pro Gln His Ser Arg 530 535 540 Arg Ala Arg Arg Gly His Arg Tyr Arg Thr Asp Thr Glu Arg Asn Val 545 550 555 560 Gly Lys Val Ala Thr Gly Pro Ser Ile Gln Ala Arg Leu Arg Ala Glu 565 570 575 Glu Ala Ser Gly Ala Gln Leu Ala Pro Gly Thr Glu Pro Ser Pro Ala 580 585 590 Pro Leu Gly Gln Pro Arg Ser Tyr Leu Ala Pro Pro Thr Arg Pro Ala 595 600 605 Pro Thr Glu Pro Pro Pro Ser Pro Ser Pro Gln Arg Asn Ser Gly Arg 610 615 620 Arg Ala Glu Arg Arg Val His Pro Asp Leu Ala Ala Gln His Ala Ala 625 630 635 640 Ala Gln Pro Asp Ser Ile Thr Ala Ala Thr Thr Gly Gly Arg Arg Arg 645 650 655 Lys Arg Ala Ala Pro Asp Leu Asp Ala Thr Gln Lys Ser Leu Arg Pro 660 665 670 Ala Ala Lys Gly Pro Lys Val Lys Lys Val Lys Pro Gln Lys Pro Lys 675 680 685 Ala Thr Lys Pro Pro Lys Val Val Ser Gln Arg Gly Trp Arg His Trp 690 695 700 Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser Pro Asp Glu Lys 705 710 715 720 Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn Pro Arg Gly Ser 725 730 735 Tyr Gln Ile Ala Val Val Gly Leu Gln Gly Gly Ala Gly Lys Thr Thr 740 745 750 Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val Arg Ala Asp Arg 755 760 765 Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn Leu Ala Asp Arg 770 775 780 Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val Leu Ala Glu Lys 785 790 795 800 Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr Ser Val Asn Ala 805 810 815 Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser Ser Ala Gln Arg 820 825 830 Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp Pro Ala Ser Arg 835 840 845 Phe Tyr Asn Leu Val Leu Ala Asp Ser Gly Ala Gly Phe Phe Asp Pro 850 855 860 Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val Val Val Val Ala 865 870 875 880 Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val Ala Leu Asp Trp 885 890 895 Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg Ala Ser Val Val 900 905 910 Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala Val Lys Asp Leu 915 920 925 Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg Val Val Val Met 930 935 940 Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile Ser Leu Asp Leu 945 950 955 960 Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu Ala Ala Ala Leu 965 970 975 Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg Thr Glu Gln Gln Trp Asn 980 985 990 Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr 995 1000 1005 Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu 1010 1015 1020 Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln 1025 1030 1035 1040 Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn 1045 1050 1055 Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu 1060 1065 1070 Gly Asn Val Thr Gly Met Phe Ala Ser Arg Ala Phe Ile Ile Asp Pro 1075 1080 1085 Thr Ile Ser Ala Ile Asp Gly Leu Tyr Asp Leu Leu Gly Ile Gly Ile 1090 1095 1100 Pro Asn Gln Gly Gly Ile Leu Tyr Ser Ser Leu Glu Tyr Phe Glu Lys 1105 1110 1115 1120 Ala Leu Glu Glu Leu Ala Ala Ala Phe Pro Gly Asp Gly Trp Leu Gly 1125 1130 1135 Ser Ala Ala Asp Lys Tyr Ala Gly Lys Asn Arg Asn His Val Asn Phe 1140 1145 1150 Phe Gln Glu Leu Ala Asp Leu Asp Arg Gln Leu Ile Ser Leu Ile His 1155 1160 1165 Asp Gln Ala Asn Ala Val Gln Thr Thr Arg Asp Ile Leu Glu Gly Ala 1170 1175 1180 Lys Lys Gly Leu Glu Phe Val Arg Pro Val Ala Val Asp Leu Thr Tyr 1185 1190 1195 1200 Ile Pro Val Val Gly His Ala Leu Ser Ala Lys Thr Leu Ile Asn Ala 1205 1210 1215 Thr Gln Leu Leu Lys Leu Leu Ala Lys Leu Ala Glu Leu Val Ala Ala 1220 1225 1230 Ala Ile Ala Asp Ile Ile Ser Asp Val Ala Asp Ile Ile Lys Gly Thr 1235 1240 1245 Leu Gly Glu Val Trp Glu Phe Ile Thr Asn Ala Leu Asn Gly Leu Lys 1250 1255 1260 Glu Leu Trp Asp Lys Leu Thr Gly Trp Val Thr Gly Leu Phe Ser Arg 1265 1270 1275 1280 Gly Trp Ser Asn Leu Glu Ser Phe Phe Ala Gly Val Pro Gly Leu Thr 1285 1290 1295 Gly Ala Thr Ser Gly Leu Ser Gln Val Thr Gly Leu Phe Gly Ala Ala 1300 1305 1310 Gly Leu Ser Ala Ser Ser Gly Leu Ala His Ala Asp Ser Leu Ala Ser 1315 1320 1325 Ser Ala Ser Leu Pro Ala Leu Ala Gly Ile Gly Gly Gly Ser Gly Phe 1330 1335 1340 Gly Gly Leu Pro Ser Leu Ala Gln Val His Ala Ala Ser Thr Arg Gln 1345 1350 1355 1360 Ala Leu Arg Pro Arg Ala Asp Gly Pro Val Gly Ala Ala Ala Glu Gln 1365 1370 1375 Val Gly Gly Gln Ser Gln Leu Val Ser Ala Gln Gly Ser Gln Gly Met 1380 1385 1390 Gly Gly Pro Val Gly Met Gly Gly Met His Pro Ser Ser Gly Ala Ser 1395 1400 1405 Lys Gly Thr Thr Thr Lys Lys Tyr Ser Glu Gly Ala Ala Ala Gly Thr 1410 1415 1420 Glu Asp Ala Glu Arg Ala Pro Val Glu Ala Asp Ala Gly Gly Gly Gln 1425 1430 1435 1440 Lys Val Leu Val Arg Asn Val Val Thr Glu Gln Gln Trp Asn Phe Ala 1445 1450 1455 Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile 1460 1465 1470 His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala 1475 1480 1485 Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys 1490 1495 1500 Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala 1505 1510 1515 1520 Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn 1525 1530 1535 Val Thr Gly Met Phe Ala Ala Pro Lys Thr Tyr Ser Glu Glu Leu Lys 1540 1545 1550 Gly Thr Asp Thr Gly Gln Ala Ser Gln Ile Gln Met Ser Asp Pro Ala 1555 1560 1565 Tyr Asn Ile Asn Ile Ser Leu Pro Ser Tyr Tyr Pro Asp Gln Lys Ser 1570 1575 1580 Leu Glu Asn Tyr Ile Ala Gln Thr Arg Asp Lys Phe Leu Ser Ala Ala 1585 1590 1595 1600 Thr Ser Ser Thr Pro Arg Glu Ala Pro Tyr Glu Leu Asn Ile Thr Ser 1605 1610 1615 Ala Thr Tyr Gln Ser Ala Ile Pro Pro Arg Gly Thr Gln Ala Val Val 1620 1625 1630 Leu Lys Val Tyr Gln Asn Ala Gly Gly Thr His Pro Thr Thr Thr Tyr 1635 1640 1645 Lys Ala Phe Asp Trp Asp Gln Ala Tyr Arg Lys Pro Ile Thr Tyr Asp 1650 1655 1660 Thr Leu Trp Gln Ala Asp Thr Asp Pro Leu Pro Val Val Phe Pro Ile 1665 1670 1675 1680 Val Gln Gly Glu Leu Ser Lys Gln Thr Gly Gln Gln Val Ser Ile Ala 1685 1690 1695 Pro Asn Ala Gly Leu Asp Pro Val Asn Tyr Gln Asn Phe Ala Val Thr 1700 1705 1710 Asn Asp Gly Val Ile Phe Phe Phe Asn Pro Gly Glu Leu Leu Pro Glu 1715 1720 1725 Ala Ala Gly Pro Thr Gln Val Leu Val Pro Arg Ser Ala Ile Asp Ser 1730 1735 1740 Met Leu Ala Gly Asp Leu Val Gly Pro Gly Ser Ala Glu Tyr Ala Ala 1745 1750 1755 1760 Ala Asn Pro Thr Gly Pro Ala Ser Val Gln Gly Met Ser Gln Asp Pro 1765 1770 1775 Val Ala Val Ala Ala Ser Asn Asn Pro Glu Leu Thr Thr Leu Thr Ala 1780 1785 1790 Ala Leu Ser Gly Gln Leu Asn Pro Gln Val Asn Leu Val Asp Thr Leu 1795 1800 1805 Asn Ser Gly Gln Tyr Thr Val Phe Ala Pro Thr Asn Ala Ala Phe Ser 1810 1815 1820 Lys Leu Pro Ala Ser Thr Ile Asp Glu Leu Lys Thr Asn Ser Ser Leu 1825 1830 1835 1840 Leu Thr Ser Ile Leu Thr Tyr His Val Val Ala Gly Gln Thr Ser Pro 1845 1850 1855 Ala Asn Val Val Gly Thr Arg Gln Thr Leu Gln Gly Ala Ser Val Thr 1860 1865 1870 Val Thr Gly Gln Gly Asn Ser Leu Lys Val Gly Asn Ala Asp Val Val 1875 1880 1885 Ser Gly Gly Val Ser Thr Ala Asn Ala Thr Val Tyr Met Ile Asp Ser 1890 1895 1900 Val Leu Met Pro Pro Ala Val Ser Gln Asp Thr Ser Pro Lys Pro Ala 1905 1910 1915 1920 Thr Ser Pro Ala Ala Pro Val Thr Thr Ala Ala Met Ala Asp Pro Ala 1925 1930 1935 Ala Asp Leu Ile Gly Arg Gly Ser Ala Gln Tyr Ala Ala Gln Asn Pro 1940 1945 1950 Thr Gly Pro Gly Ser Val Ala Gly Met Ala Gln Asp Pro Val Ala Thr 1955 1960 1965 Ala Ala Ser Asn Asn Pro Met Leu Ser Thr Leu Thr Ser Ala Leu Ser 1970 1975 1980 Gly Lys Leu Asn Pro Asp Val Asn Leu Val Asp Thr Leu Asn Gly Gly 1985 1990 1995 2000 Glu Tyr Thr Val Phe Ala Pro Thr Asn Ala Ala Phe Asp Lys Leu Pro 2005 2010 2015 Ala Ala Thr Ile Asp Gln Leu Lys Thr Asp Ala Lys Leu Leu Ser Ser 2020 2025 2030 Ile Leu Thr Tyr His Val Ile Ala Gly Gln Ala Ser Pro Ser Arg Ile 2035 2040 2045 Asp Gly Thr His Gln Thr Leu Gln Gly Ala Asp Leu Thr Val Ile Gly 2050 2055 2060 Ala Arg Asp Asp Leu Met Val Asn Asn Ala Gly Leu Val Ser Gly Gly 2065 2070 2075 2080 Val His Thr Ala Asn Ala Thr Val Tyr Met Ile Asp Thr Val Leu Met 2085 2090 2095 Pro Pro Ala Gln 2100 <210> 35 <211> 1771 <212> PRT <213> Tuberculosis <400> 35 Met Gln Ala Glu Thr Ala Val Asn Thr Leu Phe Glu Lys Leu Glu Pro 1 5 10 15 Met Ala Ser Ile Leu Asp Pro Gly Ala Ser Gln Ser Thr Thr Asn Pro 20 25 30 Ile Phe Gly Met Pro Ser Pro Gly Ser Ser Thr Pro Val Gly Gln Leu 35 40 45 Pro Pro Ala Ala Thr Gln Thr Leu Gly Gln Leu Gly Glu Met Ser Gly 50 55 60 Pro Met Gly Gly Ser Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met 65 70 75 80 Gly Leu Leu Gly Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly 85 90 95 Ser Gly Pro Ser Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro 100 105 110 Gly Ala Gly Gly Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile 115 120 125 Glu Lys Pro Val Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu 130 135 140 Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu 145 150 155 160 Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly 165 170 175 Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala 180 185 190 Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile 195 200 205 Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly 210 215 220 Met Phe Ala Trp Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser 225 230 235 240 Pro Asp Glu Lys Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn 245 250 255 Pro Arg Gly Ser Tyr Gln Ile Ala Val Val Gly Leu Gln Gly Gly Ala 260 265 270 Gly Lys Thr Thr Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val 275 280 285 Arg Ala Asp Arg Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn 290 295 300 Leu Ala Asp Arg Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val 305 310 315 320 Leu Ala Glu Lys Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr 325 330 335 Ser Val Asn Ala Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser 340 345 350 Ser Ala Gln Arg Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp 355 360 365 Pro Ala Ser Arg Phe Tyr Asn Leu Val Leu Ala Asp Ser Gly Ala Gly 370 375 380 Phe Phe Asp Pro Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val 385 390 395 400 Val Val Val Ala Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val 405 410 415 Ala Leu Asp Trp Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg 420 425 430 Ala Ser Val Val Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala 435 440 445 Val Lys Asp Leu Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg 450 455 460 Val Val Val Met Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile 465 470 475 480 Ser Leu Asp Leu Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu 485 490 495 Ala Ala Ala Leu Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg Thr Glu 500 505 510 Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln 515 520 525 Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser 530 535 540 Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr 545 550 555 560 Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn 565 570 575 Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met 580 585 590 Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Thr Glu Asn Leu 595 600 605 Thr Val Gln Pro Glu Arg Leu Gly Val Leu Ala Ser His His Asp Asn 610 615 620 Ala Ala Val Asp Ala Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly 625 630 635 640 Glu Ser Val Ala Ile Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp 645 650 655 Thr Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 660 665 670 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 675 680 685 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 690 695 700 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu 705 710 715 720 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 725 730 735 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ser 740 745 750 Arg Ala Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly Leu Tyr 755 760 765 Asp Leu Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu Tyr Ser 770 775 780 Serum Leu Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala Ala Phe 785 790 795 800 Pro Gly Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala Gly Lys 805 810 815 Asn Arg Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu Asp Arg 820 825 830 Gln Leu Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln Thr Thr 835 840 845 Arg Asp Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val Arg Pro 850 855 860 Val Ala Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala Leu Ser 865 870 875 880 Ala Lys Thr Leu Ile Asn Ala Thr Gln Leu Leu Lys Leu Leu Ala Lys 885 890 895 Leu Ala Glu Leu Val Ala Ala Ala Ile Ala Asp Ile Ile Ser Asp Val 900 905 910 Ala Asp Ile Ile Lys Gly Thr Leu Gly Glu Val Trp Glu Phe Ile Thr 915 920 925 Asn Ala Leu Asn Gly Leu Lys Glu Leu Trp Asp Lys Leu Thr Gly Trp 930 935 940 Val Thr Gly Leu Phe Ser Arg Gly Trp Ser Asn Leu Glu Ser Phe Phe 945 950 955 960 Ala Gly Val Pro Gly Leu Thr Gly Ala Thr Ser Gly Leu Ser Gln Val 965 970 975 Thr Gly Leu Phe Gly Ala Ala Gly Leu Ser Ala Ser Ser Gly Leu Ala 980 985 990 His Ala Asp Ser Leu Ala Ser Ser Ala Ser Leu Pro Ala Leu Ala Gly 995 1000 1005 Ile Gly Gly Gly Ser Gly Phe Gly Gly Leu Pro Ser Leu Ala Gln Val 1010 1015 1020 His Ala Ala Ser Thr Arg Gln Ala Leu Arg Pro Arg Ala Asp Gly Pro 1025 1030 1035 1040 Val Gly Ala Ala Ala Glu Gln Val Gly Gly Gln Ser Gln Leu Val Ser 1045 1050 1055 Ala Gln Gly Ser Gln Gly Met Gly Gly Pro Val Gly Met Gly Gly Met 1060 1065 1070 His Pro Ser Ser Gly Ala Ser Lys Gly Thr Thr Thr Lys Lys Tyr Ser 1075 1080 1085 Glu Gly Ala Ala Ala Gly Thr Glu Asp Ala Glu Arg Ala Pro Val Glu 1090 1095 1100 Ala Asp Ala Gly Gly Gly Gln Lys Val Leu Val Arg Asn Val Val Thr 1105 1110 1115 1120 Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile 1125 1130 1135 Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln 1140 1145 1150 Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala 1155 1160 1165 Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn 1170 1175 1180 Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala 1185 1190 1195 1200 Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ala Pro Lys 1205 1210 1215 Thr Tyr Ser Glu Glu Leu Lys Gly Thr Asp Thr Gly Gln Ala Ser Gln 1220 1225 1230 Ile Gln Met Ser Asp Pro Ala Tyr Asn Ile Asn Ile Ser Leu Pro Ser 1235 1240 1245 Tyr Tyr Pro Asp Gln Lys Ser Leu Glu Asn Tyr Ile Ala Gln Thr Arg 1250 1255 1260 Asp Lys Phe Leu Ser Ala Ala Thr Ser Ser Thr Pro Arg Glu Ala Pro 1265 1270 1275 1280 Tyr Glu Leu Asn Ile Thr Ser Ala Thr Tyr Gln Ser Ala Ile Pro Pro 1285 1290 1295 Arg Gly Thr Gln Ala Val Val Leu Lys Val Tyr Gln Asn Ala Gly Gly 1300 1305 1310 Thr His Pro Thr Thr Thr Tyr Lys Ala Phe Asp Trp Asp Gln Ala Tyr 1315 1320 1325 Arg Lys Pro Ile Thr Tyr Asp Thr Leu Trp Gln Ala Asp Thr Asp Pro 1330 1335 1340 Leu Pro Val Val Phe Pro Ile Val Gln Gly Glu Leu Ser Lys Gln Thr 1345 1350 1355 1360 Gly Gln Gln Val Ser Ile Ala Pro Asn Ala Gly Leu Asp Pro Val Asn 1365 1370 1375 Tyr Gln Asn Phe Ala Val Thr Asn Asp Gly Val Ile Phe Phe Phe Asn 1380 1385 1390 Pro Gly Glu Leu Leu Pro Glu Ala Ala Gly Pro Thr Gln Val Leu Val 1395 1400 1405 Pro Arg Ser Ala Ile Asp Ser Met Leu Ala Gly Asp Leu Val Gly Pro 1410 1415 1420 Gly Ser Ala Glu Tyr Ala Ala Ala Asn Pro Thr Gly Pro Ala Ser Val 1425 1430 1435 1440 Gln Gly Met Ser Gln Asp Pro Val Ala Val Ala Ala Ser Asn Asn Pro 1445 1450 1455 Glu Leu Thr Thr Leu Thr Ala Ala Leu Ser Gly Gln Leu Asn Pro Gln 1460 1465 1470 Val Asn Leu Val Asp Thr Leu Asn Ser Gly Gln Tyr Thr Val Phe Ala 1475 1480 1485 Pro Thr Asn Ala Ala Phe Ser Lys Leu Pro Ala Ser Thr Ile Asp Glu 1490 1495 1500 Leu Lys Thr Asn Ser Ser Leu Leu Thr Ser Ile Leu Thr Tyr His Val 1505 1510 1515 1520 Val Ala Gly Gln Thr Ser Pro Ala Asn Val Val Gly Thr Arg Gln Thr 1525 1530 1535 Leu Gln Gly Ala Ser Val Thr Val Thr Gly Gln Gly Asn Ser Leu Lys 1540 1545 1550 Val Gly Asn Ala Asp Val Val Ser Gly Gly Val Ser Thr Ala Asn Ala 1555 1560 1565 Thr Val Tyr Met Ile Asp Ser Val Leu Met Pro Pro Ala Val Ser Gln 1570 1575 1580 Asp Thr Ser Pro Lys Pro Ala Thr Ser Pro Ala Ala Pro Val Thr Thr 1585 1590 1595 1600 Ala Ala Met Ala Asp Pro Ala Ala Asp Leu Ile Gly Arg Gly Ser Ala 1605 1610 1615 Gln Tyr Ala Ala Gln Asn Pro Thr Gly Pro Gly Ser Val Ala Gly Met 1620 1625 1630 Ala Gln Asp Pro Val Ala Thr Ala Ala Ser Asn Asn Pro Met Leu Ser 1635 1640 1645 Thr Leu Thr Ser Ala Leu Ser Gly Lys Leu Asn Pro Asp Val Asn Leu 1650 1655 1660 Val Asp Thr Leu Asn Gly Gly Glu Tyr Thr Val Phe Ala Pro Thr Asn 1665 1670 1675 1680 Ala Ala Phe Asp Lys Leu Pro Ala Ala Thr Ile Asp Gln Leu Lys Thr 1685 1690 1695 Asp Ala Lys Leu Leu Ser Ser Ile Leu Thr Tyr His Val Ile Ala Gly 1700 1705 1710 Gln Ala Ser Pro Ser Arg Ile Asp Gly Thr His Gln Thr Leu Gln Gly 1715 1720 1725 Ala Asp Leu Thr Val Ile Gly Ala Arg Asp Asp Leu Met Val Asn Asn 1730 1735 1740 Ala Gly Leu Val Ser Gly Gly Val His Thr Ala Asn Ala Thr Val Tyr 1745 1750 1755 1760 Met Ile Asp Thr Val Leu Met Pro Pro Ala Gln 1765 1770 <210> 36 <211> 1595 <212> PRT <213> Mycobacterium tuberculosis <400> 36 Met Gln Ala Glu Thr Ala Val Asn Thr Leu Phe Glu Lys Leu Glu Pro 1 5 10 15 Met Ala Ser Ile Leu Asp Pro Gly Ala Ser Gln Ser Thr Thr Asn Pro 20 25 30 Ile Phe Gly Met Pro Ser Pro Gly Ser Ser Thr Pro Val Gly Gln Leu 35 40 45 Pro Pro Ala Ala Thr Gln Thr Leu Gly Gln Leu Gly Glu Met Ser Gly 50 55 60 Pro Met Gly Gly Ser Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met 65 70 75 80 Gly Leu Leu Gly Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly 85 90 95 Ser Gly Pro Ser Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro 100 105 110 Gly Ala Gly Gly Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile 115 120 125 Glu Lys Pro Val Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu 130 135 140 Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu 145 150 155 160 Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly 165 170 175 Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala 180 185 190 Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile 195 200 205 Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly 210 215 220 Met Phe Ala Ala Ala Asp Tyr Asp Lys Leu Phe Arg Pro His Glu Gly 225 230 235 240 Met Glu Ala Pro Asp Asp Met Ala Ala Gln Pro Phe Phe Asp Pro Ser 245 250 255 Ala Ser Phe Pro Pro Ala Pro Ala Ser Ala Asn Leu Pro Lys Pro Asn 260 265 270 Gly Gln Thr Pro Pro Pro Thr Ser Asp Asp Leu Ser Glu Arg Phe Val 275 280 285 Ser Ala Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr 290 295 300 Pro Met Pro Ile Ala Ala Gly Glu Pro Pro Ser Pro Glu Pro Ala Ala Ala 305 310 315 320 Ser Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala 325 330 335 Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro 340 345 350 Ala Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Ala 355 360 365 Pro Thr Pro Thr Glu Ser Gln Leu Ala Pro Pro Arg Pro Pro Thr Pro 370 375 380 Gln Thr Pro Thr Gly Ala Pro Gln Gln Pro Glu Ser Pro Ala Pro His 385 390 395 400 Val Pro Ser His Gly Pro His Gln Pro Arg Arg Thr Ala Pro Ala Pro 405 410 415 Pro Trp Ala Lys Met Pro Ile Gly Glu Pro Pro Pro Ala Pro Ser Arg 420 425 430 Pro Ser Ala Ser Pro Ala Glu Pro Pro Thr Arg Pro Ala Pro Gln His 435 440 445 Ser Arg Arg Ala Arg Arg Gly His Arg Tyr Arg Thr Asp Thr Glu Arg 450 455 460 Asn Val Gly Lys Val Ala Thr Gly Pro Ser Ile Gln Ala Arg Leu Arg 465 470 475 480 Ala Glu Glu Ala Ser Gly Ala Gln Leu Ala Pro Gly Thr Glu Pro Ser 485 490 495 Pro Ala Pro Leu Gly Gln Pro Arg Ser Tyr Leu Ala Pro Pro Thr Arg 500 505 510 Pro Ala Pro Thr Glu Pro Pro Pro Ser Pro Ser Pro Gln Arg Asn Ser 515 520 525 Gly Arg Arg Ala Glu Arg Arg Val His Pro Asp Leu Ala Ala Gln His 530 535 540 Ala Ala Ala Gln Pro Asp Ser Ile Thr Ala Ala Thr Thr Gly Gly Arg 545 550 555 560 Arg Arg Lys Arg Ala Ala Pro Asp Leu Asp Ala Thr Gln Lys Ser Leu 565 570 575 Arg Pro Ala Ala Lys Gly Pro Lys Val Lys Lys Val Lys Pro Gln Lys 580 585 590 Pro Lys Ala Thr Lys Pro Pro Lys Val Val Ser Gln Arg Gly Trp Arg 595 600 605 His Trp Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser Pro Asp 610 615 620 Glu Lys Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn Pro Arg 625 630 635 640 Gly Ser Tyr Gln Ile Ala Val Val Gly Leu Gln Gly Gly Ala Gly Lys 645 650 655 Thr Thr Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val Arg Ala 660 665 670 Asp Arg Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn Leu Ala 675 680 685 Asp Arg Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val Leu Ala 690 695 700 Glu Lys Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr Ser Val 705 710 715 720 Asn Ala Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser Ser Ala 725 730 735 Gln Arg Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp Pro Ala 740 745 750 Ser Arg Phe Tyr Asn Leu Val Leu Ala Asp Ser Gly Ala Gly Phe Phe 755 760 765 Asp Pro Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val Val Val 770 775 780 Val Ala Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val Ala Leu 785 790 795 800 Asp Trp Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg Ala Ser 805 810 815 Val Val Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala Val Lys 820 825 830 Asp Leu Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg Val Val 835 840 845 Val Met Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile Ser Leu 850 855 860 Asp Leu Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu Ala Ala 865 870 875 880 Ala Leu Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg Thr Glu Gln Gln 885 890 895 Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn 900 905 910 Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr 915 920 925 Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly 930 935 940 Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu 945 950 955 960 Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser 965 970 975 Thr Glu Gly Asn Val Thr Gly Met Phe Ala Thr Glu Asn Leu Thr Val 980 985 990 Gln Pro Glu Arg Leu Gly Val Leu Ala Ser His His Asp Asn Ala Ala 995 1000 1005 Val Asp Ala Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly Glu Ser 1010 1015 1020 Val Ala Ile Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp Thr Thr 1025 1030 1035 1040 Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile 1045 1050 1055 Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln 1060 1065 1070 Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala 1075 1080 1085 Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn 1090 1095 1100 Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala 1105 1110 1115 1120 Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ser Arg Ala 1125 1130 1135 Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly Leu Tyr Asp Leu 1140 1145 1150 Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu Tyr Ser Ser Leu 1155 1160 1165 Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala Ala Phe Pro Gly 1170 1175 1180 Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala Gly Lys Asn Arg 1185 1190 1195 1200 Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu Asp Arg Gln Leu 1205 1210 1215 Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln Thr Thr Arg Asp 1220 1225 1230 Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val Arg Pro Val Ala 1235 1240 1245 Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala Leu Ser Ala Lys 1250 1255 1260 Thr Leu Ile Asn Ala Thr Gln Leu Leu Lys Leu Leu Ala Lys Leu Ala 1265 1270 1275 1280 Glu Leu Val Ala Ala Ala Ile Ala Asp Ile Ile Ser Asp Val Ala Asp 1285 1290 1295 Ile Ile Lys Gly Thr Leu Gly Glu Val Trp Glu Phe Ile Thr Asn Ala 1300 1305 1310 Leu Asn Gly Leu Lys Glu Leu Trp Asp Lys Leu Thr Gly Trp Val Thr 1315 1320 1325 Gly Leu Phe Ser Arg Gly Trp Ser Asn Leu Glu Ser Phe Phe Ala Gly 1330 1335 1340 Val Pro Gly Leu Thr Gly Ala Thr Ser Gly Leu Ser Gln Val Thr Gly 1345 1350 1355 1360 Leu Phe Gly Ala Ala Gly Leu Ser Ala Ser Ser Gly Leu Ala His Ala 1365 1370 1375 Asp Ser Leu Ala Ser Ser Ala Ser Leu Pro Ala Leu Ala Gly Ile Gly 1380 1385 1390 Gly Gly Ser Gly Phe Gly Gly Leu Pro Ser Leu Ala Gln Val His Ala 1395 1400 1405 Ala Ser Thr Arg Gln Ala Leu Arg Pro Arg Ala Asp Gly Pro Val Gly 1410 1415 1420 Ala Ala Ala Glu Gln Val Gly Gly Gln Ser Gln Leu Val Ser Ala Gln 1425 1430 1435 1440 Gly Ser Gln Gly Met Gly Gly Pro Val Gly Met Gly Gly Met His Pro 1445 1450 1455 Ser Ser Gly Ala Ser Lys Gly Thr Thr Thr Lys Lys Tyr Ser Glu Gly 1460 1465 1470 Ala Ala Ala Gly Thr Glu Asp Ala Glu Arg Ala Pro Val Glu Ala Asp 1475 1480 1485 Ala Gly Gly Gly Gln Lys Val Leu Val Arg Asn Val Val Thr Glu Gln 1490 1495 1500 Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly 1505 1510 1515 1520 Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu 1525 1530 1535 Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln 1540 1545 1550 Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala 1555 1560 1565 Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala 1570 1575 1580 Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala 1585 1590 1595 <210> 37 <211> 1871 <212> PRT <213> Tuberculosis <400> 37 Met Gln Ala Glu Thr Ala Val Asn Thr Leu Phe Glu Lys Leu Glu Pro 1 5 10 15 Met Ala Ser Ile Leu Asp Pro Gly Ala Ser Gln Ser Thr Thr Asn Pro 20 25 30 Ile Phe Gly Met Pro Ser Pro Gly Ser Ser Thr Pro Val Gly Gln Leu 35 40 45 Pro Pro Ala Ala Thr Gln Thr Leu Gly Gln Leu Gly Glu Met Ser Gly 50 55 60 Pro Met Gly Gly Ser Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met 65 70 75 80 Gly Leu Leu Gly Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly 85 90 95 Ser Gly Pro Ser Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro 100 105 110 Gly Ala Gly Gly Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile 115 120 125 Glu Lys Pro Val Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu 130 135 140 Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu 145 150 155 160 Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly 165 170 175 Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala 180 185 190 Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile 195 200 205 Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly 210 215 220 Met Phe Ala Ala Ala Asp Tyr Asp Lys Leu Phe Arg Pro His Glu Gly 225 230 235 240 Met Glu Ala Pro Asp Asp Met Ala Ala Gln Pro Phe Phe Asp Pro Ser 245 250 255 Ala Ser Phe Pro Pro Ala Pro Ala Ser Ala Asn Leu Pro Lys Pro Asn 260 265 270 Gly Gln Thr Pro Pro Pro Thr Ser Asp Asp Leu Ser Glu Arg Phe Val 275 280 285 Ser Ala Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr 290 295 300 Pro Met Pro Ile Ala Ala Gly Glu Pro Pro Ser Pro Glu Pro Ala Ala Ala 305 310 315 320 Ser Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala 325 330 335 Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro 340 345 350 Ala Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Ala 355 360 365 Pro Thr Pro Thr Glu Ser Gln Leu Ala Pro Pro Arg Pro Pro Thr Pro 370 375 380 Gln Thr Pro Thr Gly Ala Pro Gln Gln Pro Glu Ser Pro Ala Pro His 385 390 395 400 Val Pro Ser His Gly Pro His Gln Pro Arg Arg Thr Ala Pro Ala Pro 405 410 415 Pro Trp Ala Lys Met Pro Ile Gly Glu Pro Pro Pro Ala Pro Ser Arg 420 425 430 Pro Ser Ala Ser Pro Ala Glu Pro Pro Thr Arg Pro Ala Pro Gln His 435 440 445 Ser Arg Arg Ala Arg Arg Gly His Arg Tyr Arg Thr Asp Thr Glu Arg 450 455 460 Asn Val Gly Lys Val Ala Thr Gly Pro Ser Ile Gln Ala Arg Leu Arg 465 470 475 480 Ala Glu Glu Ala Ser Gly Ala Gln Leu Ala Pro Gly Thr Glu Pro Ser 485 490 495 Pro Ala Pro Leu Gly Gln Pro Arg Ser Tyr Leu Ala Pro Pro Thr Arg 500 505 510 Pro Ala Pro Thr Glu Pro Pro Pro Ser Pro Ser Pro Gln Arg Asn Ser 515 520 525 Gly Arg Arg Ala Glu Arg Arg Val His Pro Asp Leu Ala Ala Gln His 530 535 540 Ala Ala Ala Gln Pro Asp Ser Ile Thr Ala Ala Thr Thr Gly Gly Arg 545 550 555 560 Arg Arg Lys Arg Ala Ala Pro Asp Leu Asp Ala Thr Gln Lys Ser Leu 565 570 575 Arg Pro Ala Ala Lys Gly Pro Lys Val Lys Lys Val Lys Pro Gln Lys 580 585 590 Pro Lys Ala Thr Lys Pro Pro Lys Val Val Ser Gln Arg Gly Trp Arg 595 600 605 His Trp Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser Pro Asp 610 615 620 Glu Lys Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn Pro Arg 625 630 635 640 Gly Ser Tyr Gln Ile Ala Val Val Gly Leu Gln Gly Gly Ala Gly Lys 645 650 655 Thr Thr Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val Arg Ala 660 665 670 Asp Arg Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn Leu Ala 675 680 685 Asp Arg Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val Leu Ala 690 695 700 Glu Lys Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr Ser Val 705 710 715 720 Asn Ala Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser Ser Ala 725 730 735 Gln Arg Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp Pro Ala 740 745 750 Ser Arg Phe Tyr Asn Leu Val Leu Ala Asp Ser Gly Ala Gly Phe Phe 755 760 765 Asp Pro Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val Val Val 770 775 780 Val Ala Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val Ala Leu 785 790 795 800 Asp Trp Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg Ala Ser 805 810 815 Val Val Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala Val Lys 820 825 830 Asp Leu Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg Val Val 835 840 845 Val Met Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile Ser Leu 850 855 860 Asp Leu Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu Ala Ala 865 870 875 880 Ala Leu Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg Thr Glu Asn Leu 885 890 895 Thr Val Gln Pro Glu Arg Leu Gly Val Leu Ala Ser His His Asp Asn 900 905 910 Ala Ala Val Asp Ala Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly 915 920 925 Glu Ser Val Ala Ile Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp 930 935 940 Thr Ser Arg Ala Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly 945 950 955 960 Leu Tyr Asp Leu Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu 965 970 975 Tyr Ser Ser Leu Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala 980 985 990 Ala Phe Pro Gly Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala 995 1000 1005 Gly Lys Asn Arg Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu 1010 1015 1020 Asp Arg Gln Leu Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln 1025 1030 1035 1040 Thr Thr Arg Asp Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val 1045 1050 1055 Arg Pro Val Ala Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala 1060 1065 1070 Leu Ser Ala Lys Thr Leu Ile Asn Ala Thr Gln Leu Leu Lys Leu Leu 1075 1080 1085 Ala Lys Leu Ala Glu Leu Val Ala Ala Ala Ile Ala Asp Ile Ile Ser 1090 1095 1100 Asp Val Ala Asp Ile Ile Lys Gly Thr Leu Gly Glu Val Trp Glu Phe 1105 1110 1115 1120 Ile Thr Asn Ala Leu Asn Gly Leu Lys Glu Leu Trp Asp Lys Leu Thr 1125 1130 1135 Gly Trp Val Thr Gly Leu Phe Ser Arg Gly Trp Ser Asn Leu Glu Ser 1140 1145 1150 Phe Phe Ala Gly Val Pro Gly Leu Thr Gly Ala Thr Ser Gly Leu Ser 1155 1160 1165 Gln Val Thr Gly Leu Phe Gly Ala Ala Gly Leu Ser Ala Ser Ser Gly 1170 1175 1180 Leu Ala His Ala Asp Ser Leu Ala Ser Ser Ala Ser Leu Pro Ala Leu 1185 1190 1195 1200 Ala Gly Ile Gly Gly Gly Ser Gly Phe Gly Gly Leu Pro Ser Leu Ala 1205 1210 1215 Gln Val His Ala Ala Ser Thr Arg Gln Ala Leu Arg Pro Arg Ala Asp 1220 1225 1230 Gly Pro Val Gly Ala Ala Ala Glu Gln Val Gly Gly Gln Ser Gln Leu 1235 1240 1245 Val Ser Ala Gln Gly Ser Gln Gly Met Gly Gly Pro Val Gly Met Gly 1250 1255 1260 Gly Met His Pro Ser Ser Gly Ala Ser Lys Gly Thr Thr Thr Lys Lys 1265 1270 1275 1280 Tyr Ser Glu Gly Ala Ala Ala Gly Thr Glu Asp Ala Glu Arg Ala Pro 1285 1290 1295 Val Glu Ala Asp Ala Gly Gly Gly Gln Lys Val Leu Val Arg Asn Val 1300 1305 1310 Val Ala Pro Lys Thr Tyr Ser Glu Glu Leu Lys Gly Thr Asp Thr Gly 1315 1320 1325 Gln Ala Ser Gln Ile Gln Met Ser Asp Pro Ala Tyr Asn Ile Asn Ile 1330 1335 1340 Ser Leu Pro Ser Tyr Tyr Pro Asp Gln Lys Ser Leu Glu Asn Tyr Ile 1345 1350 1355 1360 Ala Gln Thr Arg Asp Lys Phe Leu Ser Ala Ala Thr Ser Ser Thr Pro 1365 1370 1375 Arg Glu Ala Pro Tyr Glu Leu Asn Ile Thr Ser Ala Thr Tyr Gln Ser 1380 1385 1390 Ala Ile Pro Pro Arg Gly Thr Gln Ala Val Val Leu Lys Val Tyr Gln 1395 1400 1405 Asn Ala Gly Gly Thr His Pro Thr Thr Thr Tyr Lys Ala Phe Asp Trp 1410 1415 1420 Asp Gln Ala Tyr Arg Lys Pro Ile Thr Tyr Asp Thr Leu Trp Gln Ala 1425 1430 1435 1440 Asp Thr Asp Pro Leu Pro Val Val Phe Pro Ile Val Gln Gly Glu Leu 1445 1450 1455 Ser Lys Gln Thr Gly Gln Gln Val Ser Ile Ala Pro Asn Ala Gly Leu 1460 1465 1470 Asp Pro Val Asn Tyr Gln Asn Phe Ala Val Thr Asn Asp Gly Val Ile 1475 1480 1485 Phe Phe Phe Asn Pro Gly Glu Leu Leu Pro Glu Ala Ala Gly Pro Thr 1490 1495 1500 Gln Val Leu Val Pro Arg Ser Ala Ile Asp Ser Met Leu Ala Gly Asp 1505 1510 1515 1520 Leu Val Gly Pro Gly Ser Ala Glu Tyr Ala Ala Ala Asn Pro Thr Gly 1525 1530 1535 Pro Ala Ser Val Gln Gly Met Ser Gln Asp Pro Val Ala Val Ala Ala 1540 1545 1550 Ser Asn Asn Pro Glu Leu Thr Thr Leu Thr Ala Ala Leu Ser Gly Gln 1555 1560 1565 Leu Asn Pro Gln Val Asn Leu Val Asp Thr Leu Asn Ser Gly Gln Tyr 1570 1575 1580 Thr Val Phe Ala Pro Thr Asn Ala Ala Phe Ser Lys Leu Pro Ala Ser 1585 1590 1595 1600 Thr Ile Asp Glu Leu Lys Thr Asn Ser Ser Leu Leu Thr Ser Ile Leu 1605 1610 1615 Thr Tyr His Val Val Ala Gly Gln Thr Ser Pro Ala Asn Val Val Gly 1620 1625 1630 Thr Arg Gln Thr Leu Gln Gly Ala Ser Val Thr Val Thr Gly Gln Gly 1635 1640 1645 Asn Ser Leu Lys Val Gly Asn Ala Asp Val Val Ser Gly Gly Val Ser 1650 1655 1660 Thr Ala Asn Ala Thr Val Tyr Met Ile Asp Ser Val Leu Met Pro Pro 1665 1670 1675 1680 Ala Val Ser Gln Asp Thr Ser Pro Lys Pro Ala Thr Ser Pro Ala Ala 1685 1690 1695 Pro Val Thr Thr Ala Ala Met Ala Asp Pro Ala Ala Asp Leu Ile Gly 1700 1705 1710 Arg Gly Ser Ala Gln Tyr Ala Ala Gln Asn Pro Thr Gly Pro Gly Ser 1715 1720 1725 Val Ala Gly Met Ala Gln Asp Pro Val Ala Thr Ala Ala Ser Asn Asn 1730 1735 1740 Pro Met Leu Ser Thr Leu Thr Ser Ala Leu Ser Gly Lys Leu Asn Pro 1745 1750 1755 1760 Asp Val Asn Leu Val Asp Thr Leu Asn Gly Gly Glu Tyr Thr Val Phe 1765 1770 1775 Ala Pro Thr Asn Ala Ala Phe Asp Lys Leu Pro Ala Ala Thr Ile Asp 1780 1785 1790 Gln Leu Lys Thr Asp Ala Lys Leu Leu Ser Ser Ile Leu Thr Tyr His 1795 1800 1805 Val Ile Ala Gly Gln Ala Ser Pro Ser Arg Ile Asp Gly Thr His Gln 1810 1815 1820 Thr Leu Gln Gly Ala Asp Leu Thr Val Ile Gly Ala Arg Asp Asp Leu 1825 1830 1835 1840 Met Val Asn Asn Ala Gly Leu Val Ser Gly Gly Val His Thr Ala Asn 1845 1850 1855 Ala Thr Val Tyr Met Ile Asp Thr Val Leu Met Pro Pro Ala Gln 1860 1865 1870 <210> 38 <211> 1130 <212> PRT <213> Mycobacterium tuberculosis <400> 38 Met Gln Ala Glu Thr Ala Val Asn Thr Leu Phe Glu Lys Leu Glu Pro 1 5 10 15 Met Ala Ser Ile Leu Asp Pro Gly Ala Ser Gln Ser Thr Thr Asn Pro 20 25 30 Ile Phe Gly Met Pro Ser Pro Gly Ser Ser Thr Pro Val Gly Gln Leu 35 40 45 Pro Pro Ala Ala Thr Gln Thr Leu Gly Gln Leu Gly Glu Met Ser Gly 50 55 60 Pro Met Gly Gly Ser Gly Asn Pro Ala Asp Glu Glu Ala Ala Gln Met 65 70 75 80 Gly Leu Leu Gly Thr Ser Pro Leu Ser Asn His Pro Leu Ala Gly Gly 85 90 95 Ser Gly Pro Ser Ala Gly Ala Gly Leu Leu Arg Ala Glu Ser Leu Pro 100 105 110 Gly Ala Gly Gly Ser Leu Thr Arg Thr Pro Leu Met Ser Gln Leu Ile 115 120 125 Glu Lys Pro Val Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu 130 135 140 Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu 145 150 155 160 Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly 165 170 175 Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala 180 185 190 Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile 195 200 205 Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly 210 215 220 Met Phe Ala Ala Ala Asp Tyr Asp Lys Leu Phe Arg Pro His Glu Gly 225 230 235 240 Met Glu Ala Pro Asp Asp Met Ala Ala Gln Pro Phe Phe Asp Pro Ser 245 250 255 Ala Ser Phe Pro Pro Ala Pro Ala Ser Ala Asn Leu Pro Lys Pro Asn 260 265 270 Gly Gln Thr Pro Pro Pro Thr Ser Asp Asp Leu Ser Glu Arg Phe Val 275 280 285 Ser Ala Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr 290 295 300 Pro Met Pro Ile Ala Ala Gly Glu Pro Pro Ser Pro Glu Pro Ala Ala Ala 305 310 315 320 Ser Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro Ala 325 330 335 Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Glu Pro 340 345 350 Ala Pro Pro Lys Pro Pro Thr Pro Pro Met Pro Ile Ala Gly Pro Ala 355 360 365 Pro Thr Pro Thr Glu Ser Gln Leu Ala Pro Pro Arg Pro Pro Thr Pro 370 375 380 Gln Thr Pro Thr Gly Ala Pro Gln Gln Pro Glu Ser Pro Ala Pro His 385 390 395 400 Val Pro Ser His Gly Pro His Gln Pro Arg Arg Thr Ala Pro Ala Pro 405 410 415 Pro Trp Ala Lys Met Pro Ile Gly Glu Pro Pro Pro Ala Pro Ser Arg 420 425 430 Pro Ser Ala Ser Pro Ala Glu Pro Pro Thr Arg Pro Ala Pro Gln His 435 440 445 Ser Arg Arg Ala Arg Arg Gly His Arg Tyr Arg Thr Asp Thr Glu Arg 450 455 460 Asn Val Gly Lys Val Ala Thr Gly Pro Ser Ile Gln Ala Arg Leu Arg 465 470 475 480 Ala Glu Glu Ala Ser Gly Ala Gln Leu Ala Pro Gly Thr Glu Pro Ser 485 490 495 Pro Ala Pro Leu Gly Gln Pro Arg Ser Tyr Leu Ala Pro Pro Thr Arg 500 505 510 Pro Ala Pro Thr Glu Pro Pro Pro Ser Pro Ser Pro Gln Arg Asn Ser 515 520 525 Gly Arg Arg Ala Glu Arg Arg Val His Pro Asp Leu Ala Ala Gln His 530 535 540 Ala Ala Ala Gln Pro Asp Ser Ile Thr Ala Ala Thr Thr Gly Gly Arg 545 550 555 560 Arg Arg Lys Arg Ala Ala Pro Asp Leu Asp Ala Thr Gln Lys Ser Leu 565 570 575 Arg Pro Ala Ala Lys Gly Pro Lys Val Lys Lys Val Lys Pro Gln Lys 580 585 590 Pro Lys Ala Thr Lys Pro Pro Lys Val Val Ser Gln Arg Gly Trp Arg 595 600 605 His Trp Val His Ala Leu Thr Arg Ile Asn Leu Gly Leu Ser Pro Asp 610 615 620 Glu Lys Tyr Glu Leu Asp Leu His Ala Arg Val Arg Arg Asn Pro Arg 625 630 635 640 Gly Ser Tyr Gln Ile Ala Val Val Gly Leu Gln Gly Gly Ala Gly Lys 645 650 655 Thr Thr Leu Thr Ala Ala Leu Gly Ser Thr Leu Ala Gln Val Arg Ala 660 665 670 Asp Arg Ile Leu Ala Leu Asp Ala Asp Pro Gly Ala Gly Asn Leu Ala 675 680 685 Asp Arg Val Gly Arg Gln Ser Gly Ala Thr Ile Ala Asp Val Leu Ala 690 695 700 Glu Lys Glu Leu Ser His Tyr Asn Asp Ile Arg Ala His Thr Ser Val 705 710 715 720 Asn Ala Val Asn Leu Glu Val Leu Pro Ala Pro Glu Tyr Ser Ser Ala 725 730 735 Gln Arg Ala Leu Ser Asp Ala Asp Trp His Phe Ile Ala Asp Pro Ala 740 745 750 Ser Arg Phe Tyr Asn Leu Val Leu Ala Asp Ser Gly Ala Gly Phe Phe 755 760 765 Asp Pro Leu Thr Arg Gly Val Leu Ser Thr Val Ser Gly Val Val Val 770 775 780 Val Ala Ser Val Ser Ile Asp Gly Ala Gln Gln Ala Ser Val Ala Leu 785 790 795 800 Asp Trp Leu Arg Asn Asn Gly Tyr Gln Asp Leu Ala Ser Arg Ala Ser 805 810 815 Val Val Ile Asn His Ile Met Pro Gly Glu Pro Asn Val Ala Val Lys 820 825 830 Asp Leu Val Arg His Phe Glu Gln Gln Val Gln Pro Gly Arg Val Val 835 840 845 Val Met Pro Trp Asp Arg His Ile Ala Ala Gly Thr Glu Ile Ser Leu 850 855 860 Asp Leu Leu Asp Pro Ile Tyr Lys Arg Lys Val Leu Glu Leu Ala Ala 865 870 875 880 Ala Leu Ser Asp Asp Phe Glu Arg Ala Gly Arg Arg Thr Glu Asn Leu 885 890 895 Thr Val Gln Pro Glu Arg Leu Gly Val Leu Ala Ser His His Asp Asn 900 905 910 Ala Ala Val Asp Ala Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly 915 920 925 Glu Ser Val Ala Ile Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp 930 935 940 Thr Ser Arg Ala Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly 945 950 955 960 Leu Tyr Asp Leu Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu 965 970 975 Tyr Ser Ser Leu Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala 980 985 990 Ala Phe Pro Gly Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala 995 1000 1005 Gly Lys Asn Arg Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu 1010 1015 1020 Asp Arg Gln Leu Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln 1025 1030 1035 1040 Thr Thr Arg Asp Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val 1045 1050 1055 Arg Pro Val Ala Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala 1060 1065 1070 Leu Ser Ala Lys Thr Leu Ile Asn Ala Thr Gln Leu Leu Lys Leu Leu 1075 1080 1085 Ala Lys Leu Ala Glu Leu Val Ala Ala Ala Ile Ala Asp Ile Ile Ser 1090 1095 1100 Asp Val Ala Asp Ile Ile Lys Gly Thr Leu Gly Glu Val Trp Glu Phe 1105 1110 1115 1120 Ile Thr Asn Ala Leu Asn Gly Leu Lys Glu 1125 1130 <210> 39 <211> 760 <212> PRT <213> Mycobacterium tuberculosis <400> 39 Met Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 1 5 10 15 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 20 25 30 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 35 40 45 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu 50 55 60 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 65 70 75 80 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Lys 85 90 95 Val Lys Asn Thr Ile Ala Ala Thr Ser Phe Ala Ala Ala Gly Leu Ala 100 105 110 Ala Leu Ala Val Ala Val Ser Pro Pro Ala Ala Ala Gly Asp Leu Val 115 120 125 Gly Pro Gly Cys Ala Glu Tyr Ala Ala Ala Asn Pro Thr Gly Pro Ala 130 135 140 Ser Val Gln Gly Met Ser Gln Asp Pro Val Ala Val Ala Ala Ser Asn 145 150 155 160 Asn Pro Glu Leu Thr Thr Leu Thr Ala Ala Leu Ser Gly Gln Leu Asn 165 170 175 Pro Gln Val Asn Leu Val Asp Thr Leu Asn Ser Gly Gln Tyr Thr Val 180 185 190 Phe Ala Pro Thr Asn Ala Ala Phe Ser Lys Leu Pro Ala Ser Thr Ile 195 200 205 Asp Glu Leu Lys Thr Asn Ser Ser Leu Leu Thr Ser Ile Leu Thr Tyr 210 215 220 His Val Val Ala Gly Gln Thr Ser Pro Ala Asn Val Val Gly Thr Arg 225 230 235 240 Gln Thr Leu Gln Gly Ala Ser Val Thr Val Thr Gly Gln Gly Asn Ser 245 250 255 Leu Lys Val Gly Asn Ala Asp Val Val Cys Gly Gly Val Ser Thr Ala 260 265 270 Asn Ala Thr Val Tyr Met Ile Asp Ser Val Leu Met Pro Pro Ala Thr 275 280 285 Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile 290 295 300 Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln 305 310 315 320 Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala 325 330 335 Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn 340 345 350 Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala 355 360 365 Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ser Leu Leu 370 375 380 Asp Ala His Ile Pro Gln Leu Ile Ala Ser His Thr Ala Phe Ala Ala 385 390 395 400 Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala Glu Gln Gln Ala 405 410 415 Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ala Ala Ala Phe Gln 420 425 430 Gly Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys Val Asn Thr Leu 435 440 445 Leu Asp Ile Ala Gln Ala Asn Leu Gly Glu Ala Ala Gly Thr Tyr Val 450 455 460 Ala Ala Asp Ala Ala Ala Ser Ser Tyr Thr Gly Phe Thr Glu Gln 465 470 475 480 Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly 485 490 495 Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu 500 505 510 Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln 515 520 525 Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala 530 535 540 Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala 545 550 555 560 Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ser Gln Ile Met Tyr 565 570 575 Asn Tyr Pro Ala Met Met Ala His Ala Gly Asp Met Ala Gly Tyr Ala 580 585 590 Gly Thr Leu Gln Ser Leu Gly Ala Asp Ile Ala Ser Glu Gln Ala Val 595 600 605 Leu Ser Ser Ala Trp Gln Gly Asp Thr Gly Ile Thr Tyr Gln Gly Trp 610 615 620 Gln Thr Gln Trp Asn Gln Ala Leu Glu Asp Leu Val Arg Ala Tyr Gln 625 630 635 640 Ser Met Ser Gly Thr His Glu Ser Asn Thr Met Ala Met Leu Ala Arg 645 650 655 Asp Gly Ala Glu Ala Ala Lys Trp Gly Gly Thr Glu Gln Gln Trp Asn 660 665 670 Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr 675 680 685 Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu 690 695 700 Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln 705 710 715 720 Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn 725 730 735 Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu 740 745 750 Gly Asn Val Thr Gly Met Phe Ala 755 760 <210> 40 <211> 478 <212> PRT <213> Mycobacterium tuberculosis[[ID=3,8]] <400> 40 Met Lys Val Lys Asn Thr Ile Ala Ala Thr Ser Phe Ala Ala Ala Gly 1 5 10 15 Leu Ala Ala Leu Ala Val Ala Val Ser Pro Pro Ala Ala Ala Gly Asp 20 25 30 Leu Val Gly Pro Gly Cys Ala Glu Tyr Ala Ala Ala Asn Pro Thr Gly 35 40 45 Pro Ala Ser Val Gln Gly Met Ser Gln Asp Pro Val Ala Val Ala Ala 50 55 60 Ser Asn Asn Pro Glu Leu Thr Thr Leu Thr Ala Ala Leu Ser Gly Gln 65 70 75 80 Leu Asn Pro Gln Val Asn Leu Val Asp Thr Leu Asn Ser Gly Gln Tyr 85 90 95 Thr Val Phe Ala Pro Thr Asn Ala Ala Phe Ser Lys Leu Pro Ala Ser 100 105 110 Thr Ile Asp Glu Leu Lys Thr Asn Ser Ser Leu Leu Thr Ser Ile Leu 115 120 125 Thr Tyr His Val Val Ala Gly Gln Thr Ser Pro Ala Asn Val Val Gly 130 135 140 Thr Arg Gln Thr Leu Gln Gly Ala Ser Val Thr Val Thr Gly Gln Gly 145 150 155 160 Asn Ser Leu Lys Val Gly Asn Ala Asp Val Val Cys Gly Gly Val Ser 165 170 175 Thr Ala Asn Ala Thr Val Tyr Met Ile Asp Ser Val Leu Met Pro Pro 180 185 190 Ala Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 195 200 205 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 210 215 220 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 225 230 235 240 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu 245 250 255 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 260 265 270 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Ser 275 280 285 Leu Leu Asp Ala His Ile Pro Gln Leu Ile Ala Ser His Thr Ala Phe 290 295 300 Ala Ala Lys Ala Gly Leu Met Arg His Thr Ile Gly Gln Ala Glu Gln 305 310 315 320 Gln Ala Met Ser Ala Gln Ala Phe His Gln Gly Glu Ser Ala Ala Ala 325 330 335 Phe Gln Gly Ala His Ala Arg Phe Val Ala Ala Ala Ala Lys Val Asn 340 345 350 Thr Leu Leu Asp Ile Ala Gln Ala Asn Leu Gly Glu Ala Ala Gly Thr 355 360 365 Tyr Val Ala Ala Asp Ala Ala Ala Ala Ser Ser Tyr Thr Gly Phe Ser 370 375 380 Gln Ile Met Tyr Asn Tyr Pro Ala Met Met Ala His Ala Gly Asp Met 385 390 395 400 Ala Gly Tyr Ala Gly Thr Leu Gln Ser Leu Gly Ala Asp Ile Ala Ser 405 410 415 Glu Gln Ala Val Leu Ser Ser Ala Trp Gln Gly Asp Thr Gly Ile Thr 420 425 430 Tyr Gln Gly Trp Gln Thr Gln Trp Asn Gln Ala Leu Glu Asp Leu Val 4]]435 440 445 Arg Ala Tyr Gln Ser Met Ser Gly Thr His Glu Ser Asn Thr Met Ala 450 455 460 Met Leu Ala Arg Asp Gly Ala Glu Ala Ala Lys Trp Gly Gly 465 470 475 <210> 41 <211> 1096 <212> PRT <213> Mycobacterium tuberculosis <400> 41 Met Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 1 5 10 15 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 20 25 30 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 35 40 45 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu 50 55 60 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 65 70 75 80 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Met 85 90 95 Asp Leu Pro Gly Asn Asp Phe Asp Ser Asn Asp Phe Asp Ala Val Asp 100 105 110 Leu Trp Gly Ala Asp Gly Ala Glu Gly Trp Thr Ala Asp Pro Ile Ile 115 120 125 Gly Val Gly Ser Ala Ala Thr Pro Asp Thr Gly Pro Asp Leu Asp Asn 130 135 140 Ala His Gly Gln Ala Glu Thr Asp Thr Glu Gln Glu Ile Ala Leu Phe 145 150 155 160 Thr Val Thr Asn Pro Pro Arg Thr Val Ser Val Ser Thr Leu Met Asp 165 170 175 Gly Arg Ile Asp His Val Glu Leu Ser Ala Arg Val Ala Trp Met Ser 180 185 190 Glu Ser Gln Leu Ala Ser Glu Ile Leu Val Ile Ala Asp Leu Ala Arg 195 200 205 Gln Lys Ala Gln Ser Ala Gln Tyr Ala Phe Ile Leu Asp Arg Met Ser 210 215 220 Gln Gln Val Asp Ala Asp Glu His Arg Val Ala Leu Leu Arg Lys Thr 225 230 235 240 Val Gly Glu Thr Trp Gly Leu Pro Ser Pro Glu Glu Ala Ala Ala Ala 245 250 255 Glu Ala Glu Val Phe Ala Thr Arg Tyr Ser Asp Asp Ser Pro Ala Pro 260 265 270 Asp Asp Glu Ser Asp Pro Trp Met Thr Glu Gln Gln Trp Asn Phe Ala 275 280 285 Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile 290 295 300 His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala 305 310 315 320 Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys 325 330 335 Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala 340 345 350 Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn 355 360 365 Val Thr Gly Met Phe Ala Met Thr Glu Asn Leu Thr Val Gln Pro Glu 370 375 380 Arg Leu Gly Val Leu Ala Ser His His Asp Asn Ala Ala Val Asp Ala 385 390 395 400 Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly Glu Ser Val Ala Ile 405 410 415 Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp Thr Leu Asn Val Tyr 420 425 430 Leu Thr Ala His Asn Ala Leu Gly Ser Ser Leu His Thr Ala Gly Val 435 440 445 Asp Leu Ala Lys Ser Leu Arg Ile Ala Ala Lys Ile Tyr Ser Glu Ala 450 455 460 Asp Glu Ala Trp Arg Lys Ala Ile Asp Gly Leu Phe Thr Met Thr Glu 465 470 475 480 Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln 485 490 495 Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser 500 505 510 Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr 515 520 525 Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn 530 535 540 Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met 545 550 555 560 Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Met Thr Gly Phe 565 570 575 Leu Gly Val Val Pro Ser Phe Leu Lys Val Leu Ala Gly Met His Asn 580 585 590 Glu Ile Val Gly Asp Ile Lys Arg Ala Thr Asp Thr Val Ala Gly Ile 595 600 605 Ser Gly Arg Val Gln Leu Thr His Gly Ser Phe Thr Ser Lys Phe Asn 610 615 620 Asp Thr Leu Gln Glu Phe Glu Thr Thr Arg Ser Ser Thr Gly Thr Gly 625 630 635 640 Leu Gln Gly Val Thr Ser Gly Leu Ala Asn Asn Leu Leu Ala Ala Ala 645 650 655 Gly Ala Tyr Leu Lys Ala Asp Asp Gly Leu Ala Gly Val Ile Asp Lys 660 665 670 Ile Phe Gly Met Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala 675 680 685 Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu 690 695 700 Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly 705 710 715 720 Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr 725 730 735 Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser 740 745 750 Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met 755 760 765 Phe Ala Met Ser Thr Thr Phe Ala Ala Arg Leu Asn Arg Leu Phe Asp 770 775 780 Thr Val Tyr Pro Pro Gly Arg Gly Pro His Thr Ser Ala Glu Val Ile 785 790 795 800 Ala Ala Leu Lys Ala Glu Gly Ile Thr Met Ser Ala Pro Tyr Leu Ser 805 810 815 Gln Leu Arg Ser Gly Asn Arg Thr Asn Pro Ser Gly Ala Thr Met Ala 820 825 830 Ala Leu Ala Asn Phe Phe Arg Ile Lys Ala Ala Tyr Phe Thr Asp Asp 835 840 845 Glu Tyr Tyr Glu Lys Leu Asp Lys Glu Leu Gln Trp Leu Ser Thr Met 850 855 860 Arg Asp Asp Gly Val Arg Arg Ile Ala Gln Arg Ala His Gly Leu Pro 865 870 875 880 Ser Ala Ala Gln Gln Lys Val Leu Asp Arg Ile Asp Glu Leu Arg Arg 885 890 895 Ala Glu Gly Ile Asp Ala Met Thr Glu Gln Gln Trp Asn Phe Ala Gly 900 905 910 Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly Asn Val Thr Ser Ile His 915 920 925 Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala 930 935 940 Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp 945 950 955 960 Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg 965 970 975 Thr Ile Ser Glu Ala Gly Gln Ala Met Ala Ser Thr Glu Gly Asn Val 980 985 990 Thr Gly Met Phe Ala Met Glu Lys Met Ser His Asp Pro Ile Ala Ala 995 1000 1005 Asp Ile Gly Thr Gln Val Ser Asp Asn Ala Leu His Gly Val Thr Ala 1010 1015 1020 Gly Ser Thr Ala Leu Thr Ser Val Thr Gly Leu Val Pro Ala Gly Ala 1025 1030 1035 1040 Asp Glu Val Ser Ala Gln Ala Ala Thr Ala Phe Thr Ser Glu Gly Ile 1045 1050 1055 Gln Leu Leu Ala Ser Asn Ala Ser Ala Gln Asp Gln Leu His Arg Ala 1060 1065 1070 Gly Glu Ala Val Gln Asp Val Ala Arg Thr Tyr Ser Gln Ile Asp Asp 1075 1080 1085 Gly Ala Ala Gly Val Phe Ala Glu 1090 1095 <210> 42 <211> 1313 <212> PRT <213> Tuberculosis <400> 42 Met Gly Ser Thr Gln Ser Gln Thr Val Thr Val Asp Gln Gln Glu Ile 1 5 10 15 Leu Asn Arg Ala Asn Glu Val Glu Ala Pro Met Ala Asp Pro Pro Thr 20 25 30 Asp Val Pro Ile Thr Pro Ser Glu Leu Thr Ala Ala Lys Asn Ala Ala 35 40 45 Gln Gln Leu Val Leu Ser Ala Asp Asn Met Arg Glu Tyr Leu Ala Ala 50 55 60 Gly Ala Lys Glu Arg Gln Arg Leu Ala Thr Ser Leu Arg Asn Ala Ala 65 70 75 80 Lys Ala Tyr Gly Glu Val Asp Glu Glu Ala Ala Thr Ala Leu Asp Asn 85 90 95 Asp Gly Glu Gly Thr Val Gln Ala Glu Ser Ala Gly Ala Val Gly Gly 100 105 110 Asp Ser Ser Ala Glu Leu Thr Asp Thr Pro Arg Val Ala Thr Ala Gly 115 120 125 Glu Pro Asn Phe Met Asp Leu Lys Glu Ala Ala Arg Lys Leu Glu Thr 130 135 140 Gly Asp Gln Gly Ala Ser Leu Ala His Phe Ala Asp Gly Trp Asn Thr 145 150 155 160 Phe Asn Leu Thr Leu Gln Gly Asp Val Lys Arg Phe Arg Gly Phe Asp 165 170 175 Asn Trp Glu Gly Asp Ala Ala Thr Ala Ser Glu Ala Ser Leu Asp Gln 180 185 190 Gln Arg Gln Trp Ile Leu His Met Ala Lys Leu Ser Ala Ala Met Ala 195 200 205 Lys Gln Ala Gln Tyr Val Ala Gln Leu His Val Trp Ala Arg Arg Glu 210 215 220 His Pro Thr Tyr Glu Asp Ile Val Gly Leu Glu Arg Leu Tyr Ala Glu 225 230 235 240 Asn Pro Ser Ala Arg Asp Gln Ile Leu Pro Val Tyr Ala Glu Tyr Gln 245 250 255 Gln Arg Ser Glu Lys Val Leu Thr Glu Tyr Asn Asn Lys Ala Ala Leu 260 265 270 Glu Pro Val Asn Pro Pro Lys Pro Pro Pro Ala Ile Lys Ile Asp Pro 275 280 285 Pro Pro Pro Pro Gln Glu Gln Gly Leu Ile Pro Gly Phe Leu Met Pro 290 295 300 Pro Ser Asp Gly Ser Gly Val Thr Pro Gly Thr Gly Met Pro Ala Ala 305 310 315 320 Pro Met Val Pro Pro Thr Gly Ser Pro Gly Gly Gly Leu Pro Ala Asp 325 330 335 Thr Ala Ala Gln Leu Thr Ser Ala Gly Arg Glu Ala Ala Ala Leu Ser 340 345 350 Gly Asp Val Ala Val Lys Ala Ala Ser Leu Gly Gly Gly Gly Gly Gly 355 360 365 Gly Val Pro Ser Ala Pro Leu Gly Ser Ala Ile Gly Gly Ala Glu Ser 370 375 380 Val Arg Pro Ala Gly Ala Gly Asp Ile Ala Gly Leu Gly Gln Gly Arg 385 390 395 400 Ala Gly Gly Gly Ala Ala Leu Gly Gly Gly Gly Met Gly Met Pro Met 405 410 415 Gly Ala Ala His Gln Gly Gln Gly Gly Ala Lys Ser Lys Gly Ser Gln 420 425 430 Gln Glu Asp Glu Ala Leu Tyr Thr Glu Asp Arg Ala Trp Thr Glu Ala 435 440 445 Val Ile Gly Asn Arg Arg Arg Gln Asp Ser Lys Glu Ser Lys Gly Ser 450 455 460 Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala 465 470 475 480 Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys 485 490 495 Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu 500 505 510 Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu 515 520 525 Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln 530 535 540 Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Glu Phe 545 550 555 560 Asp Leu Pro Gly Asn Asp Phe Asp Ser Asn Asp Phe Asp Ala Val Asp 565 570 575 Leu Trp Gly Ala Asp Gly Ala Glu Gly Trp Thr Ala Asp Pro Ile Ile 580 585 590 Gly Val Gly Ser Ala Ala Thr Pro Asp Thr Gly Pro Asp Leu Asp Asn 595 600 605 Ala His Gly Gln Ala Glu Thr Asp Thr Glu Gln Glu Ile Ala Leu Phe 610 615 620 Thr Val Thr Asn Pro Pro Arg Thr Val Ser Val Ser Thr Leu Met Asp 625 630 635 640 Gly Arg Ile Asp His Val Glu Leu Ser Ala Arg Val Ala Trp Met Ser 645 650 655 Glu Ser Gln Leu Ala Ser Glu Ile Leu Val Ile Ala Asp Leu Ala Arg 660 665 670 Gln Lys Ala Gln Ser Ala Gln Tyr Ala Phe Ile Leu Asp Arg Met Ser 675 680 685 Gln Gln Val Asp Ala Asp Glu His Arg Val Ala Leu Leu Arg Lys Thr 690 695 700 Val Gly Glu Thr Trp Gly Leu Pro Ser Pro Glu Glu Ala Ala Ala Ala 705 710 715 720 Glu Ala Glu Val Phe Ala Thr Arg Tyr Ser Asp Asp Ser Pro Ala Pro 725 730 735 Asp Asp Glu Ser Asp Pro Trp Thr Glu Asn Leu Thr Val Gln Pro Glu 740 745 750 Arg Leu Gly Val Leu Ala Ser His His Asp Asn Ala Ala Val Asp Ala 755 760 765 Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly Glu Ser Val Ala Ile 770 775 780 Thr His Gly Pro Tyr Ser Ser Gln Phe Asn Asp Thr Leu Asn Val Tyr 785 790 795 800 Leu Thr Ala His Asn Ala Leu Gly Ser Ser Leu His Thr Ala Gly Val 805 810 815 Asp Leu Ala Lys Ser Leu Arg Ile Ala Ala Lys Ile Tyr Ser Glu Ala 820 825 830 Asp Glu Ala Trp Arg Lys Ala Ile Asp Gly Leu Phe Thr Ser Arg Ala 835 840 845 Phe Ile Ile Asp Pro Thr Ile Ser Ala Ile Asp Gly Leu Tyr Asp Leu 850 855 860 Leu Gly Ile Gly Ile Pro Asn Gln Gly Gly Ile Leu Tyr Ser Ser Leu 865 870 875 880 Glu Tyr Phe Glu Lys Ala Leu Glu Glu Leu Ala Ala Ala Phe Pro Gly 885 890 895 Asp Gly Trp Leu Gly Ser Ala Ala Asp Lys Tyr Ala Gly Lys Asn Arg 900 905 910 Asn His Val Asn Phe Phe Gln Glu Leu Ala Asp Leu Asp Arg Gln Leu 915 920 925 Ile Ser Leu Ile His Asp Gln Ala Asn Ala Val Gln Thr Thr Arg Asp 930 935 940 Ile Leu Glu Gly Ala Lys Lys Gly Leu Glu Phe Val Arg Pro Val Ala 945 950 955 960 Val Asp Leu Thr Tyr Ile Pro Val Val Gly His Ala Leu Asp Val Ala 965 970 975 Asp Ile Ile Lys Gly Thr Leu Gly Glu Val Trp Glu Phe Ile Thr Asn 980 985 990 Ala Leu Asn Gly Leu Lys Glu Leu Trp Asp Lys Leu Thr Gly Trp Val 995 1000 1005 Thr Gly Leu Phe Ser Arg Gly Trp Ser Asn Leu Glu Ser Phe Phe Ala 1010 1015 1020 Gly Val Pro Gly Leu Thr Gly Ala Thr Ser Gly Leu Ser Gln Val Thr 1025 1030 1035 1040 Gly Leu Phe Gly Ala Ala Gly Leu Ser Ala Ser Ser Gly Leu Ala His 1045 1050 1055 Ala Asp Ser Leu Ala Ser Ser Ala Ser Leu Pro Ala Leu Ala Gly Ile 1060 1065 1070 Gly Gly Gly Ser Gly Phe Gly Gly Leu Pro Ser Leu Ala Gln Val His 1075 1080 1085 Ala Ala Ser Thr Arg Gln Ala Leu Arg Pro Arg Ala Asp Gly Pro Val 1090 1095 1100 Gly Ala Ala Ala Glu Gln Val Gly Gly Gln Ser Gln Leu Val Ser Ala 1105 1110 1115 1120 Gln Gly Ser Gln Gly Met Gly Gly Pro Val Gly Met Gly Gly Met His 1125 1130 1135 Pro Ser Ser Gly Ala Ser Lys Gly Thr Thr Thr Lys Lys Tyr Ser Glu 1140 1145 1150 Gly Ala Ala Ala Gly Thr Glu Asp Ala Glu Arg Ala Pro Val Glu Ala 1155 1160 1165 Asp Ala Gly Gly Gly Gln Lys Val Leu Val Arg Asn Val Val Ser Thr 1170 1175 1180 Thr Phe Ala Ala Arg Leu Asn Arg Leu Phe Asp Thr Val Tyr Pro Pro 1185 1190 1195 1200 Gly Arg Gly Pro His Thr Ser Ala Glu Val Ile Ala Ala Leu Lys Ala 1205 1210 1215 Glu Gly Ile Thr Met Ser Ala Pro Tyr Leu Ser Gln Leu Arg Ser Gly 1220 1225 1230 Asn Arg Thr Asn Pro Ser Gly Ala Thr Met Ala Ala Leu Ala Asn Phe 1235 1240 1245 Phe Arg Ile Lys Ala Ala Tyr Phe Thr Asp Asp Glu Tyr Tyr Glu Lys 1250 1255 1260 Leu Asp Lys Glu Leu Gln Trp Leu Ser Thr Met Arg Asp Asp Gly Val 1265 1270 1275 1280 Arg Arg Ile Ala Gln Arg Ala His Gly Leu Pro Ser Ala Ala Gln Gln 1285 1290 1295 Lys Val Leu Asp Arg Ile Asp Glu Leu Arg Arg Ala Glu Gly Ile Asp 1300 1305 1310 Ala <210> 43 <211> 654 <212> PRT <213> Tuberculosis <400> 43 Met Ala Asp Thr Ile Gln Val Thr Pro Gln Met Leu Arg Ser Thr Ala 1 5 10 15 Asn Asp Ile Gln Ala Asn Met Glu Gln Ala Met Gly Ile Ala Lys Gly 20 25 30 Tyr Leu Ala Asn Gln Glu Asn Val Met Asn Pro Ala Thr Trp Ser Gly 35 40 45 Thr Gly Val Val Ala Ser His Met Thr Ala Thr Glu Ile Thr Asn Glu 50 55 60 Leu Asn Lys Val Leu Thr Gly Gly Thr Arg Leu Ala Glu Gly Leu Val 65 70 75 80 Gln Ala Ala Ala Leu Met Glu Gly His Glu Ala Asp Ser Gln Thr Ala 85 90 95 Phe Gln Ala Leu Phe Gly Ala Ser His Gly Ser Gly Leu Val Pro Arg 100 105 110 Gly Ser Thr Gly Met Ser Asp Gln Ile Thr Tyr Asn Pro Gly Ala Val 115 120 125 Ser Asp Phe Ala Ser Asp Val Gly Ser Arg Ala Gly Gln Leu His Met 130 135 140 Ile Tyr Glu Asp Thr Ala Ser Lys Thr Asn Ala Leu Gln Glu Phe Phe 145 150 155 160 Ala Gly His Gly Ala Gln Gly Phe Phe Asp Ala Gln Ala Gln Met Leu 165 170 175 Ser Gly Leu Gln Gly Leu Ile Glu Thr Val Gly Gln His Gly Thr Thr 180 185 190 Thr Gly His Val Leu Asp Asn Ala Ile Gly Thr Asp Gln Ala Ile Ala 195 200 205 Gly Leu Phe Leu Ile Gly Ala His Pro Arg Ala Leu Asn Val Val Lys 210 215 220 Phe Gly Gly Ala Ala Phe Leu Met Ser Leu Leu Asp Ala His Ile Pro 225 230 235 240 Gln Leu Val Ala Ser Gln Ser Ala Phe Ala Ala Lys Ala Gly Leu Met 245 250 255 Arg His Thr Ile Gly Gln Ala Glu Gln Ala Ala Met Ser Ala Gln Ala 260 265 270 Phe His Gln Gly Glu Ser Ser Ala Ala Phe Gln Ala Ala His Ala Arg 275 280 285 Phe Val Ala Ala Ala Ala Lys Val Asn Thr Leu Leu Asp Val Ala Gln 290 295 300 Ala Asn Leu Gly Glu Ala Ala Gly Thr Tyr Val Ala Ala Asp Ala Ala 305 310 315 320 Ala Ala Ser Thr Tyr Thr Gly Phe Gly Leu Val Pro Arg Gly Ser Thr 325 330 335 Gly Met Ser Gln Ile Met Tyr Asn Tyr Pro Ala Met Leu Gly His Ala 340 345 350 Gly Asp Met Ala Gly Tyr Ala Gly Thr Leu Gln Ser Leu Gly Ala Glu 355 360 365 Ile Ala Val Glu Gln Ala Ala Leu Gln Ser Ala Trp Gln Gly Asp Thr 370 375 380 Gly Ile Thr Tyr Gln Ala Trp Gln Ala Gln Trp Asn Gln Ala Met Glu 385 390 395 400 Asp Leu Val Arg Ala Tyr His Ala Met Ser Ser Thr His Glu Ala Asn 405 410 415 Thr Met Ala Met Met Ala Arg Asp Thr Ala Glu Ala Ala Lys Trp Gly 420 425 430 Gly Leu Gly Phe Gly Ala Gly Arg Leu Arg Gly Leu Phe Thr Asn Pro 435 440 445 Gly Ser Trp Arg Ile Met Thr Ser Arg Phe Met Thr Asp Pro His Ala 450 455 460 Met Arg Asp Met Ala Gly Arg Phe Glu Val His Ala Gln Thr Val Glu 465 470 475 480 Asp Glu Ala Arg Arg Met Trp Ala Ser Ala Gln Asn Ile Ser Gly Ala 485 490 495 Gly Trp Ser Gly Met Ala Glu Ala Thr Ser Leu Asp Thr Met Thr Gln 500 505 510 Met Asn Gln Ala Phe Arg Asn Ile Val Asn Met Leu His Gly Val Arg 515 520 525 Asp Gly Leu Val Arg Asp Ala Asn Asn Tyr Glu Gln Gln Glu Gln Ala 530 535 540 Ser Gln Gln Ile Leu Ser Ser Gly Leu Val Pro Arg Gly Ser Thr Gly 545 550 555 560 Met Thr Ile Asn Tyr Gln Phe Gly Asp Val Asp Ala His Gly Ala Met 565 570 575 Ile Arg Ala Gln Ala Gly Ser Leu Glu Ala Glu His Gln Ala Ile Ile 580 585 590 Ser Asp Val Leu Thr Ala Ser Asp Phe Trp Gly Gly Ala Gly Ser Ala 595 600 605 Ala Ser Gln Gly Phe Ile Thr Gln Leu Gly Arg Asn Phe Gln Val Ile 610 615 620 Tyr Glu Gln Ala Asn Ala His Gly Gln Lys Val Gln Ala Ala Gly Asn 625 630 635 640 Asn Met Ala Gln Thr Asp Ser Ala Val Gly Ser Ser Trp Ala 645 650 <210> 44 <211> 716 <212> PRT <213> Mycobacterium tuberculosis <400> 44 Met Thr Glu Gln Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser 1 5 10 15 Ala Ile Gln Gly Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly 20 25 30 Lys Gln Ser Leu Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser 35 40 45 Glu Ala Tyr Gln Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu 50 55 60 Leu Asn Asn Ala Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly 65 70 75 80 Gln Ala Met Ala Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Met 85 90 95 Asp Leu Pro Gly Asn Asp Phe Asp Ser Asn Asp Phe Asp Ala Val Asp 100 105 110 Leu Trp Gly Ala Asp Gly Ala Glu Gly Trp Thr Ala Asp Pro Ile Ile 115 120 125 Gly Val Gly Ser Ala Ala Thr Pro Asp Thr Gly Pro Asp Leu Asp Asn 130 135 140 Ala His Gly Gln Ala Glu Thr Asp Thr Glu Gln Glu Ile Ala Leu Phe 145 150 155 160 Thr Val Thr Asn Pro Pro Arg Thr Val Ser Val Ser Thr Leu Met Asp 165 170 175 Gly Arg Ile Asp His Val Glu Leu Ser Ala Arg Val Ala Trp Met Ser 180 185 190 Glu Ser Gln Leu Ala Ser Glu Ile Leu Val Ile Ala Asp Leu Ala Arg 195 200 205 Gln Lys Ala Gln Ser Ala Gln Tyr Ala Phe Ile Leu Asp Arg Met Ser 210 215 220 Gln Gln Val Asp Ala Asp Glu His Arg Val Ala Leu Leu Arg Lys Thr 225 230 235 240 Val Gly Glu Thr Trp Gly Leu Pro Ser Pro Glu Glu Ala Ala Ala Ala 245 250 255 Glu Ala Glu Val Phe Ala Thr Arg Tyr Ser Asp Asp Ser Pro Ala Pro 260 265 270 Asp Asp Glu Ser Asp Pro Trp Met Thr Glu Asn Leu Thr Val Gln Pro 275 280 285 Glu Arg Leu Gly Val Leu Ala Ser His His Asp Asn Ala Ala Val Asp 290 295 300 Ala Ser Ser Gly Val Glu Ala Ala Ala Gly Leu Gly Glu Ser Val Ala 305 310 315 320 Ile Thr His Gly Pro Tyr Ser Ser Gln Phe Asp Asp Thr Leu Asn Val 325 330 335 Tyr Leu Thr Ala His Asn Ala Leu Gly Ser Ser Leu His Thr Ala Gly 340 345 350 Val Asp Leu Ala Lys Ser Leu Arg Ile Ala Ala Lys Ile Tyr Ser Glu 355 360 365 Ala Asp Glu Ala Trp Arg Lys Ala Ile Asp Gly Leu Phe Thr Met Thr 370 375 380 Gly Phe Leu Gly Val Val Pro Ser Phe Leu Lys Val Leu Ala Gly Met 385 390 395 400 His Asn Glu Ile Val Gly Asp Ile Lys Arg Ala Thr Asp Thr Val Ala 405 410 415 Gly Ile Ser Gly Arg Val Gln Leu Thr His Gly Ser Phe Thr Ser Lys 420 425 430 Phe Asn Asp Thr Leu Gln Glu Phe Glu Thr Thr Arg Ser Ser Thr Gly 435 440 445 Thr Gly Leu Gln Gly Val Thr Ser Gly Leu Ala Asn Asn Leu Leu Ala 450 455 460 Ala Ala Gly Ala Tyr Leu Lys Ala Asp Asp Gly Leu Ala Gly Val Ile 465 470 475 480 Asp Lys Ile Phe Gly Met Ser Thr Thr Phe Ala Ala Arg Leu Asn Arg 485 490 495 Leu Phe Asp Thr Val Tyr Pro Pro Gly Arg Gly Pro His Thr Ser Ala 500 505 510 Glu Val Ile Ala Ala Leu Lys Ala Glu Gly Ile Thr Met Ser Ala Pro 515 520 525 Tyr Leu Ser Gln Leu Arg Ser Gly Asn Arg Thr Asn Pro Ser Gly Ala 530 535 540 Thr Met Ala Ala Leu Ala Asn Phe Phe Arg Ile Lys Ala Ala Tyr Phe 545 550 555 560 Thr Asp Asp Glu Tyr Tyr Glu Lys Leu Asp Lys Glu Leu Gln Trp Leu 565 570 575 Ser Thr Met Arg Asp Asp Gly Val Arg Arg Ile Ala Gln Arg Ala His 580 585 590 Gly Leu Pro Ser Ala Ala Gln Gln Lys Val Leu Asp Arg Ile Asp Glu 595 600 605 Leu Arg Arg Ala Glu Gly Ile Asp Ala Met Glu Lys Met Ser His Asp 610 615 620 Pro Ile Ala Ala Asp Ile Gly Thr Gln Val Ser Asp Asn Ala Leu His 625 630 635 640 Gly Val Thr Ala Gly Ser Thr Ala Leu Thr Ser Val Thr Gly Leu Val 645 650 655 Pro Ala Gly Ala Asp Glu Val Ser Ala Gln Ala Ala Thr Ala Phe Thr 660 665 670 Ser Glu Gly Ile Gln Leu Leu Ala Ser Asn Ala Ser Ala Gln Asp Gln 675 680 685 Leu His Arg Ala Gly Glu Ala Val Gln Asp Val Ala Arg Thr Tyr Ser 690 695 700 Gln Ile Asp Asp Gly Ala Ala Gly Val Phe Ala Glu 705 710 715 <210> 45 <211> 454 <212> PRT <213> Tuberculosis <400> 45 Met Ser Arg Pro Gly Leu Pro Val Glu Tyr Leu Gln Val Pro Ser Pro 1 5 10 15 Ser Met Gly Arg Asp Ile Lys Val Gln Phe Gln Ser Gly Gly Asn Asn 20 25 30 Ser Pro Ala Val Tyr Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Tyr 35 40 45 Asn Gly Trp Asp Ile Asn Thr Pro Ala Phe Glu Trp Tyr Tyr Gln Ser 50 55 60 Gly Leu Ser Ile Val Met Pro Val Gly Gly Gln Ser Ser Phe Tyr Ser 65 70 75 80 Asp Trp Tyr Ser Pro Ala Cys Gly Lys Ala Gly Cys Gln Thr Tyr Lys 85 90 95 Trp Glu Thr Phe Leu Thr Ser Glu Leu Pro Gln Trp Leu Ser Ala Asn 100 105 110 Arg Ala Val Lys Pro Thr Gly Ser Ala Ala Ile Gly Leu Ser Met Ala 115 120 125 Gly Ser Ser Ala Met Ile Leu Ala Ala Tyr His Pro Gln Gln Phe Ile 130 135 140 Tyr Ala Gly Ser Leu Ser Ala Leu Leu Asp Pro Ser Gln Gly Met Gly 145 150 155 160 Pro Ser Leu Ile Gly Leu Ala Met Gly Asp Ala Gly Gly Tyr Lys Ala 165 170 175 Ala Asp Met Trp Gly Pro Ser Ser Asp Pro Ala Trp Glu Arg Asn Asp 180 185 190 Pro Thr Gln Gln Ile Pro Lys Leu Val Ala Asn Asn Thr Arg Leu Trp 195 200 205 Val Tyr Cys Gly Asn Gly Thr Pro Asn Glu Leu Gly Gly Ala Asn Ile 210 215 220 Pro Ala Glu Phe Leu Glu Asn Phe Val Arg Ser Ser Asn Leu Lys Phe 225 230 235 240 Gln Asp Ala Tyr Asn Ala Ala Gly Gly His Asn Ala Val Phe Asn Phe 245 250 255 Pro Pro Asn Gly Thr His Ser Trp Glu Tyr Trp Gly Ala Gln Leu Asn 260 265 270 Ala Met Lys Gly Asp Leu Gln Ser Ser Leu Gly Ala Gly Thr Glu Gln 275 280 285 Gln Trp Asn Phe Ala Gly Ile Glu Ala Ala Ala Ser Ala Ile Gln Gly 290 295 300 Asn Val Thr Ser Ile His Ser Leu Leu Asp Glu Gly Lys Gln Ser Leu 305 310 315 320 Thr Lys Leu Ala Ala Ala Trp Gly Gly Ser Gly Ser Glu Ala Tyr Gln 325 330 335 Gly Val Gln Gln Lys Trp Asp Ala Thr Ala Thr Glu Leu Asn Asn Ala 340 345 350 Leu Gln Asn Leu Ala Arg Thr Ile Ser Glu Ala Gly Gln Ala Met Ala 355 360 365 Ser Thr Glu Gly Asn Val Thr Gly Met Phe Ala Val Ile Ala Gly Val 370 375 380 Asp Gln Ala Leu Ala Ala Thr Gly Gln Ala Ser Gln Arg Ala Ala Gly 385 390 395 400 Ala Ser Gly Gly Val Thr Val Gly Val Gly Val Gly Thr Glu Gln Arg 405 410 415 Asn Leu Ser Val Val Ala Pro Ser Gln Phe Thr Phe Ser Ser Arg Ser 420 425 430 Pro Asp Phe Val Asp Glu Thr Ala Gly Gln Ser Trp Cys Ala Ile Leu 435 440 445 Gly Leu Asn Gln Phe His 450 <210> 46 <211> 288 <212> DNA <213> Mycobacterium tuberculosis <400> 46 atgacagagc agcagtggaa tttcgcgggt atcgaggccg cggcaagcgc aatccaggga 60 aatgtcacgt ccattcattc cctccttgac gaggggaagc agtccctgac caagctcgca 120 gcggcctggg gcggtagcgg ttcggaggcg taccagggtg tccagcaaaa atgggacgcc 180 acggctaccg agctgaacaa cgcgctgcag aacctggcgc ggacgatcag cgaagccggt 240 caggcaatgg cttcgaccga aggcaacgtc actgggatgt tcgcatag 288 <210> 47 <211> 1107 <212> DNA <213> Mycobacterium tuberculosis <400> 47 atgctgtggc acgcaatgcc accggagcta aataccgcac ggctgatggc cggcgcgggt 60 ccggctccaa tgcttgcggc ggccgcggga tggcagacgc tttcggcggc tctggacgct 120 caggccgtcg agttgaccgc gcgcctgaac tctctgggag aagcctggac tggaggtggc 180 agcgacaagg cgcttgcggc tgcaacgccg atggtggtct ggctacaaac cgcgtcaaca 240 caggccaaga cccgtgcgat gcaggcgacg gcgcaagccg cggcatacac ccaggccatg 300 gccacgacgc cgtcgctgcc ggagatcgcc gccaaccaca tcacccaggc cgtccttacg 360 gccaccaact tcttcggtat caacacgatc ccgatcgcgt tgaccgagat ggattatttc 420 atccgtatgt ggaaccaggc agccctggca atggaggtct accaggccga gaccgcggtt 480 aacacgcttt tcgagaagct cgagccgatg gcgtcgatcc ttgatcccgg cgcgagccag 540 agcacgacga acccgatctt cggaatgccc tcccctggca gctcaacacc ggttggccag 600 ttgccgccgg cggctaccca gaccctcggc caactgggtg agatgagcgg cccgatgcag 660 cagctgaccc agccgctgca gcaggtgacg tcgttgttca gccaggtggg cggcaccggc 720 ggcggcaacc cagccgacga ggaagccgcg cagatgggcc tgctcggcac cagtccgctg 780 tcgaaccatc cgctggctgg tggatcaggc cccagcgcgg gcgcgggcct gctgcgcgcg 840 gagtcgctac ctggcgcagg tgggtcgttg acccgcacgc cgctgatgtc tcagctgatc 900 gaaaagccgg ttgccccctc ggtgatgccg gcggctgctg ccggatcgtc ggcgacgggt 960 ggcgccgctc cggtgggtgc gggagcgatg ggccagggtg cgcaatccgg cggctccacc 1020 aggccgggtc tggtcgcgcc ggcaccgctc gcgcaggagc gtgaagaaga cgacgaggac 1080 gactgggacg aagaggacga ctggtga 1107 <210> 48 <211> 2001 <212> DNA <213> Mycobacterium tuberculosis <400> 48 atggcggccg actacgacaa gctcttccgg ccgcacgaag gtatggaagc tccggacgat 60 atggcagcgc agccgttctt cgaccccagt gcttcgtttc cgccggcgcc cgcatcggca 120 aacctaccga agcccaacgg ccagactccg cccccgacgt ccgacgacct gtcggagcgg 180 ttcgtgtcgg ccccgccgcc gccaccccca cccccacctc cgcctccgcc aactccgatg 240 ccgatcgccg caggagagcc gccctcgccg gaaccggccg catctaaacc acccacaccc 300 cccatgccca tcgccggacc cgaaccggcc ccacccaaac cacccacacc ccccatgccc 360 atcgccggac ccgaaccggc cccacccaaa ccacccacac ctccgatgcc catcgccgga 420 cctgcaccca ccccaaccga atcccagttg gcgcccccca gaccaccgac accacaaacg 480 ccaaccggag cgccgcagca accggaatca ccggcgcccc acgtaccctc gcacgggcca 540 catcaacccc ggcgcaccgc accagcaccg ccctgggcaa agatgccaat cggcgaaccc 600 ccgcccgctc cgtccagacc gtctgcgtcc ccggccgaac caccgacccg gcctgccccc 660 caacactccc gacgtgcgcg ccggggtcac cgctatcgca cagacaccga acgaaacgtc 720 gggaaggtag caactggtcc atccatccag gcgcggctgc gggcagagga agcatccggc 780 gcgcagctcg cccccggaac ggagccctcg ccagcgccgt tgggccaacc gagatcgtat 840 ctggctccgc ccacccgccc cgcgccgaca gaacctcccc ccagcccctc gccgcagcgc 900 aactccggtc ggcgtgccga gcgacgcgtc caccccgatt tagccgccca acatgccgcg 960 gcgcaacctg attcaattac ggccgcaacc actggcggtc gtcgccgcaa gcgtgcagcg 1020 ccggatctcg acgcgacaca gaaatcctta aggccggcgg ccaaggggcc gaaggtgaag 1080 aaggtgaagc cccagaaacc gaaggccacg aagccgccca aagtggtgtc gcagcgcggc 1140 tggcgacatt gggtgcatgc gttgacgcga atcaacctgg gcctgtcacc cgacgagaag 1200 tacgagctgg acctgcacgc tcgagtccgc cgcaatcccc gcgggtcgta tcagatcgcc 1260 gtcgtcggtc tcaaaggtgg ggctggcaaa accacgctga cagcagcgtt ggggtcgacg 1320 ttggctcagg tgcgggccga ccggatcctg gctctagacg cggatccagg cgccggaaac 1380 ctcgccgatc gggtagggcg acaatcgggc gcgaccatcg ctgatgtgct tgcagaaaaa 1440 gagctgtcgc actacaacga catccgcgca cacactagcg tcaatgcggt caatctggaa 1500 gtgctgccgg caccggaata cagctcggcg cagcgcgcgc tcagcgacgc cgactggcat 1560 ttcatcgccg atcctgcgtc gaggttttac aacctcgtct tggctgattg tggggccggc 1620 ttcttcgacc cgctgacccg cggcgtgctg tccacggtgt ccggtgtcgt ggtcgtggca 1680 agtgtctcaa tcgacggcgc acaacaggcg tcggtcgcgt tggactggtt gcgcaacaac 1740 ggttaccaag atttggcgag ccgcgcatgc gtggtcatca atcacatcat gccgggagaa 1800 cccaatgtcg cagttaaaga cctggtgcgg catttcgaac agcaagttca acccggccgg 1860 gtcgtggtca tgccgtggga caggcacatt gcggccggaa ccgagatttc actcgacttg 1920 ctcgacccta tctacaagcg caaggtcctc gaattggccg cagcgctatc cgacgatttc 1980 gagagggctg gacgtcgttg a 2001 <210> 49 <211> 312 <212> DNA <213> Mycobacterium tuberculosis <400> 49 atgacggaaa acttgaccgt ccagcccgag cgtctcggtg tactggcgtc gcaccatgac 60 aacgcggcgg tcgatgcctc ctcgggcgtc gaagctgccg ctggcctagg cgaatctgtg 120 gcgatcactc acggtccgta ctgctcacag ttcaacgaca cgttaaatgt gtacttgact 180 gcccacaatg ccctgggctc gtccttgcat acggccggtg tcgatctcgc caaaagtctt 240 cgaattgcgg cgaagatata tagcgaggcc gacgaagcgt ggcgcaaggc tatcgacggg 300 ttgtttacct ga 312 <210> 50 <211> 1179 <212> DNA <213> Mycobacterium tuberculosis <400> 50 atgagcagag cgttcatcat cgatccaacg atcagtgcca ttgacggctt gtacgacctt 60 ctggggattg gaatacccaa ccaagggggt atcctttact cctcactaga gtacttcgaa 120 aaagccctgg aggagctggc agcagcgttt ccgggtgatg gctggttagg ttcggccgcg 180 gacaaatacg ccggcaaaaa ccgcaaccac gtgaattttt tccaggaact ggcagacctc 240 gatcgtcagc tcatcagcct gatccacgac caggccaacg cggtccagac gacccgcgac 300 atcctggagg gcgccaagaa aggtctcgag ttcgtgcgcc cggtggctgt ggacctgacc 360 tacatcccgg tcgtcgggca cgccctatcg gccgccttcc aggcgccgtt ttgcgcgggc 420 gcgatggccg tagtgggcgg cgcgcttgcc tacttggtcg tgaaaacgct gatcaacgcg 480 actcaactcc tcaaattgct tgccaaattg gcggagttgg tcgcggccgc cattgcggac 540 atcatttcgg atgtggcgga catcatcaag ggcaccctcg gagaagtgtg ggagttcatc 600 acaaacgcgc tcaacggcct gaaagagctt tgggacaagc tcacggggtg ggtgaccgga 660 ctgttctctc gagggtggtc gaacctggag tccttctttg cgggcgtccc cggcttgacc 720 ggcgcgacca gcggcttgtc gcaagtgact ggcttgttcg gtgcggccgg tctgtccgca 780 tcgtcgggct tggctcacgc ggatagcctg gcgagctcag ccagcttgcc cgccctggcc 840 ggcattgggg gcgggtccgg ttttgggggc ttgccgagcc tggctcaggt ccatgccgcc 900 tcaactcggc aggcgctacg gccccgagct gatggcccgg tcggcgccgc tgccgagcag 960 gtcggcgggc agtcgcagct ggtctccgcg cagggttccc aaggtatggg cggacccgta 1020 ggcatgggcg gcatgcaccc ctcttcgggg gcgtcgaaag ggacgacgac gaagaagtac 1080 tcggaaggcg cggcggcggg cactgaagac gccgagcgcg cgccagtcga agctgacgcg 1140 ggcggtgggc aaaaggtgct ggtacgaaac gtcgtctaa 1179 <210> 51 <211> 687 <212> DNA <213> Mycobacterium tuberculosis <400> 51 atgcgcatca agatcttcat gctggtcacg gctgtcgttt tgctctgttg ttcgggtgtg 60 gccacggccg cgcccaagac ctactgcgag gagttgaaag gcaccgatac cggccaggcg 120 tgccagattc aaatgtccga cccggcctac aacatcaaca tcagcctgcc cagttactac 180 cccgaccaga agtcgctgga aaattacatc gcccagacgc gcgacaagtt cctcagcgcg 240 gccacatcgt ccactccacg cgaagccccc tacgaattga atatcacctc ggccacatac 300 cagtccgcga taccgccgcg tggtacgcag gccgtggtgc tcaaggtcta ccagaacgcc 360 ggcggcacgc acccaacgac cacgtacaag gccttcgatt gggaccaggc ctatcgcaag 420 ccaatcacct atgacacgct gtggcaggct gacaccgatc cgctgccagt cgtcttcccc 480 attgtgcaag gtgaactgag caagcagacc ggacaacagg tatcgatagc gccgaatgcc 540 ggcttggacc cggtgaatta tcagaacttc gcagtcacga acgacggggt gattttcttc 600 ttcaacccgg gggagttgct gcccgaagca gccggcccaa cccaggtatt ggtcccacgt 660 tccgcgatcg actcgatgct ggcctag 687 <210> 52 <211> 582 <212> DNA <213> Mycobacterium tuberculosis <400> 52 atgaaggtaa agaacacaat tgcggcaacc agtttcgcgg cggccggcct ggcggctctg 60 gcggtggctg tctcaccgcc ggcggccgca ggcgatctgg tgggcccggg ctgcgcggaa 120 tacgcggcag ccaatcccac tgggccggcc tcggtgcagg gaatgtcgca ggacccggtc 180 gcggtggcgg cctcgaacaa tccggagttg acaacgctga cggctgcact gtcgggccag 240 ctcaatccgc aagtaaacct ggtggacacc ctcaacagcg gtcagtacac ggtgttcgca 300 ccgaccaacg cggcatttag caagctgccg gcatccacga tcgacgagct caagaccaat 360 tcgtcactgc tgaccagcat cctgacctac cacgtagtgg ccggccaaac cagcccggcc 420 aacgtcgtcg gcacccgtca gaccctccag ggcgccagcg tgacggtgac cggtcagggt 480 aacagcctca aggtcggtaa cgccgacgtc gtctgtggtg gggtgtctac cgccaacgcg 540 acggtgtaca tgattgacag cgtgctaatg cctccggcgt aa 582 <210> 53 <211> 663 <212> DNA <213> Mycobacterium tuberculosis <400> 53 atgatcaacg ttcaggccaa accggccgca gcagcgagcc tcgcagccat cgcgattgcg 60 atgatcaacg ttcaggccaa accggccgca gcagcgagcc tcgcagccat cgcgattgcg 60 ttcttagcgg gttgttcgag caccaaaccc gtgtcgcaag acaccagccc gaaaccggcg 120 ttcttagcgg gttgttcgag caccaaaccc gtgtcgcaag acaccagccc gaaaccggcg 120 accagcccgg cggcgcccgt taccacggcg gcaatggctg accccgcagc ggacctgatt 180 accagcccgg cggcgcccgt taccacggcg gcaatggctg accccgcagc ggacctgatt 180 ggtcgtgggt gcgcgcaata cgcggcgcaa aatcccaccg gtcccggatc ggtggccgga 240 ggtcgtgggt gcgcgcaata cgcggcgcaa aatcccaccg gtcccggatc ggtggccgga 240 atggcgcaag acccggtcgc taccgcggct tccaacaacc cgatgctcag taccctgacc 300 atggcgcaag acccggtcgc taccgcggct tccaacaacc cgatgctcag taccctgacc 300 tcggctctgt cgggcaagct gaacccggat gtgaatctgg tcgacaccct caacggcggc 360 tcggctctgt cgggcaagct gaacccggat gtgaatctgg tcgacaccct caacggcggc 360 gagtacaccg ttttcgcccc caccaacgcc gcattcgaca agctgccggc ggccactatc 420 gagtacaccg ttttcgcccc caccaacgcc gcattcgaca agctgccggc ggccactatc 420 gatcaactca agactgacgc caagctgctc agcagcatcc tgacctacca cgtgatagcc 480 gatcaactca agactgacgc caagctgctc agcagcatcc tgacctacca cgtgatagcc 480 ggccaggcga gtccgagcag gatcgacggc acccatcaga ccctgcaagg tgccgacctg 540 ggccaggcga gtccgagcag gatcgacggc acccatcaga ccctgcaagg tgccgacctg 540 acggtgatag gcgcccgcga cgacctcatg gtcaacaacg ccggtttggt atgtggcgga 600 acggtgatag gcgcccgcga cgacctcatg gtcaacaacg ccggtttggt atgtggcgga 600 gttcacaccg ccaacgcgac ggtgtacatg atcgatacgg tgctgatgcc cccggcacag 660 gttcacaccg ccaacgcgac ggtgtacatg atcgatacgg tgctgatgcc cccggcacag 660 taa 663 taa 663 <210> 54 <210> 54 <211> 6469 <211> 6469 <212> DNA <212> DNA <213> Mycobacterium tuberculosis <213> Mycobacterium tuberculosis <400> 54 atgcaagccg aaaccgccgt caataccctg ttcgagaaac tggagccgat ggcatctatt 60 cttgatccgg gtgcgagcca gagcactacc aatccaatct ttggcatgcc gtccccgggc 120 agctcgacgc cggtgggcca gctgccgccg gcggctaccc aaaccttagg tcagctgggt 180 gagatgagcg gcccgatggg cggctccggt aaccctgcag atgaagaggc agcacaaatg 240 ggtctgctgg gtaccagccc gctgagcaat cacccactgg ccggcggcag cggcccaagc 300 gctggtgcgg gtctgctgcg cgcggagtcc ctgccgggtg cgggtggctc cctgacgcgt 360 accccactca tgagccaact gattgagaaa ccggttgcga ccgaacaaca gtggaatttt 420 gccggtattg aagctgcggc cagcgccatc cagggcaatg ttacgagcat ccacagcctg 480 ctggatgaag gcaaacagtc gctgaccaag ctggcggcgg cgtggggtgg ctccggtagc 540 gaagcctatc agggtgttca gcagaaatgg gacgcgaccg cgactgagct gaacaatgcg 600 ctgcagaatc tggcccgcac tatttccgag gccggtcaag cgatggcaag caccgagggc 660 aacgtgaccg gtatgttcgc tgctgccgat tacgacaaac tgtttcgccc acatgaaggc 720 atggaggcac cggatgatat ggcggcgcag ccgtttttcg acccgtccgc gagctttccg 780 ccggccccag caagcgcgaa tctgcctaaa ccgaacggtc agaccccgcc gcctacgagc 840 gacgatttga gcgaacgttt tgtgagcgcg ccgccgccgc cgccaccgcc gccgcctccg 900 ccgccaccta ccccgatgcc gattgccgcg ggtgagccac cgagcccgga gccagcggcc 960 tcgaagccgc cgacgccgcc gatgccgatc gcgggccctg aaccggcgcc gccgaaacct 1020 ccgaccccgc cgatgcctat tgctggtccg gaaccggcac cgcctaaacc gccgacgccg 1080 ccgatgccaa tcgcgggtcc ggctcctacc ccgacggaaa gccagctggc tccaccgcgt 1140 ccgcctaccc cgcagacccc gaccggtgcg ccgcaacaac cggaaagccc ggcgccgcat 1200 gtcccttcgc acggccctca ccaaccgcgt cgcacggctc cggcgccgcc ttgggctaag 1260 atgccgattg gtgagccgcc gccggctccg agccgtccga gcgctagccc ggcggaaccg 1320 ccgacccgtc cggcgccaca gcattctcgt cgtgcacgcc gcggtcaccg ctaccgtacg 1380 gataccgaac gtaacgtcgg taaagtcgca accggtccgt caatccaggc acgtttgcgt 1440 gcggaagaag ccagcggtgc gcagctggcg ccgggcacgg agccgtcgcc ggcaccgctg 1500 ggtcaaccga gatcttactt ggccccgccg acccgcccgg cgccgacgga accgccgccg 1560 agcccgtccc cgcaacgcaa cagcggccgc cgtgcggagc gtcgtgttca cccggatctg 1620 gcggcgcaac acgccgcagc ccagccggac agcattaccg cggcaaccac cggcggtcgc 1680 cgccgcaagc gtgcggcgcc ggacctggac gccacccaga aaagcctgcg tccagctgca 1740 aagggcccga aagtcaaaaa ggtcaagccg caaaagccga aagcaactaa accgccgaaa 1800 gtcgttagcc agcgcggttg gcgtcactgg gtgcacgcgc tgacgcgcat taacctgggt 1860 ttgagcccgg acgaaaaata tgaactggac ttgcacgcac gtgtccgccg taacccgcgt 1920 ggttcatacc agattgccgt tgtcggtctg cagggtggcg ccggcaaaac cacgctgact 1980 gcggcactgg gttccacgct ggcgcaagtg cgtgcggatc gtattctggc actggatgct 2040 gacccgggcg cgggtaatct ggcagatcgt gtgggtcgtc agtctggtgc gaccatcgca 2100 gatgttctgg ctgagaaaga actgagccat tacaacgaca ttcgtgcgca cacgtctgtt 2160 aatgccgtta atctggaagt gctgccggca ccggagtaca gcagcgcaca gcgcgccctg 2220 agcgacgcgg actggcactt catcgcagac ccggcatctc gcttctataa cctggttttg 2280 gcggattccg gtgcgggctt tttcgatcca ctgacccgcg gtgtgctcag caccgttagc 2340 ggtgtggttg tggttgcctc cgtgagcatc gacggtgcgc aacaagcatc ggtggcgctg 2400 gactggctgc gtaataacgg ttatcaagac ttggcgagcc gcgcgagcgt tgtgattaac 2460 catatcatgc cgggcgagcc gaatgttgcg gttaaagatt tggtgcgcca ttttgagcaa 2520 caggtccaac cgggccgcgt cgttgtgatg ccttgggatc gtcatattgc agcgggcacc 2580 gaaattagcc tggacttact ggacccgatt tataagcgta aggtactgga actggccgca 2640 gcgctgagcg acgatttcga gcgtgcaggc cgtcgtaccg agcaacagtg gaatttcgct 2700 ggcattgaag ccgcagcctc tgcaattcag ggtaatgtta cctcgattca cagcctgctg 2760 gacgagggca aacaaagcct gaccaagctg gcggctgctt ggggtggttc tggtagcgaa 2820 gcgtaccagg gcgtccagca aaaatgggac gcaaccgcaa ccgagctgaa taatgctctg 2880 caaaacctgg cccgtaccat tagcgaagcg ggccaggcca tggcgagcac cgaaggcaac 2940 gtgaccggta tgtttgccac cgaaaacttg accgttcagc cggaacgtct gggtgtgctg 3000 gcgagccatc atgacaacgc agcagttgat gcgagcagcg gtgtcgaggc agccgcgggt 3060 ttgggcgaga gcgttgccat cacccatggt ccgtattcat ctcagttcaa cgacaccacg 3120 gaacagcagt ggaacttcgc aggcatcgaa gccgcggcgt ccgcgatcca gggtaacgtg 3180 actagcattc attcgttgct ggacgaaggc aagcaaagcc tgaccaaact ggcagctgca 3240 tggggtggct cgggctctga ggcgtaccaa ggcgtccaac agaagtggga cgcgactgcg 3300 accgagttaa acaatgcttt acagaacctg gctcgtacta tcagcgaggc tggtcaggcc 3360 atggcgagca ccgaaggtaa tgttaccggc atgttcgcga gccgcgcttt tattatcgac 3420 ccgacgatca gcgccatcga tggtctgtac gatctgttgg gtattggcat tccgaaccaa 3480 ggcggcatcc tgtatagctc cctggaatac ttcgagaaag ccttggaaga actggcggcc 3540 gcgttcccgg gcgatggctg gctgggtagc gcggctgaca aatatgcggg caaaaaccgt 3600 aaccatgtca actttttcca agagttggcg gatttggatc gtcaattgat tagcctgatc 3660 cacgatcagg cgaatgcagt gcaaaccact cgcgatatcc tcgaaggtgc caaaaagggt 3720 ctggagtttg tacgtccagt cgcagtcgac ctgacctaca ttccagttgt cggtcatgcg 3780 ttgtcggcaa agactctgat taatgcgacg caactgttaa aactgttggc gaaattggca 3840 gagctggtcg cggcagcgat cgccgacatc atcagcgatg tggcagatat cattaagggt 3900 acgctgggcg aagtgtggga atttattact aatgcgctca acggtctgaa ggaactgtgg 3960 gataaattga cgggttgggt taccggcttg tttagccgtg gttggtccaa tttggagagc 4020 tttttcgccg gtgttccggg cctgaccggc gcaacgagcg gtttgtccca ggtcacgggc 4080 ttgtttggtg ctgcgggtct gagcgcatcc agcggtctgg cgcacgcaga ttccttggcg 4140 tctagcgcaa gcctgccggc gctggcaggc atcggcggcg gctctggctt cggtggcttg 4200 ccaagcctgg cccaagtcca tgctgcttcc acgcgccagg ctctgcgccc acgtgcagat 4260 ggtccggttg gtgccgcagc cgagcaagtt ggtggccaga gccaactggt cagcgctcag 4320 ggctctcagg gcatgggcgg tccggttggt atgggtggta tgcacccgag ctccggcgca 4380 tccaagggta ctaccacgaa aaagtatagc gagggtgcag cggcgggtac tgaagatgcc 4440 gaacgtgcac cggttgaggc tgatgcgggt ggcggccaaa aagttttggt ccgtaatgtc 4500 gtgactgagc agcagtggaa ttttgccggc atcgaagcgg cggcgagcgc gatccaaggc 4560 aacgtcactt cgattcactc tctgctggat gagggcaaac agtccctgac gaaactggca 4620 gccgcgtggg gtggtagcgg ttccgaggca taccaaggtg ttcagcagaa gtgggacgcg 4680 acggccacgg aactgaacaa cgcgctgcag aacctggcgc gtaccatctc ggaagcaggc 4740 caggcaatgg ctagcactga aggcaacgtg acgggtatgt tcgcagcacc taagacctat 4800 agcgaagagt tgaagggtac tgatactggt caagcgagcc agattcagat gagcgacccg 4860 gcgtacaata tcaacatctc cctgccaagc tactatccag accaaaaaag cctggagaac 4920 tacatcgcgc agactcgcga taagttcctg tccgcagcga cgagcagcac cccacgcgaa 4980 gcaccgtatg agctgaacat tactagcgcg acgtaccaaa gcgcgattcc gccgcgtggt 5040 acccaagccg ttgtcctgaa agtttatcag aacgccggtg gcactcatcc gacgacgacc 5100 tataaggcct ttgactggga ccaggcatat cgtaagccaa tcacgtacga caccctgtgg 5160 caggccgaca cggatccact gccagttgtg tttccgatcg tgcaaggtga actgtcaaaa 5220 caaactggtc aacaggtgag cattgcccca aacgccggtc tggatccggt gaattatcaa 5280 aattttgctg ttacgaacga cggcgttatc tttttcttca acccgggtga actgctgccg 5340 gaagcggcgg gtccgaccca agtgctggtt ccgcgtagcg cgattgacag catgctggcg 5400 ggcgatctgg ttggcccggg tagcgcagaa tacgccgcgg ccaacccgac cggtccggca 5460 tccgtccaag gtatgagcca ggatccggtg gctgttgcag cctctaacaa tccggaactg 5520 accaccttga cggcagcgct gagcggtcag ctgaatcctc aagtgaatct ggttgatacg 5580 ctgaatagcg gccaatatac cgtctttgca ccgaccaacg ctgcgttcag caaactgccg 5640 gccagcacca ttgacgaatt gaaaaccaat tccagcctgt tgacgagcat tctgacgtat 5700 cacgttgtgg cgggccagac gtccccagcg aacgtcgtcg gtacccgtca gacgctgcag 5760 ggtgcgtctg taaccgttac gggccaaggc aactctttga aagtcggtaa tgccgacgtc gtcagcggtg gcgttagcac ggctaatgca accgtctaca tgattgacag cgttttgatg ccgcctgccg tcagccagga tacgtccccg aaaccggcta cgagcccggc agccccggtt 5940 accactgcag caatggcgga ccctgcggct gatttgatcg gtcgcggctc tgcacagtat gctgcccaaa acccgacggg tccgggctct gttgccggca tggcgcagga cccggtcgca 6060 accgcggcta gcaataatcc aatgctgtcg accgctgactt ccgctctgtc cggtaaactg aatccggacg tcaacctggt ggataccctg aacggtggcg agtataccgt ctttgccccg 6180. accaacgctg cgttcgacaa gctgccggca gccaccatcg accaactga aaccgacgcc aagctgctga gcagcatcct gacctatcac gtgattgcgg gtcaagctag cccgagccgc 6300. atcgacggca cccaccagac cttgcagggc gctgacctga cggttattgg tgctcgcgat 6360. gatctgatgg tgaataacgc cggtctggtt tccggcggcg ttcatacggc taatgcaacg gtgtacatga ttgatacggt ccttatgcct ccggcgcagt gataagctt 6469 <210> 55 <211> 978 <212> DNA <213> Mycobacterium tuberculosis <400> 55 atgacagacg tgagccgaaa gattcgagct tggggacgcc gattgatgat cggcacggca 60 gcggctgtag tccttccggg cctggtgggg cttgccggcg gagcggcaac cgcgggcgcg 120 ttctcccggc cggggctgcc ggtcgagtac ctgcaggtgc cgtcgccgtc gatgggccgc 180 gacatcaagg ttcagttcca gagcggtggg aacaactcac ctgcggttta tctgctcgac 240 ggcctgcgcg cccaagacga ctacaacggc tgggatatca acaccccggc gttcgagtgg 300 tactaccagt cgggactgtc gatagtcatg ccggtcggcg ggcagtccag cttctacagc 360 gactggtaca gcccggcctg cggtaaggct ggctgccaga cttacaagtg ggaaaccttc 420 ctgaccagcg agctgccgca atggttgtcc gccaacaggg ccgtgaagcc caccggcagc 480 gctgcaatcg gcttgtcgat ggccggctcg tcggcaatga tcttggccgc ctaccacccc 540 cagcagttca tctacgccgg ctcgctgtcg gccctgctgg acccctctca ggggatgggg 600 cctagcctga tcggcctcgc gatgggtgac gccggcggtt acaaggccgc agacatgtgg 660 ggtccctcga gtgacccggc atgggagcgc aacgacccta cgcagcagat ccccaagctg 720 gtcgcaaaca acacccggct atgggtttat tgcgggaacg gcaccccgaa cgagttgggc 780 ggtgccaaca tacccgccga gttcttggag aacttcgttc gtagcagcaa cctgaagttc 840 caggatgcgt acaacgccgc gggcgggcac aacgccgtgt tcaacttccc gcccaacggc 900 acgcacagct gggagtactg gggcgctcag ctcaacgcca tgaagggtga cctgcagagt 960 tcgttaggcg ccggctga 978 <210> 56 <211> 1017 <212> DNA <213> Mycobacterium tuberculosis <400> 56 atgcagcttg ttgacagggt tcgtggcgcc gtcacgggta tgtcgcgtcg actcgtggtc 60 ggggccgtcg gcgcggccct agtgtcgggt ctggtcggcg ccgtcggtgg cacggcgacc 120 gcgggggcat tttcccggcc gggcttgccg gtggagtacc tgcaggtgcc gtcgccgtcg 180 atgggccgtg acatcaaggt ccaattccaa agtggtggtg ccaactcgcc cgccctgtac 240 ctgctcgacg gcctgcgcgc gcaggacgac ttcagcggct gggacatcaa caccccggcg 300 ttcgagtggt acgaccagtc gggcctgtcg gtggtcatgc cggtgggtgg ccagtcaagc 360 ttctactccg actggtacca gcccgcctgc ggcaaggccg gttgccagac ttacaagtgg 420 gagaccttcc tgaccagcga gctgccgggg tggctgcagg ccaacaggca cgtcaagccc 480 accggaagcg ccgtcgtcgg tctttcgatg gctgcttctt cggcgctgac gctggcgatc 540 tatcaccccc agcagttcgt ctacgcggga gcgatgtcgg gcctgttgga cccctcccag 600 gcgatgggtc ccaccctgat cggcctggcg atgggtgacg ctggcggcta caaggcctcc 660 gacatgtggg gcccgaagga ggacccggcg tggcagcgca acgacccgct gttgaacgtc 720 gggaagctga tcgccaacaa cacccgcgtc tgggtgtact gcggcaacgg caagccgtcg 780 gatctgggtg gcaacaacct gccggccaag ttcctcgagg gcttcgtgcg gaccagcaac 840 atcaagttcc aagacgccta caacgccggt ggcggccaca acggcgtgtt cgacttcccg 900 gacagcggta cgcacagctg ggagtactgg ggcgcgcagc tcaacgctat gaagcccgac 960 ctgcaacggg cactgggtgc cacgcccaac accgggcccg cgccccaggg cgcctag 1017 <210> 57 <211> 291 <212> DNA <213> Mycobacterium tuberculosis <400> 57 atgtcgcaaa tcatgtacaa ctaccccgcg atgttgggtc acgccgggga tatggccgga 60 tatgccggca cgctgcagag cttgggtgcc gagatcgccg tggagcaggc cgcgttgcag 120 agtgcgtggc agggcgatac cgggatcacg tatcaggcgt ggcaggcaca gtggaaccag 180 gccatggaag atttggtgcg ggcctatcat gcgatgtcca gcacccatga agccaacacc 240 atggcgatga tggcccgcga cacggccgaa gccgccaaat ggggcggcta g 291 <210> 58 <211> 294 <212> DNA <213> Mycobacterium tuberculosis <400> 58 atgagccttt tggatgctca tatcccacag ttggtggcct cccagtcggc gtttgccgcc 60 aaggcggggc tgatgcggca cacgatcggt caggccgagc aggcggcgat gtcggctcag 120 gcgtttcacc agggggagtc gtcggcggcg tttcaggccg cccatgcccg gtttgtggcg 180 gcggccgcca aagtcaacac cttgttggat gtcgcgcagg cgaatctggg tgaggccgcc 240 ggtacctatg tggccgccga tgctgcggcc gcgtcgacct ataccgggtt ctga 294 <210> 59 <211> 1182 <212> DNA <213> Mycobacterium tuberculosis <400> 59 gtggtggatt tcggggcgtt accaccggag atcaactccg cgaggatgta cgccggcccg 60 ggttcggcct cgctggtggc cgccgcgaag atgtgggaca gcgtggcgag tgacctgttt 120 tcggccgcgt cggcgtttca gtcggtggtc tggggtctga cggtggggtc gtggataggt 180 tcgtcggcgg gtctgatggc ggcggcggcc tcgccgtatg tggcgtggat gagcgtcacc 240 gcggggcagg cccagctgac cgccgcccag gtccgggttg ctgcggcggc ctacgagaca 300 gcgtataggc tgacggtgcc cccgccggtg atcgccgaga accgtaccga actgatgacg 360 ctgaccgcga ccaacctctt ggggcaaaac acgccggcga tcgaggccaa tcaggccgca 420 tacagccaga tgtggggcca agacgcggag gcgatgtatg gctacgccgc cacggcggcg 480 acggcgaccg aggcgttgct gccgttcgag gacgccccac tgatcaccaa ccccggcggg 540 ctccttgagc aggccgtcgc ggtcgaggag gccatcgaca ccgccgcggc gaaccagttg 600 atgaacaatg tgccccaagc gctgcaacag ctggcccagc cagcgcaggg cgtcgtacct 660 tcttccaagc tgggtgggct gtggacggcg gtctcgccgc atctgtcgcc gctcagcaac 720 gtcagttcga tagccaacaa ccacatgtcg atgatgggca cgggtgtgtc gatgaccaac 780 accttgcact cgatgttgaa gggcttagct ccggcggcgg ctcaggccgt ggaaaccgcg 840 gcggaaaacg gggtctgggc gatgagctcg ctgggcagcc agctgggttc gtcgctgggt 900 tcttcgggtc tgggcgctgg ggtggccgcc aacttgggtc gggcggcctc ggtcggttcg ⑨60 ttgtcggtgc cgccagcatg ggccgcggcc aaccaggcgg tcaccccggc ggcgcgggcg 1020 ctgccgctga ccagcctgac cagcgccgcc caaaccgccc ccggacacat gctgggcggg 1080 ctaccgctgg ggcactcggt caacgccggc agcggtatca acaatgcgct gcgggtgccg 1140 gcacgggcct acgcgatacc ccgcacaccg gccgccggat ag 1182 <210> 60 <211> 600 <212> DNA <213> Mycobacterium tuberculosis<00​ atggctgaaa actcgaacat tgatgacatc aaggctccgt tgcttgccgc gcttggagcg 60Methionine-alanine-glutamic acid-threonine-glutamic acid-histidine-aspartic acid-aspartic acid-isoleucine-lysine-alanine-proline-valine-alanine-alanine-alanine-leucine-glutamic acid-serine 60 gccgacctgg ccttggccac tgtcaacgag ttgatcacga acctgcgtga gcgtgcggag 120Alanine-aspartic acid-leucine-alanine-leucine-alanine-histidine-cysteine-asparagine-glutamic acid-leucine-isoleucine-histidine-glutamic acid-leucine-cysteine-glutamic acid-valine-cysteine-glutamic acid 120 gagactcgta cggacacccg cagccgggtc gaggagagcc gtgctcgcct gaccaagctg 180Glutamic acid-threonine-valine-arginine-aspartic acid-threonine-proline-alanine-proline-glycine-serine-glutamic acid-glutamic acid-serine-alanine-leucine-alanine-leucine-threonine-lysine-leucine 180 caggaagatc tgcccgagca gctcaccgag ctgcgtgaga agttcaccgc cgaggagctg 240Glutamine-glutamic acid-aspartic acid-leucine-proline-glutamic acid-alanine-serine-threonine-glutamic acid-leucine-cysteine-glutamic acid-lysine-phenylalanine-threonine-alanine-glutamic acid-glutamic acid-leucine 240 cgtaaggccg ccgagggcta cctcgaggcc gcgactagcc ggtacaacga gctggtcgag 300 Arginine-lysine-alanine-alanine-glutamic acid-glycine-tyrosine-leucine-glutamic acid-alanine-alanine-leucine-alanine-glycine-tyrosine-asparagine-glutamic acid-leucine-valine-glutamic acid 300 cgcggtgagg ccgctctaga gcggctgcgc agccagcaga gcttcgagga agtgtcggcg 360 Arginine-arginine-valine-glutamic acid-alanine-leucine-stop codon-serine-alanine-cysteine-alanine-glutamine-glutamine-alanine-serine-glutamic acid-glutamic acid-valine-serine-alanine 360 cgcgccgaag gctacgtgga ccaggcggtg gagttgaccc aggaggcgtt gggtacggtc 420 Arginine-alanine-glutamic acid-alanine-tyrosine-valine-aspartic acid-glutamine-alanine-valine-glutamic acid-leucine-aspartic acid-proline-glutamic acid-alanine-leucine-glycine-tyrosine-glycine-serine 420 gcatcgcaga cccgcgcggt cggtgagcgt gccgccaagc tggtcggcat cgagctgcct 480 Glycine-isoleucine-alanine-glutamine-threonine-alanine-arginine-valine-serine-valine-alanine-alanine-lysine-leucine-valine-glycine-isoleucine-glutamic acid-leucine-proline 480 aagaaggctg ctccggccaa gaaggccgct ccggccaaga aggccgctcc ggccaagaag 540 Lysine-lysine-alanine-alanine-serine-alanine-lysine-lysine-alanine-alanine-serine-alanine-lysine-lysine-alanine-alanine-serine-alanine-lysine-lysine 540 gcggcggcca agaaggcgcc cgcgaagaag gcggcggcca agaaggtcac ccagaagtag 600 Alanine-alanine-alanine-lysine-lysine-alanine-alanine-arginine-glutamic acid-glutamic acid-alanine-alanine-alanine-lysine-lysine-valine-threonine-glutamic acid-valine 600 <210> 61<210> 61 <211> 288<211> 288<atgtcagatc aaatcacgta taacccggga gccgtatccg acttcgcttc cgacgtgggc 60 tcgcgcgccg gccagctcca catgatttac gaagacaccg ccagcaaaac aaatgcgctg 120 caagagtttt tcgcgggcca cggcgcgcaa gggtttttcg acgcccaggc gcagatgctg 180 tcggggctgc aggggctcat tgagacggtg ggtcagcatg ggactaccac cggccacgtg 240 ctggacaacg cgatcggaac cgaccaggcc atcgcgggct tgttctaa 288 <210> 62 <211> 324 <212> DNA <213> Mycobacterium tuberculosis <400> 62 gtggcagaca caattcaggt aacaccgcag atgctgcgca gcaccgccaa cgatatccag 60 gcgaatatgg agcaagccat gggaatcgcc aagggctacc tagccaacca ggaaaacgtc 120 atgaaccccg ccacctggtc tggtaccggc gtcgttgctt cgcatatgac agccaccgag 180 atcaccaatg aattgaacaa ggtccttacc gggggcacgc gcctggccga gggcctcgtg 240 caggccgcag ccctgatgga gggacacgag gcggactcgc agacagcgtt tcaggcgctg 300 ttcggcgcta gccacggatc ctga 324 <210> 63 <211> 492 <212> DNA <213> Mycobacterium tuberculosis <400> 63 gtgacggtta ccgacgacta cctggccaac aacgtggact acgcgagcgg tttcaagggc 60 ccgctaccga tgccgccgag caaacacatc gcaatcgtgg cgtgcatgga cgcccggctg 120 gacgtctacc gcatgctggg catcaaggag ggcgaggcac acgtcatccg caacgccgga 180 tgcgtggtca ccgacgatgt gatccgttca ctggccatca gccagcggct gctgggaacc 240 cgcgaaatca tcctgctgca ccacaccgac tgtgggatgc tgactttcac cgacgacgac 300 ttcaagcgcg ccatccagga cgagaccggc atcagaccca cgtggtcgcc cgagtcgtac 360 cccgacgccg tcgaggacgt ccgtcagtcg ctgcgccgca tcgaggtcaa cccgttcgtc 420 accaagcaca cgtcgctgcg cggcttcgtc ttcgatgtcg ccaccggcaa actcaacgag 480 gtcacgccct ag 492 <210> 64 <211> 291 <212> DNA <213> Mycobacterium tuberculosis <400> 64 atgtcgcaga ttatgtacaa ctatccggcg atgatggctc atgccgggga catggccggt 60 tatgcgggca cgctgcagag cttgggggcc gatatcgcca gtgagcaggc cgtgctgtcc 120 agtgcttggc agggtgatac cgggatcacg tatcagggct ggcagaccca gtggaaccag 180 gccctagagg atctggtgcg ggcctatcag tcgatgtctg gcacccatga gtccaacacc 240 atggcgatgt tggctcgaga tggggccgaa gccgccaagt ggggcggcta g 291 <210> 65 <211> 294 <212> DNA <213> Mycobacterium tuberculosis <400> 65 atgagtttgt tggatgccca tattccgcag ttgatcgctt cgcatacggc gtttgccgct 60 aaggcggggt tgatgcggca tacgatcggt caggccgagc agcaggcgat gtcggcgcag 120 gcgtttcatc agggagagtc cgcggcggcg tttcagggtg cgcatgcccg gtttgtggcc 180 gcggccgcca aggtcaatac cttgctggat atcgcgcaag ccaatttggg tgaggccgcg 240 ggcacgtatg tggccgccga tgccgccgcc gcgtccagct acaccgggtt ttaa 294 <210> 66 <211> 363 <212> DNA <213> Mycobacterium tuberculosis <400> 66 gtgtcgcaga gtatgtacag ctacccggcg atgacggcca atgtcggaga catggccggt 60 tatacgggca cgacgcagag cttgggggcc gatatcgcca gtgagcgcac cgcgccgtcg 120 cgtgcttgcc aaggtgatct cgggatgagt catcaggact ggcaggccca gtggaatcag 180 gccatggagg ctctcgcgcg ggcctaccgt cggtgccggc gagcactacg ccagatcggg 240 gtgctggaaa ggccggtagg cgattcgtca gactgcggaa cgattagggt ggggtcgttc 300 cggggtcggt ggctggaccc gcgccatgcg ggtccagcca cggccgccga cgccggagac 360 taa 363 <210> 67 <211> 435 <212> DNA <213> Mycobacterium tuberculosis <400> 67 atggccacca cccttcccgt tcagcgccac ccgcggtccc tcttccccga gttttctgag 60 ctgttcgcgg ccttcccgtc attcgccgga ctccggccca ccttcgacac ccggttgatg 120 cggctggaag acgagatgaa agaggggcgc tacgaggtac gcgcggagct tcccggggtc 180 gaccccgaca aggacgtcga cattatggtc cgcgatggtc agctgaccat caaggccgag 240 cgcaccgagc agaaggactt cgacggtcgc tcggaattcg cgtacggttc cttcgttcgc 300 acggtgtcgc tgccggtagg tgctgacgag gacgacatta aggccaccta cgacaagggc 360 attcttactg tgtcggtggc ggtttcggaa gggaagccaa ccgaaaagca cattcagatc 420 cggtccacca actga 435 <210> 68 <211> 1395 <212> DNA; <213> Mycobacterium tuberculosis <400> 68 atggccaagt tggcccgagt agtgggccta gtacaggaag agcaacctag cgacatgacg 60 aatcacccac ggtattcgcc accgccgcag cagccgggaa ccccaggtta tgctcagggg 120 cagcagcaaa cgtacagcca gcagttcgac tggcgttacc caccgtcccc gcccccgcag 180 ccaacccagt accgtcaacc ctacgaggcg ttgggtggta cccggccggg tctgatacct 240 ggcgtgattc cgaccatgac gccccctcct gggatggttc gccaacgccc tcgtgcaggc 300 atgttggcca tcggcgcggt gacgatagcg gtggtgtccg ccggcatcgg cggcgcggcc 360 gcatccctgg tcgggttcaa ccgggcaccc gccggcccca gcggcggccc agtggctgcc 420 ; It should be noted that this is a technical patent text translation, and the accuracy of the translation needs to be further verified and adjusted according to the specific context and relevant professional knowledge.agcgcggcgc caagcatccc cgcagcaaac atgccgccgg ggtcggtcga acaggtggcg 480 gccaaggtgg tgcccagtgt cgtcatgttg gaaaccgatc tgggccgcca gtcggaggag 540 ggctccggca tcattctgtc tgccgagggg ctgatcttga ccaacaacca cgtgatcgcg 600 gcggccgcca agcctcccct gggcagtccg ccgccgaaaa cgacggtaac cttctctgac 660 gggcggaccg cacccttcac ggtggtgggg gctgacccca ccagtgatat cgccgtcgtc 720 cgtgttcagg gcgtctccgg gctcaccccg atctccctgg gttcctcctc ggacctgagg 780 gtcggtcagc cggtgctggc gatcgggtcg ccgctcggtt tggagggcac cgtgaccacg 840 gggatcgtca gcgctctcaa ccgtccagtg tcgacgaccg gcgaggccgg caaccagaac 900 accgtgctgg acgccattca gaccgacgcc gcgatcaacc ccggtaactc cgggggcgcg 960 ctggtgaaca tgaacgctca actcgtcgga gtcaactcgg ccattgccac gctgggcgcg 1020 gactcagccg atgcgcagag cggctcgatc ggtctcggtt ttgcgattcc agtcgaccag 1080 gccaagcgca tcgccgacga gttgatcagc accggcaagg cgtcacatgc ctccctgggt 1140 gtgcaggtga ccaatgacaa agacaccctg ggcgccaaga tcgtcgaagt agtggccggt 1200 gtgcaggtga ccaatgacaa agacaccctg ggcgccaaga tcgtcgaagt agtggccggt 1200 ggtgctgccg cgaacgctgg agtgccgaag ggcgtcgttg tcaccaaggt cgacgaccgc 1260 ggtgctgccg cgaacgctgg agtgccgaag ggcgtcgttg tcaccaaggt cgacgaccgc 1260 ccgatcaaca gcgcggacgc gttggttgcc gccgtgcggt ccaaagcgcc gggcgccacg 1320 ccgatcaaca gcgcggacgc gttggttgcc gccgtgcggt ccaaagcgcc gggcgccacg 1320 gtggcgctaa cctttcagga tccctcgggc ggtagccgca cagtgcaagt caccctcggc 1380 gtggcgctaa cctttcagga tccctcgggc ggtagccgca cagtgcaagt caccctcggc 1380 aaggcggagc agtga 1395 aaggcggagc agtga 1395 <210> 69<210> 69 <211> 1176<211> 1176 <212> DNA<212> DNA <213> 结核分枝杆菌<213> Mycobacterium tuberculosis <400> 69 <400> 69 atggtggatt tcggggcgtt accaccggag atcaactccg cgaggatgta cgccggcccg 60 atggtggatt tcggggcgtt accaccggag atcaactccg cgaggatgta cgccggcccg 60 ggttcggcct cgctggtggc cgcggctcag atgtgggaca gcgtggcgag tgacctgttt 120 ggttcggcct cgctggtggc cgcggctcag atgtgggaca gcgtggcgag tgacctgttt 120 tcggccgcgt cggcgtttca gtcggtggtc tggggtctga cggtggggtc gtggataggt 180 tcggccgcgt cggcgtttca gtcggtggtc tggggtctga cggtggggtc gtggataggt 180 tcgtcggcgg gtctgatggt ggcggcggcc tcgccgtatg tggcgtggat gagcgtcacc 240 tcgtcggcgg gtctgatggt ggcggcggcc tcgccgtatg tggcgtggat gagcgtcacc 240 gcggggcagg ccgagctgac cgccgcccag gtccgggttg ctgcggcggc ctacgagacg 300 gcggggcagg ccgagctgac cgccgcccag gtccgggttg ctgcggcggc ctacgagacg 300 gcgtatgggc tgacggtgcc cccgccggtg atcgccgaga accgtgctga actgatgatt 360 gcgtatgggc tgacggtgcc cccgccggtg atcgccgaga accgtgctga actgatgatt 360 ctgatagcga ccaacctctt ggggcaaaac accccggcga tcgcggtcaa cgaggccgaa 420 tacggcgaga tgtgggccca agacgccgcc gcgatgtttg gctacgccgc ggcgacggcg 480 acggcgacgg cgacgttgct gccgttcgag gaggcgccgg agatgaccag cgcgggtggg 540 ctcctcgagc aggccgccgc ggtcgaggag gcctccgaca ccgccgcggc gaaccagttg 600 atgaacaatg tgccccaggc gctgcaacag ctggcccagc ccacgcaggg caccacgcct 660 tcttccaagc tgggtggcct gtggaagacg gtctcgccgc atcggtcgcc gatcagcaac 720 atggtgtcga tggccaacaa ccacatgtcg atgaccaact cgggtgtgtc gatgaccaac 780 accttgagct cgatgttgaa gggctttgct ccggcggcgg ccgcccaggc cgtgcaaacc 840 gcggcgcaaa acggggtccg ggcgatgagc tcgctgggca gctcgctggg ttcttcgggt 900 ctgggcggtg gggtggccgc caacttgggt cgggcggcct cggtcggttc gttgtcggtg 960 ccgcaggcct gggccgcggc caaccaggca gtcaccccgg cggcgcgggc gctgccgctg 1020 accagcctga ccagcgccgc ggaaagaggg cccgggcaga tgctgggcgg gctgccggtg 1080 gggcagatgg gcgccagggc cggtggtggg ctcagtggtg tgctgcgtgt tccgccgcga 1140 ccctatgtga tgccgcattc tccggcggcc ggctag 1176 <210> 70 <211> 1743 <212> DNA <213> Mycobacterium tuberculosis <400> 70 atgaatttcg ccgttttgcc gccggaggtg aattcggcgc gcatattcgc cggtgcgggc 60 ctgggcccaa tgctggcggc ggcgtcggcc tgggacgggt tggccgagga gttgcatgcc 120 gcggcgggct cgttcgcgtc ggtgaccacc gggttggcgg gcgacgcgtg gcatggtccg 180 gcgtcgctgg cgatgacccg cgcggccagc ccgtatgtgg ggtggttgaa cacggcggcg 240 ggtcaggccg cgcaggcggc cggccaggcg cggctagcgg cgagcgcgtt cgaggcgacg 300 ctggcggcca ccgtgtctcc agcgatggtc gcggccaacc ggacacggct ggcgtcgctg 360 gtggcagcca acttgctggg ccagaacgcc ccggcgatcg cggccgcgga ggctgaatac 420 gagcagatat gggcccagga cgtggccgcg atgttcggct atcactccgc cgcgtcggcg 480 gtggccacgc agctggcgcc tattcaagag ggtttgcagc agcagctgca aaacgtgctg 540 gcccagttgg ctagcgggaa cctgggcagc ggaaatgtgg gcgtcggcaa catcggcaac 600 gacaacattg gcaacgcaaa catcggcttc ggaaatcgag gcgacgccaa catcggcatc 660 gggaatatcg gcgacagaaa cctcggcatt gggaacaccg gcaattggaa tatcggcatc 720 ggcatcaccg gcaacggaca aatcggcttc ggcaagcctg ccaaccccga cgtcttggtg 780 gtgggcaacg gcggcccggg agtaaccgcg ttggtcatgg gcggcaccga cagcctactg 840 ccgctgccca acatcccctt actcgagtac gctgcgcggt tcatcacccc cgtgcatccc 900 ggatacaccg ctacgttcct ggaaacgcca tcgcagtttt tcccattcac cgggctgaat 960 agcctgacct atgacgtctc cgtggcccag ggcgtaacga atctgcacac cgcgatcatg 1020 gcgcaactcg cggcgggaaa cgaagtcgtc gtcttcggca cctcccaaag cgccacgata 1080 gccaccttcg aaatgcgcta tctgcaatcc ctgccagcac acctgcgtcc gggtctcgac 1140 gaattgtcct ttacgttgac cggcaatccc aaccggcccg acggtggcat tcttacgcgt 1200 tttggcttct ccataccgca gttgggtttc acattgtccg gcgcgacgcc cgccgacgcc 1260 taccccaccg tcgattacgc gttccagtac gacggcgtca acgacttccc caaatacccg 1320 ctgaatgtct tcgcgaccgc caacgcgatc gcgggcatcc ttttcctgca ctccgggttg 1380 attgcgttgc cgcccgatct tgcctcgggc gtggttcaac cggtgtcctc accggacgtc 1440 ctgaccacct acatcctgct gcccagccaa gatctgccgc tgctggtccc gctgcgtgct 1500 atccccctgc tgggaaaccc gcttgccgac ctcatccagc cggacttgcg ggtgctcgtc 1560 gagttgggtt atgaccgcac cgcccaccag gacgtgccca gcccgttcgg actgtttccg 1620 gacgtcgatt gggccgaggt ggccgcggac ctgcagcaag gcgccgtgca aggcgtcaac 1680 gacgccctgt ccggactggg gctgccgccg ccgtggcagc cggcgctacc ccgacttttc 1740 taa 1743 <210> 71 <211> 285 <212> DNA <213> Mycobacterium tuberculosis <400> 71 atgaccatca actatcaatt cggggacgtc gacgctcacg gcgccatgat ccgcgctcag 60 gccgggtcgc tggaggccga gcatcaggcc atcatttctg atgtgttgac cgcgagtgac 120 ttttggggcg gcgccggttc ggcggcctgc caggggttca ttacccagct gggccgtaac 180 ttccaggtga tctacgagca ggccaacgcc cacgggcaga aggtgcaggc tgccggcaac 240 aacatggcac aaaccgacag cgccgtcggc tccagctggg cctaa 285 <210> 72 <211> 297 <212> DNA <213> Mycobacterium tuberculosis <400> 72 atgacctcgc gttttatgac ggatccgcac gcgatgcggg acatggcggg ccgttttgag 60 gtgcacgccc agacggtgga ggacgaggct cgccggatgt gggcgtccgc gcaaaacatt 120 tccggcgcgg gctggagtgg catggccgag gcgacctcgc tagacaccat gacccagatg 180 aatcaggcgt ttcgcaacat cgtgaacatg ctgcacgggg tgcgtgacgg gctggttcgc 240 gacgccaaca actacgaaca gcaagagcag gcctcccagc agatcctcag cagctga 297 <210> 73 <211> 228 <212> DNA <213> Mycobacterium tuberculosis <400> 73 gtgatagcgg gcgtcgacca ggcgcttgca gcaacaggcc aggctagcca gcgggcggca 60 ggcgcatctg gtggggtcac cgtcggtgtc ggcgtgggca cggaacagag gaacctttcg 120 gtggttgcac cgagtcagtt cacatttagt tcacgcagcc cagattttgt ggatgaaacc 180 gcaggtcaat cgtggtgcgc gatactggga ttgaaccagt ttcactag 228 <210> 74 <211> 555 <212> DNA <213> Mycobacterium tuberculosis <400> 74 gtggacttgc ccggaaatga ctttgacagc aacgatttcg acgccgtgga tctctggggt 60 gccgacggcg cggagggctg gactgcggat ccgattattg gcgtcgggtc ggcggcgacc 120 ccggacaccg gacccgacct ggacaatgcc cacggtcagg cggagacgga caccgaacaa 180 gagatcgcgc tttttaccgt gacgaatccc ccacgcacgg tgtcggtatc gacgctgatg 240 gacggccgga ttgaccatgt cgagctgtcg gccagggtgg cctggatgag tgagtcgcag 300 ctcgcttctg agatcctggt gattgccgac ctggcgcggc agaaggcgca gtcggcccag 360 tacgccttca tccttgacag gatgagtcaa caggtcgatg cagatgaaca ccgcgtcgca 420 ctgctacgta agaccgtggg cgaaacctgg gggttaccat cgccggaaga agccgcggca 480 gcagaagctg aggtgttcgc gacgcgctac agcgacgatt gtccagcacc agacgacgag 540 agcgatccat ggtga 555 <210> 75 <211> 312 <212> DNA <213> Mycobacterium tuberculosis <400> 75 atgaccggat ttctcggtgt cgtgccttcg ttcctgaagg tgctggcggg catgcacaac 60 gagatcgtgg gtgatatcaa aagggcgacc gatacggtcg ccgggattag cggacgagtt 120 cagcttaccc atggttcgtt cacgtcgaaa ttcaatgaca cgctgcaaga gtttgagacc 180 acccgtagca gcacgggcac gggtttgcag ggagtcacca gcggactggc caataatctg 240 ctcgcagccg ccggcgccta cctcaaggcc gacgatggcc tagccggtgt tatcgacaag 300 attttcggtt ga 312 <210> 76 <211> 399 <212> DNA <213> Mycobacterium tuberculosis <400> 76 atgagcacga cgttcgctgc ccgcctgaac cgcctgttcg acacggttta tccgcccgga 60 cgcgggccac atacctccgc ggaggtgatc gcggcgctca aggcagaggg catcacgatg 120 tcggctccct acctatcaca gctacgctca ggaaaccgta cgaacccatc gggggcgacc 180 atggccgccc tggccaactt cttccgcatc aaggcggcct acttcaccga cgacgagtac 240 tacgaaaagc tcgacaagga attgcagtgg ctgtgcacga tgcgcgacga cggcgtgcgc 300 cggatcgcgc agcgggccca cgggttgccc tccgcggcgc agcagaaggt gttggaccgg 360 atcgacgagc tgcggcgtgc cgaagggatc gacgcttag 399 <210> 77 <211> 300 <212> DNA <213> Mycobacterium tuberculosis <400> 77 atggaaaaaa tgtcacatga tccgatcgct gccgacattg gcacgcaagt gagcgacaac 60 gctctgcacg gcgtgacggc cggctcgacg gcgctgacgt cggtgaccgg gctggttccc 120 gcgggggccg atgaggtctc cgcccaagcg gcgacggcgt tcacatcgga gggcatccaa 180 ttgctggctt ccaatgcatc ggcccaagac cagctccacc gtgcgggcga agcggtccag 240 gacgtcgccc gcacctattc gcaaatcgac gacggcgccg ccggcgtctt cgccgaatag 300 <210> 78 <211> 1383 <212> DNA <213> Mycobacterium tuberculosis <400> 78 atgacgcagt cgcagaccgt gacggtggat cagcaagaga ttttgaacag ggccaacgag 60 gtggaggccc cgatggcgga cccaccgact gatgtcccca tcacaccgtg cgaactcacg 120 gcggctaaaa acgccgccca acagctggta ttgtccgccg aacacatgcg ggaatacctg 180 gcggccggtg ccaaagagcg gcagcgtctg gcgacctcgc tgcgcaacgc ggccaaggcg 240 tatggcgagg ttgatgagga ggctgcgacc gcgctggaca acgacggcga aggaactgtg 300 caggcagaat cggccggggc cgtcggaggg gacagttcgg ccgaactaac cgatacgccg 360 agggtggcca cggccggtga acccaacttc atggatctca aagaagcgg aaagagctc 420 gaaacgggcg accaaggcgc atcgctcgcg cactttgcgg atgggtggaa cactttcaac 480 540 gctaccgctt gcgaggcttc gctcgatcaa caacggcaat ggatactcca catggccaaa 600 ttgagcgctg cgatggccaa gcaggctcaa tatgtcgcgc agctgcacgt gtgggctagg 660 cgggaacatc cgacttatga agaacagtc gggctcgaac ggctttacgc ggaaaaccct 720 tcggcccgcg accaaattct cccggtgtac gcggagtatc agcagaggtc ggagaaggtg 780 ctgaccgaat acaacaacaa ggcagccctg gaaccggtaa acccgccgaa gcctcccccc 840 gccatcaaga tcgacccgcc cccgcctcg caagagcagg gattgatccc tggcttctg 900 atgccgccgt ctgacggctc cggtgtgact cccggtaccg ggatgccagc cgcaccgatg 960 gttccgccta ccggatcgcc gggtggtggc ctcccggctg acacggcggc gcagctgacg 1020 tcggctgggc gggaagccgc agcgctgtcg ggcgacgtgg cggtcaaagc ggcatcgctc 1080 ggtggcggtg gaggcggcgg ggtgccgtcg gcgccgttgg gatccgcgat cgggggcgcc 1140 gaatcggtgc ggcccgctgg cgctggtgac attgccggct taggccaggg aagggccggc 1200 ggcggcgccg cgctgggcgg cggtggcatg ggaatgccga tggtgccgc gcatcaggga 1260 caagggggcg ccaagtccaa gggttctcag caggaagacg aggcgccta caccgaggat 1320 cgggcatgga ccgaggccgt cattggtaac cgtcggcgcc aggacagtaa ggagtcgaag 1380 is 1383 <210> 79 <211> 6303 <212> DNA <213> Mycobacterium tuberculosis <400> 79 atgaccgagc agcaatggaa cttcgcgggc attgaagcgg cagcgagcgc aatccagggt 60 aatgtgacca gcatccatag cctgcttgac gagggtaagc aaagcctgac caaacttgcg 120 gcggcctggg gtggcagcgg ttcggaagcg taccagggtg tgcagcaaaa atgggatgcg 180 accgcaaccg agctgaataa cgcgttacaa aacctcgcgc gcaccatctc tgaggccggt 240 caagcgatgg cgagcaccga aggcaatgta accggtatgt tcgcgcaagc cgaaaccgcc 300 gtcaataccc tgttcgagaa actggagccg atggcatcta ttcttgatcc gggtgcgagc 360 cagagcacta ccaatccaat ctttggcatg ccgtccccgg gcagctcgac gccggtgggc 420 cagctgccgc cggcggctac ccaaacctta ggtcagctgg gtgagatgag cggcccgatg 480 ggcggctccg gtaaccctgc agatgaagag gcagcacaaa tgggtctgct gggtaccagc 540 ccgctgagca atcacccact ggccggcggc agcggcccaa gcgctggtgc gggtctgctg 600 cgcgcggagt ccctgccggg tgcgggtggc tccctgacgc gtaccccact catgagccaa 660[[ID=2,6]] ctgattgaga aaccggttgc gaccgaacaa cagtggaatt ttgccggtat tgaagctgcg 720 gccagcgcca tccagggcaa tgttacgagc atccacagcc tgctggatga aggcaaacag 780 tcgctgacca agctggcggc ggcgtggggt ggctccggta gcgaagccta tcagggtgtt 840 cagcagaaat gggacgcgac cgcgactgag ctgaacaatg cgctgcagaa tctggcccgc 900 actatttccg aggccggtca agcgatggca agcaccgagg gcaacgtgac cggtatgttc 960 gctgctgccg attacgacaa actgtttcgc ccacatgaag gcatggaggc accggatgat 1020 atggcggcgc agccgttttt cgacccgtcc gcgagctttc cgccggcccc agcaagcgcg 1080 aatctgccta aaccgaacgg tcagaccccg ccgcctacga gcgacgattt gagcgaacgt 1140 tttgtgagcg cacctccacc accgccgccg ccaccaccgc ctccgcctcc tacgccgatg 1200 ccaattgccg ccggtgaacc gcctagcccg gaaccggctg caagcaaacc gccaactcct 1260 ccgatgccga ttgccggtcc agaaccggca ccgccgaaac caccgacgcc tccgatgcct 1320 atcgcgggtc ctgaaccggc gccaccgaaa ccgccgaccc caccgatgcc gattgctggt 1380 ccggcgccaa ccccaacgga gtctcaattg gcaccacctc gtccaccgac cccacaaacg 1440 cctaccggtg cgccgcaaca gccagaatca ccggccccgc atgttcctag ccacggtccg 1500 caccaaccgc gtcgtaccgc gcctgcaccg ccgtgggcga aaatgccgat tggtgaaccg 1560 cctccggctc cttctcgtcc gtcggcttca ccggcggaac cgcctactcg tccagcgccg 1620 caacactccc gtcgtgcgcg tcgtggtcac cgctaccgta cggacacgga acgtaacgtc 1680 ggtaaagtcg ctaccggccc gagcatccag gctcgtctgc gcgcagaaga agcatcaggt 1740 gcgcaactgg cccctggtac cgagccttct ccggctccgc tgggtcaacc gagatcttac 1800 ctggcaccgc ctacccgtcc ggcgcctacc gaaccaccgc ctagccctag cccgcagcgt 1860 aactccggtc gccgtgcaga gcgtcgtgtc cacccggatc tggcggcgca acacgctgcg 1920 gctcagccag attctattac tgccgccact accggtggca gacgccgcaa acgtgctgca 1980 ccggacttgg acgccaccca gaaaagcctg cgtccagctg caaagggccc gaaagtcaaa 2040 aaggtcaagc cgcaaaagcc gaaagcaact aaaccgccga aagtcgttag ccagcgcggt 2100 tggcgtcact gggtgcacgc gctgacgcgc attaacctgg gtttgagccc ggacgaaaaa 2160 tatgaactgg acttgcacgc acgtgtccgc cgtaacccgc gtggttcata ccagattgcc 2220 gttgtcggtc tgcagggtgg cgccggcaaa accacgctga ctgcggcact gggttccacg 2280 ctggcgcaag tgcgtgcgga tcgtattctg gcactggatg ctgacccggg cgcgggtaat 2340 ctggcagatc gtgtgggtcg tcagtctggt gcgaccatcg cagatgttct ggctgagaaa 2400 gaactgagcc attacaacga cattcgtgcg cacacgtctg ttaatgccgt taatctggaa 2460 gtgctgccgg caccggagta cagcagcgca cagcgcgccc tgagcgacgc ggactggcac 2520 ttcatcgcag acccggcatc tcgcttctat aacctggttt tggcggattc cggtgcgggc 2580 tttttcgatc cactgacccg cggtgtgctc agcaccgtta gcggtgtggt tgtggttgcc 2640 tccgtgagca tcgacggtgc gcaacaagca tcggtggcgc tggactggct gcgtaataac 2700 ggttatcaag acttggcgag ccgcgcgagc gttgtgatta accatatcat gccgggcgag 2760 ccgaatgttg cggttaaaga tttggtgcgc cattttgagc aacaggtcca accgggccgc 2820 gtcgttgtga tgccatggga tcgtcatatt gcagcgggca ccgaaattag cctggactta 2880 ctggacccga tttataagcg taaggtactg gaactggccg cagcgctgag cgacgatttc 2940 gagcgtgcag gccgtcgtac cgagcaacag tggaatttcg ctggcattga agccgcagcc 3000 tctgcaattc agggtaacgt gactagcatt cattcgttgc tggacgaagg caagcaaagc 3060 ctgaccaaac tggcagctgc atggggtggc tcgggctctg aggcgtacca aggcgtccaa 3120 cagaagtggg acgcgactgc gaccgagtta aacaatgctt tacagaacct ggctcgtact 3180 atcagcgagg ctggtcaggc aatggcgagc accgaaggta atgttaccgg catgttcgcg 3240 agccgcgctt ttattatcga cccgacgatc agcgccatcg atggtctgta cgatctgttg 3300 ggtattggca ttccgaacca aggcggcatc ctgtatagct ccctggaata cttcgagaaa 3360 gccttggaag aactggcggc cgcgttcccg ggcgatggct ggctgggtag cgcggctgac 3420 aaatatgcgg gcaaaaaccg taaccatgtc aactttttcc aagagttggc ggatttggat 3480 cgtcaattga ttagcctgat ccacgatcag gcgaatgcag tgcaaaccac tcgcgatatc 3540 ctcgaaggtg ccaaaaaggg tctggagttt gtacgtccag tcgcagtcga cctgacctac 3600 attccagttg tcggtcatgc gttgtcggca aagactctga ttaatgcgac gcaactgtta 3660 aaactgttgg cgaaattggc agagctggtc gcggcagcga tcgccgacat catcagcgat 3720 gtggcagata tcattaaggg tacgctgggc gaagtgtggg aatttattac taatgcgctc 3780 aacggtctga aggaactgtg ggataaattg acgggttggg ttaccggctt gtttagccgt 3840 ggttggtcca atttggagag ctttttcgcc ggtgttccgg gcctgaccgg cgcaacgagc 3900 ggtttgtccc aggtcacggg cttgtttggt gctgcgggtc tgagcgcatc cagcggtctg 3960 gcgcacgcag attccttggc gtctagcgca agcctgccgg cgctggcagg catcggcggc 4020 ggctctggct tcggtggctt gccaagcctg gcccaagtcc atgctgcttc cacgcgccag 4080 gctctgcgcc cacgtgcaga tggtccggtt ggtgccgcag ccgagcaagt tggtggccag 4140 agccaactgg tcagcgctca gggctctcag ggcatgggcg gtccggttgg tatgggtggt 4200 atgcacccga gctccggcgc atccaagggt actaccacga aaaagtatag cgagggtgca 4260 gcggcgggta ctgaagatgc cgaacgtgca ccggttgagg ctgatgcggg tggcggccaa 4320 aaagttttgg tccgtaatgt cgtgactgag cagcagtgga attttgccgg catcgaagcg 4380 gcggcgagcg cgatccaagg caacgtcact tcgattcact ctctgctgga tgagggcaaa 4440 cagtccctga cgaaactggc agccgcgtgg ggtggtagcg gttccgaggc ataccaaggt 4500 gttcagcaga agtgggacgc gacggccacg gaactgaaca acgcgctgca gaacctggcg 4560 cgtaccatct cggaagcagg ccaggcaatg gctagcactg aaggcaacgt gacgggtatg 4620 ttcgcagcac ctaagaccta tagcgaagag ttgaagggta ctgatactgg tcaagcgagc 4680 cagattcaga tgagcgaccc ggcgtacaat atcaacatct ccctgccaag ctactatcca 4740 gaccaaaaaa gcctggagaa ctacatcgcg cagactcgcg ataagttcct gtccgcagcg 4800 acgagcagca ccccacgcga agcaccgtat gagctgaaca ttactagcgc gacgtaccaa 4860 agcgcgattc cgccgcgtgg tacccaagcc gttgtcctga aagtttatca gaacgccggt 4920 ggcactcatc cgacgacgac ctataaggcc tttgactggg accaggcata tcgtaagcca 4980 atcacgtacg acaccctgtg gcaggccgac acggatccac tgccagttgt gtttccgatc 5040 gtgcaaggtg aactgtcaaa acaaactggt caacaggtga gcattgcccc aaacgccggt 5100 ctggatccgg tgaattatca aaattttgct gttacgaacg acggcgttat ctttttcttc 5160 aacccgggtg aactgctgcc ggaagcggcg ggtccgaccc aagtgctggt tccgcgtagc 5220 gcgattgaca gcatgctggc gggcgatctg gttggcccgg gtagcgcaga atacgccgcg 5280 gccaacccga ccggtccggc atccgtccaa ggtatgagcc aggatccggt ggctgttgca 5340 gcctctaaca atccggaact gaccaccttg acggcagcgc tgagcggtca gctgaatcct 5400 caagtgaatc tggttgatac gctgaatagc ggccaatata ccgtctttgc accgaccaac 5460 gctgcgttca gcaaactgcc ggccagcacc attgacgaat tgaaaaccaa ttccagcctg 5520 ttgacgagca ttctgacgta tcacgttgtg gcgggccaga cgtccccagc gaacgtcgtc 5580 ggtacccgtc agacgctgca gggtgcgtct gtaaccgtta cgggccaagg caactctttg 5640 aaagtcggta atgccgacgt cgtcagcggt ggcgttagca cggctaatgc aaccgtctac 5700 atgattgaca gcgttttgat gccgcctgcc gtcagccagg atacgtcccc gaaaccggct 5760 acgagcccgg cagccccggt taccactgca gcaatggcgg accctgcggc tgatttgatc 5820 ggtcgcggct ctgcacagta tgctgcccaa aacccgacgg gtccgggctc tgttgccggc 5880 atggcgcagg acccggtcgc aaccgcggct agcaataatc caatgctgtc tacgctgact 5940 tccgctctgt ccggtaaact gaatccggac gtcaacctgg tggataccct gaacggtggc 6000 gagtataccg tctttgcccc gaccaacgct gcgttcgaca agctgccggc agccaccatc 6060 gaccaactga aaaccgacgc caagctgctg agcagcatcc tgacctatca cgtgattgcg 6120 ggtcaagcta gcccgagccg catcgacggc acccaccaga ccttgcaggg cgctgacctg 6180 acggttattg gtgctcgcga tgatctgatg gtgaataacg ccggtctggt ttccggcggc 6240 gttcatacgg ctaatgcaac ggtgtacatg attgatacgg tccttatgcc tccggcgcag 6300 tga 6303 <210> 80 <211> 5313 <212> DNA <213> Mycobacterium tuberculosis <400> 80 atgcaggcgg aaaccgcggt gaacaccctg tttgaaaaac tggaaccgat ggcgagcatt 60 ctggatccgg gcgcgagcca gagcaccacc aacccgattt ttggcatgcc gagcccgggc 120 agcagcaccc cggtgggcca gctgccgccg gcggcgaccc agaccctggg ccagctgggc 180 gaaatgagcg gcccgatggg cggcagcggc aacccggcgg atgaagaagc ggcgcagatg 240 ggcctgctgg gcaccagccc gctgagcaac catccgctgg cgggcggcag cggcccgagc 300 gcgggcgcgg gcctgctgcg cgcggaaagc ctgccgggcg cgggcggcag cctgacccgc 360 acccccgctga tgagccagct gattgaaaaa ccggtggcga ccgaacagca gtggaacttt 420 gcgggcattg aagcggcggc gagcgcgatt cagggcaacg tgaccagcat tcatagcctg 480 ctggatgaag gcaaacagag cctgaccaaa ctggcggcgg cgtggggcgg cagcggcagc 540 gaagcgtatc agggcgtgca gcagaaatgg gatgcgaccg cgaccgaact gaacaacgcg 600 ctgcagaacc tggcgcgcac cattagcgaa gcgggccagg cgatggcgag caccgaaggc 660 aacgtgaccg gcatgtttgc gtgggtgcat gcgctgaccc gcattaacct gggcctgagc 720 ccggatgaaa aatatgaact ggatctgcat gcgcgcgtgc gccgcaaccc gcgcggcagc 780 tatcagattg cggtggtggg cctgcagggc ggcgcgggca aaaccaccct gaccgcggcg 840 ctgggcagca ccctggcgca ggtgcgcgcg gatcgcattc tggcgctgga tgcggatccg 900 ggcgcgggca acctggcgga tcgcgtgggc cgccagagcg gcgcgaccat tgcggatgtg 960 ctggcggaaa aagaactgag ccattataac gatattcgcg cgcataccag cgtgaacgcg 1020 gtgaacctgg aagtgctgcc ggcgccggaa tatagcagcg cgcagcgcgc gctgagcgat 1080 gcggattggc attttattgc ggatccggcg agccgctttt ataacctggt gctggcggat 1140 agcggcgcgg gcttttttga tccgctgacc cgcggcgtgc tgagcaccgt gagcggcgtg 1200 gtggtggtgg cgagcgtgag cattgatggc gcgcagcagg cgagcgtggc gctggattgg 1260 ctgcgcaaca acggctatca ggatctggcg agccgcgcga gcgtggtgat taaccatatt 1...

Claims

1. A fusion protein, wherein the amino acid sequence of the fusion protein is selected from: SEQ ID NO:9, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:41 or SEQ ID NO:

91.

2. A vaccine or immunogenic composition comprising the fusion protein according to claim 1.

3. The vaccine or immunogenic composition according to claim 2, wherein the vaccine or immunogenic composition further comprises one or more adjuvants.

4. The vaccine or immunogenic composition according to claim 3, wherein the adjuvant is selected from cationic liposomes, Quil A, QS21, poly I:C, aluminum hydroxide, Freund's incomplete adjuvant, IFN-γ, IL-2, IL-12, monophosphoryl lipid A (MPL), trehalose dimycoate (TDM), trehalose dibehenate (TDB), muramyl dipeptide (MDP), monobranched acylglycerol (MMG), CpG and "IC31", or a combination thereof.

5. The vaccine or immunogenic composition of claim 4, wherein the cationic liposome is dimethyldioctadecyl ammonium bromide (DDA).

6. The vaccine or immunogenic composition of claim 2, wherein the vaccine or immunogenic composition further comprises BCG.

7. Use of the fusion protein according to claim 1 in the preparation of a medicament for vaccinating or immunizing a subject against infection and / or disease caused by Mycobacterium tuberculosis.

8. Use of the vaccine or immunogenic composition according to claim 2 in the preparation of a medicament for vaccinating or immunizing a subject against infection and / or disease caused by Mycobacterium tuberculosis.

9. The use according to claim 7 or 8, wherein the subject is a mammal.

10. The use according to claim 9, wherein the mammal is a human.

11. A kit comprising: i) the fusion protein according to claim 1 or the vaccine or immunogenic composition according to any one of claims 2 to 6, and ii)BCG.

12. The kit according to claim 11, wherein i) and ii) are for simultaneous, separate or sequential administration.

13. A nucleic acid comprising a sequence encoding the fusion protein according to claim 1.

14. A recombinant expression vector comprising the nucleic acid sequence according to claim 13, said nucleic acid sequence being operably linked to one or more control sequences suitable for directing the production of a fusion protein in a suitable host. A recombinant host cell comprising the expression vector according to claim 14 .

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