MYCOBACTERIAL ANTIGEN COMPOSITION

Abstract

The present invention provides an antigenic composition for use as a mycobacterial vaccine, said composition comprising (i) a first mycobacterial antigenic polypeptide, wherein said first mycobacterial antigenic polypeptide comprises a polypeptide sequence having at least 70% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 6, 12, 2, 18, 8, 10, 6, 4, or a fragment thereof having at least 50 consecutive amino acids thereof; or (ii) a first mycobacterial polynucleotide, wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide, or 10 wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence selected from SEQ ID NO: 5, 11, 1, 17, 7, 9, 15, 3.

Description

[0001] The present invention relates to mycobacterial polynucleotides and polypeptides, to fragments or variants thereof, to antibodies that bind thereto, to vectors and microbial carriers, to therapeutic compositions such as vaccines against mycobacterial infections, and to compositions and methods for detecting mycobacterial infection.

[0002] Microorganisms such as species of Salmonella, Yersinia, Shigella, Campylobacter, Chlamydia and Mycobacterium are capable of causing intracellular infections. These infections may be exclusively intracellular, or may contain both intracellular and extracellular components. Generally, these microorganisms do not circulate freely in the body (e.g. in the bloodstream) and are often not amenable to drug treatment regimes.

[0003] The difficulties associated with treating intracellular infection have been exacerbated by the development of multiple drug-resistant microorganisms. Vaccine therapies have not proved effective against intracellular microorganisms because of the difficulties in the ability of the host defenses to access the pathogen.

[0004] Mycobacterium tuberculosis (MTB) and closely related species make up a small group of mycobacteria known as the Mycobacterium tuberculosis complex (MTC). This group comprises five distinct species: M. tuberculosis, M. microti, M. bovis, M. caneti, and M. africanum. M. avium subsp. paratuberculosis causes Johne's disease in ruminants, M. bovis causes tuberculosis in cattle, M. avium and M. intracellulare cause tuberculosis in immunocompromised patients (eg. AIDS patients, and bone marrow transplant patients), and M. leprae causes leprosy in humans. Another important mycobacterial species is M. vaccae.

[0005] As the aetiological agent of tuberculosis infection, Mycobacterium tuberculosis (M. tuberculosis) is the leading cause of death by bacterial infectious disease worldwide--latent infection affecting as much as one third of the world's population. The World Health Organisation (WHO) estimates that over eight million new cases of TB, and over one million deaths, occur globally each year. The largest number of new TB cases in 2005 occurred in South-East Asia (34% of incident cases globally), and the estimated incidence rate in sub-Saharan Africa is nearly 350 cases per 100,000 population. However, TB infection is not limited to the developing world: the UK has seen a resurgence of tuberculosis since the late 1980s and there are currently over 8000 new cases each year--a rate of 14.0 per 100,000 population. About 40% of these new cases occur in the London region, where the rate of infection is 44.8 per 100,000 population.

[0006] Optimal patient management requires early initiation of drug therapy and isolation of infectious individuals as soon as possible. Left untreated, each person with active TB disease will infect on average between 10 and 15 people every year. TB infection can normally be treated by a 6 month course of antibiotics; however, patient compliance to long-term drug treatment is varied, with patients often stopping therapy when their symptoms cease. Failure to complete the treatment can promote the development of multiple drug-resistant mycobacteria.

[0007] The term `latency` is synonymous with `persistence`, and describes a reversible state of low metabolic activity in which mycobacterial cells can survive for extended periods with limited or no cell division. During latency (ie. latent infection), the clinical symptoms associated with a mycobacterial infection do not become manifest. However, re-activation of latent mycobacteria may be induced by environmental stimuli. During active infection, mycobacteria demonstrate high metabolic activity and replicate rapidly, resulting in the development of active infection with clinical symptoms.

[0008] In vitro studies have demonstrated that mycobacteria such as M. tuberculosis are able to adapt to and survive under nutrient- and oxygen-depleted conditions, and can grow over a range of nutrient availabilities and oxygen tensions. Adaptation to carbon starvation and/or to a low dissolved oxygen tension in vitro triggers transition to a non-replicating persistent state that may be analogous to latency in vivo. Intracellular survival and multiplication of mycobacteria is suspected to be a main supportive factor for mycobacterial disease progression. The presence of a large reservoir of asymptomatic individuals latently-infected with mycobacteria is a major problem for the control of mycobacterial infections, especially M. tuberculosis infections. In addition, conventional methods for the detection of a latent mycobacterial infection by skin testing may be compromised by BCG vaccination and by exposure to environmental mycobacteria.

[0009] The effectiveness of vaccine prevention against M. tuberculosis has varied widely. The current M. tuberculosis vaccine, BCG, is an attenuated strain of M. bovis. It is effective against severe complications of TB in children, but it varies greatly in its effectiveness in adults in different countries, particularly across ethnic groups. BCG vaccination has been used to prevent tuberculous meningitis and helps prevent the spread of M. tuberculosis to extra-pulmonary sites, but does not prevent infection. The limited efficacy of BCG and the global prevalence of TB has led to an international effort to generate new, more effective vaccines. This, in turn, requires the identification of new vaccine candidates. In view of the increasing threat and global prevalence of mycobacterial infection, new strategies are required for more effective prevention, treatment, and diagnosis of mycobacterial infection.

[0010] The invention provides an antigenic composition comprising a mycobacterial antigen, wherein said antigen comprises:

[0011] a polypeptide sequence having at least 70% amino acid sequence identity to the amino acid sequence of a polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10. 12, 14, 16 and 18, or a fragment thereof having at least 50 consecutive amino acids thereof; or

[0012] (ii) a polynucleotide sequence encoding a polypeptide sequence according to (i); or a polynucleotide sequence having at least 70% nucleotide sequence identity to the nucleic acid sequence of a polynucleotide selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15 and 17, or a fragment thereof having at least 150 consecutive nucleotides thereof.

[0013] As used herein, the term "mycobacterial" or "mycobacterium" embraces the species M. phlei, M. smegmatis, M. africanum, M. caneti, M. fortuitum, M. marinum, M. ulcerans, M. tuberculosis, M. bovis, M. microti, M. avium, M. paratuberculosis, M. leprae, M. lepraemurium, M. intracellulare, M. scrofulaceum, M. xenopi, M. genavense, M. kansasii, M. simiae, M. szulgai, M. haemophilum, M. asiaticum, M. malmoense, M. vaccae, M. caneti, and M. shimoidei. Of particular interest are the members of the MTC, such as M. tuberculosis.

[0014] The term antigen means any substance that can be recognized by the immune system and/or that stimulates an immune response. For example, an antigen may stimulate a cell mediated immune response and/or may stimulate the generation of antibodies.

[0015] In one embodiment, a mycobacterial antigen of the invention provides a cell mediated response to infection involving immune cells such as T cells (CD4+ and/or CD8+ T cells) and/or the ability to respond with Th1-type cytokines such as IFN-.gamma.. In one embodiment, a mycobacterial antigen induces IFN-.gamma.-secreting cells (eg. predominantly CD4+ T cells). In this regard, recent studies suggest that immune cell responses (particularly T cell immune responses in, for example, the lung) may be critical for protection against pulmonary mycobacterial disease.

[0016] In one embodiment, a mycobacterial antigen of the invention provides protection (such as long term protection) against challenge by mycobacteria such as M. tuberculosis. By way of example, a mycobacterial antigen of the invention may induce `memory T cells`, which can continue to stimulate protective immunity in the long term (eg. for decades). Memory immune responses have been attributed to the reactivation of long-lived, antigen-specific T lymphocytes that arise directly from differentiated effector T-cells and persist in a quiescent state. Memory T cells are heterogeneous; at least two subsets have been identified, having different migratory capacity and effector function. Memory T cells of the first subset are known as `effector memory T cells` (TEM) because they resemble the effector T cells generated in the primary response, in that they lack the lymph node-homing receptors for migration into inflamed tissues. Upon re-encounter with antigen, the TEM rapidly produce IFN-.gamma. or IL-4, or release pre-stored perforin. Memory T cells of the second subset (known as `central memory cells` (TCM)) express L-selectin and CCR7 and lack immediate effector function. The TCM have a low activation threshold and proliferate and differentiate to effectors when re-stimulated in secondary lymphoid organs.

[0017] In one embodiment, a mycobacterial antigen provides a neutralizing antibody response to mycobacterial (eg. M. tuberculosis) infection. In one embodiment, each mycobacterial antigen in the antigenic composition of the present invention independently induces an effective immune response (eg. a cell mediated immune response or antibody response). Thus, in accordance with this embodiment, following administration of the antigenic composition to a subject, an immune response is induced in the subject to each mycobacterial antigen in the antigenic composition.

[0018] In one embodiment, a mycobacterial antigen comprises (eg. consists of) a polypeptide sequence. Alternatively, or in addition, a mycobacterial antigen comprises a polynucleotide (e.g. DNA or RNA) sequence.

[0019] The specific sub-set of mycobacterial polypeptides represented by SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16 and 18 are `latency-regulated polypeptides`. The specific subset of mycobacterial polynucleotides represented by SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15 and 17 are `latency-regulated polynucleotides`.

[0020] In one embodiment, a `latency-regulated polypeptide` is encoded by a `latency-regulated polynucleotide`. By way of example, the latency-regulated polypeptide SEQ ID NO: 2 is encoded by latency-regulated polynucleotide SEQ ID NO: 1; SEQ ID NO: 4 is encoded by SEQ ID NO: 3; SEQ ID NO: 6 is encoded by SEQ ID NO: 5; SEQ ID NO: 8 is encoded by SEQ ID NO: 7; SEQ ID NO: 10 is encoded by SEQ ID NO: 9; SEQ ID NO: 12 is encoded by SEQ ID NO: 11; SEQ ID NO: 14 is encoded by SEQ ID NO: 13; SEQ ID NO: 16 is encoded by SEQ ID NO: 15; and SEQ ID NO: 18 is encoded by SEQ ID NO: 17.

[0021] The expression or activity of a latency-regulated polypeptide or polynucleotide is modulated in response to mycobacterial latency--eg. in response to growth of mycobacteria (eg. MTB) under conditions that induce or maintain mycobacterial latency.

[0022] In one embodiment, "modulation" of expression or activity of the latency-regulated polypeptide or polynucleotide in response to conditions of mycobacterial latency means that the expression or activity is induced or upregulated in response to latency. Thus, the latency-regulated polypeptide or polynucleotide may be a `latency-induced` or `latency-upregulated` polypeptide or polynucleotide.

[0023] For example, the expression or activity of a latency-upregulated polypeptide or polynucleotide may be up-regulated by at least 1.5-fold, 2-fold, 5-fold, 10-fold, 20-fold or 50-fold under latency conditions as compared to non-latency conditions. Alternatively, vaccine efficacy of the polypeptides/polynucleotides of the present invention may be measured in terms of murine splenocyte interferon-gamma (IFN-.gamma.) release--for example, said polypeptide/polynucleotide demonstrate at least 380, at least 400, at least 420, at least 450, at least 500, at least 550, at least 600 or higher spot forming units/10.sup.6 (murine splenocyte IFN-.gamma. release)--see FIG. 1. Alternatively, vaccine efficacy of the polypeptides/polynucleotides of the present invention may be measured in terms of protective efficacy (%) relative to BCG alone (i.e. when administered as a boost to a BCG prime vaccine)--for example, said polypeptide/polynucleotide demonstrates at least 120, at least 150, at least 180, at least 200, at least 220, at least 250, at least 300% increase in protective efficacy, such as measured by bacterial load (eg. in murine spleen and/or lung)--see FIGS. 2 & 3.

[0024] The expression or activity of latency-induced and latency-upregulated polypeptides and polynucleotides may be induced or upregulated in vivo during latency in the mycobacterium's natural environment. As such, the present inventors believe that latency-induced or latency-upregulated mycobacterial polypeptides and polynucleotides represent good vaccine candidates for preventing the establishment, spread and reactivation of disease and/or make good diagnostic tools for latent infection.

[0025] In one embodiment, the mycobacterial antigen comprises (or consists of) a polypeptide sequence having at least 70% amino acid sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% amino acid sequence identity) to the amino acid sequence of a polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16 and 18, or a fragment thereof having at least 50 consecutive amino acids thereof. SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16 and 18 are defined in Table 1, below:

TABLE-US-00001 TABLE 1 SEQ Polypeptide ID No. name 2 Rv0982 4 Rv1462 6 Rv1937 8 Rv2500c 10 Rv2504c 12 Rv3270 14 Rv3537 16 Rv3608c 18 Rv3879c

[0026] Thus, in the context of the present application, an "Rv0982 polypeptide antigen" comprises or consists of SEQ ID NO: 2 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv1462 polypeptide antigen" comprises or consists of SEQ ID NO: 4 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv1937 polypeptide antigen" comprises or consists of SEQ ID NO: 6 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv2500c polypeptide antigen" comprises or consists of SEQ ID NO: 8 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv2504c polypeptide antigen" comprises or consists of SEQ ID NO: 10 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv3270 polypeptide antigen" comprises or consists of SEQ ID NO: 12 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv3537 polypeptide antigen" comprises or consists of SEQ ID NO: 14 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv3608c polypeptide antigen" comprises or consists of SEQ ID NO: 16 (or a sequence `variant` or `fragment` thereof as defined herein); and an "Rv3879c polypeptide antigen" comprises or consists of SEQ ID NO: 18 (or a sequence `variant` or `fragment` thereof as defined herein).

[0027] In one embodiment, the amino acid sequence identity exists over a region of the polypeptide sequences that is at least 50 consecutive amino acid residues in length (eg. at least 75, 100, 150, 200, 250, 300 consecutive amino acid residues in length). Conventional methods for determining amino acid sequence identity are discussed in more detail later in the specification.

[0028] In the context of the first mycobacterial antigen, a fragment of a polypeptide comprises (or consists of) at least 50 consecutive amino acid residues of said polypeptide (eg. at least 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 consecutive amino acid residues of said polypeptide). Said fragment includes at least one epitope of the polypeptide. A fragment of a polypeptide has a sequence length that is at least 25%, 50%, 60%, 70%, 80%, or 90% of that of the sequence of the full-length polypeptide.

[0029] In one embodiment, in the context of the first mycobacterial antigen, a fragment of a polypeptide comprises (or consists of) a truncated form of said polypeptide. For example, a fragment of a polypeptide may have an N-terminal truncation (as compared with the polypeptide), or a fragment of a polypeptide may have a C-terminal truncation (as compared with the polypeptide).

[0030] In one embodiment, in the context of the first mycobacterial antigen, a fragment of a polypeptide comprises (or consists of) a mature form of the polypeptide. For example, the polypeptide may comprise a signal sequence (ie. a secretion/targeting sequence) (eg. at the N-terminus), and a fragment of the polypeptide may lack this signal sequence. In one embodiment, the fragment is formed by cleavage of a signal sequence from the polypeptide. In one embodiment, a fragment of polypeptide SEQ ID NO: 2 is an N-terminally truncated form of SEQ ID NO: 2. In one embodiment, a fragment of polypeptide SEQ ID NO: 2 has an N-terminal truncation of 50, 100, 150, 200 or 250 amino acid residues as compared with the amino acid sequence of SEQ ID NO: 2. In one embodiment, a fragment of SEQ ID NO: 2 comprises at least the C-terminal 50, 100, 150, 200 or 250 amino acid sequence of SEQ ID NO: 2. Similarly, in one embodiment, a fragment of polypeptide SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 is an N-terminally truncated form of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively). In one embodiment, a fragment of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 is a mature polypeptide sequence, which differs from the sequence of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively) by removal of an N-terminal signal sequence. In one embodiment, a fragment of polypeptide SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 has an N-terminal truncation of 5, 10, 15, 20, 25, 30, 35, 40, 50, 100, 150, 200 or 250 amino acid residues as compared with the amino acid sequence of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively). In one embodiment, a fragment of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 comprises at least the C-terminal 50, 100, 150, 200 or 250 amino acid sequence of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively).

[0031] In one embodiment, the mycobacterial antigen of the invention comprises a polypeptide or fragment thereof that has a common antigenic cross-reactivity and/or substantially the same in vivo biological activity as a polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18. As used herein, `common antigenic cross-reactivity` means that the mycobacterial polypeptide or fragment of the invention and the latency-regulated polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18 share a common ability to induce a "recall response" of an immune cell such as a T-lymphocyte (eg. CD4+, CD8+, effector T cell or memory T cell such as a TEM or TCM), which has been previously exposed to an antigenic component of a mycobacterial infection. New immunological assays for measuring and quantifying immune cell responses (eg. T cell responses) have been established over the last 10+ years. For example, the interferon-gamma (IFN-.gamma.) ELISPOT assay is useful as an immunological readout because the secretion of IFN-.gamma. from antigen-specific immune cells such as T cells is a good correlate of protection against M. tuberculosis. Furthermore, the ELISPOT assay is a very reproducible and sensitive method of quantifying the number of IFN-.gamma. secreting antigen-specific immune cells such as T cells. Alternatively, or in addition, `common antigenic cross-reactivity` means that an antibody capable of binding to the mycobacterial polypeptide or fragment of the invention would also be capable of binding to the corresponding latency-regulated polypeptide (SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18).

[0032] In one embodiment, the mycobacterial antigen comprises, or consists of, a polynucleotide sequence that encodes the corresponding mycobacterial polypeptide as defined above.

[0033] Thus, in one embodiment, the first mycobacterial antigen comprises (or consists of) a polynucleotide sequence that encodes a polypeptide that comprises (or consists of) an amino acid sequence having at least 70% amino acid sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% amino acid sequence identity) to the amino acid sequence of a latency-regulated polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, or a fragment thereof having at least 50 consecutive amino acids thereof (eg. as defined above). In one embodiment, the mycobacterial antigen comprises (or consists of) a polynucleotide sequence having at least 70% nucleotide sequence identity (at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% nucleotide sequence identity) to the nucleic acid sequence of a latency-regulated polynucleotide selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, or a fragment thereof having at least 150 consecutive nucleotides thereof. In use, said polynucleotide is in a form (e.g. vector) that provides corresponding mycobacterial antigenic peptide/protein. Thus, in one embodiment, the mycobacterial antigen is a `mycobacterial polynucleotide` (or fragment), as defined above. SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15 and 17 are defined in Table 2, below:

TABLE-US-00002 TABLE 2 SEQ Polynucleotide ID No. name 1 Rv0982 3 Rv1462 5 Rv1937 7 Rv2500c 9 Rv2504c 11 Rv3270 13 Rv3537 15 Rv3608c 17 Rv3879c

[0034] Thus, in the context of the present application, an "Rv0982 polynucleotide antigen" comprises or consists of SEQ ID NO: 1 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv1462 polynucleotide antigen" comprises or consists of SEQ ID NO: 3 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv1937 polynucleotide antigen" comprises or consists of SEQ ID NO: 5 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv2500c polynucleotide antigen" comprises or consists of SEQ ID NO: 7 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv2504c polynucleotide antigen" comprises or consists of SEQ ID NO: 9 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv3270 polynucleotide antigen" comprises or consists of SEQ ID NO: 11 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv3537 polynucleotide antigen" comprises or consists of SEQ ID NO: 13 (or a sequence `variant` or `fragment` thereof as defined herein); an "Rv3608c polynucleotide antigen" comprises or consists of SEQ ID NO: 15 (or a sequence `variant` or `fragment` thereof as defined herein); and an "Rv3879c polynucleotide antigen" comprises or consists of SEQ ID NO: 17 (or a sequence `variant` or `fragment` thereof as defined herein).

[0035] In one embodiment, the nucleotide sequence identity exists over a region of the polynucleotide sequences that is at least 150 consecutive nucleotide residues in length (eg. at least 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700 or 750 consecutive nucleotide residues in length). Conventional methods for determining nucleotide sequence identity are discussed in more detail later in the specification.

[0036] In the context of the mycobacterial antigen, a fragment of said polynucleotide comprises (or consists of) at least 150 consecutive nucleotide residues of said polynucleotide (eg. at least 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700 or 750 consecutive nucleotide residues of said polynucleotide). In one embodiment, the length of the sequence of the polynucleotide fragment is at least 25%, 50%, 60%, 70%, 80%, or 90% that of the polynucleotide.

[0037] In one embodiment, in the context of the mycobacterial antigen, a fragment of a polynucleotide comprises (or consists of) a truncated form of said polynucleotide. In one embodiment, a fragment of a polynucleotide is truncated at the 5' end and/or the 3' end, as compared with the full-length polynucleotide sequence. In one embodiment, a fragment of a polynucleotide encodes a truncated form of said polypeptide. For example, a fragment of a polynucleotide may encode a polypeptide that is N-terminally truncated and/or C-terminally truncated polypeptide (as compared with the polypeptide encoded by the full-length polynucleotide). In one embodiment, in the context of the first mycobacterial antigen, a fragment of a polynucleotide encodes a polypeptide that comprises (or consists of) a mature polypeptide. For example, the full-length polypeptide comprises a signal sequence (ie. a secretion/targeting sequence) (eg. at the N-terminus), and the polynucleotide fragment encodes a mature polypeptide that lacks this signal sequence.

[0038] In one embodiment, a fragment of polynucleotide SEQ ID NO: 1 is a 5' truncated form of SEQ ID NO: 1. In one embodiment, a fragment of polynucleotide SEQ ID NO: 1 has a 5' truncation of 100, 200 or 300 nucleotide residues as compared with the nucleotide sequence of SEQ ID NO: 1. In one embodiment, a fragment of polynucleotide SEQ ID NO: 1 encodes an N-terminally truncated form of SEQ ID NO: 2. In one embodiment, a fragment of polynucleotide SEQ ID NO: 1 encodes a polypeptide having an N-terminal truncation of 50, 100, 150, 200 or 250 amino acid residues as compared with the amino acid sequence of SEQ ID NO: 2. In one embodiment, a fragment of SEQ ID NO: 1 comprises the 3' terminal 100, 200 or 300 nucleotide residues as compared with the nucleotide sequence of SEQ ID NO: 1. In one embodiment, a fragment of polynucleotide SEQ ID NO: 1 encodes a polypeptide comprising at least the C-terminal 50, 100, 150, 200, 250 or 300 amino acid sequence of SEQ ID NO: 2.

[0039] In one embodiment, a fragment of polynucleotide SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 is a 5' truncated form of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 has a 5' truncation of 25, 50, 75, 100 or 125 nucleotide residues as compared with the nucleotide sequence of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 encodes an N-terminally truncated form of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 encodes a mature polypeptide sequence, which differs from the sequence of SEQ ID NO: SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively) by removal of an N-terminal signal sequence. In one embodiment, a fragment of polynucleotide SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 encodes a polypeptide that has an N-terminal truncation of 5, 10, 15, 20, 25, 30, 35, 40, 50, 100, 150, 200 or 250 amino acid residues as compared with the amino acid sequence of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively). In one embodiment, a fragment of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 comprises the 3' terminal 150, 300, 450, 600 or 750 nucleotide residues as compared with the nucleotide sequence of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 (respectively). In one embodiment, a fragment of SEQ ID NO: 3, 5, 7, 9, 11, 13, 15, 17 encodes a polypeptide that comprises at least the C-terminal 50, 100, 150, 200 or 250 amino acid sequence of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18 (respectively).

[0040] In one embodiment, said mycobacterial polynucleotide, or fragment thereof, encodes a polypeptide that has a common antigenic cross-reactivity and/or substantially the same in vivo biological activity as a latency-regulated polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, or 18. For example, said first mycobacterial antigen may comprise (or consist of) a polynucleotide sequence that encodes a polypeptide sequence that is capable of evoking a protective immune cell response (eg. T-cell response) against mycobacterial infection. By way of example, the polypeptide encoded by the first mycobacterial polynucleotide or fragment shares, with the latency-regulated polypeptide, a common ability to induce a "recall response" of an immune cell such as a T-lymphocyte (eg. CD4+, CD8+, effector T cell or memory T cell such as TEM or TCM) that has previously been exposed to an antigenic component of a mycobacterial infection. In this regard, the secretion of IFN-.gamma. from antigen-specific immune cells such as T cells is a good correlate of protection against M. tuberculosis. Accordingly, the interferon-gamma (IFN-.gamma.) ELISPOT assay is a useful immunological readout, and enables reproducible and sensitive quantification of IFN-.gamma. secreting antigen-specific immune cells such as T cells. Alternatively, or in addition, an antibody capable of binding to a polypeptide encoded by the mycobacterial polynucleotide or fragment of the invention would also be capable of binding to the latency-regulated polypeptide (SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16 or 18).

[0041] The antigenic composition of the invention may comprise at least a second mycobacterial antigen, in addition to the first mycobacterial antigen. The second mycobacterial antigen may be any one of the aforementioned mycobacterial antigens, and is preferably different from said first mycobacterial antigen.

[0042] In one embodiment, the second mycobacterial antigen is capable of evoking a protective immune response (eg. a T-cell response) against mycobacterial infection. In one embodiment, the second mycobacterial antigen comprises (eg. consists of) a polypeptide sequence. In one embodiment, the second mycobacterial antigen comprises (eg. consists of) a polynucleotide sequence such as a DNA or RNA sequence, or a mycobacterial glycolipid, such as a mycobacterial sulphoglycolipid. In one embodiment, the second mycobacterial antigen comprises (eg. consists of) a mycobacterial carbohydrate antigen such as a mycobacterial saccharide or polysaccharide. Optionally, the saccharide may be linked (eg. chemically conjugated) to a carrier (eg. a polypeptide) to enhance immunogenicity.

[0043] In one embodiment, the `difference` between the second mycobacterial antigen and the first mycobacterial antigen is defined by the specificity of the immune response to the first and second mycobacterial antigens. For example, in one embodiment, each of the first and second antigens induces an immune response that is substantially specific to that antigen. The `difference` between the second mycobacterial antigen and the first mycobacterial antigen may be defined in terms of a substantial lack (eg. an absence) of common antigenic cross-reactivity between the first and second mycobacterial antigens. The `difference` between the second mycobacterial antigen and the first mycobacterial antigen may alternatively (or in addition) be defined as a substantial lack (eg. an absence) of common in vivo biological activity between the first and second mycobacterial antigens.

[0044] For example, in one embodiment, the first and second mycobacterial antigens may exhibit (substantially) no common antigenic cross-reactivity. In one embodiment, the first and second mycobacterial antigens may exhibit (substantially) no common in vivo biological activity. For example, the first and second mycobacterial antigens induce different immune responses and/or have different in vivo biological activities.

[0045] In one embodiment, the first and second mycobacterial antigens comprise polypeptides (as defined herein), and the second mycobacterial antigen has substantially no common antigenic cross-reactivity with the first mycobacterial antigen and/or has a substantially different in vivo biological activity from the first mycobacterial antigen. In one embodiment, the first and second mycobacterial antigens comprise polynucleotides (as defined herein), and the second mycobacterial antigen encodes a polypeptide that has substantially no common antigenic cross-reactivity with the polypeptide encoded by the first mycobacterial antigen. In one embodiment, the first and second mycobacterial antigens comprise polynucleotides (as defined herein), and the second mycobacterial antigen has a substantially different in vivo biological activity from the first mycobacterial antigen and/or encodes a polypeptide that has a substantially different in vivo biological activity from the polypeptide encoded by the first mycobacterial antigen.

[0046] In one embodiment, the first mycobacterial antigen comprises a polypeptide and the second mycobacterial antigen comprises a polynucleotide (as defined herein), and the second mycobacterial antigen or polypeptide encoded thereby has substantially no common antigenic cross-reactivity with the first mycobacterial antigen and/or has a substantially different in vivo biological activity from the first mycobacterial antigen.

[0047] In one embodiment, the first mycobacterial antigen comprises a polynucleotide and the second mycobacterial antigen comprises a polypeptide (as defined herein), and the second mycobacterial antigen has substantially no common antigenic cross-reactivity with the first mycobacterial antigen or polypeptide encoded thereby, and/or has a substantially different in vivo biological activity from the first mycobacterial antigen or polypeptide encoded thereby. By way of example, in one embodiment, the first and second mycobacterial antigens do not share a common ability to induce a "recall response" of an immune cell such as a T-lymphocyte (eg. CD4+, CD8+, effector or memory T cell such as TEM or TCM) that has previously been exposed to an antigenic component of a mycobacterial infection. In other words, in one embodiment, the first and second mycobacterial antigens are `different` because they induce recall responses in different immune cells (eg. T cells).

[0048] In one embodiment, the second mycobacterial polypeptide comprises (or consists of) an antigenic mycobacterial polypeptide--ie. a mycobacterial polypeptide that is capable of evoking a protective T-cell response against mycobacterial infection.

[0049] In one embodiment, the second mycobacterial antigen comprises a polypeptide that is selected from the same group of polypeptides as discussed above in connection with the first mycobacterial antigen (preferably the second mycobacterial polypeptide is different from the first mycobacterial polypeptide).

[0050] Thus, in one embodiment, the second mycobacterial antigen comprises (or consists of) a polypeptide sequence having at least 70% amino acid sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% amino acid sequence identity) to the amino acid sequence of a latency-regulated polypeptide selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, or a fragment thereof having at least 50 consecutive amino acids thereof (such as at least 75, 100, 125, 150, 175, 200, 225 or 250 consecutive amino acid residues thereof).

[0051] In one embodiment, the second mycobacterial antigen comprises a polypeptide that is not selected from the same group of polypeptides as discussed above in connection with the first mycobacterial antigen. For example, in one embodiment, the second mycobacterial antigen comprises (or consists of) a polypeptide sequence having at least 70% amino acid sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% amino acid sequence identity) to an amino acid sequence selected from SEQ ID NOs: 19-41, or a fragment thereof having at least 50 consecutive amino acids thereof. SEQ ID NOs: 19-41 are illustrated in Table 3, below:

TABLE-US-00003 TABLE 3 SEQ Polypeptide ID NO: name: 19 Ag85A/Rv3804c 20 Ag85B/Rv1886c 21 ESAT-6/Rv3875 22 TB10.4/Rv0288 23 Rv0125 24 PPE18/Rv1196 25 P27/Rv1411c 26 Hsp65/Rv0440 27 HBHA/Rv0475 28 Rv2659c 29 Rv2660c 30 HspX/Rv2031c 31 RPFA/Rv0867c 32 RPFB/Rv1009 33 RPFC/Rv1884c 34 RPFD/Rv2389c 35 RPFE/Rv2450c 36 Rv1733 37 Rv2029c 38 Rv2032 39 Rv2626c 40 Rv2627c 41 Rv2628

[0052] The polypeptide "Ag85A" represented by SEQ ID NO: 19 of the present application (Accession Nos. CAA17868 and BX842584) is a member of a family of proteins ("the Ag85 complex"), which also comprises Ag85B (SEQ ID NO: 20 of the present application) and Ag85C. This family of proteins is secreted by M. tuberculosis, M. bovis BCG, and many other species of mycobacteria. Ag85A is highly conserved amongst all mycobacterial species and is immunodominant in animal and human studies.

[0053] The polypeptides represented by SEQ ID NOs: 30 and 36-41 are comprised within the DosR regulon (also known as the DevR regulon), which includes the polypeptides represented by Rv2623-2631 and Rv3126-3134. The expression of these polypeptides is regulated via DosR (DevR). The polypeptides represented by SEQ ID NOs: 31-35 are members of the RPF family of polypeptides (RPFA, RPFB, RPFC, RPFD and RPFE, respectively).

[0054] In one embodiment, the amino acid sequence identity exists over a region of the polypeptide sequences that is at least 50 consecutive amino acid residues in length (eg. at least 75, 100, 125, 150, 175, 200, 225 or 250 consecutive amino acid residues in length). In one embodiment, in the context of the second mycobacterial antigen, a fragment of said polypeptide comprises at least 50 consecutive amino acid residues of said polypeptide sequence. In one embodiment, the fragment comprises (or consists of) at least 75, 100, 125, 150, 175, 200, 225 or 250 consecutive amino acid residues of said polypeptide sequence. In one embodiment, a fragment of a polypeptide is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the length of the mycobacterial polypeptide. A fragment of a polypeptide includes at least one epitope of the polypeptide.

[0055] In one embodiment, in the context of the second mycobacterial antigen, a fragment of a polypeptide comprises (or consists of) a truncated form of said polypeptide. For example, a fragment of a polypeptide may have an N-terminal truncation (as compared with the polypeptide), or a fragment of a polypeptide may have a C-terminal truncation (as compared with the polypeptide). In one embodiment, in the context of the second mycobacterial antigen, a fragment of a polypeptide comprises (or consists of) a mature form of the polypeptide. For example, the polypeptide may comprise a signal sequence (ie. a secretion/targeting sequence) (eg. at the N-terminus), and a fragment of the polypeptide may lack this signal sequence. In one embodiment, the fragment is formed by cleavage of a signal sequence from the polypeptide.

[0056] In one embodiment, a fragment of polypeptide SEQ ID NO: 19-41 is an N-terminally truncated form of SEQ ID NO: 19-41. In one embodiment, a fragment of SEQ ID NO: 19-41 is a mature polypeptide sequence, which differs from the sequence of SEQ ID NO: 19-41 by removal of an N-terminal signal sequence. In one embodiment, a fragment of polypeptide SEQ ID NO: 19-41 has an N-terminal truncation of 10, 20, 30 or 40 amino acid residues as compared with the amino acid sequence of SEQ ID NO: 19-41. In one embodiment, a fragment of SEQ ID NO: 19-41 comprises at least the C-terminal 50, 100, 150, 200 or 250 amino acid sequence of SEQ ID NO: 19-41. In one embodiment, the second mycobacterial polypeptide or fragment thereof has a common antigenic cross-reactivity and/or substantially the same in vivo biological activity as the polypeptide selected from SEQ ID NOs: 19-41. In one embodiment, `common antigenic cross-reactivity` means that the second mycobacterial polypeptide, or fragment, shares a common ability, with the polypeptide selected from SEQ ID NOs: 19-41, to induce a "recall response" of an immune cell such as a T-lymphocyte which has been previously exposed to an antigenic component of a mycobacterial infection. For example, the interferon-gamma (IFN-.gamma.) ELISPOT assay is useful as an immunological readout because the secretion of IFN-.gamma. from antigen-specific immune cells such as T cells is a good correlate of protection against M. tuberculosis. Furthermore, the ELISPOT assay is a very reproducible and sensitive method of quantifying the number of IFN-.gamma. secreting antigen-specific immune cells such as T cells. Alternatively, or in addition, `common antigenic cross-reactivity` means that an antibody capable of binding to the second mycobacterial polypeptide, or fragment, would also be capable of binding to the polypeptide selected from SEQ ID NOs: 19-41.

[0057] In one embodiment, the second mycobacterial polynucleotide comprises (or consists of) an antigenic mycobacterial polynucleotide that is capable (following translation) of evoking a protective immune cell response (eg. T-cell response) against mycobacterial infection. In one embodiment, the second mycobacterial polynucleotide encodes an antigenic mycobacterial polypeptide that is capable of evoking a protective immune cell response (eg. T-cell response) against mycobacterial infection. Thus, in one embodiment, the second mycobacterial antigen is a `second mycobacterial polynucleotide` (or fragment), as defined above. In one embodiment, the second mycobacterial antigen comprises a polynucleotide selected from the polynucleotides discussed above in connection with the first mycobacterial antigen (though preferably different from the first mycobacterial polynucleotide).

[0058] Thus, in one embodiment, the second mycobacterial antigen comprises (or consists of) a polynucleotide sequence that encodes a polypeptide selected from the polypeptides discussed above in connection with the first mycobacterial antigen (though the polypeptide encoded by the second mycobacterial polynucleotide is different from the polypeptide encoded by the first mycobacterial polynucleotide). Thus, said second mycobacterial polynucleotide comprises a polynucleotide sequence (e.g. as a vector or plasmid) encoding a second mycobacterial polypeptide of the invention, as defined above.

[0059] In one embodiment, said encoded second mycobacterial polypeptide comprises a polypeptide sequence having at least 70% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 2. 4. 6, 8, 10, 12, 14, 16, 18, or a fragment thereof having at least 50 consecutive amino acids thereof. In one embodiment, the second mycobacterial antigen comprises (or consists of) a polynucleotide sequence having at least 70% nucleotide sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% nucleotide sequence identity) to the nucleic acid sequence of a latency-regulated polynucleotide selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, or a fragment thereof having at least 150 consecutive nucleotides thereof (such as at least 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700 or 750 consecutive nucleotides thereof). In one embodiment, the second mycobacterial polypeptide comprises (or consists of) a polynucleotide sequence having at least 70% nucleotide sequence identity to the nucleic acid sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, or a fragment thereof having at least 150 consecutive nucleotides thereof. In one embodiment, the limitations discussed above with respect to the first mycobacterial polypeptide apply equally to this embodiment of the second mycobacterial polypeptide.

[0060] In one embodiment, the second mycobacterial antigen comprises a polynucleotide that is not selected from the same group of polynucleotides as discussed above in connection with the first mycobacterial antigen. In one embodiment, the second mycobacterial antigen comprises a polynucleotide that encodes a polypeptide that is not selected from the same group of polypeptides as discussed above in connection with the first mycobacterial antigen. In one embodiment, the second mycobacterial antigen comprises a polynucleotide sequence that encodes a second mycobacterial polypeptide as defined above.

[0061] Thus, in one embodiment, the second mycobacterial antigen comprises (or consists of) a polynucleotide sequence, wherein said polynucleotide sequence encodes a polypeptide that comprises (or consists of) an amino acid sequence having at least 70% amino acid sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% amino acid sequence identity) to an amino acid sequence selected from SEQ ID NOs: 19-41, or a fragment thereof having at least 50 consecutive amino acid residues thereof. In one embodiment, the second mycobacterial antigen comprises (or consists of) a polynucleotide sequence having at least 70% nucleotide sequence identity (such as at least 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% nucleotide sequence identity) to a nucleic acid sequence selected from SEQ ID NOs: 42-64, or a fragment thereof having at least 150 consecutive nucleotides thereof. SEQ ID NOs: 42-64 are illustrated in Table 4, below:

TABLE-US-00004 TABLE 4 SEQ Polynucleotide ID NO: name: 42 Ag85A/Rv3804c 43 Ag85B/Rv1886c 44 ESAT-6/Rv3875 45 TB10.4/Rv0288 46 Rv0125 47 PPE18/Rv1196 48 P27/Rv1411c 49 Hsp65/Rv0440 50 HBHA/Rv0475 51 Rv2659c 52 Rv2660c 53 HspX/Rv2031c 54 RPFA/Rv0867c 55 RPFB/Rv1009 56 RPFC/Rv1884c 57 RPFD/Rv2389c 58 RPFE/Rv2450c 59 Rv1733 60 Rv2029c 61 Rv2032 62 Rv2626c 63 Rv2627c 64 Rv2628

[0062] The polynucleotide "Ag85A" represented by SEQ ID NO: 42 of the present application (Accession Nos. CAA17868 and BX842584) is a member of a family of genes ("the Ag85 complex"), which also comprises Ag85B (SEQ ID NO: 43 of the present application) and Ag85C. This family of genes encodes proteins that are secreted by M. tuberculosis, M. bovis BCG, and many other species of mycobacteria. Ag85A is highly conserved amongst all mycobacterial species and is immunodominant in animal and human studies.

[0063] The polynucleotides represented by SEQ ID NOs: 53 and 59-64 are comprised within the DosR regulon (also known as the DevR regulon), which includes the polynucleotides represented by Rv2623-2631 and Rv3126-3134. The expression of these polynucleotides is regulated via DosR (DevR). The polynucleotides represented by SEQ ID NOs: 54-49 are members of the RPF family of polynucleotides (RPFA, RPFB, RPFC, RPFD and RPFE, respectively).

[0064] In one embodiment, the nucleotide sequence identity exists over a region of the polynucleotide sequences that is at least 150 consecutive nucleotide residues in length (eg. at least 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700 or 750 consecutive nucleotide residues in length).

[0065] In one embodiment, in the context of the second mycobacterial antigen, a fragment of a polynucleotide comprises at least 150 consecutive nucleotide residues of said polynucleotide sequence. In one embodiment, the fragment comprises (or consists of) at least 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700 or 750 consecutive nucleotide residues of said polynucleotide sequence. In one embodiment, a fragment of said polynucleotide is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the length of the polynucleotide.

[0066] In one embodiment, in the context of the second mycobacterial antigen, a fragment of a polynucleotide comprises (or consists of) a truncated form of said polynucleotide. For example, a fragment of a polynucleotide may have a 5' truncation and/or 3' truncation as compared with the polynucleotide. In one embodiment, in the context of the second mycobacterial antigen, a fragment of a polynucleotide encodes a polypeptide that is truncated as compared with the polypeptide sequence encoded by the full-length polynucleotide. For example, the polynucleotide fragment may encode a polypeptide that is N-terminally truncated and/or C-terminally truncated, as compared with the polypeptide encoded by the full-length polynucleotide. In one embodiment, in the context of the second mycobacterial antigen, a fragment of a polynucleotide encodes a mature polypeptide. For example, the polypeptide may comprise a signal sequence (ie. a secretion/targeting sequence) (eg. at the N-terminus), and the polynucleotide fragment may encode a polypeptide fragment that lacks this signal sequence.

[0067] In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 is a 5' truncated form of SEQ ID NO: 42-64 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 has an N-terminal truncation of 25, 50, 75, 100 or 125 nucleotide residues as compared with the nucleotide sequence of SEQ ID NO: 42-64 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 comprises at least the C-terminal 150, 300, 450, 600 or 750 nucleotide residues of SEQ ID NO: 42-64 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 encodes an N-terminally truncated form of SEQ ID NO: 42-64 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 encodes a mature polypeptide sequence, which differs from the sequence of SEQ ID NO: 42-64 (respectively) by removal of an N-terminal signal sequence. In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 encodes a polypeptide fragment of SEQ ID NO: 42-64 (respectively) that has an N-terminal truncation of 10, 20, 30, 40, 50, 100, 150 amino acid residues as compared with the amino acid sequence of SEQ ID NO: 42-64 (respectively). In one embodiment, a fragment of polynucleotide SEQ ID NO: 42-64 encodes a polypeptide fragment of SEQ ID NO: 42-64 (respectively) that comprises at least the C-terminal 50, 100, 150, 200 or 250 amino acid residues of SEQ ID NO: 42-64 (respectively). In one embodiment, a polypeptide encoded by the second mycobacterial polynucleotide or fragment has a common antigenic cross-reactivity and/or substantially the same in vivo biological activity as the polypeptide selected from SEQ ID NOs: 19-41. By way of example, the polypeptide encoded by the second mycobacterial polynucleotide, or fragment, shares a common ability, with the polypeptide selected from SEQ ID NOs: 19-41, to induce a "recall response" of an immune cell such as a T-lymphocyte which has been previously exposed to an antigenic component of a mycobacterial infection. For example, the interferon-gamma (IFN-.gamma.) ELISPOT assay is useful as an immunological readout because the secretion of IFN-.gamma. from antigen-specific immune cells such as T cells is a good correlate of protection against M. tuberculosis. Furthermore, the ELISPOT assay is a very reproducible and sensitive method of quantifying the number of IFN-.gamma. secreting antigen-specific immune cells such as T cells. Alternatively, or in addition, an antibody capable of binding to a polypeptide encoded by the second mycobacterial polynucleotide, or fragment, would also be capable of binding to the polypeptide selected from SEQ ID NOs: 19-41.

[0068] In one embodiment, the antigenic composition comprises both an Rv0111 antigen (antigenic polypeptide or polynucleotide) and an Rv0198 antigen (antigenic polypeptide or polynucleotide).

[0069] In one embodiment, where there are multiple additional mycobacterial antigens (eg. 2 or more additional mycobacterial antigens, as well as the first and second mycobacterial antigens), each of said additional mycobacterial antigens is different from each other. In one embodiment, the `difference` between the additional mycobacterial antigen(s) and the first and second mycobacterial antigens is defined by the specificity of the immune response to the mycobacterial antigens. For example, in one embodiment, each of the first, second and additional antigens induce an immune response that is substantially specific to that antigen. The `difference` between the first, second and additional mycobacterial antigens may be defined in terms of a substantial lack (eg. an absence) of common antigenic cross-reactivity between the mycobacterial antigens. The `difference` between the first, second and additional mycobacterial antigens may be alternatively (or in addition) be defined as a substantial lack (eg. an absence) of common in vivo biological activity between the mycobacterial antigens. For example, in one embodiment, the first, second and additional mycobacterial antigens (eg. first, second and additional mycobacterial antigenic polypeptides, or first, second and additional mycobacterial antigenic polynucleotides or polypeptide encoded thereby) may exhibit (substantially) no common antigenic cross-reactivity.

[0070] In one embodiment, the first, second and additional mycobacterial antigens (eg. first, second and additional mycobacterial antigenic polypeptides, or first, second and additional mycobacterial antigenic polynucleotides or polypeptide encoded thereby) exhibit (substantially) no common in vivo biological activity. For example, the first, second and additional mycobacterial antigens (eg. first, second and additional mycobacterial antigenic polypeptides, or first, second and additional mycobacterial antigenic polynucleotides or polypeptide encoded thereby) may each induce different immune responses and/or each have different in vivo biological activities. By way of example, in one embodiment, the first, second and additional mycobacterial antigens (eg. first, second and additional mycobacterial antigenic polypeptides, or first, second and additional mycobacterial antigenic polynucleotides or polypeptide encoded thereby) do not share a common ability to induce a "recall response" of an immune cell such as a T-lymphocyte (eg. CD4+, CD8+, effector or memory T cell--TEM or TCM) that has previously been exposed to an antigenic component of a mycobacterial infection. In other words, in one embodiment, the first, second and additional mycobacterial antigens are `different` because they induce recall responses in different immune cells (eg. different T cells).

[0071] In one embodiment, the one or more additional mycobacterial antigen(s) is expressed or up-regulated under different culture conditions and/or mycobacterial infection states as compared with the first and/or second mycobacterial antigens. In one embodiment, the activity of the one or more additional mycobacterial antigen(s) is up-regulated under different culture conditions and/or mycobacterial infection states as compared with the first and/or second mycobacterial antigens. Thus, in one embodiment, the expression or activity of first mycobacterial antigen is up-regulated during conditions of mycobacterial latency, whereas the expression or activity of the second and/or additional mycobacterial antigen is up-regulated during active mycobacterial infection or upon re-activation from a latent state (and/or down-regulated during conditions of mycobacterial latency). In one embodiment, where there are multiple additional mycobacterial antigens (eg. 2 or more additional mycobacterial antigens, as well as the first and second mycobacterial antigens), each additional mycobacterial antigen is expressed/up-regulated at different stages of mycobacterial infection, or the activity of each additional mycobacterial antigen is up-regulated at different stages of mycobacterial infection.

[0072] In one embodiment, the one or more additional mycobacterial antigens are from a mycobacterium other than M. tuberculosis. For example, the one or more additional mycobacterial antigens may be from another member of the MTC, such as M. microti, M. bovis, M. canetti or M. africanum, or a non-MTC mycobacterium such as M. avium-intracellulare, M. kansasii, M. marinum or M. ulcerans.

[0073] In one embodiment, the antigenic composition comprises at least 1, 2, 3, 4 or 5 further mycobacterial antigens, in addition to the first and second mycobacterial antigens discussed above. In one embodiment, each of said at least 1, 2, 3, 4 or 5 additional mycobacterial antigens is different from each other and from the first and second mycobacterial antigens. In one embodiment, the antigenic composition comprises up to about 10 different mycobacterial antigens (eg. including the first and second mycobacterial antigens discussed above). In one embodiment, the antigenic composition comprises 1 additional mycobacterial antigen, and thus comprises a total of 3 different mycobacterial antigens (ie. the antigenic composition is trimeric). In one embodiment, the antigenic composition comprises 2 additional mycobacterial antigens, and thus comprises a total of 4 different mycobacterial antigens (ie. the antigenic composition is tetrameric). In one embodiment, the antigenic composition comprises 3 additional mycobacterial antigens, and thus comprises a total of 5 different mycobacterial antigens (ie. the antigenic composition is pentameric). In one embodiment, the antigenic composition comprises up to 8 additional mycobacterial antigens, and thus comprises up to a total of 10 different mycobacterial antigens (ie. the antigenic composition is up to decameric).

[0074] In one embodiment, the one or more additional mycobacterial antigens comprises (or consists of) a polypeptide sequence having at least 70% amino acid sequence identity to the amino acid sequence of a latency-regulated polypeptide selected from SEQ ID NOs: 1, 3, 5, 7 and 56, or a fragment thereof having at least 150 consecutive amino acids thereof, as defined above with respect to the first mycobacterial antigen (so long as the one or more additional mycobacterial antigens is different from the first mycobacterial antigen). Alternatively, or in addition, the one or more additional mycobacterial antigens may comprise (or consist of) a polypeptide sequence having at least 70% amino acid sequence identity to an amino acid sequence selected from SEQ ID NOs: 19-41, or a fragment thereof having at least 50 consecutive amino acids thereof, as defined above with respect to the second mycobacterial antigen (so long as the one or more additional mycobacterial antigens is different from the second mycobacterial antigen).

[0075] In one embodiment, the one or more additional mycobacterial antigens comprises (or consists of) a polynucleotide sequence that encodes a polypeptide sequence as described above with respect to the first mycobacterial antigenic polypeptide (preferably different from the first mycobacterial antigen). In one embodiment, the one or more additional mycobacterial antigens comprises (or consists of) a polynucleotide sequence that encodes a polypeptide sequence as described above with respect to the second mycobacterial antigenic polypeptide (preferably different from the second mycobacterial antigen).

[0076] In one embodiment, the one or more additional mycobacterial antigens comprises (or consists of) a polynucleotide sequence having at least 70% nucleotide sequence identity to the nucleic acid sequence of a latency-regulated polynucleotide selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, or a fragment thereof having at least 150 consecutive nucleotides thereof, as described above with respect to the first mycobacterial antigen (so long as the one or more additional mycobacterial antigens is different from the first mycobacterial antigen). Alternatively, or in addition, the one or more additional mycobacterial antigens may comprise (or consist of) a polynucleotide sequence having at least 70% nucleotide sequence identity to a nucleic acid sequence selected from SEQ ID NOs: 42-64, or a fragment thereof having at least 150 consecutive nucleotides thereof, as described above with respect to the second mycobacterial antigen (so long as the one or more additional mycobacterial antigens is different from the second mycobacterial antigen).

[0077] In one embodiment, at least two of the mycobacterial antigens in the antigenic composition comprise (or consist of) a polypeptide sequence, and said at least two polypeptide sequences are optionally joined together to form a fusion protein. By way of example, in one embodiment, the first mycobacterial antigen and second mycobacterial antigen each comprise (or consist of) a polypeptide sequence, as defined above, and said first and second polypeptide sequences are optionally joined together to form a fusion protein.

[0078] In one embodiment, said fusion protein further comprises at least one additional mycobacterial antigenic polypeptide sequence, optionally joined to said first and/or second polypeptide sequences, wherein each of said further mycobacterial antigens is different from each other and from the first and second mycobacterial antigens. For example, the fusion protein may comprise at least 1, 2, 3, 4 or 5 further mycobacterial antigens, in addition to said first and second mycobacterial antigens, wherein each of said further mycobacterial antigens is different from each other and from the first and second mycobacterial antigens. In one embodiment, the fusion protein may comprise up to about 10 different mycobacterial antigens (eg. including the first and second mycobacterial antigens).

[0079] In one embodiment, the antigenic composition comprises at least one additional mycobacterial antigen (eg. at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 additional mycobacterial antigens) and the first mycobacterial antigen and said at least one additional mycobacterial antigen each comprise (or consist of) a polypeptide sequence, as defined above, and said polypeptide sequences are optionally joined together to form a fusion protein. In one embodiment, the antigenic composition comprises at least one additional mycobacterial antigen (eg. at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 additional mycobacterial antigens), and the second mycobacterial antigen and said at least one additional mycobacterial antigen each comprise (or consist of) a polypeptide sequence, as defined above, and said polypeptide sequences are optionally joined together to form a fusion protein. Alternatively, in one embodiment, the antigenic composition comprises at least two additional mycobacterial antigens (eg. at least 2, 3, 4, 5, 6, 7, 8, 9 or 10 additional mycobacterial antigens), and said at least two additional mycobacterial antigens each comprise (or consist of) a polypeptide sequence, as defined above, and said polypeptide sequences are optionally joined together to form a fusion protein.

[0080] In one embodiment, a recombinant fusion protein may be generated by expression of a recombinant polynucleotide sequence that encodes said fusion protein. By way of example, polynucleotide sequences encoding mycobacterial antigenic polypeptides of the invention may be positioned in the same reading frame downstream of a promoter in an expression vector, thereby allowing transcription through the polynucleotide sequences and translation as one protein product. In one embodiment, intervening `linker` sequences are located between the polynucleotide sequence for each polypeptide antigen, arising from the inclusion of restriction sites. In general, the amino acids encoded by these linker sequences are not deleterious to the immunogenicity of the resultant fusion protein, and may even be beneficial to immunogenicity. Alternatively, a fusion protein of the invention may be produced as an epitope string, by expression of polynucleotide sequences that are linked without intervening nucleotides. The absence of intervening linker sequence avoids the presence of unnecessary nucleic acid and/or amino acid material. Alternatively, a fusion protein of the invention may be prepared by chemically conjugating the mycobacterial antigenic polypeptides of the invention. By way of example, the first and/or second and/or additional mycobacterial polypeptides of the invention may be coupled to each other using conventional chemical conjugation techniques.

[0081] In one embodiment, at least two of the mycobacterial antigens in the antigenic composition comprise (or consist of) a polynucleotide sequence, and said at least two polynucleotide sequences are optionally joined together to form a polycistronic nucleic acid sequence. By way of example, in one embodiment, the first mycobacterial antigen and second mycobacterial antigen each comprise (or consist of) a polynucleotide sequence, as defined above, and said first and second polynucleotide sequences are optionally joined together to form a polycistronic nucleic acid sequence.

[0082] In one embodiment, said polycistronic nucleic acid sequence comprises or consists of (in any order from the 5' to 3' end):

[0083] (i) a first mycobacterial polynucleotide, wherein said first mycobacterial polynucleotide comprises (or consists of) a polynucleotide sequence as hereinbefore defined; and

[0084] (ii) a second mycobacterial polynucleotide, wherein said second mycobacterial polynucleotide comprises (or consists of) a polynucleotide sequence as hereinbefore defined.

[0085] In one embodiment, said polycistronic sequence further comprises at least one additional mycobacterial antigenic polynucleotide sequence, joined to said first and second polynucleotide sequences. Alternatively, in one embodiment, the antigenic composition comprises at least one additional mycobacterial antigen, and the first mycobacterial antigen and at least one additional mycobacterial antigen each comprise (or consist of) a polynucleotide sequence, as defined above, and said polynucleotide sequences are joined together to form a polycistronic nucleic acid sequence. Alternatively, in one embodiment, the antigenic composition comprises at least one additional mycobacterial antigen, and the second mycobacterial antigen and at least one additional mycobacterial antigen each comprise (or consist of) a polynucleotide sequence, as defined above, and said polynucleotide sequences are joined together to form a polycistronic nucleic acid sequence. Alternatively, in one embodiment, the antigenic composition comprises at least two additional mycobacterial antigens, and said at least two additional mycobacterial antigens each comprise (or consist of) a polynucleotide sequence, as defined above, and said polynucleotide sequences are joined together to form a polycistronic nucleic acid sequence.

[0086] In one embodiment, the polycistronic nucleic acid sequence of the invention is positioned downstream of a promoter in frame in a vector (eg. an expression vector or viral vector as discussed below), thereby allowing transcription through the polynucleotide sequences and optional translation as one `fusion protein` product. Accordingly, in one embodiment, the polycistronic nucleic acid sequence encodes a fusion protein as discussed above. Alternatively, in one embodiment, the polycistronic nucleic acid sequence encodes separate mycobacterial antigenic polypeptide sequences, as discussed above. In one embodiment, the polycistronic nucleic acid sequence is operably linked to a nucleic acid sequence encoding a tag polypeptide, such that the encoded tag is covalently linked to the encoded antigenic polypeptide(s) upon translation. The tag may facilitate detection of antigenic polypeptide expression, or detection of clones that express the antigen, and/or may lead to increases in antigen efficacy. Suitable tag polypeptides include a PK tag, FLAG tag, MYC tag, polyhistidine tag or any detectable tag (eg. a tag that can be detected by an antibody such as a monoclonal antibody). Other examples of tags will be clear to skilled persons in the art. The nucleic acid sequence encoding the tag polypeptide may be positioned such that, following translation, the tag is located at the C-terminus of the expressed antigenic polypeptide (ie. in the order: antigenic polypeptide-tag). Alternatively, the nucleic acid sequence encoding the tag polypeptide may be positioned such that, following translation, the tag is located at the N-terminus of the expressed antigenic polypeptide (ie. in the order: tag-antigenic polypeptide). Alternatively, the nucleic acid sequence encoding the tag polypeptide may be positioned such that, following translation, the tag is located internally to the expressed antigenic polypeptide, or between the expressed antigenic polypeptides of an encoded fusion protein.

[0087] Nucleotides encoding a linker sequence may be inserted between the polycistronic nucleic acid sequence encoding the antigenic polypeptide(s) and the nucleic acid sequence encoding the tag polypeptide. In one embodiment, the linker sequence encodes the amino acid sequence Gly-Ser-Ile. In one embodiment, the encoded linker sequence is located between an expressed antigenic polypeptide and a tag polypeptide (ie. in the order: antigenic polypeptide-linker-tag, or tag-linker-antigenic polypeptide). In one embodiment, the nucleic acid sequence encoding the tag polypeptide and the nucleotides encoding the linker sequence are positioned such that, following translation, the linker sequence (eg. Gly-Ser-Ile) is located at the C-terminus of the expressed antigenic polypeptide and the tag is located at the C-terminus of the expressed linker sequence (ie. in the order antigenic polypeptide-linker-tag). Intervening `linker` sequences may alternatively (or additionally) be located between the mycobacterial polynucleotide sequences of the polycistronic sequence, arising from the inclusion of restriction sites (eg. in the form: mycobacterial polynucleotide-linker-mycobacterial polynucleotide). However, to avoid the presence of unnecessary nucleic acid and/or amino acid material, the polynucleotide sequences may be linked without intervening nucleotides.

[0088] In one embodiment, the polycistronic nucleic acid sequence is operably linked to a leader sequence. For example, the leader sequence may be fused to the N-terminus of the polycistronic sequence (ie. in the form: leader-polycistronic sequence) or to the C-terminus of the polycistronic sequence (ie. in the form: polycistronic sequence-leader). A leader sequence may affect processing of a primary DNA transcript to mRNA, and/or may affect mRNA stability or translation efficiency. In one embodiment, a leader sequence ensures that the encoded polypeptide antigen is directed to the secretory machinery of a host cell. In one embodiment, a leader sequence enhances expression and/or immunogenicity of the antigen. Enhanced expression may be determined by a conventional assay, such as using an antibody (eg. monoclonal antibody) to detect the amount of protein produced. Enhanced immunogenicity may be determined using a conventional assay such as a cultured or ex vivo ELISPOT assay. In one embodiment, the presence of a leader sequence enhances the expression and/or immunogenicity of the mycobacterial antigenic polypeptide by 2-fold, 3-fold or more when compared with antigenic polypeptide expressed without the leader sequence. An example of a suitable leader sequence is t-PA (tissue plasminogen activator). Accordingly, in one embodiment, the polycistronic nucleic acid sequence encoding said mycobacterial antigenic polypeptides is operably linked to a leader sequence and a tag sequence. For example, the leader sequence may be fused to the N-terminus of the polycistronic sequence and the tag sequence may be fused to the C-terminus of the polycistronic sequence (ie. in the form: leader-polycistronic sequence-tag. In one embodiment, a linker sequence is located between the polycistronic sequence and the nucleic acid sequence encoding the tag (ie. in the form leader-polycistronic sequence-linker-tag). In one embodiment, the leader sequence is a t-PA leader sequence and/or the tag sequence is a PK tag sequence (ie. in the form: t-PA leader-polycistronic sequence-PK tag). In one embodiment, a linker sequence is located between the polycistronic sequence and the nucleic acid sequence encoding the tag (ie. in the form t-PA leader-polycistronic sequence-linker-PK tag). In one embodiment, intervening leader sequences are located between one or more of the mycobacterial polynucleotide sequences of the polycistronic sequence (ie. in the form: mycobacterial polynucleotide-leader-mycobacterial polynucleotide). In one embodiment, the polycistronic nucleic acid sequence encoding the mycobacterial antigenic polypeptides is operably linked to an N-terminal leader sequence, internal leader sequence and a tag sequence (ie. in the form: leader-first mycobacterial polynucleotide-leader-second mycobacterial polynucleotide-tag). In one embodiment, a linker sequence is located between the polycistronic sequence and the nucleic acid sequence encoding the tag (ie. in the form: leader-first mycobacterial polynucleotide-leader--second mycobacterial polynucleotide-linker-tag). In one embodiment, the leader sequence is a t-PA leader sequence and/or the tag sequence is a PK tag sequence (ie. in the form: t-PA leader-first mycobacterial polynucleotide-t-PA leader-second mycobacterial polynucleotide-PK tag). In one embodiment, a linker sequence is located between the polycistronic sequence and the nucleic acid sequence encoding the tag (ie. in the form t-PA leader-first mycobacterial polynucleotide-t-PA leader-second mycobacterial polynucleotide-linker-PK tag). In one embodiment, the polycistronic nucleic acid sequence further comprises a polyadenylation signal, such as a bovine growth hormone (BGH) polyadenylation signal.

[0089] In one embodiment, the antigenic composition comprises one or more cells, wherein said cells comprise at least one of the mycobacterial antigens. In one embodiment, said one or more cells comprise a first mycobacterial antigen and/or a second mycobacterial antigen, as defined above. In one embodiment, said one or more cells comprises one or more of said additional mycobacterial antigens, as defined above. In one embodiment, one or more of said additional mycobacterial antigens comprises a polypeptide sequence as defined above. In one embodiment, one or more of said additional mycobacterial antigens comprises a polynucleotide sequence as filed above.

[0090] In one embodiment, said at least one mycobacterial antigen (eg. polypeptide) is at least partially exposed at the surface of the cell(s). In an alternative embodiment, the cell becomes degraded in vivo so that at least part of the mycobacterial antigen (eg. polypeptide) becomes exposed to a host's immune system. In an alternative embodiment, the cell at least partially releases (eg. secretes or exports) the mycobacterial antigen (eg. polypeptide) to the outside of the cell, so that it is exposed to a host's immune system. In one embodiment, said antigenic composition comprises an individual cell, wherein said cell comprises at least two of said mycobacterial antigens. By way of example, in one embodiment, said antigenic composition comprises an individual cell, wherein said cell comprises both said first mycobacterial antigen and said second mycobacterial antigen. In one embodiment, said individual cell further comprises one or more of said additional mycobacterial antigens.

[0091] In one embodiment, said antigenic composition comprises an individual cell, wherein said cell comprises said first mycobacterial antigen and said one or more additional mycobacterial antigens. In one embodiment, said antigenic composition comprises an individual cell, wherein said cell comprises said second mycobacterial antigen and said one more additional mycobacterial antigens. In one embodiment, said antigenic composition comprises an individual cell, wherein said cell comprises said at least two of said additional mycobacterial antigens. In an alternative embodiment, the antigenic composition comprises at least first and second cells, wherein said first cell comprises said first mycobacterial antigen (as defined above) and wherein said second cell comprises said second mycobacterial antigen (as defined above). In this embodiment, the first and second mycobacterial antigens are not present in the same cell; rather, the first and second mycobacterial antigens are in different cells. In one embodiment, said antigenic composition further comprises at least a third cell, wherein said cell comprises an additional mycobacterial antigen, as defined above.

[0092] In one embodiment, said at least one cell is an attenuated microbial carrier. An attenuated carrier is a cell (such as a bacterial cell) that is incapable of causing a significant pathological effect in an animal subject, typically a mammalian subject such as a human, bovine, porcine or equine subject. Suitable examples of attenuated microbial carriers include attenuated salmonella, attenuated M. tuberculosis, or attenuated M. bovis (eg. BCG strain).

[0093] In one embodiment, the antigenic composition comprises one or more vectors, wherein said vectors comprise at least one of the mycobacterial antigens. In one embodiment, said one or more vectors comprises a first mycobacterial antigen, as defined above. In one embodiment, said first mycobacterial antigen comprises a polypeptide sequence as defined above. In one embodiment, said one or more vectors comprises a second mycobacterial antigen, as defined above. In one embodiment, said vector comprises said first mycobacterial antigen and said second (and optionally additional) mycobacterial antigen. By way of example, said vector may comprise a first mycobacterial polynucleotide as defined herein and a second (and optionally additional) mycobacterial polynucleotide as defined herein. In one embodiment, said vector comprises:

[0094] (i) a first mycobacterial polynucleotide, wherein said first mycobacterial polynucleotide comprises (or consists of) a polynucleotide sequence as hereinbefore defined; and optionally

[0095] (ii) a second mycobacterial polynucleotide, wherein said second mycobacterial polynucleotide comprises (or consists of) a polynucleotide sequence as hereinbefore defined.

[0096] In one embodiment, said one or more vectors comprises one or more of said additional mycobacterial antigens, as defined above. In one embodiment, one or more of said additional mycobacterial antigens comprises a polypeptide sequence as defined above. In one embodiment, one or more of said additional mycobacterial antigens comprises a polynucleotide sequence as filed above.

[0097] Examples of vectors include DNA vectors (e.g. Vaccinia virus vectors, such as MVA) and RNA vectors (e.g. Sinbis or Semiliki Forest virus vectors). The term `vector` embraces expression vectors (which may be useful for preparation of mycobacterial antigens of the invention), and viral vectors (which may be useful for replication and/or delivery of mycobacterial antigens of the invention). The vectors optionally include appropriate control sequences such as a promoter and/or terminator. In one embodiment, the vector comprises one or more polynucleotide sequence(s) encoding said mycobacterial antigen(s). Said polynucleotide sequence may be operably linked to a nucleic acid sequence encoding a tag polypeptide, such that the encoded tag is covalently linked to the antigen upon translation. The tag may facilitate detection of antigen expression, or of clones that express the antigen, and/or may lead to increases in antigen efficacy. Suitable tag polypeptides include a PK tag, FLAG tag, MYC tag, polyhistidine tag or any detectable tag (eg. a tag that can be detected by an antibody such as a monoclonal antibody). The nucleic acid sequence encoding the tag polypeptide may be positioned such that, following translation, the tag is located at the C-terminus of the expressed antigen. Alternatively, the nucleic acid sequence encoding the tag polypeptide may be positioned such that, following translation, the tag is located at the N-terminus of the expressed antigen. Alternatively, the nucleic acid sequence encoding the tag polypeptide may be positioned such that, following translation, the tag is located internally to the expressed antigen. Nucleotides encoding a linker sequence may be inserted between the polynucleotide encoding the expressed antigen and the nucleic acid sequence encoding the tag polypeptide. In one embodiment, the encoded linker sequence is located between an expressed antigen polypeptide and a tag polypeptide. In one embodiment, the nucleic acid sequence encoding the tag polypeptide and the nucleotides encoding the linker sequence are positioned such that, following translation, the linker sequence is located at the C-terminus of the expressed antigen and the tag is located at the C-terminus of the expressed linker sequence.

[0098] In one embodiment, the vector comprises one or more polynucleotide sequences encoding mycobacterial antigenic polypeptide(s), wherein said polynucleotide sequence is operably linked to a leader sequence. A leader sequence may affect processing of the primary transcript to mRNA, and/or may affect mRNA stability or translation efficiency. In one embodiment, a leader sequence ensures that the encoded polypeptide antigen is directed to the secretory machinery of a host cell. In one embodiment, a leader sequence enhances expression and/or immunogenicity of the antigen. Enhanced immunogenicity may be determined using a conventional assay such as a cultured or ex vivo ELISPOT assay. Enhanced expression may be determined by a conventional assay, such as using an antibody (eg. monoclonal antibody) to detect the amount of protein produced. In one embodiment, the presence of a leader sequence enhances the expression and/or immunogenicity of the mycobacterial antigen by 2-fold, 3-fold or more when compared with antigen expressed without the leader sequence. An example of a suitable leader sequence is t-PA (tissue plasminogen activator). In one embodiment, the vector comprises a C-terminally truncated polynucleotide encoding said mycobacterial antigen fused to a t-PA leader sequence. In one embodiment, the vector comprises a C-terminally truncated polynucleotide encoding said mycobacterial antigen fused to a t-PA leader sequence and a PK tag sequence. For example, the leader sequence may be fused to the N-terminus of the polynucleotide encoding the antigen and the tag sequence may be fused to the C-terminus of the polynucleotide encoding the antigen. In one embodiment, a linker sequence (eg. Gly-Ser-Ile) is located between the polynucleotide encoding the antigen and the nucleic acid sequence encoding the tag.

[0099] In one embodiment, said antigenic composition comprises an individual vector, wherein said vector comprises both said first mycobacterial antigen and said second mycobacterial antigen. In one embodiment, said individual vector further comprises one or more of said additional mycobacterial antigens. In one embodiment, said antigenic composition comprises an individual vector, wherein said vector comprises said first mycobacterial antigen and said one or more additional mycobacterial antigens. In one embodiment, said antigenic composition comprises an individual vector, wherein said vector comprises said second mycobacterial antigen and said one more additional mycobacterial antigens. In one embodiment, said antigenic composition comprises an individual vector, wherein said cell comprises said one or more additional mycobacterial antigens. In an alternative embodiment, the antigenic composition comprises at least first and second vectors, wherein said first vector comprises said first mycobacterial antigen (as defined above) and wherein said second vector comprises said second mycobacterial antigen (as defined above). In this embodiment, the first and second mycobacterial antigens are not present in the same vector; rather, first and second mycobacterial antigens are in different vectors. In one embodiment, said antigenic composition further comprises at least a third vector, wherein said third vector comprises an (one or more) additional mycobacterial antigen(s), as defined above.

[0100] In one embodiment, the vector (or at least one of said vectors) is a viral vector. Viral vectors are usually non-replicating or replication-impaired vectors, which means that the viral vector cannot replicate to any significant extent in normal cells (eg. normal human cells), as measured by conventional means--eg. via measuring DNA synthesis and/or viral titre. Non-replicating or replication-impaired vectors may have become so naturally (ie. they have been isolated as such from nature) or artificially (eg. by breeding in vitro or by genetic manipulation). There will generally be at least one cell-type in which the replication-impaired viral vector can be grown--for example, modified vaccinia Ankara (MVA) can be grown in CEF cells. Typically, the viral vector is incapable of causing a significant infection in an animal subject, typically in a mammalian subject such as a human, bovine, porcine or equine patient. Examples of viral vectors that are useful in this context include attenuated vaccinia virus vectors such as modified vaccinia Ankara (MVA) and NYVAC, or strains derived therefrom. Other suitable viral vectors include poxvirus vectors, such as avipox vectors, for example attenuated fowlpox vectors (eg. FP9) or canarypox vectors (eg. ALVAC and strains derived therefrom). Alternative viral vectors useful in the present invention include adenoviral vectors (eg. non-human adenovirus vectors), alphavirus vectors, flavivirus vectors, herpes viral vectors, influenza virus vectors and retroviral vectors.

[0101] In one embodiment, the vector (or at least one of said vectors) is an expression vector. Expression vectors are nucleic acid molecules (linear or circular) that comprise one or more polynucleotide sequences encoding a polypeptide(s) of interest, operably linked to additional regulatory elements required for its expression. In this regard, expression vectors generally include promoter and terminator sequences, and optionally one or more enhancer sequences, polyadenylation signals, and the like. Expression vectors may also include suitable translational regulatory elements, including ribosomal binding sites, and translation initiation and termination sequences. The transcriptional and translational regulatory elements employed in the expression vectors of the invention are functional in the host cell used for expression, and may include those naturally associated with mycobacterial genes.

[0102] The selection of suitable promoters, terminators, selectable markers and other elements is a matter of routine design within the level of ordinary skill in the art. Promoters such as the trp, lac and phage promoters, tRNA promoters and glycolytic enzyme promoters may be used in prokaryotic hosts. Useful yeast promoters include the promoter regions for metallothionein, 3-phosphoglycerate kinase or other glycolytic enzymes such as enolase or glyceraldehyde-3-phosphate dehydrogenase, enzymes responsible for maltose and galactose utilization, and others. Appropriate non-native mammalian promoters may include the early and late promoters from SV40 or promoters derived from murine moloney leukemia virus, mouse mammary tumour virus, avian sarcoma viruses, adenovirus II, bovine papilloma virus or polyoma. In one embodiment, the expression vector comprises a CMV promoter.

[0103] Generally, "operably linked" means that the nucleic acid sequences being linked are contiguous and arranged so that they function in concert for their intended purposes--for example, transcription initiates in the promoter and proceeds through the coding polynucleotide segment to the terminator. Where necessary to join two protein coding regions, the polynucleotide coding sequences should be contiguous and in reading frame.

[0104] In one embodiment, the invention provides a host cell comprising an antigenic composition of the invention, as defined above. The host cell thus comprises the first mycobacterial antigen and second mycobacterial antigen of the invention, wherein said mycobacterial antigens may comprise polypeptide and/or polynucleotide sequences, as discussed above.

[0105] Accordingly, in one embodiment, a host cell comprises an antigenic composition comprising a first mycobacterial antigen and a second mycobacterial antigen; wherein said first mycobacterial antigen comprises:

[0106] (i) a first mycobacterial polypeptide sequence as hereinbefore defined; and optionally

[0107] (ii) a second (and optionally additional) mycobacterial polynucleotide sequence as hereinbefore defined;

[0108] and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0109] In one embodiment, said host cell comprises either:

[0110] (i) a first mycobacterial antigenic polypeptide as herein before defined; or

[0111] (ii) a first mycobacterial polynucleotide, wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0112] and optionally either:

[0113] (iii) a second mycobacterial antigenic polypeptide as hereinbefore defined; or

[0114] (iv) a second mycobacterial polynucleotide, wherein said second mycobacterial polynucleotide comprises a polynucleotide sequence encoding said second mycobacterial polypeptide.

[0115] The antigenic compositions, polynucleotides or polypeptides of the present invention may be prepared by expressing the polynucleotide sequences of the invention in vectors or other expression vehicles in compatible prokaryotic or eukaryotic host cells using standard molecular biology methods (e.g., Sambrook et al. 1989, Molecular Cloning a Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; incorporated herein by reference).

[0116] The most commonly used prokaryotic hosts are strains of E. coli, although other prokaryotes, such as B. subtilis or Pseudomonas may be used. Mammalian or other eukaryotic host cells, such as those of yeast, filamentous fungi, plant, insect, amphibian or avian species, may also be useful in the present invention. Propagation of mammalian cells in culture is per se well known. Examples of commonly used mammalian host cell lines are VERO and HeLa cells, Chinese hamster ovary (CHO) cells, and WI38, BHK, and COS cell lines, although other cell lines may be appropriate, e.g., to provide higher expression. As used herein, "recombinant host cells", "host cells", "cells", "cell lines", "cell cultures", and other such terms denoting microorganisms or higher eukaryotic cell lines cultured as unicellular entities refer to cells which can be, or have been, used as recipients for recombinant vector or other transfer DNA, and include the progeny of the original cell which has been transformed. It is understood that the progeny of a single parental cell may not necessarily be completely identical in morphology or in genomic or total DNA complement as the original parent, due to natural, accidental or deliberate mutation.

[0117] Polynucleotide sequences of interest can be transcribed in vitro and the resulting RNA introduced into the host cell (eg. by injection), or the polynucleotide sequences can be introduced directly into host cells by methods which vary depending on the type of cellular host, including electroporation; transfection employing calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances; microprojectile bombardment; lipofection; infection (where the vector is an infectious agent, such as a retroviral genome). "Transformation" refers to the insertion of an exogenous polynucleotide into a host cell, irrespective of the method used for the insertion, for example, direct uptake, transduction, f-mating or electroporation.

[0118] Vectors may replicate autonomously, or may replicate by being inserted into the genome of a host cell, in which case they include an insertion sequence. Expression and cloning vectors may contain a selectable marker, a gene encoding a protein necessary for the survival or growth of a host cell transformed with the vector. This gene ensures the growth of only those host cells which express the inserts. Conventional selection genes encode proteins that (a) confer resistance to antibiotics or other toxic substances, eg. ampicillin, neomycin, methotrexate, etc.; (b) complement auxotrophic deficiencies; or (c) supply critical nutrients not available from complex media, e.g. the gene encoding D-alanine racemase for bacilli. The choice of appropriate selectable marker will depend on the host cell. The transformed host cell can be cultured in accordance with known methods, and the expressed polypeptide may be recovered and isolated (eg. from the culture medium) using conventional protocols.

[0119] In one aspect, the present invention provides a method for producing an antigenic composition comprising (at least) a first mycobacterial antigen and optionally a second mycobacterial antigen, as defined above; said method comprising:

[0120] (a) expressing a polynucleotide sequence as defined above that encode said at least first and optionally second mycobacterial antigens; or

[0121] (b) culturing a host cell as described above, whereby said cell produces said at least first and optionally second mycobacterial antigens;

[0122] and recovering the expressed antigen(s).

[0123] The invention also relates to antibodies that bind a first mycobacterial antigen (eg. polypeptide) as defined above and a second mycobacterial antigen (eg. polypeptide) as defined above. Thus, in one embodiment, the invention provides an immunogenic composition comprising a first antibody and optionally a second antibody, wherein said first antibody binds a first mycobacterial antigen and said second antibody binds a second mycobacterial antigen;

[0124] wherein said first mycobacterial antigen comprises:

[0125] (i) a polypeptide sequence as hereinbefore defined; or

[0126] (ii) a polynucleotide sequence encoding a polypeptide sequence (i) as hereinbefore defined;

[0127] and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0128] In one embodiment, the immunogenic composition comprises a first antibody and a second antibody, wherein said first antibody binds a first mycobacterial antigenic polypeptide and said second antibody binds a second mycobacterial antigenic polypeptide as hereinbefore defined. Optionally, said immunogenic composition further comprises at least a third antibody (eg. at least a 3, 4, 5, 6, 7, 8 additional antibodies), wherein said third antibody binds a third mycobacterial antigen that is different from the first and second mycobacterial antigens.

[0129] The term `antibody` encompasses any polypeptide that comprises an antigen binding fragment or an antigen-binding domain. Examples include, but are not limited to, polyclonal, monoclonal, specific, monospecific, polyspecific, non specific, humanized, human, single chain, chimeric, affibodies, synthetic, recombinant, hybrid, mutated, grafted, and in vitro generated antibodies. Unless preceded by the word "intact", the term "antibody" includes antibody fragments such as Fab, F(ab')2, Fv, scFv, Fd, dAb, and other antibody fragments that retain antigen binding function. In one embodiment the antibody belongs to the IgG, IgM or IgA isotype families. Reference to the IgA isotype includes the secretory form of this antibody (ie. sIgA). The secretory component (SC) of sIgA may be added in vitro or in vivo. In the latter case, the use of a patient's natural SC labeling machinery may be employed. In one embodiment, the antibody specifically binds the mycobacterial antigen in question. "Specific binding" is intended to mean the formation of a complex between two or more molecules that is relatively stable under physiologic conditions. Specific binding is characterized by a high affinity and a low to moderate capacity, as distinguished from nonspecific binding which usually has a low affinity with a moderate to high capacity. Typically, binding between an antibody and an antigen is considered to be specific when the association constant KA is higher than 106 M 1. If necessary, nonspecific binding can be reduced without substantially affecting specific binding by varying the binding conditions. The appropriate binding conditions, such as antibody concentration, ionic strength of the solution, temperature, time allowed for binding, concentration of a blocking agent (e.g., serum albumin, milk casein), etc., may be optimized by a skilled person using routine techniques. In one embodiment, said first and second antibodies have been raised against the first and second mycobacterial antigens of the invention, as described herein, respectively. In one embodiment, said first and second antibodies have been raised against the first and second mycobacterial antigenic polypeptides of the invention, as described herein, respectively. In one embodiment, the invention provides antisera isolated from animals that have been immunized with an antigenic composition of the invention. As used herein, the term `antisera` refers to antibodies in serum that possess detectable binding, e.g., by ELISA or flow cytometry, for a particular antigen. Methods of preparing immune sera are known in the art. For example, the first and second antibodies (and optional additional antibodies) of the invention, or immunogenic composition of the invention, can be administered to an animal (such as a mammal--eg. a horse or a human) until an antibody response (for instance, neutralizing antibody response) is generated to the first and second mycobacterial antigens.

[0130] Antibodies raised against antigenic fragments disclosed herein (eg. polypeptide fragments) may have the property of recognizing the full-length antigen (eg. full-length polypeptide) from which they are derived. In this regard, polypeptide fragments bear antigenic determinants that are detectable by conventional immunoassays. One or more antigenic determinants is shared by full-length antigens of the invention and fragments thereof, thus antibodies raised against an antigenic fragment may also bind corresponding full-length antigens of the invention. In one embodiment, the antibodies are provided in an isolated form. The antibodies may be tagged with a detectable or functional label. These labels include radiolabels (eg. 1311 or 99Tc), enzymatic labels (eg. horseradish peroxidase or alkaline phosphatase), and other chemical moieties (eg. biotin).

[0131] The above-described antibodies may provide improved survival when administered to a mammal, such as a human, prior to or shortly after exposure to mycobacteria such as M. tuberculosis. Accordingly, the first and second antibodies (and optional additional antibodies) of the invention (or immunogenic, antibody-containing composition of the invention) can be used as a passive immune serum to prevent mycobacterial infection, or to treat patients exposed to mycobacteria (such as M. tuberculosis). In one embodiment, binding of the antibodies to the mycobacterial antigens of the invention may initiate coating of a mycobacterium expressing said antigen. Coating of the mycobacterium preferably leads to opsonization thereof, which leads to the bacterium being destroyed. Opsonization by antibodies may influence cellular entry and spread of mycobacteria in phagocytic and non-phagocytic cells by preventing or modulating receptor-mediated entry and replication in macrophages. Without being bound by any theory, the inventors believe that macrophage lysis may result in bacilli release. It is at this stage that the mycobacteria are considered to be most vulnerable to antibody attack. Thus, the antibodies of the present invention may act on released bacilli, and thereby exert a post-infection effect. It is therefore possible that passive protection (ie. delivery of antibodies of the present invention) is facilitated by enhanced accessibility of the antibodies of the present invention to antigens on mycobacterial bacilli, and that antibody binding may block macrophage infection by steric hindrance or disruption of its oligomeric structure. Thus, antibodies acting on mycobacterial bacilli released from killed, infected macrophages may interfere with the spread of re-infection to fresh macrophages. This hypothesis involves a synergistic action between antibodies and cytotoxic T cells, acting early after infection, eg. NK T cells, but could later involve also CD8 and CD4 cytotoxic T cells.

[0132] In the context of the therapeutic uses and methods discussed below, a `subject` is any animal subject that would benefit from stimulation of an immune response against mycobacteria, such as M. tuberculosis. Typical animal subjects are mammals, for example, human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subjects. In one embodiment, the subject is human, bovine, porcine or equine.

[0133] According to one aspect of the present invention, there is provided the use of a first mycobacterial antigen and (optionally) a second mycobacterial antigen for the manufacture of a medicament for stimulating an immune response in a subject, such as a mammalian subject, (eg. a human, bovine, porcine or equine subject); wherein said first mycobacterial antigen comprises:

[0134] (i) a polypeptide sequence as hereinbefore defined; or

[0135] (ii) a polynucleotide sequence as hereinbefore defined encoding a polypeptide sequence according to (i); and wherein said optional second mycobacterial antigen is different from said first mycobacterial antigen.

[0136] The invention also provides a first mycobacterial antigen and (optionally) a second mycobacterial antigen for use in stimulating an immune response in a subject, such as a mammalian subject, (eg. a human, bovine, porcine or equine subject); wherein said first mycobacterial antigen comprises:

[0137] (i) a polypeptide sequence as hereinbefore defined; or

[0138] (ii) a polynucleotide sequence as hereinbefore defined encoding a polypeptide sequence according to (i); and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0139] In one embodiment, said second mycobacterial antigen comprises or consists of a mycobacterial antigenic polypeptide or polynucleotide sequence, such as a mycobacterial antigenic polypeptide or polynucleotide sequence as defined in (i) or (ii) (wherein said second mycobacterial antigen is different from said first mycobacterial antigen).

[0140] In one embodiment, the invention provides (a) a first mycobacterial antigenic polypeptide or a first mycobacterial polynucleotide, and optionally (b) a second mycobacterial antigenic polypeptide or a second mycobacterial polynucleotide, for use in stimulating an immune response in a subject; wherein

[0141] (i) said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined;

[0142] (ii) said first mycobacterial polynucleotide sequence as hereinbefore defined comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0143] (iii) said optional second mycobacterial antigenic polypeptide as hereinbefore defined; and

[0144] (iv) said second mycobacterial polynucleotide sequence as hereinbefore defined comprises a polynucleotide sequence encoding said second mycobacterial antigenic polypeptide.

[0145] In one embodiment, the invention provides (a) a first mycobacterial antigenic polypeptide or a first mycobacterial polynucleotide, and optionally (b) a second mycobacterial antigenic polypeptide or a second mycobacterial polynucleotide, for use in stimulating an immune response in a subject; wherein

[0146] (i) said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined;

[0147] (ii) said first mycobacterial polynucleotide sequence as hereinbefore comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0148] (iii) said optional second mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; and

[0149] (iv) said second mycobacterial polynucleotide sequence as hereinbefore defined comprises a polynucleotide sequence encoding said second mycobacterial antigenic polypeptide.

[0150] In one embodiment, immune stimulation is measured by a protective effect in an in vivo survival assay. In one embodiment, immune stimulation is measured by an increased frequency in immune cells such as T lymphocytes specific for the antigen in the vaccine--ie. an immune cell response (eg. T cell immune response). In one embodiment, the immune stimulation is a memory T cell immune response, such as a central memory T cell response (eg. a CCR7+ response). In one embodiment, immune stimulation is measured by an increase in antibody titer that is specific for the antigen in the vaccine.

[0151] In one embodiment, said medicament further comprises one or more additional mycobacterial antigens, as described herein. In one embodiment, one or more additional mycobacterial antigens, as described herein, are also for use with said first and second mycobacterial antigens. In one embodiment of this therapeutic use, said first and optional second (and optional additional mycobacterial antigen(s)) are provided in the form of an antigenic composition as described herein. In one embodiment, one or more of said first, second and/or optional additional mycobacterial antigens may be comprised within one or more vectors or cells as described herein. In one embodiment of this therapeutic use, said first and optional second mycobacterial antigens (and optional additional mycobacterial antigen(s)) are for administration to the subject substantially simultaneously, or sequentially. Simultaneous and sequential administration regimes are discussed in more detail below.

[0152] The present invention also provides the use of a first mycobacterial antigen and optionally a second mycobacterial antigen for the manufacture of a medicament for treating or preventing a mycobacterial infection (eg. M. tuberculosis infection) in a subject, such as a mammalian subject (eg. a human, bovine, porcine or equine subject); wherein said first mycobacterial antigen comprises:

[0153] (i) a polypeptide sequence as hereinbefore defined; or

[0154] (ii) a polynucleotide sequence as hereinbefore defined encoding a polypeptide sequence according to (i); and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0155] The invention also provides a first mycobacterial antigen and optionally a second mycobacterial antigen for use in treating or preventing a mycobacterial infection (eg. M. tuberculosis infection) in a subject, such as a mammalian subject (eg. a human, bovine, porcine or equine subject); wherein said first mycobacterial antigen comprises:

[0156] (i) a polypeptide sequence as hereinbefore defined; or

[0157] (ii) a polynucleotide sequence encoding a polypeptide sequence as hereinbefore according to (i; and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0158] In one embodiment, said second mycobacterial antigen comprises or consists of a mycobacterial antigenic polypeptide or polynucleotide sequence, such as a mycobacterial antigenic polypeptide or polynucleotide sequence as defined in (i) or (ii) (wherein said second mycobacterial antigen is different from said first mycobacterial antigen).

[0159] In one embodiment, the invention provides (a) a first mycobacterial antigenic polypeptide or a first mycobacterial polynucleotide, and optionally (b) a second mycobacterial antigenic polypeptide or a second mycobacterial polynucleotide, for use in treating or preventing a mycobacterial infection (eg. M. tuberculosis infection) in a subject; wherein:

[0160] (i) said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined;

[0161] (ii) said first mycobacterial polynucleotide sequence as hereinbefore defined comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0162] (iii) said optional second mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; and

[0163] (iv) said second mycobacterial polynucleotide sequence as hereinbefore defined comprises a polynucleotide sequence encoding said second mycobacterial antigenic polypeptide.

[0164] For example, said use or medicament may protect the subject against infection with mycobacteria, such as M. tuberculosis. For example, suitable subjects include human, bovine, porcine or equine subjects. In one embodiment, said medicament further comprises one or more additional mycobacterial antigens, as described herein. In one embodiment, one or more additional mycobacterial antigens, as described herein, are also for use with said first and second mycobacterial antigens.

[0165] In one embodiment of this therapeutic use, said first and second (and optional additional mycobacterial antigen(s)) are provided in the form of an antigenic composition as described herein. In one embodiment, one or more of said first, second and/or optional additional mycobacterial antigens may be comprised within one or more vectors or cells as described herein. In one embodiment of this therapeutic use, said first and second mycobacterial antigens (and optional additional mycobacterial antigen(s)) are for administration to the subject substantially simultaneously, or sequentially.

[0166] A related aspect includes a method for stimulating an immune response in a subject, comprising administering to a subject, such as a mammal (eg. a human, bovine, porcine or equine subject) an effective amount of a first mycobacterial antigen and optionally a second mycobacterial antigen;

[0167] wherein said first mycobacterial antigen comprises:

[0168] (i) a polypeptide sequence has hereinbefore defined; or

[0169] (ii) a polynucleotide sequence encoding a polypeptide sequence as hereinbefore defined according to (i); and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0170] In one embodiment, said optional second mycobacterial antigen comprises or consists of a mycobacterial antigenic polypeptide or polynucleotide sequence, such as a mycobacterial antigenic polypeptide or polynucleotide sequence as defined in (i) or (ii) (wherein said second mycobacterial antigen is different from said first mycobacterial antigen).

[0171] In one embodiment, the invention provides a method of stimulating an immune response in a subject, comprising administrating to said subject: (a) a first mycobacterial antigenic polypeptide or a first mycobacterial polynucleotide, and optionally (b) a second mycobacterial antigenic polypeptide or a second mycobacterial polynucleotide; wherein:

[0172] (i) said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined;

[0173] (ii) said first mycobacterial polynucleotide as hereinbefore defined comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0174] (iii) said second mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; and

[0175] (iv) said second mycobacterial polynucleotide as hereinbefore defined comprises a polynucleotide sequence encoding said second mycobacterial polypeptide.

[0176] In one embodiment, immune stimulation is measured by a protective effect in an in vivo survival assay. In one embodiment, immune stimulation is measured by an increased frequency in immune cells such as T lymphocytes specific for the antigen in the vaccine--ie. an immune cell response (eg. a T cell immune response). In one embodiment, the immune stimulation is a memory T cell immune response, such as a central memory T cell response (eg. a CCR7+ response). In one embodiment, immune stimulation is measured by an increase in antibody titre that is specific for the antigen in the vaccine. In one embodiment, said method further comprises administering one or more additional mycobacterial antigens, as described herein. In one embodiment of this therapeutic method, said first and optional second (and optional additional mycobacterial antigen(s)) are provided in the form of an antigenic composition or formulation as described herein. In one embodiment, one or more of said first, second and/or optional additional mycobacterial antigens may be comprised within one or more vectors or cells as described herein. In one embodiment of this therapeutic method, any of the limitations described herein with respect to said first and/or second mycobacterial antigens (and/or optional additional mycobacterial antigens) apply equally to the therapeutic uses thereof. In one embodiment, the method comprises administering said first and second mycobacterial antigens to the subject substantially simultaneously, or sequentially. Simultaneous and sequential administration regimes are discussed in more detail below.

[0177] In a related aspect, there is provided a method of treating or preventing a mycobacterial infection (eg. an M. tuberculosis infection), comprising administering to a subject, such as a mammal (eg. a human, bovine, porcine or equine subject) an effective amount of a first mycobacterial antigen and optionally a second mycobacterial antigen;

[0178] wherein said first mycobacterial antigen comprises:

[0179] (i) a polypeptide sequence as hereinbefore defined; or

[0180] (ii) a polynucleotide sequence as hereinbefore defined encoding a polypeptide sequence according to (i); and wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0181] In one embodiment, said second mycobacterial antigen comprises or consists of a mycobacterial antigenic polypeptide or polynucleotide sequence, such as a mycobacterial antigenic polypeptide or polynucleotide sequence as defined in (i) or (ii) (wherein said second mycobacterial antigen is different from said first mycobacterial antigen). In one embodiment, the invention provides a method of treating or preventing a mycobacterial infection (eg. M. tuberculosis infection) in a subject; comprising administering to said subject: (a) a first mycobacterial antigenic polypeptide or a first mycobacterial polynucleotide, and optionally (b) a second mycobacterial antigenic polypeptide or a second mycobacterial polynucleotide; wherein:

[0182] (i) said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined;

[0183] (ii) said first mycobacterial polynucleotide as hereinbefore defined comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0184] (iii) said second mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; and

[0185] (iv) said second mycobacterial polynucleotide as hereinbefore defined.

[0186] For example, said method may protect the subject against infection with mycobacteria, such as M. tuberculosis. For example, said method may treat TB in the subject. In one embodiment, said method may protect the subject against an early stage infection with mycobacteria, such as M. tuberculosis. Early stage mycobacterial infection is defined above. In one embodiment, said method further comprises administering one or more additional mycobacterial antigens, as described herein. In one embodiment of this therapeutic method, said first and optional second (and optional additional mycobacterial antigen(s)) are provided in the form of an antigenic composition as described herein. In one embodiment, one or more of said first, second and/or optional additional mycobacterial antigens may be comprised within one or more vectors or cells as described herein. In one embodiment, the method comprises administering said first and second mycobacterial antigens to the subject substantially simultaneously, or sequentially. In a related aspect, the first and second (and optional additional) mycobacterial antigens, antigenic composition, antibodies, immunogenic composition or medicament of the present invention, as defined herein, may be useful in therapies (including preventative treatments) for a range of mycobacterial diseases not limited to tuberculosis (TB), leprosy, M. avium infection, M. bovis infection, M. paratuberculosis infection, M. ulcerans infection (eg. Buruli ulcer), or other non-tuberculosis mycobacterial infection.

[0187] The first and second (and optional additional) mycobacterial antigens, antigenic composition, antibodies, immunogenic composition or medicament of the present invention may be useful for inducing a range of immune responses and may therefore be useful in methods for treating a range of diseases. In one embodiment, the first and optional second (and optional additional) mycobacterial antigens, antigenic composition or medicament of the present invention is useful for treating or preventing a range of non-mycobacterial diseases in which mycobacteria are implicated. For example, diseases that may benefit from the medicament of the invention include inflammatory diseases such as autoimmune disease, cancer (eg. bladder cancer), inflammatory bowel disease, Crohn's Disease, Johne's Disease, Hansen's Disease, osteomyelitis, lymphadenitis, smallpox or monkeypox.

[0188] As used herein, the term "treatment" or "treating" embraces therapeutic or preventative/prophylactic measures, and includes post-infection therapy and amelioration/suppression of a mycobacterial infection. As used herein, the term "preventing" includes preventing the initiation of a mycobacterial infection and/or reducing the severity or intensity of a mycobacterial infection. As used herein, the term "vaccine efficacy" describes the ability of a vaccine to protect a subject (typically a mammalian subject eg. a human, bovine, porcine or equine subject) from challenge with mycobacteria such as M. tuberculosis. By way of example, "vaccine efficacy" may refer to the efficacy of a vaccine in preventing the initiation of a mycobacterial infection and/or reducing the severity/intensity of a mycobacterial infection.

[0189] A therapeutic/prophylactic composition or medicament may be administered to a subject (typically a mammalian subject such as a human, bovine, porcine or equine subject) already having a mycobacterial infection, condition or symptoms associated with a mycobacterial infection, to treat or prevent said mycobacterial infection. In one embodiment, the subject is suspected of having come in contact with mycobacteria, or has had known contact with mycobacteria, but is not yet showing symptoms of exposure. In one embodiment, the subject has an early-stage infection. When administered to a subject (eg. a mammal such as a human, bovine, porcine or equine subject) that already has a mycobacterial infection or disease, or is showing symptoms associated with a mycobacterial infection, the therapeutic composition/medicament can cure, delay, reduce the severity of, or ameliorate one or more symptoms, and/or prolong the survival of a subject beyond that expected in the absence of such treatment. Alternatively, a therapeutic/prophylactic composition or medicament may be administered to a subject (eg. a mammal such as a human, bovine, porcine or equine subject) who ultimately may acquire a mycobacterial infection, in order to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of said mycobacterial infection, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. In one embodiment, the subject has previously been exposed to mycobacteria. For example, the subject may have had a mycobacterial infection in the past (but is optionally not currently infected with mycobacteria). The subject may be latently infected with mycobacteria. Alternatively, or in addition, the subject may have been vaccinated against mycobacterial infection in the past (eg. the subject has previously received a BCG vaccination).

[0190] The treatments and preventative therapies of the present invention are applicable to a variety of different subjects of different ages. In the context of humans, the therapies are applicable to children (eg. infants, children under 5 years old, older children or teenagers) and adults. In the context of other animal subjects (eg. mammals such as bovine, porcine or equine subjects), the therapies are applicable to immature subjects (eg. calves, piglets, foals) and mature/adult subjects. The treatments and preventative therapies of the present invention are applicable to subjects who are immunocompromised or immunosuppressed (eg. human patients who have HIV or AIDS, or other animal patients with comparable immunodeficiency diseases), subjects who have undergone an organ transplant, bone marrow transplant, or who have genetic immuno-deficiencies.

[0191] The invention provides therapeutic formulations, medicaments and prophylactic formulations (eg. vaccines) comprising a pharmaceutically acceptable carrier, a first mycobacterial antigen of the invention as defined above, and a second mycobacterial antigen of the invention, as defined above (and optionally one or more additional mycobacterial antigens of the invention, as described above). In one embodiment, the invention provides a therapeutic or prophylactic formulation (eg. vaccine), comprising pharmaceutically acceptable carrier and:

[0192] (a) a first mycobacterial antigen, wherein said first mycobacterial antigen comprises:

[0193] (i) a polypeptide sequence as hereinbefore defined; or

[0194] (ii) a polynucleotide sequence as hereinbefore defined encoding a polypeptide sequence according to (i); and optionally

[0195] (b) a second mycobacterial antigen, wherein said second mycobacterial antigen is different from said first mycobacterial antigen; wherein said formulation is for simultaneous or sequential administration of said first and second mycobacterial antigens.

[0196] In one embodiment, said second mycobacterial antigen comprises or consists of a mycobacterial antigenic polypeptide or polynucleotide sequence, such as a mycobacterial antigenic polypeptide or polynucleotide sequence as defined in (i) or (ii) (wherein said second mycobacterial antigen is different from said first mycobacterial antigen).

[0197] In one embodiment, said therapeutic or prophylactic formulation (eg. vaccine), comprises (a) a pharmaceutically acceptable carrier; (b) a first mycobacterial antigenic polypeptide or a first mycobacterial polynucleotide; and optionally (c) a second mycobacterial antigenic polypeptide or a second mycobacterial polynucleotide; wherein:

[0198] (i) said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined;

[0199] (ii) said first mycobacterial polynucleotide as hereinbefore defined comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide;

[0200] (iii) said second mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; and

[0201] (iv) said second mycobacterial polynucleotide as hereinbefore defined comprises a polynucleotide sequence encoding said second mycobacterial polypeptide;

[0202] wherein said formulation is for simultaneous or sequential administration of said first mycobacterial antigenic polypeptide or polynucleotide and said second mycobacterial antigenic polypeptide or polynucleotide.

[0203] In one embodiment, said therapeutic formulation, medicament or prophylactic formulation (eg. vaccine) of the invention comprises an antigenic composition of the invention, as defined above. In one embodiment, said therapeutic formulation, medicament or prophylactic formulation (eg. vaccine) comprises an antigenic composition comprising one or more vectors or cells, as described above, wherein said vectors or cells comprise at least one of the mycobacterial antigens. In one embodiment of said therapeutic formulation, medicament or prophylactic formulation (eg. vaccine), any of the limitations described herein with respect to said first and/or second (or additional) mycobacterial antigens apply equally to said therapeutic formulation, medicament or prophylactic formulation (eg. vaccine). In one embodiment, a vaccine of the invention is a "vectored vaccine" comprising one or more vectors as described above.

[0204] In one embodiment, the therapeutic formulations, medicaments or prophylactic formulations (eg. vaccines) of the invention are for simultaneous administration of said first and second (and/or optional additional) mycobacterial antigens. In an alternative embodiment, the therapeutic formulations, medicaments or prophylactic formulations (eg. vaccines) of the invention are for sequential administration of said first and second (and/or optional additional) mycobacterial antigens. Therapeutic formulations, medicaments and prophylactic formulations (eg. vaccines) of the invention comprise a pharmaceutically acceptable carrier, and optionally one or more of a salt, excipient, diluent and/or adjuvant. In one embodiment, the therapeutic formulation, medicament or prophylactic formulation (eg. vaccine) of the invention may comprise one or more immunoregulatory agents selected from, for example, immunoglobulins, antibiotics, interleukins (eg. IL-2, IL-12), and/or cytokines (eg. IFN-.gamma.). In one embodiment, the therapeutic formulation, medicament or prophylactic formulation (eg. vaccine) of the invention may comprise one or more antimicrobial compounds, such as conventional anti-tuberculosis drugs (eg. rifampicin, isoniazid, ethambutol or pyrazinamide).

[0205] Accordingly, in one aspect, the invention provides a method for producing a therapeutic or prophylactic formulation (eg. vaccine), the method comprising combining a pharmaceutically acceptable carrier with a first mycobacterial antigen of the invention, as defined above; and a second mycobacterial antigen of the invention, as defined above (and optionally one or more additional mycobacterial antigens, as defined above).

[0206] Thus, in one embodiment, the invention provides a method for producing a therapeutic or prophylactic formulation (eg. vaccine), the method comprising combining a pharmaceutically acceptable carrier with:

[0207] (a) a first mycobacterial antigen, wherein said first mycobacterial antigen comprises:

[0208] (i) a polypeptide sequence as hereinbefore defined; or

[0209] (ii) a polynucleotide sequence as hereinbefore defined encoding a polypeptide sequence according to (i); and optionally

[0210] (b) a second mycobacterial antigen, wherein said second mycobacterial antigen is different from said first mycobacterial antigen.

[0211] In one embodiment, said second mycobacterial antigen comprises or consists of a mycobacterial antigenic polypeptide or polynucleotide sequence, such as a mycobacterial antigenic polypeptide or polynucleotide sequence as defined in (i) or (ii) (wherein said second mycobacterial antigen is different from said first mycobacterial antigen).

[0212] In one embodiment, the invention provides a method for producing a therapeutic or prophylactic formulation (eg. vaccine), the method comprising:

combining a pharmaceutically acceptable carrier with either:

[0213] (i) a first mycobacterial antigenic polypeptide, wherein said first mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; or

[0214] (ii) a first mycobacterial polynucleotide, wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence as hereinbefore defined encoding said first mycobacterial antigenic polypeptide; and optionally with either:

[0215] (iii) a second mycobacterial antigenic polypeptide, wherein said second mycobacterial antigenic polypeptide comprises a polypeptide sequence as hereinbefore defined; or

[0216] (iv) a second mycobacterial polynucleotide, wherein said second mycobacterial polynucleotide as hereinbefore defined comprises a polynucleotide sequence encoding said second mycobacterial polypeptide.

[0217] In one embodiment, said mycobacterial antigens are in the form of an antigenic composition of the invention, as defined above. In one embodiment, the method further comprises combining said pharmaceutically acceptable carrier and mycobacterial antigens (or antigenic composition) with one or more of a salt, excipient, diluent, adjuvant, immunoregulatory agent and/or antimicrobial compound.

[0218] As used, herein, a "vaccine" is a formulation that, when administered to an animal subject such as a mammal (eg. a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject), stimulates a protective immune response against mycobacterial infection. The immune response may be a humoral and/or cell-mediated immune response (eg. a T cell response). A vaccine of the invention can be used, for example, to protect an animal from the effects of mycobacterial infection (eg. M. tuberculosis infection), such as an early-stage infection. The immunogenicity of the epitopes of the first and second mycobacterial antigens (eg. polypeptides) of the invention may be enhanced by preparing them in mammalian or yeast systems fused with or assembled with particle-forming proteins such as, for example, that associated with hepatitis B surface antigen. In one embodiment, the vaccine comprises at least one mycobacterial polypeptide that has been treated with a chemical modifying agent (such as formaldehyde) to give a vaccine of improved efficacy.

[0219] The polypeptides and/or polynucleotides of the invention may be formulated into a vaccine as neutral or salt forms. Pharmaceutically acceptable salts include acid addition salts formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or with organic acids such as acetic, oxalic, tartaric, maleic, and the like. Salts formed with the free carboxyl groups may also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

[0220] Administration of therapeutic formulations, medicaments and prophylactic formulations (eg. vaccines) is generally by conventional routes e.g. intravenous, subcutaneous, intraperitoneal, or mucosal (eg. intranasal) routes. The administration may be by parenteral injection, for example, a subcutaneous or intramuscular injection. Formulations comprising neutralizing antibodies may be particularly suited to administration intravenously, intramuscularly, intradermally, or subcutaneously. Accordingly, the therapeutic formulations, medicaments and prophylactic formulations (eg. vaccines) of the invention are typically prepared as injectables, either as liquid solutions or suspensions. Solid forms suitable for solution in, or suspension in, liquid prior to injection may alternatively be prepared. The preparation may also be emulsified, or the peptide encapsulated in liposomes or microcapsules. The active immunogenic ingredients are often mixed with excipients which are pharmaceutically acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, or the like and combinations thereof. In addition, if desired, the vaccine may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and/or adjuvants which enhance the effectiveness of the vaccine. Generally, the carrier is a pharmaceutically-acceptable carrier. Non-limiting examples of pharmaceutically acceptable carriers include water, saline, and phosphate-buffered saline. In some embodiments, however, the composition is in lyophilized form, in which case it may include a stabilizer, such as BSA. In some embodiments, it may be desirable to formulate the composition with a preservative, such as thiomersal or sodium azide, to facilitate long term storage. Examples of adjuvants which may be effective include but are not limited to: complete Freunds adjuvant (CFA), Incomplete Freunds adjuvant (IVA), Saponin, a purified extract fraction of Saponin such as Quil A, a derivative of Saporin such as QS-21, lipid particles based on Saponin such as ISCOM/ISCOMATIX, E. coli heat labile toxin (LT) mutants such as LTK63 and/or LTK72, aluminium hydroxide, N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-nor-muramyl-L-alanyl-D-isoglutamine (CGP 11637, referred to as nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(1'-2'-dip- almitoyl-sn-glycero-3-hydroxyphosphoryl oxy)-ethylamine (CGP 19835A, referred to as MTP-PE), and RIBI, which contains three components extracted from bacteria, monophosphoryl lipid A, trehalose dimycolate and cell wall skeleton (MPL+TDM+CWS) in a 2% squalene/Tween 80 emulsion. Examples of buffering agents include, but are not limited to, sodium succinate (pH 6.5), and phosphate buffered saline (PBS; pH 6.5 and 7.5). Additional formulations which are suitable for other modes of administration include suppositories and, in some cases, oral formulations or formulations suitable for distribution as aerosols. For suppositories, traditional binders and carriers may include, for example, polyalkylene glycols or triglycerides; such suppositories may be formed from mixtures containing the active ingredient in the range of 0.5% to 10%, preferably 1%-2%. Oral formulations include such normally employed excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders.

[0221] In the case of a mycobacterial respiratory infection (eg. a M. tuberculosis infection), efficient transmission of the therapeutic/prophylactic composition or medicament to the site of infection in the lungs may be achieved by oral or intra-nasal administration (i.n.). These modes of delivery correspond to the route of delivery of a M. tuberculosis infection. In the case of antibody-based compositions, these modes of delivery ensure that antibodies are present at the site of infection to combat the bacterium before it becomes intracellular and also during the period when it spreads between cells. Formulations for intranasal administration may in the form of nasal droplets or a nasal spray. An intranasal formulation may comprise droplets having approximate diameters in the range of 100-5000 .mu.m, such as 500-4000 .mu.m, 1000-3000 .mu.m or 100-1000 .mu.m. Alternatively, in terms of volume, the droplets may be in the range of about 0.001-100 .mu.l, such as 0.1-50 .mu.l or 1.0-25 .mu.l, or such as 0.001-1 .mu.l. Alternatively, the therapeutic/prophylactic formulation or medicament may be an aerosol formulation. The aerosol formulation may take the form of a powder, suspension or solution. The size of aerosol particles is relevant to the delivery capability of an aerosol. Smaller particles may travel further down the respiratory airway towards the alveoli than would larger particles. In one embodiment, the aerosol particles have a diameter distribution to facilitate delivery along the entire length of the bronchi, bronchioles, and alveoli. Alternatively, the particle size distribution may be selected to target a particular section of the respiratory airway, for example the alveoli. In the case of aerosol delivery of the medicament, the particles may have diameters in the approximate range of 0.1-50 .mu.m, preferably 1-25 .mu.m, more preferably 1-5 .mu.m. Aerosol particles may be for delivery using a nebulizer (eg. via the mouth) or nasal spray. An aerosol formulation may optionally contain a propellant and/or surfactant. It is possible that, following i.n. delivery of mycobacterial antigens or antibodies, their passage to the lungs is facilitated by a reverse flow of mucosal secretions, although mucociliary action in the respiratory tract is thought to take particles within the mucus out of the lungs. The relatively long persistence in lung lavage, fast clearance from the bile and lack of transport to the saliva of some antibodies suggests the role of mucosal site-specific mechanisms. By controlling the size of the droplets/particles to within the defined range of the present invention, it is possible to avoid (or minimize) inadvertent antigen delivery to the alveoli and thus avoid alveoli-associated pathological problems such as inflammation and fibrotic scarring of the lungs. I.n. vaccination engages both T and B cell mediated effector mechanisms in nasal and bronchus associated mucosal tissues, which differ from other mucosae-associated lymphoid tissues. The protective mechanisms invoked by the intranasal route of administration may include: the activation of T lymphocytes with preferential lung homing; up-regulation of co-stimulatory molecules (eg. B7.2); and/or activation of macrophages or secretory IgA antibodies. Intranasal delivery of antigens may facilitate the invoking of a mucosal antibody response, which is favoured by a shift in the T cell response toward the Th2 phenotype which helps antibody production. A mucosal response is characterised by enhanced IgA production, and a Th2 response is characterised by enhanced IL-4 production. Intranasal delivery of mycobacterial antigens of the invention allows targeting of the antigens to sub-mucosal B cells of the respiratory system. These B cells are the major local IgA-producing cells in mammals and intranasal delivery facilitates a rapid increase in IgA production by these cells against the mycobacterial antigens. In one embodiment, the therapeutic/prophylactic formulation or medicament of the invention stimulates a mucosal and/or Th2 immune response. In another embodiment, IgA antibody production is stimulated, and the IgA antibody binds to the mycobacterial antigen.

[0222] In one embodiment, the first and second (and optional additional) mycobacterial antigens or antibodies of the invention are for simultaneous administration. Thus, in one embodiment, the methods/uses of the invention comprise simultaneous administration of the first and second (and optional additional) mycobacterial antigens. Simultaneous administration means administration at (substantially) the same time. For example, in one embodiment the first and second (and optional additional) mycobacterial antigens are administered to the subject within 5 minutes of each other, such as within 4, 3, 2 or 1 minute of each other, for example within 30 seconds of each other. In one embodiment of `simultaneous administration`, at least two components (eg. antigens) of the invention are combined into one composition (eg. a single antigenic composition or immunogenic composition of the invention as defined herein). This composition is administered to the subject (such as a mammal--eg. a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject) thereby providing both components to the subject simultaneously. In an alternative embodiment of `simultaneous administration`, at least two of the components (eg. antigens) of the invention are provided separately from each other, but are administered to the subject (such as a mammal--eg. a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject) at (substantially) the same time. The concurrent/parallel administration of said separate compositions provides both components to the subject at (substantially) the same time. By way of example, the therapeutic or prophylactic formulation (eg. vaccine) of the invention may comprise a first mycobacterial antigen in a first composition and the second mycobacterial antigen in a second composition. In one embodiment, the first and second (and optional additional) mycobacterial antigens of the invention are for simultaneous administration at (substantially) the same site. Thus, in one embodiment, the methods/uses of the invention comprise simultaneous administration of the first and second (and optional additional) mycobacterial antigens at (substantially) the same site. In this regard, it is considered advantageous to administer each different antigenic component of conventional multivalent vaccines at different sites of the subject's body, in order to stimulate different lymph nodes. Administration of different antigenic components of conventional multivalent vaccines at different sites is also considered advantageous in order to reduce or avoid undesirable antigenic competition.

[0223] In one embodiment, the present invention advantageously avoids the need to administer each different antigenic component to different sites/locations of the subject's body. In this regard, in one embodiment, the first and second (and optional additional) antigens of the present invention (substantially) do not compete with each other, or are associated with relatively low levels of antigenic competition, as compared with the competitive effect that might have been expected in view of known multivalent vaccine compositions. If at least two components (eg. antigens) of the invention are combined into a single composition (eg. a single antigenic composition or immunogenic composition of the invention as defined herein), it is evident that all components of the invention are administered to the subject at the same site. In one embodiment, if the first and second (and optional additional) mycobacterial antigens of the invention are provided separately from each other, for simultaneous, parallel administration to the subject at (substantially) the same time, the separate compositions are administered at the same (or substantially the same) site on/in the subject.

[0224] In one embodiment, administration at (substantially) the same site on/in the subject means that the site at which each mycobacterial antigen of the invention is administered is in the vicinity of, or in close proximity to, the site at which the other mycobacterial antigens of the invention are administered. Alternatively, administration at (substantially) the same site on/in the subject means that the site at which the each mycobacterial antigen of the invention is administered is at the precise site at which the other mycobacterial antigens of the invention are administered. By way of example, the first and second (and optional additional) mycobacterial antigens of the invention may be for administration to the same vein, artery or muscle of the subject, or via the same nostril of the subject; or to the same limb (eg. arm) of the subject (eg. to the same upper arm of the subject); or the first and second (and optional additional) mycobacterial antigens of the invention may all be for oral or sublingual administration. In one embodiment, the first and second (and optional additional) mycobacterial antigens of the invention may all be for administration at or in close proximity to the same lymph node. Alternatively, the mycobacterial antigens of the invention are for administration to the subject (eg. a mammal such as a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject) sequentially (ie. one after the other). In this embodiment, at least two of the components (eg. antigens) of the invention are provided separately from each other, and are administered sequentially to the subject. By way of example, the therapeutic or prophylactic formulation (eg. vaccine) of the invention may comprise a first mycobacterial antigen in a first composition and the second mycobacterial antigen in a second composition. The sequential administration of said first and second compositions provides both components to the subject one after the other. Thus, in one embodiment, the methods of the invention comprise administration of the first mycobacterial antigen, and then administration of the second mycobacterial antigen. Alternatively, the second mycobacterial antigen may be administered and then the first mycobacterial antigen is administered. Any additional mycobacterial antigens may be administered together with the first and/or second mycobacterial antigens. Alternatively, any additional mycobacterial antigens may be administered before or after the first and/or second mycobacterial antigens.

[0225] In one embodiment, each sequential administration of antigen is made immediately one after the other. In one embodiment, there is a time-gap or pause between one or more (eg. between each) of the administrations. A time-gap or pause between sequential administrations may be at least 5, 10, 15, or 30 minutes, or may be at least 1, 2, 5, 12, 18 or 24 hours, or may be at least 1, 2, or 5 days, or may be at least 1 or 2 weeks. In one embodiment, the first and second (and optional additional) mycobacterial antigens of the invention are for sequential administration at (substantially) the same site. Thus, in one embodiment, the methods/uses of the invention comprise sequential administration of the first and second (and optional additional) mycobacterial antigens at (substantially) the same site. In one embodiment, administration at (substantially) the same site on/in the subject means that the site at which the each mycobacterial antigen of the invention is administered is in the vicinity of, or in close proximity to, the site at which the other mycobacterial antigens of the invention are administered. Alternatively, administration at (substantially) the same site on/in the subject means that the site at which each mycobacterial antigen of the invention is administered is at the precise site at which the other mycobacterial antigens of the invention are administered. By way of example, the first and second (and optional additional) mycobacterial antigens of the invention may be for administration to the same vein, artery or muscle of the subject, or via the same nostril of the subject; or to the same limb (eg. arm) of the subject (eg. to the same upper arm of the subject); or the first and second (and optional additional) mycobacterial antigens of the invention may all be for oral or sublingual administration. In one embodiment, the first and second (and optional additional) mycobacterial antigens of the invention may all be for administration at or in close proximity to the same lymph node.

[0226] The therapeutic formulation, medicament or prophylactic formulation (eg. a vaccine) of the invention may be given in a single dose schedule (ie. the full dose is given at substantially one time). Alternatively, the therapeutic formulation, medicament or prophylactic formulation (eg. a vaccine) of the invention may be given in a multiple dose schedule. A multiple dose schedule is one in which a primary course of treatment (eg. vaccination) may be with 1-6 separate doses, followed by other doses given at subsequent time intervals required to maintain and or reinforce the immune response, for example (for human subjects), at 1-4 months for a second dose, and if needed, a subsequent dose(s) after a further 1-4 months. The dosage regimen will be determined, at least in part, by the need of the individual and be dependent upon the judgment of the practitioner (eg. doctor or veterinarian). In one embodiment, the vaccine of the present invention may be administered as part of a `prime-boost` vaccination regime.

[0227] Prime-boost vaccination regimes involve: Priming--ie. exposing a subject to one or more antigens or a vaccine; and subsequently: Boosting--ie. exposing the subject to one or more antigens or a vaccine. The `boost` antigens/vaccine is typically different from the `primer` antigens/vaccine (known as "heterologous" prime-boost). In this regard, heterologous prime-boost immunization strategies have been shown to induce higher levels of immune cell responses (eg. effector T cell responses) in subjects as compared with homologous boosting with the same vaccine. For example, repeated vaccination with conventional vaccines such as BCG does not appear to further enhance protection against TB. However, incorporating BCG into a heterologous prime-boost regime may retain the protective effects of BCG. Thus, in one embodiment the invention provides a method of vaccination against mycobacterial infection comprising `priming` a subject's immune system by administration of a heterologous conventional vaccine (eg. BCG vaccine) and then `boosting` the subject's immune system by administration of the vaccine of the present invention. In one embodiment, the invention provides a method of vaccination against mycobacterial infection comprising administering the vaccine of the present invention to a subject that has been pre-exposed to a heterologous conventional vaccine such as BCG. Alternatively, a subject's immune system may be `primed` by administration of the vaccine of the present invention, and then `boosted` by administration of a heterologous conventional vaccine (eg. BCG vaccine). Accordingly, in one embodiment, the vaccine is administered to a subject that is subsequently to be exposed to a heterologous conventional vaccine such as BCG. The `priming` step may be carried out on the subject at any age--in the case of mammalian subjects (eg. human, bovine, porcine, ovine, caprine, equine, cervine, canine or feline subjects), priming with BCG is conventionally carried out neonatally, or during infancy, adolescence or adulthood. The `boosting` step may be carried out at any time after the `priming` step. In the case of mammalian subjects (eg. human, bovine, porcine, ovine, caprine, equine, cervine, canine or feline subjects), a boosting step may be carried out at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks after the priming step, or at least about 3, 6, 8 or 12 months after the priming step, or at least about 2, 5, 10, 15, 20, 25, 30, 35, or 40 or more years after the boosting step. In one embodiment, for a human subject, the priming step is carried out during infancy and the boosting step is carried out during adolescence. In one embodiment, the therapeutic formulation, medicament or prophylactic formulation (eg. a vaccine) of the invention can be administered to a subject such as a mammal (eg. a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject) in conjunction with (simultaneously or sequentially) one or more immunoregulatory agents selected from, for example, immunoglobulins, antibiotics, interleukins (eg. IL-2, IL-12), and/or cytokines (eg. IFN.gamma.). In one embodiment, the therapeutic formulation, medicament or prophylactic formulation (eg. vaccine) of the invention can be administered to a subject such as a mammal (eg. a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject) in conjunction with (simultaneously or sequentially) one or more antimicrobial compounds, such as conventional anti-tuberculosis drugs (eg. rifampicin, isoniazid, ethambutol or pyrazinamide).

[0228] The therapeutic formulation, medicament or prophylactic formulation (eg. vaccine) may contain 5% to 95% of active ingredient, such as at least 10% or 25% of active ingredient, or at least 40% of active ingredient or at least 50, 55, 60, 70 or 75% active ingredient. The therapeutic formulation, medicament or prophylactic formulation (eg. a vaccine) is administered in a manner compatible with the dosage formulation, and in such amount as will be prophylactically and/or therapeutically effective. In this regard, as used herein, an "effective amount" is a dosage or amount that is sufficient to achieve a desired biological outcome. As used herein, a "therapeutically effective amount" is an amount which is effective, upon single or multiple dose administration to a subject (such as a mammal--eg. a human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subject) for treating, preventing, curing, delaying, reducing the severity of, ameliorating at least one symptom of a disorder or recurring disorder, or prolonging the survival of the subject beyond that expected in the absence of such treatment. Accordingly, the quantity of active ingredient to be administered, which is generally in the range of 5 micrograms to 250 micrograms of antigen per dose (or higher if delivered orally or in the form of viral vectors), depends on the subject to be treated, capacity of the subject's immune system to generate a protective immune response, and the degree of protection desired. Precise amounts of active ingredient required to be administered may depend on the judgment of the practitioner and may be particular to each subject.

[0229] According to a further aspect of the invention, the first and second mycobacterial antigens (and optional additional mycobacterial antigens) of the invention, as described herein, are useful in immunoassays to detect the presence in a test sample of antibodies to said first and second mycobacterial antigens. In one embodiment, said first and second mycobacterial antigens (and optional additional antigens) are used in the form of an antigenic composition, as described herein. According to another aspect of the invention, the first and second antibodies (and optional additional antibodies) of the invention, as described herein, are useful in immunoassays to detect the presence in a test sample of said first and second mycobacterial antigens. In one embodiment, said first and second antibodies (and optional additional antibodies) are used in the form of an immunogenic antibody-containing composition, as described herein.

[0230] A test sample may be a biological sample such as a clinical sample or environmental sample. As used herein, a `clinical sample` refers to a sample of tissue or fluid isolated from an individual, including but not limited to, for example, plasma, serum, spinal fluid, lymph fluid, the external sections of the skin, respiratory, intestinal, and genitourinary tracts, tears, saliva, milk, blood cells, tumours, organs, and also samples of in vitro cell culture constituents (including but not limited to conditioned medium resulting from the growth of cells in cell culture medium, putatively infected cells, recombinant cells, and cell components). In the context of the diagnostic methods discussed below, a `subject` is any animal subject that would benefit from detection of mycobacterial infection, such as M. tuberculosis infection. Typical animal subjects are mammals, for example, human, bovine, porcine, ovine, caprine, equine, corvine, canine or feline subjects. In one embodiment, the subject is human, bovine, porcine or equine.

[0231] Design of immunoassays is subject to a great deal of variation, and many formats are known in the art. Protocols may be based, for example, upon competition, direct reaction, or sandwich type assays. Protocols may also, for example, use solid supports, or may employ immuno-precipitation. Most assays involve the use of labeled antibodies or polypeptides; the labels may be, for example, enzymatic, fluorescent, chemiluminescent, radioactive, or dye molecules. Assays that comprise signal amplification are also known; for example, assays that utilize biotin and avidin, or enzyme-labeled and mediated immunoassays, such as ELISA assays. In one aspect of the invention, the first and second mycobacterial antigens (or antigenic composition) of the invention are useful for detecting the presence of a T-lymphocyte that has been previously exposed to an antigenic component of a mycobacterial infection in a patient. Accordingly, in one embodiment, the invention provides an in vitro method of diagnosing a mycobacterial infection, such as an early stage mycobacterial infection, comprising incubating (`challenging`) a test sample containing an immune cell such as a T-lymphocyte from a subject (eg. a mammal such as a human, bovine, porcine or equine subject) with a first mycobacterial antigen of the invention and a second mycobacterial antigen of the invention, as defined herein; or an antigenic composition of the invention, as defined herein; and detecting activation of said immune cell (eg. T-lymphocyte). Activation of said immune cell is indicative of a mycobacterial infection in the subject.

[0232] In one embodiment of said in vitro method, said first mycobacterial antigen is selected from (i) a first mycobacterial antigenic polypeptide comprising a polypeptide sequence having at least 70% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 1 or 7, or a fragment thereof having at least 50 consecutive amino acids thereof; or (ii) a first mycobacterial polynucleotide sequence comprising a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide. In one embodiment of said in vitro method, said second mycobacterial antigen is selected from (iii) a second mycobacterial antigenic polypeptide comprising a polypeptide sequence having at least 70% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 5, or a fragment thereof having at least 7 consecutive amino acids thereof; or (iv) a second mycobacterial polynucleotide sequence comprising a polynucleotide sequence encoding said second mycobacterial antigenic polypeptide.

[0233] An immune cell, such as a T-lymphocyte, that has been previously exposed to one or both of the first and second mycobacterial antigens will become `activated` on subsequent challenge by the same antigen. As such, activation of said immune cell (eg. T-lymphocyte) is indicative of a mycobacterial infection in the subject, and provides a means for identifying a positive diagnosis of mycobacterial infection. In contrast, the same activation is not achieved by an immune cell (eg. T-lymphocyte) that has not been previously exposed to the particular antigen. The above-described `activation` of an immune cell (eg. T-lymphocyte) is sometimes referred to as a `recall response` and may be measured, for example, by determining the release of interferon (eg. IFN-.gamma.) from the activated immune cell (eg. T-lymphocyte). Thus, the presence of a mycobacterial infection in a patient may be determined by detecting activation of immune cell (eg. T-lymphocyte) in response to in vitro challenge with the first and second mycobacterial antigens (or antigenic composition) of the present invention--eg. by detecting the release of a minimum concentration of interferon from immune cell (eg. T-lymphocyte) after a defined time period following the challenge. The above immune cell (eg. T-lymphocyte) diagnostic assay may further include an antigen presenting cell (APC) expressing at least one major histocompatibility complex (MHC) class II molecule expressed by the patient in question. The APC may be inherently provided in the biological sample, or may be added exogenously. In one embodiment, the T-lymphocyte is a CD4 T-lymphocyte.

[0234] Alternative immunoassays for diagnosing mycobacterial infection depend upon detection of antibodies to the first and second mycobacterial antigens (eg. polypeptides) of the invention. Such assays may comprise the step of incubating a test sample (eg. a biological sample) suspected of containing the antibodies with said first and second antigens (or antigenic composition) of the invention. Accordingly, the invention also provides an in vitro method of diagnosing a mycobacterial infection, such as an early stage mycobacterial infection, comprising incubating a test sample from a subject (eg. a mammal such as a human, bovine, porcine or equine subject) with a first mycobacterial antigen and a second mycobacterial antigen of the invention, as defined herein; or an antigenic composition of the invention, as defined herein; wherein said incubating is performed under conditions that allow binding of said first and second mycobacterial antigens with antibodies in the sample to form antigen-antibody complexes; and then detecting the formation of such complexes. The presence of antigen-antibody complexes is indicative of a mycobacterial infection in the subject.

[0235] In one embodiment of said in vitro method, said first mycobacterial antigen is selected from (i) a first mycobacterial antigenic polypeptide comprising a polypeptide sequence as hereinbefore defined; or (ii) a first mycobacterial polynucleotide sequence comprising a polynucleotide sequence as hereinbefore defined encoding said first mycobacterial antigenic polypeptide; and optionally said second mycobacterial antigen is selected from (iii) a second mycobacterial antigenic polypeptide comprising a polypeptide sequence as hereinbefore defined; and (iv) a second mycobacterial polynucleotide sequence as hereinbefore defined comprising a polynucleotide sequence encoding said second mycobacterial antigenic polypeptide. Antigen-antibody complexes (or, in the case of competitive assays, the amount of competing antibody) may be detected by any of a number of known techniques, depending on the format. For example, unlabelled antibodies in the complex may be detected using a conjugate of anti-xenogeneic Ig complexed with a label (eg. an enzyme label). The immunoassay may be of a standard or competitive type. In one embodiment, the first and second mycobacterial antigens are bound to one or more solid supports to facilitate separation of the sample from the antigens after incubation. Examples of solid supports that can be used are nitrocellulose (eg. in membrane or microtitre well form), polyvinyl chloride (eg. in sheets or microtiter wells), polystyrene latex (eg. in beads or microtiter plates, polyvinylidine fluoride (known as Immulon), diazotized paper, nylon membranes, activated beads, and Protein A beads. For example, Dynatech Immulon microtiter plates or 60 mm diameter polystyrene beads (Precision Plastic Ball) may be used. The solid support(s) containing the first and second mycobacterial antigens is typically washed after separating it from the test sample, and prior to detection of bound antibodies. The invention also embraces immunoassays for detecting the presence of the first and second mycobacterial antigens (eg. polypeptides) of the invention in a test sample (eg. a biological sample). In such methods, a test sample suspected of containing said mycobacterial antigens may be incubated with antibodies directed against the first and second mycobacterial antigens.

[0236] Accordingly, the invention provides an in vitro method of diagnosing a mycobacterial infection, such as an early stage mycobacterial infection, comprising incubating a test sample from a subject (eg. a mammal such as a human, bovine, porcine or equine subject) with a first antibody and a second antibody of the invention, as defined herein; or an immunogenic composition of the invention, as defined herein; wherein said incubating is performed under conditions that allow binding of said first and second antibodies with antigens in the sample to form antigen-antibody complexes; and then detecting the formation of such complexes, wherein the presence of antigen-antibody complexes is indicative of a mycobacterial infection in the subject. In one embodiment of said in vitro method, said first antibody binds a first mycobacterial antigenic polypeptide; and said optional second antibody binds a second mycobacterial antigenic polypeptide. It may be desirable to treat the biological sample prior to testing, to release putative bacterial components. Various formats can be employed. For example, a "sandwich assay" may be employed, where antibodies bound to a solid support are incubated with the test sample; washed; incubated with second, labeled antibodies to the first and second antigens, and the support is washed again. The first and second mycobacterial antigens are detected by determining if the second antibody is bound to the support. In a competitive format, a test sample is usually incubated with antibodies and a labeled, competing antigen is also incubated, either sequentially or simultaneously.

[0237] In one aspect, the invention provides an immunoassay kit, comprising an antigenic composition of the invention, or antibodies to said first and second mycobacterial antigens. The immunoassay kit may further comprise a buffer. The term "polypeptide" throughout this specification is synonymous with the terms "oligopeptide", "peptide" and "protein". These terms are used interchangeably and do not refer to a specific length of the product. These terms embrace post-translational modifications such as glycosylation, acetylation and phosphorylation. In one embodiment, the isolated polypeptides of the invention are substantially free from other proteins with which they are co-produced as well as from other contaminants. For instance, an isolated polypeptide is substantially free of material or other proteins from the cell, bacterial, or tissue source from which it was derived.

[0238] The present invention encompasses polypeptides that are substantially homologous to a polypeptide based on any one of the reference SEQ ID NOs identified in this application (including fragments thereof). The terms "sequence identity" and "sequence homology" are considered synonymous in this specification. By way of example, a polypeptide of interest may comprise an amino acid sequence having at least 70, 75, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 99 or 100% amino acid sequence identity with the amino acid sequence of a reference polypeptide.

[0239] There are many established algorithms available to align two amino acid sequences. Typically, one sequence acts as a reference sequence, to which test sequences may be compared. The sequence comparison algorithm calculates the percentage sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters. Alignment of amino acid sequences for comparison may be conducted, for example, by computer implemented algorithms (eg. GAP, BESTFIT, FASTA or TFASTA), BLAST and BLAST 2.0 algorithms, or BLOSUM62--Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915-10919, 1992; incorporated herein by reference). In a homology comparison, the identity may exist over a region of the sequences that is at least 50 amino acid residues in length (eg. at least 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600 or 650 amino acid residues in length--eg. up to the entire length of the reference sequence. Substantially homologous polypeptides have one or more amino acid substitutions, deletions, or additions. In many embodiments, those changes are of a minor nature, for example, involving only conservative amino acid substitutions. Conservative substitutions are those made by replacing one amino acid with another amino acid within the following groups: Basic: arginine, lysine, histidine; Acidic: glutamic acid, aspartic acid; Polar: glutamine, asparagine; Hydrophobic: leucine, isoleucine, valine; Aromatic: phenylalanine, tryptophan, tyrosine; Small: glycine, alanine, serine, threonine, methionine. Substantially homologous polypeptides also encompass those comprising other substitutions that do not significantly affect the folding or activity of the polypeptide; small deletions, typically of 1 to about 30 amino acids (such as 1-10, or 1-5 amino acids); and small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or an affinity tag.

[0240] The polypeptides of the present invention may also comprise non-naturally occurring amino acid residues. In this regard, in addition to the 20 standard amino acids, non-standard amino acids (such as 4-hydroxyproline, 6-N-methyl lysine, 2-aminoisobutyric acid, isovaline and .alpha.-methyl serine) may be substituted for amino acid residues of the mycobacterial polypeptides of the present invention. A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code, and unnatural amino acids may be substituted for mycobacterial polypeptide amino acid residues. Non-naturally occurring amino acids include, without limitation, trans-3-methylproline, 2,4-methano-proline, cis-4-hydroxyproline, trans-4-hydroxy-proline, N-methylglycine, allo-threonine, methyl-threonine, hydroxy-ethylcysteine, hydroxyethylhomo-cysteine, nitro-glutamine, homoglutamine, pipecolic acid, tert-leucine, norvaline, 2-azaphenylalanine, 3-azaphenyl-alanine, 4-azaphenyl-alanine, and 4-fluorophenylalanine.

[0241] Routine deletion analyses of nucleic acid molecules can be performed to obtain functional fragments of a nucleic acid molecule that encodes a polypeptide of the invention. As an illustration, DNA molecules can be digested with Bal31 nuclease to obtain a series of nested deletions. These DNA fragments are then inserted into expression vectors in proper reading frame, and the expressed polypeptides are isolated and tested for the desired activity. An alternative to exonuclease digestion is to use oligonucleotide-directed mutagenesis to introduce deletions, or stop codons to specify production of a desired fragment. Alternatively, particular polynucleotide fragments can be synthesized using the polymerase chain reaction. A mutant of a polypeptide of the invention may contain one or more analogs of an amino acid (eg. an unnatural amino acid), or a substituted linkage, as compared with the sequence of the reference polypeptide. In a further embodiment, a polypeptide of interest may be a mimic of the reference polypeptide, which mimic reproduces at least one epitope of the reference polypeptide. Mutants of the disclosed polynucleotide and polypeptide sequences of the invention can be generated through DNA shuffling. Briefly, mutant DNAs are generated by in vitro homologous recombination by random fragmentation of a parent DNA followed by reassembly using PCR, resulting in randomly introduced point mutations. This technique can be modified by using a family of parent DNAs, to introduce additional variability into the process. Selection or screening for the desired activity, followed by additional iterations of mutagenesis and assay provides for rapid "evolution" of sequences by selecting for desirable mutations while simultaneously selecting against detrimental changes. Mutagenesis methods as disclosed above can be combined with high-throughput screening methods to detect activity of cloned mutant polypeptides. Mutagenized nucleic acid molecules that encode polypeptides of the invention, or fragments thereof, can be recovered from the host cells and rapidly sequenced using modern equipment. These methods allow the rapid determination of the importance of individual amino acid residues in a polypeptide of interest, and can be applied to polypeptides of unknown structure.

[0242] A "fragment" of a polypeptide of interest comprises a series of consecutive amino acid residues from the sequence of said polypeptide. By way of example, a "fragment" of a polypeptide of interest may comprise (or consist of) at least 50 consecutive amino acid residues from the sequence of said polypeptide (eg. at least 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 consecutive amino acid residues of said polypeptide). A fragment includes at least one epitope of the polypeptide of interest.

[0243] As used herein, the terms "nucleic acid sequence" and "polynucleotide" are used interchangeably and do not imply any length restriction. As used herein, the terms "nucleic acid" and "nucleotide" are used interchangeably. The terms "nucleic acid sequence" and "polynucleotide" embrace DNA (including cDNA) and RNA sequences. The polynucleotide sequences of the present invention include nucleic acid sequences that have been removed from their naturally occurring environment, recombinant or cloned DNA isolates, and chemically synthesized analogues or analogues biologically synthesized by heterologous systems. The natural or synthetic DNA fragments coding for a desired fragment may be incorporated into recombinant nucleic acid constructs, typically DNA constructs, capable of introduction into and replication in a prokaryotic or eukaryotic cell. Usually the DNA constructs will be suitable for autonomous replication in a unicellular host, such as yeast or bacteria, but may also be intended for introduction to and integration within the genome of a cultured insect, mammalian, plant or other eukaryotic cell lines. The term "recombinant" as used herein intends a polynucleotide of genomic, cDNA, semi-synthetic, or synthetic origin which, by virtue of its origin or manipulation: (1) is not associated with all or a portion of a polynucleotide with which it is associated in nature; or (2) is linked to a polynucleotide other than that to which it is linked in nature; and (3) does not occur in nature. When applied to a nucleic acid sequence, the term "isolated" in the context of the present invention denotes that the polynucleotide sequence has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences (but may include naturally occurring 5' and 3' untranslated regions such as promoters and terminators), and is in a form suitable for use within genetically engineered protein production systems. Such isolated molecules are those that are separated from their natural environment. A "variant" nucleic acid sequence has substantial homology or substantial similarity to a reference nucleic acid sequence (or a fragment thereof). A nucleic acid sequence or fragment thereof is "substantially homologous" (or "substantially identical") to a reference sequence if, when optimally aligned (with appropriate nucleotide insertions or deletions) with the other nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 70%, 75%, 80%, 82, 84, 86, 88, 90, 92, 94, 96, 98 or 99% of the nucleotide bases. Homology determination is performed as described supra for polypeptides. Alternatively, a "variant" nucleic acid sequence is substantially homologous with (or substantially identical to) a reference sequence (or a fragment thereof) if the "variant" and the reference sequence they are capable of hybridizing under stringent (eg. highly stringent) hybridization conditions. Nucleic acid sequence hybridization will be affected by such conditions as salt concentration (eg. NaCl), temperature, or organic solvents, in addition to the base composition, length of the complementary strands, and the number of nucleotide base mismatches between the hybridizing nucleic acids, as will be readily appreciated by those skilled in the art. Stringent temperature conditions are preferably employed, and generally include temperatures in excess of 30.degree. C., typically in excess of 37.degree. C. and preferably in excess of 45.degree. C. Stringent salt conditions will ordinarily be less than 1000 mM, typically less than 500 mM, and preferably less than 200 mM. The pH is typically between 7.0 and 8.3. The combination of parameters is much more important than any single parameter. A "fragment" of a polynucleotide of interest comprises a series of consecutive amino acid residues from the sequence of said full-length polynucleotide. By way of example, a "fragment" of a polynucleotide of interest may comprise (or consist of) at least 150 consecutive nucleic acid residues from the sequence of said polypeptide (eg. at least 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700 or 750 consecutive nucleic acid residues of said polynucleotide). A fragment encodes at least one antigenic epitope of the corresponding polypeptide of interest.

TABLE-US-00005 SEQ ID No. 1 atgtggtggttccgccgccgagaccgggcgccgctgcgcgccaccagctcattatccctgcggtggcgggtcat- gctgctggcgatgtccatg gtcgcgatggtggttgtgctgatgtcgttcgccgtctatgcggtgatctcggccgcgctctacagcgacatcga- caaccaactgcagagccggg cgcaactgctcatcgccagtggctcgctggcagctgatccgggtaaggcaatcgagggtaccgcctattcggat- gtcaacgcgatgctggtca accccggccagtccatctacaccgctcaacagccgggccagacgctgccggtcggtgctgccgagaaggcggtg- atccgtggcgagttgtt catgtcgcggcgcaccaccgccgaccaacgggtgcttgccatccgtctgaccaacggtagttcgctgctgatct- ccaaaagtctcaagccca ccgaagcagtcatgaacaagctgcgttgggtgctattgatcgtgggtgggatcggggtggcggtcgccgcggtg- gccggggggatggtcac ccgggccgggctgaggccggtgggccgcctcaccgaagcggccgagcgggtggcgcgaaccgacgacctgcggc- ccatccccgtcttc ggcagcgacgaattggccaggctgacagaggcattcaatttaatgctgcgggcgctggccgagtcacgggaacg- gcaggcaaggctggtt accgacgccggacatgaattgcgtaccccgctaacgtcgctgcgcaccaatgtcgaactcttgatggcctcgat- ggccccgggggctccgcg gctacccaagcaggagatggtcgacctgcgtgccgatgtgctggctcaaatcgaggaattgtccacactggtag- gcgatttggtggacctgtc ccgaggcgacgccggagaagtggtgcacgagccggtcgacatggctgacgtcgtcgaccgcagcctggagcggg- tcaggcggcggcgc aacgatatccttttcgacgtcgaggtgattgggtggcaggtttatggcgataccgctggattgtcgcggatggc- gcttaacctgatggacaacgc cgcgaagtggagcccgccgggcggccacgtgggtgtcaggctgagccagctcgacgcgtcgcacgctgagctgg- tggtttccgaccgcgg cccgggcattcccgtgcaggagcgccgtctggtgtttgaacggttttaccggtcggcatcggcacgggcgttgc- cgggttcgggcctcgggttg gcgatcgtcaaacaggtggtgctcaaccacggcggattgctgcgcatcgaagacaccgacccaggcggccagcc- ccctggaacgtcgatt tacgtgctgctccccggccgtcggatgccgattccgcagcttcccggtgcgacggctggcgctcggagcacgga- catcgagaactctcgggg ttcggcgaacgttatctcagtggaatctcagtccacgcgcgcaacctag SEQ ID No. 2 MWWFRRRDRAPLRATSSLSLRWRVMLLAMSMVAMVVVLMSFAVYAVISAALYSDIDNQLQSRAQLL IASGSLAADPGKAIEGTAYSDVNAMLVNPGQSIYTAQQPGQTLPVGAAEKAVIRGELFMSRRTTADQ RVLAIRLTNGSSLLISKSLKPTEAVMNKLRWVLLIVGGIGVAVAAVAGGMVTRAGLRPVGRLTEAAER VARTDDLRPIPVFGSDELARLTEAFNLMLRALAESRERQARLVTDAGHELRTPLTSLRTNVELLMASM APGAPRLPKQEMVDLRADVLAQIEELSTLVGDLVDLSRGDAGEVVHEPVDMADVVDRSLERVRRRR NDILFDVEVIGWQVYGDTAGLSRMALNLMDNAAKWSPPGGHVGVRLSQLDASHAELVVSDRGPGIP VQERRLVFERFYRSASARALPGSGLGLAIVKQVVLNHGGLLRIEDTDPGGQPPGTSIYVLLPGRRMPI PQLPGATAGARSTDIENSRGSANVISVESQSTRAT SEQ ID No. 3 atgacggctccgggactgacagcagccgtcgaggggatcgcacacaacaagggcgagctgttcgcctcctttga- cgtggacgcgttcgagg ttccgcacggccgcgacgagatctggcggttcaccccgttgcggcggctgcgtggcctgcacgacggctccgcg- cgggccaccggtagcg ccacgatcacggtcagcgagcggccgggcgtatacacccagaccgtgcgccgcggcgatccacgactgggcgag- ggcggcgtacccac cgaccgcgttgccgcccaagcgttttcgtcgttcaactccgcgactctggtcaccgtcgagcgcgacacccagg- tcgtcgagccggtaggcat caccgtgaccgggccgggggagggcgcggtggcctatgggcacctgcaggtgcgtatcgaggagcttggcgagg- cggtcgtggtcatcga ccaccggggcggcggaacctacgccgacaacgtcgagttcgttgtcgacgacgccgctcggctgaccgccgtgt- ggatcgccgactgggc cgacaacaccgttcacctcagcgcgcaccatgctcggatcggcaaggacgcggtgctgcgccacgtcaccgtca- tgttgggcggcgacgtg gtgcgaatgtcggcgggcgtgcggttctgcggtgcgggtggggacgcggaactgctggggctgtatttcgccga- cgacggccagcacctgg agtcgcggctgctggtggaccacgcccaccccgactgcaagtcgaacgtgctgtataagggtgcactgcaaggt- gatccggcgtcgtcgttg cccgacgcacacacggtctgggtgggtgacgtgctgatccgtgcgcaggccaccggcaccgacaccttcgaggt- gaaccggaacctggtg ctcaccgacggcgcgcgtgccgactcggtgcccaacctggagatcgagaccggcgagatcgtcggcgccggaca- cgccagcgccaccg gtcgcttcgacgatgagcaattgttctacctgcgttcgcgcggtattcccgaagcacaggcccgccggctggtg- gtccgcggcttcttcggtgag atcatcgccaagatcgcggtgcccgaggtacgcgagcgcctgaccgcagccatcgaacacgagctggaaatcac- ggaatcaacggaaa agacaacagtctcatga SEQ ID No. 4 MTAPGLTAAVEGIAHNKGELFASFDVDAFEVPHGRDEIWRFTPLRRLRGLHDGSARATGSATITVSE RPGVYTQTVRRGDPRLGEGGVPTDRVAAQAFSSFNSATLVTVERDTQVVEPVGITVTGPGEGAVAY GHLQVRIEELGEAVVVIDHRGGGTYADNVEFVVDDAARLTAVWIADWADNTVHLSAHHARIGKDAVL RHVTVMLGGDVVRMSAGVRFCGAGGDAELLGLYFADDGQHLESRLLVDHAHPDCKSNVLYKGALQ GDPASSLPDAHTVWVGDVLIRAQATGTDTFEVNRNLVLTDGARADSVPNLEIETGEIVGAGHASATG RFDDEQLFYLRSRGIPEAQARRLVVRGFFGEIIAKIAVPEVRERLTAAIEHELEITESTEKTTVS SEQ ID No. 5 atggcggttcgtcaggtcaccgtcggctattcggacggcacgcacaagacgatgccggtgcggtgcgaccagac- ggtcctggatgccgccg aggaacacggcgtggccatcgtcaacgaatgccaaagcgggatatgtggcacctgcgtggccacctgcaccgcc- ggccgctaccagatg ggacgcaccgagggactgtccgatgtcgagcgggcggcgcgaaagatcctcacctgccagacgtttgttacctc- cgattgccggatcgagct gcagtatccggtcgacgacaacgccgccctgctggtcaccggtgacggtgtggtgaccgcggtcgagttggtgt- cgcccagcaccgccatcc tgcgggtggacacctctggcatggccggcgcgctgagataccgggccggccagttcgcccaattgcaggttccc- ggtaccaacgtatggcg caactactcctacgcccatccggccgacggccgcggtgagtgcgagttcatcatcaggttgctgccggacggcg- tgatgtcgaattatcttcgc gaccgcgcccagcccggtgaccatatcgcgctgcgctgcagcaagggcagcttttatctgcgcccgatcgtgcg- accggtgatcctggtcgc cggaggaaccggcctgtcagcgatcctggcgatggcccagagcctggatgccgatgtcgctcacccggtctacc- tgctctacggggtcgagc gcaccgaagacctgtgcaagctcgacgaactcaccgagctgcgccgccgcgttggccgcctggaggtgcacgtc- gtcgtcgctcgcccgg accccgactgggatgggcgcaccgggctggtcaccgacctgctcgacgagcggatgctggcgagcggtgacgcc- gacgtgtatctgtgcg gtccggtcgccatggtcgacgcagcccgaacctggctggaccacaatggctttcaccgtgtcgggttgtactac- gagaagttcgtggccagcg gggcggcgcgccgccgcaccccggctcggctggattacgcgggcgtggacattgccgaggtgtgccgccgcggc- cgcggcaccgcggtg gtcatcggcggcagcatcgcgggcatcgcggcggcgaaaatgctcagcgagaccttcgatcgcgtcatcgtgct- ggagaaggacggcccg caccgtcgccgcgagggcaggccgggcgcggcacagggttggcacctgcaccacctgctgaccgccgggcagat- cgagctggagcgca tcttccctggcatcgtcgacgacatggtgcgcgagggagcgttcaaggtcgacatggccgcgcagtaccgtatc- cggctgggcggcacctgg aagaagcccggcactagtgacatcgagatcgtctgcgcgggaaggccgctgctcgaatggtgtgtgcgccgccg- gctcgacgacgaaccg cgcatcgacttccgctacgaatcggaggtggccgatctcgccttcgaccgcgccaacaatgccatcgtcggcgt- cgccgtggacaatggcga cgccgacggaggcgacggtttgcaggtggtgcccgccgagttcgtcgtggacgcgtcgggcaagaacacccgcg- tgccggagttcttggag cgtctcggtgttggcgctcccgaggccgagcaggacatcatcaactgcttctactccacgatgcagcaccgggt- tccgccggagcggcggtg gcaggacaaggtgatggtgatctgctatgcgtaccgccctttcgaggatacctacgccgcgcagtactacaccg- acagctcccgcaccatcct gtccacctcactggtggcctacaactgctattcgccgccgcgtaccgcccgagaattccgcgcgttcgccgacc- tgatgccgtccccggtcatc ggggagaacatcgacgggctggagccggcatcgcccatctacaatttccgctatcccaacatgctgcggctgcg- ctacgagaagaagcgc aacctgccgcgggctttgctggcggtgggcgatgcctacaccagcgccgacccggtgtcgggtctgggtatgag- cctggcgctcaaggaagt tcgggagatgcaggcgctgctggctaaatacggcgccggtcaccgggatctgccgcgccggtactaccgggcga- tcgccaagatggccga cacggcctggttcgtgatccgcgagcagaacctgcgcttcgactggatgaaggacgtcgacaagaagcgcccgt- tctatttcggtgtgctgac ctggtacatggaccgcgtgctggagctggtgcatgacgatctcgacgcgtaccgggaattcttggccgtcgtcc- atctggtcaagccgccgtcg gcgctgatgcgacccaggatcgccagccgcgtcctcggcaaatgggcacgaacccgattgtcgggccagaagac- gttgattgcccgcaac tacgaaaatcatccgataccagccgaacccgcggaccaacttgtaaacgcttag SEQ ID No. 6 MAVRQVTVGYSDGTHKTMPVRCDQTVLDAAEEHGVAIVNECQSGICGTCVATCTAGRYQMGRTEG LSDVERAARKILTCQTFVTSDCRIELQYPVDDNAALLVTGDGVVTAVELVSPSTAILRVDTSGMAGAL RYRAGQFAQLQVPGTNVWRNYSYAHPADGRGECEFIIRLLPDGVMSNYLRDRAQPGDHIALRCSKG SFYLRPIVRPVILVAGGTGLSAILAMAQSLDADVAHPVYLLYGVERTEDLCKLDELTELRRRVGRLEVH VVVARPDPDWDGRTGLVTDLLDERMLASGDADVYLCGPVAMVDAARTWLDHNGFHRVGLYYEKFV ASGAARRRTPARLDYAGVDIAEVCRRGRGTAVVIGGSIAGIAAAKMLSETFDRVIVLEKDGPHRRREG RPGAAQGWHLHHLLTAGQIELERIFPGIVDDMVREGAFKVDMAAQYRIRLGGTWKKPGTSDIEIVCA GRPLLEWCVRRRLDDEPRIDFRYESEVADLAFDRANNAIVGVAVDNGDADGGDGLQVVPAEFVVDA SGKNTRVPEFLERLGVGAPEAEQDIINCFYSTMQHRVPPERRWQDKVMVICYAYRPFEDTYAAQYY TDSSRTILSTSLVAYNCYSPPRTAREFRAFADLMPSPVIGENIDGLEPASPIYNFRYPNMLRLRYEKKR NLPRALLAVGDAYTSADPVSGLGMSLALKEVREMQALLAKYGAGHRDLPRRYYRAIAKMADTAWFVI REQNLRFDWMKDVDKKRPFYFGVLTWYMDRVLELVHDDLDAYREFLAVVHLVKPPSALMRPRIASR VLGKWARTRLSGQKTLIARNYENHPIPAEPADQLVNA SEQ ID No. 7 atgaccacaacgactacaacgatttctggggggatattacccaaggaataccaagatcttcgggatacggtggc- cgattttgcgcgcaccgtg gtcgcgccggtatcggccaaacacgatgcggaacacagcttcccatacgaaattgtcgccaagatgggagagat- gggcctgttcgggctgc cgtttccggaggagtacggcggcatgggcggcgactacttcgcgctgtcgctggtacttgaggagctgggcaag- gttgaccaatcggtagcg atcacgctggaggccgcggtgggcctgggtgcgatgccgatctaccggttcggtaccgaggagcagaaacagaa- gtggttgcccgacttga cgtctggccgtgcgctcgccggttttggtctcaccgagccgggagcgggatcggacgcgggcagcacccgcacc- acggcgcgtctcgaag

gtgacgagtggatcatcaacggctccaagcaatttatcaccaactcgggcaccgacatcacatcgctggtcacc- gtcactgcggttaccggg accaccggaaccgctgcggatgccaagaaagagatttcgacgatcatcgtgcccagcggcacaccgggattcac- cgtggaaccggtctat aacaaggtcggctggaacgcctcggacacccacccactgacatttgccgatgcgcgggtcccgagggagaacct- gctgggagcccgggg gagcggctatgccaacttcttgtccatcctggacgagggccggattgcgattgcagcgctggccaccggcgcgg- cgcagggctgtgttgacg agagcgtcaagtacgccaaccagcgtcagtcgtttggccagccgatcggcgcttatcaggcgatcggcttcaag- atcgcgcggatggaggc acgcgcccatgttgcccgcacagcgtactatgatgccgccgcaaagatgttggcgggcaagcccttcaagaagg- aggcggcgatcgcgaa gatgatctcctcggaggcggcgatggacaactcccgcgatgccacccagatacacggcggatacggctttatga- acgaatatccggtggcg cgtcattaccgcgacagcaaggtgctcgagattggtgagggcaccacggaagtgcagctgatgcttatcgcgcg- atcgttgggactgcagtg a SEQ ID No. 8 MTTTTTTISGGILPKEYQDLRDTVADFARTVVAPVSAKHDAEHSFPYEIVAKMGEMGLFGLPFPEEYG GMGGDYFALSLVLEELGKVDQSVAITLEAAVGLGAMPIYRFGTEEQKQKWLPDLTSGRALAGFGLTE PGAGSDAGSTRTTARLEGDEWIINGSKQFITNSGTDITSLVTVTAVTGTTGTAADAKKEISTIIVPSGTP GFTVEPVYNKVGWNASDTHPLTFADARVPRENLLGARGSGYANFLSILDEGRIAIAALATGAAQGCV DESVKYANQRQSFGQPIGAYQAIGFKIARMEARAHVARTAYYDAAAKMLAGKPFKKEAAIAKMISSEA AMDNSRDATQIHGGYGFMNEYPVARHYRDSKVLEIGEGTTEVQLMLIARSLGLQ SEQ ID No. 9 atggacaaggtggtggccaccgccgcggaggcggtcgcagacatagccaacgggtcgtcgcttgcggttggtgg- attcgggctttgcggcat ccccgaagcactgatcgcagcgttggtggatagcggtgtcaccgacctggaaacagtctcgaacaactgcggaa- tcgacggtgttggtctgg gactattgttgcaacacaagcgaattcgccggacagtctcctcctacgtgggggagaacaaggagttcgcccgc- cagttcctcgcgggcgag ctcgaggtggaactgaccccgcagggcacgctggccgagcggttgcgggccggagggatgggcataccggcctt- ctatacaccggcagg ggtcggtacccaggtcgccgacggcgggttgccgtggcgctacgacgcctcgggcggggtggcggtggtgtcgc- cggccaaggagactcg ggagttcgatggtgtcacctatgtcctcgagcgggggatccggaccgacttcgcactggtgcatgcctggcagg- gggaccggcacggcaac ctgatgtaccgccacgccgcggccaacttcaacccggagtgcgcatccgcaggcaggatcacgatcgccgaggt- cgagcacttggtcgag ccgggtgagatcgaccctgccaccgtacacaccccgggcgtgtttgtgcaccgggtggttcatgtgcccaaccc- cgccaagaagatcgaga gggagacggtgcggcaatga SEQ ID No. 10 MDKVVATAAEAVADIANGSSLAVGGFGLCGIPEALIAALVDSGVTDLETVSNNCGIDGVGLGLLLQHK RIRRTVSSYVGENKEFARQFLAGELEVELTPQGTLAERLRAGGMGIPAFYTPAGVGTQVADGGLPW RYDASGGVAVVSPAKETREFDGVTYVLERGIRTDFALVHAWQGDRHGNLMYRHAAANFNPECASA GRITIAEVEHLVEPGEIDPATVHTPGVFVHRVVHVPNPAKKIERETVRQ SEQ ID No. 11 atgaccctggaagtggtatcggacgcggccggacgcatgcgggtcaaagtcgactgggtccgttgcgattcccg- gcgcgcggtcgcggtcg aagaggccgttgccaagcagaacggtgtgcgcgtcgtgcacgcctacccgcgcaccgggtccgtggtcgtgtgg- tattcacccagacgcgc cgaccgcgcggcggtgctggcggcgatcaagggcgccgcgcacgtcgccgccgaactgatccccgcgcgtgcgc- cgcactcggccgag atccgcaacaccgacgtgctccggatggtcatcggcggggtggcactggccttgctcggggtgcgccgctacgt- gttcgcgcggccaccgct gctcggaaccaccgggcggacggtggccaccggtgtcaccattttcaccgggtatccgttcctgcgtggcgcgc- tgcgctcgctgcgctccgg aaaggccggcaccgatgccctggtctccgcggcgacggtggcaagcctcatcctgcgcgagaacgtggtcgcac- tcaccgtcctgtggttgc tcaacatcggtgagtacctgcaggatctgacgctgcggcggacccggcgggccatctcggagctgctgcgcggc- aaccaggacacggcct gggtgcgcctcaccgatccttctgcaggctccgacgcggccaccgaaatccaggtcccgatcgacaccgtgcag- atcggtgacgaggtggt ggtccacgagcacgtcgcgataccggtcgacggtgaggtggtcgacggcgaagcgatcgtcaatcagtccgcga- tcaccggggaaaacct gccggtcagcgtcgtggtcggaacgcgcgtgcacgccggttcggtcgtggtgcgcggacgcgtggtggtgcgcg- cccacgcggtaggcaa ccaaaccaccatcggtcgcatcattagcagggtcgaagaggctcagctcgaccgggcacccatccagacggtgg- gcgagaacttctcccg ccgcttcgttcccacctcgttcatcgtctcggccatcgcgttgctgatcaccggcgacgtgcggcgcgcgatga- ccatgttgttgatcgcatgccc gtgcgcggtgggactgtccaccccgaccgcgatcagcgcagcgatcggcaacggcgcgcgccgtggcatcctga- tcaagggcggatccc acctcgagcaggcgggccgcgtcgacgccatcgtgttcgacaagaccgggacgttgaccgtgggccgccccgtg- gtcaccaatatcgttgc catgcataaagattgggagcccgagcaagtgctggcctatgccgccagctcggagatccactcacgtcatccgc- tggccgaggcggtgatc cgctcgacggaggaacgccgcatcagcatcccaccacacgaggagtgcgaggtgctggtcggcctgggcatgcg- gacctgggccgacg gtcggaccctgctgctgggcagtccgtcgttgctgcgcgccgaaaaagttcgggtgtccaagaaggcgtcggag- tgggtcgacaagctgcg ccgccaggcggagaccccgctgctgctcgcggtggacggcacgctggtcggcctgatcagcctgcgcgacgagg- tgcgtccggaggcgg cccaggtgctgacgaagctgcgggccaatgggattcgccggatcgtcatgctcaccggcgaccacccggagatc- gcccaggttgtcgccga cgaactggggattgatgagtggcgcgccgaggtcatgccggaggacaagctcgcggcggtgcgcgagctgcagg- acgacggctacgtcg tcgggatggtcggcgacggcatcaacgacgccccggcgctggccgccgccgatatcgggatcgccatgggcctt- gccggaaccgacgtcg ccgtcgagaccgccgatgtcgcgctggccaacgacgacctgcaccgcctgctcgacgttggggacctgggcgag- cgggcagtggatgtaa tccggcagaactacggcatgtccatcgccgtcaacgcggccgggctgctgatcggcgcgggcggtgcgctctcg- ccggtgctggcggcgat cctgcacaacgcgtcgtcggtggcggtggtggccaacagttcccggttgatccgctaccgcctggaccgctag SEQ ID No. 12 MTLEVVSDAAGRMRVKVDWVRCDSRRAVAVEEAVAKQNGVRVVHAYPRTGSVVVWYSPRRADRA AVLAAIKGAAHVAAELIPARAPHSAEIRNTDVLRMVIGGVALALLGVRRYVFARPPLLGTTGRTVATGV TIFTGYPFLRGALRSLRSGKAGTDALVSAATVASLILRENVVALTVLWLLNIGEYLQDLTLRRTRRAISE LLRGNQDTAWVRLTDPSAGSDAATEIQVPIDTVQIGDEVVVHEHVAIPVDGEVVDGEAIVNQSAITGE NLPVSVVVGTRVHAGSVVVRGRVVVRAHAVGNQTTIGRIISRVEEAQLDRAPIQTVGENFSRRFVPTS FIVSAIALLITGDVRRAMTMLLIACPCAVGLSTPTAISAAIGNGARRGILIKGGSHLEQAGRVDAIVFDKT GTLTVGRPVVTNIVAMHKDWEPEQVLAYAASSEIHSRHPLAEAVIRSTEERRISIPPHEECEVLVGLG MRTWADGRTLLLGSPSLLRAEKVRVSKKASEWVDKLRRQAETPLLLAVDGTLVGLISLRDEVRPEAA QVLTKLRANGIRRIVMLTGDHPEIAQVVADELGIDEWRAEVMPEDKLAAVRELQDDGYVVGMVGDGI NDAPALAAADIGIAMGLAGTDVAVETADVALANDDLHRLLDVGDLGERAVDVIRQNYGMSIAVNAAGL LIGAGGALSPVLAAILHNASSVAVVANSSRLIRYRLDR SEQ ID No. 13 atgactgtgcaggagttcgacgtcgtggtggtcggcagcggcgccgccggcatggttgctgcgctggtcgccgc- tcaccgaggtctctcgacg gtagtcgtcgagaaggccccgcactacggcggctccaccgcacgctcgggcggcggcgtctggatccccaacaa- cgaggtcctcaagcg ccgcggcgttcgagatacaccggaggcggcacgcacctatctgcacggcatcgtcggcgaaatcgtcgagccgg- aacgcatcgatgctta cctcgaccgcgggcccgagatgctgtcgttcgtgctgaagcacacgccgctgaagatgtgctgggtacccggct- actccgactactaccccg aggctccgggcggccgcccgggcggacgttcgatcgagccgaaaccgttcaacgcgcgcaagcttggtgccgac- atggccgggctggag cccgcgtatggcaaggttccgctcaatgtggttgtgatgcagcaggactacgttcgcctcaatcagctcaaacg- tcacccccgtggcgtgctgc gcagcatgaaggtcggcgcccgcacgatgtgggcgaaggcaacaggtaagaacctggtcggcatgggtcgagcc- ctcattgggccgttgc ggatcgggttgcagcgcgccggagtgccggtcgaactcaacaccgccttcaccgatcttttcgtcgaaaatggc- gtcgtgtccggggtatacgt ccgcgattcccacgaggcggaatccgctgagccgcagctgatccgggctcgccgcggcgtgatcctggcctgtg- gtggtttcgagcataacg agcagatgcgaatcaagtaccagcgggcacccatcaccaccgagtggaccgtgggcgccagcgccaataccggt- gacggcattctcgcc gccgaaaagctcggcgcagcactggatctgatggatgacgcttggtggggcccgacggtaccgctggtcggcaa- accatggttcgcgctctc ggagcgcaactctcccggttcgatcatcgtcaacatgtcaggcaagcgattcatgaacgaatcgatgccatacg- tcgaagcctgtcatcatatg tacggcggcgaacacggccaggggcccggaccgggcgagaacattccggcgtggctggtgttcgaccagcgata- ccgggaccgctacat cttcgcgggactacaaccagggcaacgcattccgagcaggtggctggattccggcgtcatcgtccaggccgata- cccttgcggagctggcc ggcaaggccggtctacccgcggacgaactcactgccaccgtccagcgtttcaacgcattcgcccggtccggtgt- cgacgaggactaccacc gcggggaaagtgcctacgatcgctactacggcgacccgagcaacaagcccaatccgaacctcggcgaggtcggc- cacccgccctattatg gcgccaagatggttccgggcgacctggggaccaagggcggtatccgcaccgatgtcaacggacgtgctctgcgg- gacgacggcagcatc atcgacggcctttacgctgcaggcaatgtcagtgccccagtgatgggacacacctaccccggtccgggcggcag- ataggcccggcgatg acgttcgggtacctggcggcgctgcacattgccgatcaggcgggaaagcgctga SEQ ID No. 14 MTVQEFDVVVVGSGAAGMVAALVAAHRGLSTVVVEKAPHYGGSTARSGGGVWIPNNEVLKRRGVR DTPEAARTYLHGIVGEIVEPERIDAYLDRGPEMLSFVLKHTPLKMCWVPGYSDYYPEAPGGRPGGRS IEPKPFNARKLGADMAGLEPAYGKVPLNVVVMQQDYVRLNQLKRHPRGVLRSMKVGARTMWAKAT GKNLVGMGRALIGPLRIGLQRAGVPVELNTAFTDLFVENGVVSGVYVRDSHEAESAEPQLIRARRGVI LACGGFEHNEQMRIKYQRAPITTEWTVGASANTGDGILAAEKLGAALDLMDDAWWGPTVPLVGKPW FALSERNSPGSIIVNMSGKRFMNESMPYVEACHHMYGGEHGQGPGPGENIPAWLVFDQRYRDRYIF AGLQPGQRIPSRWLDSGVIVQADTLAELAGKAGLPADELTATVQRFNAFARSGVDEDYHRGESAYD RYYGDPSNKPNPNLGEVGHPPYYGAKMVPGDLGTKGGIRTDVNGRALRDDGSIIDGLYAAGNVSAP VMGHTYPGPGGTIGPAMTFGYLAALHIADQAGKR SEQ ID No. 15 gtgagtccggcgcccgtgcaggtgatgggggttctaaacgtcacggacgactctttctcggacggcgggtgtta- tctcgatctcgacgatgcgg tgaagcacggtctggcgatggcagccgcaggtgcgggcatcgtcgacgtcggtggtgagtcgagccggcccggt-

gccactcgggttgaccc ggcggtggagacgtctcgtgtcatacccgtcgtcaaagagcttgcagcacaaggcatcaccgtcagcatcgata- ccatgcgcgcggatgtcg ctcgggcggcgttgcagaacggtgcccagatggtcaacgacgtgtcgggtgggcgggccgatccggcgatgggg- ccgctgttggccgagg ccgatgtgccgtgggtgttgatgcactggcgggcggtatcggccgataccccgcatgtgcctgtgcgctacggc- aacgtggtggccgaggtcc gtgccgacctgctggccagcgtcgccgacgcggtggccgcaggcgtcgacccggcaaggctggtgctcgatccc- gggcttggattcgccaa gacggcgcaacataattgggcgatcttgcatgcccttccggaactggtcgcgaccggaatcccagtgctggtgg- gtgcttcgcgcaagcgctt cctcggtgcgttgttggccgggcccgacggcgtgatgcggccaaccgatgggcgtgacaccgcgacggcggtga- tttccgcgctggccgca ctgcacggggcctggggtgtgcgggtgcatgatgtgcgggcctcggtcgatgccatcaaggtggtcgaagcgtg- gatgggagcggaaagg atagaacgcgatggctga SEQ ID No. 16 VSPAPVQVMGVLNVTDDSFSDGGCYLDLDDAVKHGLAMAAAGAGIVDVGGESSRPGATRVDPAVE TSRVIPVVKELAAQGITVSIDTMRADVARAALQNGAQMVNDVSGGRADPAMGPLLAEADVPWVLMH WRAVSADTPHVPVRYGNVVAEVRADLLASVADAVAAGVDPARLVLDPGLGFAKTAQHNWAILHALP ELVATGIPVLVGASRKRFLGALLAGPDGVMRPTDGRDTATAVISALAALHGAWGVRVHDVRASVDAI KVVEAWMGAERIERDG SEQ ID No. 17 atgagtattaccaggccgacgggcagctatgccagacagatgctggatccgggcggctgggtggaagccgatga- agacactttctatgacc gggcccaggaatatagccaggttttgcaaagggtcaccgatgtattggacacctgccgccagcagaaaggccac- gtcttcgaaggcggcct atggtccggcggcgccgccaatgctgccaacggcgccctgggtgcaaacatcaatcaattgatgacgctgcagg- attatctcgccacggtga ttacctggcacaggcatattgccgggttgattgagcaagctaaatccgatatcggcaataatgtggatggcgct- caacgggagatcgatatcct ggagaatgaccctagcctggatgctgatgagcgccataccgccatcaattcattggtcacggcgacgcatgggg- ccaatgtcagtctggtcgc cgagaccgctgagcgggtgctggaatccaagaattggaaacctccgaagaacgcactcgaggatttgcttcagc- agaagtcgccgccacc cccagacgtgcctaccctggtcgtgccatccccgggcacaccgggcacaccgggaaccccgatcaccccgggaa- ccccgatcaccccgg gaaccccaatcacacccatcccgggagcgccggtaactccgatcacaccaacgcccggcactcccgtcacgccg- gtgaccccgggcaa gccggtcaccccggtgaccccggtcaaaccgggcacaccaggcgagccaaccccgatcacgccggtcacccccc- cggtcgccccggcc acaccggcaaccccggccacgcccgttaccccagctcccgctccacacccgcagccggctccggcaccggcgcc- atcgcctgggcccca gccggttacaccggccactcccggtccgtctggtccagcaacaccgggcaccccagggggcgagccggcgccgc- acgtcaaacccgcg gcgttggcggagcaacctggtgtgccgggccagcatgcgggcggggggacgcagtcggggcctgcccatgcgga- cgaatccgccgcgtc ggtgacgccggctgcggcgtccggtgtcccgggcgcacgggcggcggccgccgcgccgagcggtaccgccgtgg- gagcgggcgcgcgt tcgagcgtgggtacggccgcggcctcgggcgcggggtcgcatgctgccactgggcgggcgccggtggctacctc- ggacaaggcggcggc accgagcacgcgggcggcctcggcgcggacggcacctcctgcccgcccgccgtcgaccgatcacatcgacaaac- ccgatcgcagcgag tctgcagatgacggtacgccggtgtcgatgatcccggtgtcggcggctcgggcggcacgcgacgccgccactgc- agctgccagcgcccgc cagcgtggccgcggtgatgcgctgcggttggcgcgacgcatcgcggcggcgctcaacgcgtccgacaacaacgc- gggcgactacgggtt cttctggatcaccgcggtgaccaccgacggttccatcgtcgtggccaacagctatgggctggcctacatacccg- acgggatggaattgccga ataaggtgtacttggccagcgcggatcacgcaatcccggttgacgaaattgcacgctgtgccacctacccggtt- ttggccgtgcaagcctggg cggctttccacgacatgacgctgcgggcggtgatcggtaccgcggagcagttggccagttcggatcccggtgtg- gccaagattgtgctggag ccagatgacattccggagagcggcaaaatgacgggccggtcgcggctggaggtcgtcgacccctcggcggcggc- tcagctggccgacac taccgatcagcgtttgctcgacttgttgccgccggcgccggtggatgtcaatccaccgggcgatgagcggcaca- tgctgtggttcgagctgatg aagcccatgaccagcaccgctaccggccgcgaggccgctcatctgcgggcgttccgggcctacgctgcccactc- acaggagattgccctgc accaagcgcacactgcgactgacgcggccgtccagcgtgtggccgtcgcggactggctgtactggcaatacgtc- accgggttgctcgaccg ggccctggccgccgcatgctga SEQ ID No. 18 MSITRPTGSYARQMLDPGGWVEADEDTFYDRAQEYSQVLQRVTDVLDTCRQQKGHVFEGGLWSG GAANAANGALGANINQLMTLQDYLATVITWHRHIAGLIEQAKSDIGNNVDGAQREIDILENDPSLDADE RHTAINSLVTATHGANVSLVAETAERVLESKNWKPPKNALEDLLQQKSPPPPDVPTLVVPSPGTPGT PGTPITPGTPITPGTPITPIPGAPVTPITPTPGTPVTPVTPGKPVTPVTPVKPGTPGEPTPITPVTPPVAP ATPATPATPVTPAPAPHPQPAPAPAPSPGPQPVTPATPGPSGPATPGTPGGEPAPHVKPAALAEQP GVPGQHAGGGTQSGPAHADESAASVTPAAASGVPGARAAAAAPSGTAVGAGARSSVGTAAASGA GSHAATGRAPVATSDKAAAPSTRAASARTAPPARPPSTDHIDKPDRSESADDGTPVSMIPVSAARAA RDAATAAASARQRGRGDALRLARRIAAALNASDNNAGDYGFFWITAVTTDGSIVVANSYGLAYIPDG MELPNKVYLASADHAIPVDEIARCATYPVLAVQAWAAFHDMTLRAVIGTAEQLASSDPGVAKIVLEPD DIPESGKMTGRSRLEVVDPSAAAQLADTTDQRLLDLLPPAPVDVNPPGDERHMLWFELMKPMTSTA TGREAAHLRAFRAYAAHSQEIALHQAHTATDAAVQRVAVADWLYWQYVTGLLDRALAAAC SEQ ID NO: 19 MQLVDRVRGAVTGMSRRLVVGAVGAALVSGLVGAVGGTATAGAFSRPGLPVEYLQVPSPSMGRDI KVQFQSGGANSPALYLLDGLRAQDDFSGWDINTPAFEWYDQSGLSVVMPVGGQSSFYSDWYQPA CGKAGCQTYKWETFLTSELPGWLQANRHVKPTGSAVVGLSMAASSALTLAIYHPQQFVYAGAMSGL LIDPSQAMGPTLIGLAMGDAGGYKASDMWGPKEDPAWQRNDPLLNVGKLIANNTRVWVYCGNGKP SDLGGNNLPAKFLEGFVRTSNIKFQDAYNAGGGHNGVFDFPDSGTHSWEYWGAQLNAMKPDLQRA LGATPNTGPAPQGA SEQ ID NO: 20 MTDVSRKIRAWGRRLMIGTAAAVVLPGLVGLAGGAATAGAFSRPGLPVEYLQVPSPSMGRDIKVQF QSGGNNSPAVYLLDGLRAQDDYNGWDINTPAFEWYYQSGLSIVMPVGGQSSFYSDWYSPACGKAG CQTYKWETFLTSELPQWLSANRAVKPTGSAAIGLSMAGSSAMILAAYHPQQFIYAGSLSALLDPSQG MGPSLIGLAMGDAGGYKAADMWGPSSDPAWERNDPTQQIPKLVANNTRLWVYCGNGTPNELGGA NIPAEFLENFVRSSNLKFQDAYNAAGGHNAVFNFPPNGTHSVVEYWGAQLNAMKGDLQSSLGAG SEQ ID NO: 21 MTEQQWNFAGIEAAASAIQGNVTSIHSLLDEGKQSLTKLAAAWGGSGSEAYQGVQQKWDATATELN NALQNLARTISEAGQAMASTEGNVTGMFA SEQ ID NO: 22 MSQIMYNYPAMLGHAGDMAGYAGTLQSLGAEIAVEQAALQSAWQGDTGITYQAWQAQWNQAMED LVRAYHAMSSTHEANTMAMMARDTAEAAKWGG SEQ ID NO: 23 MSNSRRRSLRWSWLLSVLAAVGLGLATAPAQAAPPALSQDRFADFPALPLDPSAMVAQVGPQVVNI NTKLGYNNAVGAGTGIVIDPNGVVLTNNHVIAGATDINAFSVGSGQTYGVDVVGYDRTQDVAVLQLR GAGGLPSAAIGGGVAVGEPVVAMGNSGGQGGTPRAVPGRVVALGQTVQASDSLTGAEETLNGLIQ FDAAIQPGDSGGPVVNGLGQVVGMNTAASDNFQLSQGGQGFAIPIGQAMAIAGQIRSGGGSPTVHI GPTAFLGLGVVDNNGNGARVQRVVGSAPAASLGISTGDVITAVDGAPINSATAMADALNGHHPGDVI SVTWQTKSGGTRTGNVTLAEGPPA SEQ ID NO: 24 MVDFGALPPEINSARMYAGPGSASLVAAAQMWDSVASDLFSAASAFQSVVWGLTVGSWIGSSAGL MVAAASPYVAWMSVTAGQAELTAAQVRVAAAAYETAYGLTVPPPVIAENRAELMILIATNLLGQNTPA IAVNEAEYGEMWAQDAAAMFGYAAATATATATLLPFEEAPEMTSAGGLLEQAAAVEEASDTAAANQ LMNNVPQALQQLAQPTQGTTPSSKLGGLWKTVSPHRSPISNMVSMANNHMSMTNSGVSMTNTLSS MLKGFAPAAAAQAVQTAAQNGVRAMSSLGSSLGSSGLGGGVAANLGRAASVGSLSVPQAWAAAN QAVTPAARALPLTSLTSAAERGPGQMLGGLPVGQMGARAGGGLSGVLRVPPRPYVMPHSPAAG SEQ ID NO: 25 MRTPRRHCRRIAVLAAVSIAATVVAGCSSGSKPSGGPLPDAKPLVEEATAQTKALKSAHMVLTVNGKI PGLSLKTLSGDLTTNPTAATGNVKLTLGGSDIDADFVVFDGILYATLTPNQWSDFGPAADIYDPAQVL NPDTGLANVLANFADAKAEGRDTINGQNTIRISGKVSAQAVNQIAPPFNATQPVPATVWIQETGDHQL AQAQLDRGSGNSVQMTLSKWGEKVQVTKPPVS SEQ ID NO: 26 MAKTIAYDEEARRGLERGLNALADAVKVTLGPKGRNVVLEKKWGAPTITNDGVSIAKEIELEDPYEKIG AELVKEVAKKTDDVAGDGTTTATVLAQALVREGLRNVAAGANPLGLKRGIEKAVEKVTETLLKGAKEV ETKEQIAATAAISAGDQSIGDLIAEAMDKVGNEGVITVEESNTFGLQLELTEGMRFDKGYISGYFVTDP ERQEAVLEDPYILLVSSKVSTVKDLLPLLEKVIGAGKPLLIIAEDVEGEALSTLVVNKIRGTFKSVAVKAP GFGDRRKAMLQDMAILTGGQVISEEVGLTLENADLSLLGKARKVVVTKDETTIVEGAGDTDAIAGRVA QIRQEIENSDSDYDREKLQERLAKLAGGVAVIKAGAATEVELKERKHRIEDAVRNAKAAVEEGIVAGG GVTLLQAAPTLDELKLEGDEATGANIVKVALEAPLKQIAFNSGLEPGVVAEKVRNLPAGHGLNAQTGV YEDLLAAGVADPVKVTRSALQNAASIAGLFLTTEAVVADKPEKEKASVPGGGDMGGMDF SEQ ID NO: 27 MAENSNIDDIKAPLLAALGAADLALATVNELITNLRERAEETRTDTRSRVEESRARLTKLQEDLPEQLT ELREKFTAEELRKAAEGYLEAATSRYNELVERGEAALERLRSQQSFEEVSARAEGYVDQAVELTQEA LGTVASQTRAVGERAAKLVGIELPKKAAPAKKAAPAKKAAPAKKAAAKKAPAKKAAAKKVTQK SEQ ID NO: 28 VTQTGKRQRRKFGRIRQFNSGRWQASYTGPDGRVYIAPKTFNAKIDAEAWLTDRRREIDRQLWSPA SGQEDRPGAPFGEYAEGVVLKQRGIKDRTRAHYRKLLDNHILATFADTDLRDITPAAVRRWYATTAVG TPTMRAHSYSLLRAIMQTALADDLIDSNPCRISGASTARRVHKIRPATLDELETITKAMPDPYQAFVLM AAWLAMRYGELTELRRKDIDLHGEVARVRRAVVRVGEGFKVTTPKSDAGVRDISIPPHLIPAIEDHLH KHVNPGRESLLFPSVNDPNRHLAPSALYRMFYKARKAAGRPDLRVHDLRHSGAVLAASTGATLAEL MQRLGHSTAGAALRYQHAAKGRDREIAALLSKLAENQEM SEQ ID NO: 29 VIAGVDQALAATGQASQRAAGASGGVTVGVGVGTEQRNLSVVAPSQFTFSSRSPDFVDETAGQSW CAILGLNQFH SEQ ID NO: 30 MATTLPVQRHPRSLFPEFSELFAAFPSFAGLRPTFDTRLMRLEDEMKEGRYEVRAELPGVDPDKDV DIMVRDGQLTIKAERTEQKDFDGRSEFAYGSFVRTVSLPVGADEDDIKATYDKGILTVSVAVSEGKPT EKHIQIRSTN

SEQ ID NO: 31 MSGRHRKPTTSNVSVAKIAFTGAVLGGGGIAMAAQATAATDGEWDQVARCESGGNWSINTGNGYL GGLQFTQSTWAAHGGGEFAPSAQLASREQQIAVGERVLATQGRGAWPVCGRGLSNATPREVLPAS AAMDAPLDAAAVNGEPAPLAPPPADPAPPVELAANDLPAPLGEPLPAAPADPAPPADLAPPAPADVA PPVELAVNDLPAPLGEPLPAAPADPAPPADLAPPAPADLAPPAPADLAPPAPADLAPPVELAVNDLPA PLGEPLPAAPAELAPPADLAPASADLAPPAPADLAPPAPAELAPPAPADLAPPAAVNEQTAPGDQPA TAPGGPVGLATDLELPEPDPQPADAPPPGDVTEAPAETPQVSNIAYTKKLWQAIRAQDVCGNDALDS LAQPYVIG SEQ ID NO: 32 MLRLVVGALLLVLAFAGGYAVAACKTVTLTVDGTAMRVTTMKSRVIDIVEENGFSVDDRDDLYPAAG VQVHDADTIVLRRSRPLQISLDGHDAKQVWTTASTVDEALAQLAMTDTAPAAASRASRVPLSGMALP VVSAKTVQLNDGGLVRTVHLPAPNVAGLLSAAGVPLLQSDHVVPAATAPIVEGMQIQVTRNRIKKVTE RLPLPPNARRVEDPEMNMSREVVEDPGVPGTQDVTFAVAEVNGVETGRLPVANVVVTPAHEAVVR VGTKPGTEVPPVIDGSIWDAIAGCEAGGNWAINTGNGYYGGVQFDQGTWEANGGLRYAPRADLAT REEQIAVAEVTRLRQGWGAWPVCAARAGAR SEQ ID NO: 33 VHPLPADHGRSRCNRHPISPLSLIGNASATSGDMSSMTRIAKPLIKSAMAAGLVTASMSLSTAVAHAG PSPNWDAVAQCESGGNWAANTGNGKYGGLQFKPATWAAFGGVGNPAAASREQQIAVANRVLAEQ GLDAWPTCGAASGLPIALWSKPAQGIKQIINEIIWAGIQASIPR SEQ ID NO: 34 MTPGLLTTAGAGRPRDRCARIVCTVFIETAVVATMFVALLGLSTISSKADDIDWDAIAQCESGGNWAA NTGNGLYGGLQISQATWDSNGGVGSPAAASPQQQIEVADNIMKTQGPGAWPKCSSCSQGDAPLGS LTHILTFLAAETGGCSGSRDD SEQ ID NO: 35 LKNARTTLIAAAIAGTLVTTSPAGIANADDAGLDPNAAAGPDAVGFDPNLPPAPDAAPVDTPPAPEDA GFDPNLPPPLAPDFLSPPAEEAPPVPVAYSVNWDAIAQCESGGNWSINTGNGYYGGLRFTAGTWRA NGGSGSAANASREEQIRVAENVLRSQGIRAWPVCGRRG SEQ ID NO: 36 MIATTRDREGATMITFRLRLPCRTILRVFSRNPLVRGTDRLEAVVMLLAVTVSLLTIPFAAAAGTAVQD SRSHVYAHQAQTRHPATATVIDHEGVIDSNTTATSAPPRTKITVPARWVVNGIERSGEVNAKPGTKS GDRVGIWVDSAGQLVDEPAPPARAIADAALAALGLWLSVAAVAGALLALTRAILIRVRNASWQHDIDS LFCTQR SEQ ID NO: 37 MTEPAAWDEGKPRIITLTMNPALDITTSVDVVRPTEKMRCGAPRYDPGGGGINVARIVHVLGGCSTAL FPAGGSTGSLLMALLGDAGVPFRVIPIAASTRESFTVNESRTAKQYRFVLPGPSLTVAEQEQCLDELR GAAASAAFVVASGSLPPGVAADYYQRVADICRRSSTPLILDTSGGGLQHISSGVFLLKASVRELRECV GSELLTEPEQLAAAHELIDRGRAEVVVVSLGSQGALLATRHASHRFSSIPMTAVSGVGAGDAMVAAIT VGLSRGWSLIKSVRLGNAAGAAMLLTPGTAACNRDDVERFFELAAEPTEVGQDQYVWHPIVNPEAS P SEQ ID NO: 38 MPDTMVTTDVIKSAVQLACRAPSLHNSQPWRWIAEDHTVALFLDKDRVLYATDHSGREALLGCGAVL DHFRVAMAAAGTTANVERFPNPNDPLHLASIDFSPADFVTEGHRLRADAILLRRTDRLPFAEPPDWD LVESQLRTTVTADTVRIDVIADDMRPELAAASKLTESLRLYDSSYHAELFWWTGAFETSEGIPHSSLV SAAESDRVTFGRDFPVVANTDRRPEFGHDRSKVLVLSTYDNERASLLRCGEMLSAVLLDATMAGLA TCTLTHITELHASRDLVAALIGQPATPQALVRVGLAPEMEEPPPATPRRPIDEVFHVRAKDHR SEQ ID NO: 39 MTTARDIMNAGVTCVGEHETLTAAAQYMREHDIGALPICGDDDRLHGMLTDRDIVIKGLAAGLDPNTA TAGELARDSIYYVDANASIQEMLNVMEEHQVRRVPVISEHRLVGIVTEADIARHLPEHAIVQFVKAICS PMALAS SEQ ID NO: 40 MASSASDGTHERSAFRLSPPVLSGAMGPFMHTGLYVAQSWRDYLGQQPDKLPIARPTIALAAQAFR DEIVLLGLKARRPVSNHRVFERISQEVAAGLEFYGNRRWLEKPSGFFAQPPPLTEVAVRKVKDRRRS FYRIFFDSGFTPHPGEPGSQRWLSYTANNREYALLLRHPEPRPWLVCVHGTEMGRAPLDLAVFRAW KLHDELGLNIVMPVLPMHGPRGQGLPKGAVFPGEDVLDDVHGTAQAVWDIRRLLSWIRSQEEESLIG LNGLSLGGYIASLVASLEEGLACAILGVPVADLIELLGRHCGLRHKDPRRHTVKMAEPIGRMISPLSLT PLVPMPGRFIYAGIADRLVHPREQVTRLWEHWGKPEIVWYPGGHTGFFQSRPVRRFVQAALEQSGL LDAPRTQRDRSA SEQ ID NO: 41 MSTQRPRHSGIRAVGPYAWAGRCGRIGRWGVHQEAMMNLAIVVHPRKVQSATIYQVTDRSHDGRTA RVPGDEITSTVSGWLSELGTQSPLADELARAVRIGDWPAAYAIGEHLSVEIAVAV SEQ ID NO: 42 atgcagcttgttgacagggttcgtggcgccgtcacgggtatgtcgcgtcgactcgtggtcggggccgtcggcgc- ggccctagtgtcgggtctgg tcggcgccgtcggtggcacggcgaccgcgggggcattttcccggccgggcttgccggtggagtacctgcaggtg- ccgtcgccgtcgatggg ccgtgacatcaaggtccaattccaaagtggtggtgccaactcgcccgccctgtacctgctcgacggcctgcgcg- cgcaggacgacttcagcg gctgggacatcaacaccccggcgttcgagtggtacgaccagtcgggcctgtcggtggtcatgccggtgggtggc- cagtcaagcttctactccg actggtaccagcccgcctgcggcaaggccggttgccagacttacaagtgggagaccttcctgaccagcgagctg- ccggggtggctgcagg ccaacaggcacgtcaagcccaccggaagcgccgtcgtcggtctttcgatggctgcttcttcggcgctgacgctg- gcgatctatcacccccagc agttcgtctacgcgggagcgatgtcgggcctgttggacccctcccaggcgatgggtcccaccctgatcggcctg- gcgatgggtgacgctggc ggctacaaggcctccgacatgtggggcccgaaggaggacccggcgtggcagcgcaacgacccgctgttgaacgt- cgggaagctgatcgc caacaacacccgcgtctgggtgtactgcggcaacggcaagccgtcggatctgggtggcaacaacctgccggcca- agttcctcgagggcttc gtgcggaccagcaacatcaagttccaagacgcctacaacgccggtggcggccacaacggcgtgttcgacttccc- ggacagcggtacgcac agctgggagtactggggcgcgcagctcaacgctatgaagcccgacctgcaacgggcactgggtgccacgcccaa- caccgggcccgcgc cccagggcgcctag SEQ ID NO: 43 atgacagacgtgagccgaaagattcgagcttggggacgccgattgatgatcggcacggcagcggctgtagtcct- tccgggcctggtggggct tgccggcggagcggcaaccgcgggcgcgttctcccggccggggctgccggtcgagtacctgcaggtgccgtcgc- cgtcgatgggccgcga catcaaggttcagttccagagcggtgggaacaactcacctgcggtttatctgctcgacggcctgcgcgcccaag- acgactacaacggctggg atatcaacaccccggcgttcgagtggtactaccagtcgggactgtcgatagtcatgccggtcggcgggcagtcc- agcttctacagcgactggt acagcccggcctgcggtaaggctggctgccagacttacaagtgggaaaccttcctgaccagcgagctgccgcaa- tggttgtccgccaacag ggccgtgaagcccaccggcagcgctgcaatcggcttgtcgatggccggctcgtcggcaatgatcttggccgcct- accacccccagcagttca tctacgccggctcgctgtcggccctgctggacccctctcaggggatggggcctagcctgatcggcctcgcgatg- ggtgacgccggcggttaca aggccgcagacatgtggggtccctcgagtgacccggcatgggagcgcaacgaccctacgcagcagatccccaag- ctggtcgcaaacaa cacccggctatgggtttattgcgggaacggcaccccgaacgagttgggcggtgccaacatacccgccgagttct- tggagaacttcgttcgtag cagcaacctgaagttccaggatgcgtacaacgccgcgggcgggcacaacgccgtgttcaacttcccgcccaacg- gcacgcacagctggg agtactggggcgctcagctcaacgccatgaagggtgacctgcagagttcgttaggcgccggctga SEQ ID NO: 44 atgacagagcagcagtggaatttcgcgggtatcgaggccgcggcaagcgcaatccagggaaatgtcacgtccat- tcattccctccttgacga ggggaagcagtccctgaccaagctcgcagcggcctggggcggtagcggttcggaggcgtaccagggtgtccagc- aaaaatgggacgcc acggctaccgagctgaacaacgcgctgcagaacctggcgcggacgatcagcgaagccggtcaggcaatggcttc- gaccgaaggcaacg tcactgggatgttcgcatag SEQ ID NO: 45 atgtcgcaaatcatgtacaactaccccgcgatgttgggtcacgccggggatatggccggatatgccggcacgct- gcagagcttgggtgccga gatcgccgtggagcaggccgcgttgcagagtgcgtggcagggcgataccgggatcacgtatcaggcgtggcagg- cacagtggaaccagg ccatggaagatttggtgcgggcctatcatgcgatgtccagcacccatgaagccaacaccatggcgatgatggcc- cgcgacacggccgaag ccgccaaatggggcggctag SEQ ID NO: 46 atgagcaattcgcgccgccgctcactcaggtggtcatggttgctgagcgtgctggctgccgtcgggctgggcct- ggccacggcgccggccca ggcggccccgccggccttgtcgcaggaccggttcgccgacttccccgcgctgcccctcgacccgtccgcgatgg- tcgcccaagtggggcca caggtggtcaacatcaacaccaaactgggctacaacaacgccgtgggcgccgggaccggcatcgtcatcgatcc- caacggtgtcgtgctg accaacaaccacgtgatcgcgggcgccaccgacatcaatgcgttcagcgtcggctccggccaaacctacggcgt- cgatgtggtcgggtatg accgcacccaggatgtcgcggtgctgcagctgcgcggtgccggtggcctgccgtcggcggcgatcggtggcggc- gtcgcggttggtgagcc cgtcgtcgcgatgggcaacagcggtgggcagggcggaacgccccgtgcggtgcctggcagggtggtcgcgctcg- gccaaaccgtgcagg cgtcggattcgctgaccggtgccgaagagacattgaacgggttgatccagttcgatgccgcgatccagcccggt- gattcgggcgggcccgtc gtcaacggcctaggacaggtggtcggtatgaacacggccgcgtccgataacttccagctgtcccagggtgggca- gggattcgccattccgat cgggcaggcgatggcgatcgcgggccagatccgatcgggtggggggtcacccaccgttcatatcgggcctaccg- ccttcctcggcttgggtg ttgtcgacaacaacggcaacggcgcacgagtccaacgcgtggtcgggagcgctccggcggcaagtctcggcatc- tccaccggcgacgtg atcaccgcggtcgacggcgctccgatcaactcggccaccgcgatggcggacgcgcttaacgggcatcatcccgg- tgacgtcatctcggtga cctggcaaaccaagtcgggcggcacgcgtacagggaacgtgacattggccgagggacccccggcctga SEQ ID NO: 47 atggtggatttcggggcgttaccaccggagatcaactccgcgaggatgtacgccggcccgggttcggcctcgct- ggtggccgcggctcagat gtgggacagcgtggcgagtgacctgttttcggccgcgtcggcgtttcagtcggtggtctggggtctgacggtgg- ggtcgtggataggttcgtcgg cgggtctgatggtggcggcggcctcgccgtatgtggcgtggatgagcgtcaccgcggggcaggccgagctgacc- gccgcccaggtccggg ttgctgcggcggcctacgagacggcgtatgggctgacggtgcccccgccggtgatcgccgagaaccgtgctgaa- ctgatgattctgatagcg

accaacctcttggggcaaaacaccccggcgatcgcggtcaacgaggccgaatacggcgagatgtgggcccaaga- cgccgccgcgatgtt tggctacgccgcggcgacggcgacggcgacggcgacgttgctgccgttcgaggaggcgccggagatgaccagcg- cgggtgggctcctcg agcaggccgccgcggtcgaggaggcctccgacaccgccgcggcgaaccagttgatgaacaatgtgccccaggcg- ctgcaacagctggc ccagcccacgcagggcaccacgccttcttccaagctgggtggcctgtggaagacggtctcgccgcatcggtcgc- cgatcagcaacatggtgt cgatggccaacaaccacatgtcgatgaccaactcgggtgtgtcgatgaccaacaccttgagctcgatgttgaag- ggctttgctccggcggcgg ccgcccaggccgtgcaaaccgcggcgcaaaacggggtccgggcgatgagctcgctgggcagctcgctgggttct- tcgggtctgggcggtg gggtggccgccaacttgggtcgggcggcctcggtcggttcgttgtcggtgccgcaggcctgggccgcggccaac- caggcagtcaccccggc ggcgcgggcgctgccgctgaccagcctgaccagcgccgcggaaagagggcccgggcagatgctgggcgggctgc- cggtggggcagat gggcgccagggccggtggtgggctcagtggtgtgctgcgtgttccgccgcgaccctatgtgatgccgcattctc- cggcggccggctag SEQ ID NO: 48 atgcggacccccagacgccactgccgtcgcatcgccgtcctcgccgccgttagcatcgccgccactgtcgttgc- cggctgctcgtcgggctcg aagccaagcggcggaccacttccggacgcgaagccgctggtcgaggaggccaccgcgcagaccaaggctctcaa- gagcgcgcacatg gtgctgacggtcaacggcaagatcccgggactgtctctgaagacgctgagcggcgatctcaccaccaaccccac- cgccgcgacgggaaa cgtcaagctcacgctgggtgggtctgatatcgatgccgacttcgtggtgttcgacgggatcctgtacgccaccc- tgacgcccaaccagtggag cgatttcggtcccgccgccgacatctacgaccccgcccaggtgctgaatccggataccggcctggccaacgtgc- tggcgaatttcgccgacg caaaagccgaagggcgggataccatcaacggccagaacaccatccgcatcagcgggaaggtatcggcacaggcg- gtgaaccagatag cgccgccgttcaacgcgacgcagccggtgccggcgaccgtctggattcaggagaccggcgatcatcaactggca- caggcccagttggacc gcggctcgggcaattccgtccagatgaccttgtcgaaatggggcgagaaggtccaggtcacgaagcccccggtg- agctga SEQ ID NO: 49 atggccaagacaattgcgtacgacgaagaggcccgtcgcggcctcgagcggggcttgaacgccctcgccgatgc- ggtaaaggtgacattg ggccccaagggccgcaacgtcgtcctggaaaagaagtggggtgcccccacgatcaccaacgatggtgtgtccat- cgccaaggagatcga gctggaggatccgtacgagaagatcggcgccgagctggtcaaagaggtagccaagaagaccgatgacgtcgccg- gtgacggcaccacg acggccaccgtgctggcccaggcgttggttcgcgagggcctgcccaacgtcgcggccggcgccaacccgctcgg- tctcaaacgcggctc gaaaaggccgtggagaaggtcaccgagaccctgctcaagggcgccaaggaggtcgagaccaaggagcagattgc- ggccaccgcagc gatttcggcgggtgaccagtccatcggtgacctgatcgccgaggcgatggacaaggtgggcaacgagggcgtca- tcaccgtcgaggagtc caacacctttgggctgcagctcgagctcaccgagggtatgcggttcgacaagggctacatctcggggtacttcg- tgaccgacccggagcgtc aggaggcggtcctggaggacccctacatcctgctggtcagctccaaggtgtccactgtcaaggatctgctgccg- ctgctcgagaaggtcatcg gagccggtaagccgctgctgatcatcgccgaggacgtcgagggcgaggcgctgtccaccctggtcgtcaacaag- atccgcggcaccttca agtcggtggcggtcaaggctcccggcttcggcgaccgccgcaaggcgatgctgcaggatatggccattctcacc- ggtggtcaggtgatcagc gaagaggtcggcctgacgctggagaacgccgacctgtcgctgctaggcaaggcccgcaaggtcgtggtcaccaa- ggacgagaccaccat cgtcgagggcgccggtgacaccgacgccatcgccggacgagtggcccagatccgccaggagatcgagaacagcg- actccgactacgac cgtgagaagctgcaggagcggctggccaagctggccggtggtctcgcggtgatcaaggccggtgccgccaccga- ggtcgaactcaagga gcgcaagcaccgcatcgaggatgcggttcgcaatgccaaggccgccgtcgaggagggcatcgtcgccggtgggg- gtgtgacgctgttgca agcggccccgaccctggacgagctgaagctcgaaggcgacgaggcgaccggcgccaacatcgtgaaggtggcgc- tggaggccccgct gaagcagatcgccttcaactccgggctggagccgggcgtggtggccgagaaggtgcgcaacctgccggctggcc- acggactgaacgctc agaccggtgtctacgaggatctgctcgctgccggcgttgctgacccggtcaaggtgacccgttcggcgctgcag- aatgcggcgtccatcgcg gggctgttcctgaccaccgaggccgtcgttgccgacaagccggaaaaggagaaggcttccgttcccggtggcgg- cgacatgggtggcatg gatttctga SEQ ID NO: 50 atggctgaaaactcgaacattgatgacatcaaggctccgttgcttcccgcgcttggagcggccgacctggcctt- ggccactgtcaacgagttga tcacgaacctgcgtgagcgtgcggaggagactcgtacggacacccgcagccgggtcgaggagagccgtgctcgc- ctgaccaagctgcag gaagatctgcccgagcagctcaccgagctgcgtgagaagttcaccgccgaggagctgcgtaaggccgccgaggg- ctacctcgaggccgc gactagccggtacaacgagctggtcgagcgcggtgaggccgctctagagcggctgcgcagccagcagagcttcg- aggaagtgtcggcgc gcgccgaaggctacgtggaccaggcggtggagttgacccaggaggcgttgggtacggtcgcatcgcagacccgc- gcggtcggtgagcgt gccgccaagctggtcggcatcgagctgcctaagaaggctgctccggccaagaaggccgctccggccaagaaggc- cgctccggccaaga aggcggcggccaagaaggcgcccgcgaagaaggcggcggccaagaaggtcacccagaagtag SEQ ID NO: 51 gtgacgcaaaccggcaagcgtcagagacgcaaattcggtcgcatccgacagttcaactccggccgctggcaagc- cagctacaccggccc cgacggccgcgtgtacatcgcccccaaaaccttcaacgccaagatcgacgccgaagcatggctcaccgaccgcc- gccgcgaaatcgacc gacaactatggtccccggcatcgggtcaggaagaccgccccggagccccattcggtgagtacgccgaaggatgg- ctgaagcagcgtgga atcaaggaccgcacccgcgcccactatcgcaaactgctggacaaccacatcctggccaccttcgctgacaccga- cctacgcgacatcacc ccggccgccgtgcgccgctggtacgccaccaccgccgtgggcacaccgaccatgcgggcacactcctacagctt- gctgcgcgcaatcatg cagaccgccttggccgacgacctgatcgactccaacccctgccgcatctcaggcgcgtccaccgcccgccgcgt- ccacaagatcaggccc gccaccctcgacgagctggaaaccatcaccaaagccatgcccgacccctaccaggcgttcgtgctgatggcggc- atggctggccatgcgct acggcgagctgaccgaattacgccgcaaagacatcgacctgcacggcgaggttgcgcgggtgcggcgggctgtc- gttcgggtgggcgaa ggcttcaaggtgacgacaccgaaaagcgatgcgggagtgcgcgacataagtatcccgccacatctgatacccgc- catcgaagaccacctt cacaaacacgtcaaccccggccgggagtccctgctgttcccatcggtcaacgaccccaaccgtcacctagcacc- ctcggcgctgtaccgca tgttctacaaggcccgaaaagccgccggccgaccagacttacgggtgcacgaccttcgacactccggcgccgtg- ttggctgcatccaccgg cgccacactggccgaactgatgcagcggctaggacacagcacagccggcgccgcactccgctaccagcacgccg- ccaagggccggga ccgcgaaatcgccgcactgttaagcaaactggccgagaaccaggagatgtga SEQ ID NO: 52 gtgatagcgggcgtcgaccaggcgcttgcagcaacaggccaggctagccagcgggcggcaggcgcatctggtgg- ggtcaccgtcggtgtc ggcgtgggcacggaacagaggaacctttcggtggttgcaccgagtcagttcacatttagttcacgcagcccaga- ttttgtggatgaaaccgca ggtcaatcgtggtgcgcgatactgggattgaaccagtttcactag SEQ ID NO: 53 atggccaccacccttcccgttcagcgccacccgcggtccctcttccccgagttttctgagctgttcgcggcctt- cccgtcattcgccggactccgg cccaccttcgacacccggttgatgcggctggaagacgagatgaaagaggggcgctacgaggtacgcgcggagct- tcccggggtcgaccc cgacaaggacgtcgacattatggtccgcgatggtcagctgaccatcaaggccgagcgcaccgagcagaaggact- tcgacggtcgctcgga attcgcgtacggttccttcgttcgcacggtgtcgctgccggtaggtgctgacgaggacgacattaaggccacct- acgacaagggcattcttactg tgtcggtggcggtttcggaagggaagccaaccgaaaagcacattcagatccggtccaccaactga SEQ ID NO: 54 atgagtggacgccaccgtaagcccaccacatccaacgtcagcgtcgccaagatcgcctttaccggcgcagtact- cggtggcggcggcatcg ccatggccgctcaggcgaccgcggccaccgacggggaatgggatcaggtggcccgctgcgagtcgggcggcaac- tggtcgatcaacac cggcaacggttacctcggtggcttgcagttcactcaaagcacctgggccgcacatggtggcggcgagttcgccc- cgtcggctcagctggcca gccgggagcagcagattgccgtcggtgagcgggtgctggccacccagggtcgcggcgcctggccggtgtgcggc- cgcgggttatcgaacg caacaccccgcgaagtgcttcccgcttcggcagcgatggacgctccgttggacgcggccgcggtcaacggcgaa- ccagcaccgctggccc cgccgcccgccgacccggcgccacccgtggaacttgccgctaacgacctgcccgcaccgctgggtgaacccctc- ccggcagctcccgcc gacccggcaccacccgccgacctggcaccacccgcgcccgccgacgtcgcgccacccgtggaacttgccgtaaa- cgacctgcccgcac cgctgggtgaacccctcccggcagctcccgccgacccggcaccacccgccgacctggcaccacccgcgcccgcc- gacctggcgccacc cgcgcccgccgacctggcgccacccgcgcccgccgacctggcaccacccgtggaacttgccgtaaacgacctgc- ccgcgccgctgggtg aacccctcccggcagctcccgccgaactggcgccacccgccgatctggcacccgcgtccgccgacctggcgcca- cccgcgcccgccgac ctggcgccacccgcgcccgccgaactggcgccacccgcgcccgccgacctggcaccacccgctgcggtgaacga- gcaaaccgcgccg ggcgatcagcccgccacagctccaggcggcccggttggccttgccaccgatttggaactccccgagcccgaccc- ccaaccagctgacgca ccgccgcccggcgacgtcaccgaggcgcccgccgaaacgccccaagtctcgaacatcgcctatacgaagaagct- gtggcaggcgattcg ggcccaggacgtctgcggcaacgatgcgctggactcgctcgcacagccgtacgtcatcggctga SEQ ID NO: 55 atgttgcgcctggtagtcggtgcgctgctgctggtgttggcgttcgccggtggctatgcggtcgccgcatgcaa- aacggtgacgttgaccgtcga cggaaccgcgatgcgggtgaccacgatgaaatcgcgggtgatcgacatcgtcgaagagaacgggttctcagtcg- acgaccgcgacgacc tgtatcccgcggccggcgtgcaggtccatgacgccgacaccatcgtgctgcggcgtagccgtccgctgcagatc- tcgctggatggtcacgac gctaagcaggtgtggacgaccgcgtcgacggtggacgaggcgctggcccaactcgcgatgaccgacacggcgcc- ggccgcggcttctcg cgccagccgcgtcccgctgtccgggatggcgctaccggtcgtcagcgccaagacggtgcagctcaacgacggcg- ggttggtgcgcacggt

gcacttgccggcccccaatgtcgcggggctgctgagtgcggccggcgtgccgctgttgcaaagcgaccacgtgg- tgcccgccgcgacggc cccgatcgtcgaaggcatgcagatccaggtgacccgcaatcggatcaagaaggtcaccgagcggctgccgctgc- cgccgaacgcgcgtc gtgtcgaggacccggagatgaacatgagccgggaggtcgtcgaagacccgggggttccggggacccaggatgtg- acgttcgcggtagctg aggtcaacggcgtcgagaccggccgtttgcccgtcgccaacgtcgtggtgaccccggcccacgaagccgtggtg- cgggtgggcaccaagc ccggtaccgaggtgcccccggtgatcgacggaagcatctgggacgcgatcgccggctgtgaggccggtggcaac- tgggcgatcaacacc ggcaacgggtattacggtggtgtgcagtttgaccagggcacctgggaggccaacggcgggctgcggtatgcacc- ccgcgctgacctcgcca cccgcgaagagcagatcgccgttgccgaggtgacccgactgcgtcaaggttggggcgcctggccggtatgtgct- gcacgagcgggtgcgc gctga SEQ ID NO: 56 gtgcatcctttgccggccgaccacggccggtcgcggtgcaatagacacccgatctcaccactctctctaatcgg- taacgcttcggccacttccg gcgatatgtcgagcatgacaagaatcgccaagccgctcatcaagtccgccatggccgcaggactcgtcacggca- tccatgtcgctctccacc gccgttgcccacgccggtcccagcccgaactgggacgccgtcgcgcagtgcgaatccgggggcaactgggcggc- caacaccggaaacg gcaaatacggcggactgcagttcaagccggccacctgggccgcattcggcggtgtcggcaacccagcagctgcc- tctcgggaacaacaa atcgcagttgccaatcgggttctcgccgaacagggattggacgcgtggccgacgtgcggcgccgcctctggcct- tccgatcgcactgtggtcg aaacccgcgcagggcatcaagcaaatcatcaacgagatcatttgggcaggcattcaggcaagtattccgcgctg- a SEQ ID NO: 57 atgacaccgggtttgcttactactgcgggtgctggccgaccacgtgacaggtgcgccaggatcgtatgcacggt- gttcatcgaaaccgccgttg tcgcgaccatgtttgtcgcgttgttgggtctgtccaccatcagctcgaaagccgacgacatcgattgggacgcc- atcgcgcaatgcgaatccgg cggcaattgggcggccaacaccggtaacgggttatacggtggtctgcagatcagccaggcgacgtgggattcca- acggtggtgtcgggtcg ccggcggccgcgagtccccagcaacagatcgaggtcgcagacaacattatgaaaacccaaggcccgggtgcgtg- gccgaaatgtagttct tgtagtcagggagacgcaccgctgggctcgctcacccacatcctgacgttcctcgcggccgagactggaggttg- ttcggggagcagggacg attga SEQ ID NO: 58 ttgaagaacgcccgtacgacgctcatcgccgccgcgattgccgggacgttggtgaccacgtcaccagccggtat- cgccaatgccgacgacg cgggcttggacccaaacgccgcagccggcccggatgccgtgggctttgacccgaacctgccgccggccccggac- gctgcacccgtcgata ctccgccggctccggaggacgcgggctttgatcccaacctccccccgccgctggccccggacttcctgtccccg- cctgcggaggaagcgcct cccgtgcccgtggcctacagcgtgaactgggacgcgatcgcgcagtgcgagtccggtggaaactggtcgatcaa- caccggtaacggttact acggcggcctgcggttcaccgccggcacctggcgtgccaacggtggctcggggtccgcggccaacgcgagccgg- gaggagcagatccg ggtggctgagaacgtgctgcgttcgcagggtatccgcgcctggccggtctgcggccgccgcggctga SEQ ID NO: 59 atgatcgccacaacccgcgatcgtgaaggagccaccatgatcacgtttaggctgcgcttgccgtgccggacgat- actgcgggtgttcagccg caatccgctggtgcgtgggacggatcgactcgaggcggtcgtcatgctgctggccgtcacggtctcgctgctga- ctatcccgttcgccgccgcg gccggcaccgcagtccaggattcccgcagccacgtctatgcccaccaggcccagacccgccatcccgcaaccgc- gaccgtgatcgatcac gagggggtgatcgacagcaacacgaccgccacgtcagcgccgccgcgcacgaagatcaccgtgcctgcccgatg- ggtcgtgaacggaa tagaacgcagcggtgaggtcaacgcgaagccgggaaccaaatccggtgaccgcgtcggcatttgggtcgacagt- gccggtcagctggtcg atgaaccagctccgccggcccgtgccattgcggatgcggccctggccgccttgggactctggttgagcgtcgcc- gcggttgcgggcgccctg ctggcgctcactcgggcgattctgatccgcgttcgcaacgccagttggcaacacgacatcgacagcctgttctg- cacgcagcggtga SEQ ID NO: 60 atgacggagccagcggcgtgggacgaaggcaagccgcgaatcatcactttgaccatgaaccccgccttggacat- cacgacgagcgtcga cgtggtgcgcccgaccgagaaaatgcgttgtggcgcacctcgctacgatcccggcggcggcggtatcaatgtcg- cccgcattgtgcatgtcct cggcggttgctcgacagcactgttcccggccggcgggtcgaccgggagcctgctgatggcgctgctcggtgatg- cgggagtgccatttcgcgt cattccgatcgcggcctcgacgcgggagagcttcacggtcaacgagtccaggaccgccaagcagtatcgtttcg- tgcttccggggccgtcgct gaccgtcgcggagcaggagcaatgcctcgacgaactgcgcggtgcggcggcttcggccgcctttgtggtggcca- gtggcagcctgccgcc aggtgtggctgccgactactatcagcgggttgccgacatctgccgccgatcgagcactccgctgatcctggata- catctggtggcgggttgcag cacatttcgtccggggtgtttcttctcaaggcgagcgtgcgggaactgcgcgagtgcgtcggatccgaactgct- gaccgagcccgaacaactg gccgccgcacacgaactcattgaccgtgggcgcgccgaggtcgtggtggtctcgcttggatctcagggcgcgct- attggccacacgacatgc gagccatcgattttcgtcgattccgatgaccgcggttagcggtgtcggcgccggcgacgcgatggtggccgcga- ttaccgtgggcctcagccg tggctggtcgctcatcaagtccgttcgcttgggaaacgcggcaggtgcagccatgctgctgacgccaggcaccg- cggcctgcaatcgcgac gatgtggagaggttcttcgagctggcggccgaacccaccgaagtcgggcaggatcaatacgtttggcacccgat- cgttaacccggaagcctc gccatga SEQ ID NO: 61 atgccggacaccatggtgaccaccgatgtcatcaagagcgcggtgcagttggcctgccgcgcaccgtcgctcca- caacagccagccctgg cgctggatagccgaggaccacacggttgcgctgttcctcgacaaggatcgggtgctttacgcgaccgaccactc- cggccgggaagcgctgc tggggtgcggcgccgtactcgaccactttcgggtggcgatggcggccgcgggtaccaccgccaatgtggaacgg- tttcccaaccccaacga tcctttgcatctggcgtcaattgacttcagcccggccgatttcgtcaccgagggccaccgtctaagggcggatg- cgatcctactgcgccgtaccg accggctgcctttcgccgagccgccggattgggacttggtggagtcgcagttgcgcacgaccgtcaccgccgac- acggtgcgcatcgacgtc atcgccgacgatatgcgtcccgaactggcggcggcgtccaaactcaccgaatcgctgcggctctacgattcgtc- gtatcatgccgaactcttttg gtggacaggggcttttgagacttctgagggcataccgcacagttcattggtatcggcggccgaaagtgaccggg- tcaccttcggacgcgactt cccggtcgtcgccaacaccgataggcgcccggagtttggccacgaccgctctaaggtcctggtgctctccacct- acgacaacgaacgcgcc agcctactgcgctgcggcgagatgctttccgccgtattgcttgacgccaccatggctgggcttgccacctgcac- gctgacccacatcaccgaa ctgcacgccagccgagacctggtcgcagcgctgattgggcagcccgcaactccgcaagccttggttcgcgtcgg- tctggccccggagatgg aagagccgccaccggcaacgcctcggcgaccaatcgatgaagtgtttcacgttcgggctaaggatcaccggtag SEQ ID NO: 62 atgaccaccgcacgcgacatcatgaacgcaggtgtgacctgtgttggcgaacacgagacgctaaccgctgccgc- tcaatacatgcgtgagc acgacatcggcgcgttgccgatctgcggggacgacgaccggctgcacggcatgctcaccgaccgcgacattgtg- atcaaaggcctggctg cgggcctagacccgaataccgccacggctggcgagttggcccgggacagcatctactacgtcgatgcgaacgca- agcatccaggagatg ctcaacgtcatggaagaacatcaggtccgccgtgttccggtcatctcagagcaccgcttggtcggaatcgtcac- cgaagccgacatcgcccg acacctgcccgagcacgccattgtgcagttcgtcaaggcaatctgctcgcccatggccctcgccagctag SEQ ID NO: 63 atggcaagttctgcgagcgacggcacccacgaacgctcggcttttcgcctgagtccaccggtcttgagcggcgc- catgggaccgttcatgcac accggtctgtacgtcgctcaatcgtggcgcgactatctgggtcaacagcccgataaactgccgatcgcacggcc- cactattgccttagcggcg caagcctttcgagacgaaatcgtcctgctgggcctcaaggcacgacgtccggtcagcaatcatcgagtgttcga- gcgcatcagccaagaagt ggccgctggactggagttctatgggaatcgcagatggctggagaagcctagcggattttttgcccagcccccac- cgctcaccgaggtcgcggt ccgaaaggtcaaggaccgcagacgctccttttatcgcatcttcttcgacagtgggtttacgccgcatccgggtg- aaccgggcagccaacggtg gctctcatacactgcgaacaatcgcgagtacgccctgttactgcggcacccagagccgcgtccctggctggttt- gtgtacacggcaccgagat gggcagggccccgttggatctcgcggtgttccgcgcctggaagctgcatgacgaactcggcctgaacattgtca- tgccggttcttccgatgcat ggtccccgcgggcaaggtctgccgaagggcgccgtttttcccggagaagatgttctcgacgatgtgcatgggac- ggctcaagcggtgtggga tatccggcggctgttgtcctggatacgatcgcaggaggaggagtcgctgatcgggttgaacggtctctcgctgg- gcggctacatcgcgtcattg gtcgccagcctcgaagaaggtctcgcctgcgcgattctcggtgtcccagtggctgatctgatcgagttgttggg- ccgccactgcggtcttcggca caaagacccccgccgccacaccgtcaagatggccgaaccgatcggccgaatgatctcgccgctctcacttacgc- cactggtgcccatg ccgggccgctttatctacgcgggcattgccgaccgactcgtgcatccacgcgaacaggtgactcgcctctggga- gcactggggcaaacccg aaatcgtgtggtatccaggcggtcacactggcttcttccagtcgcggccggtacgacggtttgtccaggctgcg- ctggagcagtcgggcctgttg gacgcgccacggacacagcgcgaccgttccgcctaa SEQ ID NO: 64 atgtccacgcaacgaccgaggcactccggtattcgggctgttggcccctacgcatgggccggccgatgtggtcg- gataggcaggtgggggg tgcaccaggaggcgatgatgaatctagcgatatggcacccgcgcaaggtgcaatccgccaccatctatcaggtg- accgatcgctcgcacga cgggcgcacagcacgggtgcctggtgacgagatcactagcaccgtgtccggttggttgtcggagttgggcaccc- aaagcccgttggccgatg agcttgcgcgtgcggtgcggatcggcgactggcccgctgcgtacgcaatcggtgagcacctgtccgttgagatt- gccgttgcggtctaa SEQ ID NO: 65 LDFATLPPEINSARMYSGAGSAPMLAAASAWHGLSAELRASALSYSSVLSTLTGEEWHGPASASMT AAAAPYVAWMSVTAVRAEQAGAQAEAAAAAYEAAFAATVPPPVIEANRAQLMALIATNVLGQNAPAI AATEAQYAEMWSQDAMAMYGYAGASAAATQLTPFTEPVQTTNASGLAAQSAAIAHATGASAGAQQ TTLSQLIAAIPSVLQGLSSSTAATFASGPSGLLGIVGSGSSWLDKLWALLDPNSNFWNTIASSGLFLPS NTIAPFLGLLGGVAAADAAGDVLGEATSGGLGGALVAPLGSAGGLGGTVAAGLGNAATVGTLSVPPS

WTAAAPLASPLGSALGGTPMVAPPPAVAAGMPGMPFGTMGGQGFGRAVPQYGFRPNFVARPPAA G

EXAMPLES

[0244] The invention will be further clarified by the following examples, which are intended to be purely exemplary of the invention and in no way limiting.

Example 1

Sub-Unit Vaccines Containing Polypeptides of the Invention

[0245] To prepare sub-unit vaccines comprising polypeptides it is first of all necessary to obtain a supply of polypeptide to prepare the vaccine. This can be achieved by purifying proteins of interest from TB culture, or by cloning the gene of interest and producing a recombinant protein. The coding sequences for the genes of interest are amplified by PCR with restriction sites inserted at the N terminus and C terminus to permit cloning in-frame into a protein expression vector such as pET-15b. The genes are inserted behind an inducible promoter such as lacZ. The vector is then transformed into E. coli which is grown in culture. The recombinant protein is over-expressed and is purified. One of the common purification methods is to produce a recombinant protein with an N-terminal tag for purification eg a His-tag. The protein can then be purified on the basis of the affinity of the His-tag for metal ions on a Ni-NTA column after which the His-tag is cleaved. The purified protein is then administered to animals in a suitable adjuvant.

Example 2

Use of BCG as a Microbial Carrier

[0246] The polynucleotide sequence of interest is amplified by PCR. The amplified product is purified and cloned into a plasmid (pMV306) that integrates site specifically into the mycobacterial genome at the attachment site (attB) for mycobacteriophage L5. BCG is transformed with the plasmid by electroporation, which involves damaging the cell envelope with high voltage electrical pulses, resulting in uptake of the DNA. The plasmid integrates into the BCG chromosome at the attB site, generating stable recombinants. Recombinants are selected and are checked by PCR or Southern blotting to ensure that the gene has been integrated. The recombinant strain is then used for protection studies.

Example 3

Viral Vectors (Eg. Attenuated Vaccinia Virus) Expressing Mycobacterial Genes

##STR00001##

[0248] Methodologies permitting recombination of foreign or target genes into the genome of MVA are well known in the art [1,2]. Insertion of the target gene(s) is mediated by transfer DNA with features similar to those shown above. The transfer DNA may be in the form of a plasmid that can be propagated in a bacterial strain optimised for routine cloning procedures. The target gene(s) is introduced to the cassette downstream of a promoter such as mH5, p7.5 or another. The target gene(s) may comprise one or more of nucleotide Seq IDs 1-18 and/or fragments thereof. The target gene(s) may also comprise adjuvanting cofactors such as B7-1 or IL-12 as is well described in the art [3]. The target gene(s) would be positioned downstream and in frame with an optimised Kozak sequence e.g. GCCACCATGG. The target gene(s) may also be positioned downstream and in frame with a leader sequence e.g. tPA. The target gene(s) may be positioned upstream of an in-frame tag e.g. V5, HIS or another. Transfer of the cassette into the genome of MVA is mediated by homologous flanking regions well known in the art e.g. Del I-VI.

[0249] 1. Generation of recombinant vaccinia viruses. (2001) Earl P L et al. Current Protocols in Protein Science

[0250] 2. Preparation of cell cultures and vaccine virus stocks. (2001) Earl P L et al. Current Protocols in Protein Science

[0251] 3. Construction and characterisation of a triple-recombinant vaccine virus encoding B7-1, Interleukin 12 and a model tumor antigen. (1998) Carroll M W et al. Journal of the National Cancer Institute Dec. 16, 1998; 90(24): 1881-1887

Example 4

Preparation of DNA Expression Vectors

[0252] DNA vaccines consist of a nucleic acid sequence of interest cloned into a bacterial plasmid. The plasmid vector pVAX1 is commonly used in the preparation of DNA vaccines. The vector is designed to facilitate high copy number replication in E. coli and high level transient expression of the peptide of interest in most mammalian cells (for details see manufacturer's protocol for pVAX1 (catalog No. V260-20 www.invitrogen.com).

[0253] The vector contains the following elements:

[0254] Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a variety of mammalian cells

[0255] T7 promoter/priming site to allow in vitro transcription in the sense orientation and sequencing through the insert

[0256] Bovine growth hormone (BGH) polyadenylation signal for efficient transcription termination and polyadenylation of mRNA

[0257] Kanamycin resistance gene for selection in E. coli

[0258] A multiple cloning site

[0259] pUC origin for high-copy number replication and growth in E. coli

[0260] BGH reverse priming site to permit sequencing through the insert

[0261] Vectors may be prepared by means of standard recombinant techniques that are known in the art, for example Sambrook et al. (1989). Key stages in preparing the vaccine are as follows:

[0262] The polynucleotide of interest is ligated into pVAX1 via one of the multiple cloning sites

[0263] The ligation mixture is then transformed into a competent E. coli strain (e.g. TOP10) and LB plates containing 50 .mu.g/ml kanamycin are used to select transformants.

[0264] Clones are selected and may be sequenced to confirm the presence and orientation of the gene of interest.

[0265] Once the presence of the gene has been verified, the vector can be used to transfect a mammalian cell line to check for protein expression. Methods for transfection are known in the art and include, for example, electroporation, calcium phosphate, and lipofection.

[0266] Once polypeptide expression has been confirmed, large quantities of the vector can be produced and purified from the appropriate cell host, eg. E. coli.

[0267] pVAX1 does not integrate into the host chromosome. All non-essential sequences have been removed to minimise the possibility of integration. When constructing a specific vector, a leader sequence may be included to direct secretion of the encoded protein when expressed inside the eukaryotic cell. Other examples of vectors that can be used include V1Jns.tPA and pCMV4. Expression vectors may be used that integrate into the genome of the host, however, it is more common and more preferable to use a vector that does not integrate. Integration would lead to the generation of a genetically modified host which raises other issues.

Example 5

Plasmid DNA Vaccines Carrying Mycobacterial Polynucleotides

[0268] A polynucleotide sequence of interest is amplified by PCR, purified and inserted into specialized vectors developed for vaccine development, such as pVAX1. As above (Example 4), these vectors contain promoter sequences (eg. CMV or SV40 promoters), which direct strong expression of the introduced polynucleotide (encoding the candidate antigen) in eukaryotic cells; and polyadenylation signals (eg. SV40 or bovine growth hormone) to stabilize the mRNA transcript. The target gene(s) would be positioned downstream and in frame with an optimised Kozak sequence e.g. GCCACCATGG. The target gene(s) may also be positioned downstream and in frame with a leader sequence e.g. tPA. The target gene(s) may be positioned upstream of an in-frame tag e.g. V5. The vector is transformed into E. coli and transformants are selected using a marker, such as kanamycin resistance, encoded by the plasmid. The plasmid is then recovered from transformed colonies and is sequenced to check that the polynucleotide of interest is present and encoded properly without PCR generated mutations. Large quantities of the plasmid are then produced in E. coli and the plasmid is recovered and purified using commercially available kits (e.g. Qiagen Endofree-plasmid preparation). The vaccine is then administered to animals (e.g. by intramuscular injection) in the presence or absence of an adjuvant.

Example 6

RNA Vaccine

[0269] RNA can be introduced directly into the host. Thus, a vector construct may be used to generate RNA in vitro and the purified RNA is then injected into the host. The RNA then serves as a template for translation in the host cell. In this embodiment, integration would not normally occur.

[0270] An alternative option is to use an infectious agent such as the retroviral genome carrying RNA corresponding to the gene of interest. In this embodiment, integration into the host genome will occur. Another option is the use of RNA replicon vaccines which can be derived from virus vectors such as Sindbis virus or Semliki Forest virus. These vaccines are self-replicating and self-limiting and may be administered as either RNA or DNA which is then transcribed into RNA replicons in vivo. The vector eventually causes lysis of the transfected cells thereby reducing concerns about integration into the host genome.

Example 7

Diagnostic Assays Based on Assessing T Cell Responses

[0271] For a diagnostic assay based on assessing T cell responses it would be sufficient to obtain a sample of blood from the patient. Mononuclear cells (monocytes, T and B lymphocytes) can be separated from the blood using density gradients such as Ficoll gradients.

[0272] Both monocytes and B-lymphocytes are both able to present antigen, although less efficiently than professional antigen presenting cells (APCs) such as dendritic cells. The latter are more localized in lymphoid tissue.

[0273] The simplest approach would be to add antigen to the separated mononuclear cells and incubate for a week and then assess the amount of proliferation. If the individual had been exposed to the antigen previously through infection, then T-cell closes specific to the antigen should be more prevalent in the sample and should respond. It is also possible to separate the different cellular populations should it be desired to control the ratio of T cells to APCs. Another variation of this type of assay is to measure cytokine production by the responding lymphocytes as a measure of response. The ELISPOT assay is a suitable example of this assay.

Example 8

Detection of Latent Mycobacteria

[0274] The presence of latent mycobacteria-associated antigen may be detected either by detecting antigen-specific antibody, or by detecting T-cells in blood samples.

[0275] A 96 well plate is coated with cytokine (e.g. interferon-, IL-2)-specific antibody. Peripheral blood monocytes are then isolated from patient whole blood and are applied to the wells. Antigen is added to stimulate specific T cells that may be present and the plates are incubated for 24 h. The antigen stimulates the T-cells to produce cytokines, which bind a specific antibody. The plates are washed leaving a footprint where antigen-specific T cells were present. A second antibody coupled with a suitable detection system, e.g. enzyme, is then added and the number of spots is enumerated after the appropriate substrate has been added. The number of spots, each corresponding to a single antigen-specific T cell, is related to the total number of cells originally added. The above-described assay may also be used to distinguish TB-infected individuals from BCG-vaccinated individuals.

Example 9

Antigenic Activity

[0276] Mice are immunised with a mycobacterial antigen. Delivery systems include, but are not restricted to DNA vaccines, recombinant MVA, adjuvanted protein. Delivery routes include, but are not restricted to sub-cutaneous, intra-dermal, intra-muscular or aerosol administration. The immunisation regimen sometimes involves heterologous prime-boosting e.g. DNA vaccine followed by MVA vaccine. The immunisation regimen may also involve multiple doses.

[0277] After vaccination e.g. 2 weeks later, splenocytes are removed from the vaccinated animals and stimulated with polypeptide representative of the immunising antigen. An immune response is measurable through antigen-specific induction of cytokine release e.g. IFN-.gamma., and is evidence for immunisation against the target antigen.

Example 10

Demonstrating Vaccine Efficacy in an Experimental Model

[0278] Vaccine candidate efficacy in guinea pigs or mice may be assessed on the basis of reducing the bacterial burden of M. tuberculosis in the lungs and/or spleens at 6-24 weeks post-aerosol challenge--see FIGS. 1 & 2.

[0279] The mycobacterial antigens are delivered as sub-unit DNA vaccines or protein in a Th1-inducing adjuvant such as DDA/MPL, or by expression vectors such as recombinant viruses or BCG (see examples 1-4). The mycobacterial antigens may be delivered as a boost to an initial prime provided by BCG. There may be additional boosts provided by repeat inoculation of either DNA, polypeptide or viral vector or (less commonly) recombinant BCG. Groups of six to eight animals are immunised and then rested for 6 weeks prior to challenge. A group of positive control animals are inoculated sub-cutaneously with 5.times.10.sup.4 colony forming units (CFU) of BCG Danish (1331), and a group of negative control animals remain unvaccinated. Six weeks following the final vaccination, fine particle aerosols of M. tuberculosis (2 .mu.m mean diameter; generated in a Collison nebuliser), are delivered directly to the animal snout using a contained Henderson apparatus. 6 weeks post-aerosol challenge, the animals are euthanised and the lungs and spleen removed for CFU determination. Homogenised samples are serially diluted and plated on Middlebrook 7H11 selective agar and the mean CFU for each treatment group is determined. Vaccine efficacy is assessed in terms of reduction in bacterial counts in lungs or spleens compared to the unvaccinated control group. The reduction in bacterial load of test groups can be expressed as a proportion of the reduction achieved by BCG alone. Protective efficacy in animal models is indicative of the ability of the mycobacterial antigen to protect humans and animals from pathogenic mycobacterial infection.

Example 11

Demonstrating Vaccine Efficacy in an Experimental Model

[0280] Vaccine candidate efficacy in guinea pigs or mice may be assessed on the basis of reducing the bacterial burden of M. tuberculosis in the lungs and/or spleens at 6-24 weeks post-aerosol challenge--see FIG. 3.

[0281] The mycobacterial antigens are delivered as sub-unit DNA vaccines or protein in a Th1-inducing adjuvant such as DDA/MPL, or by expression vectors such as recombinant viruses or BCG (see examples 1-4). The mycobacterial antigens may be delivered as a boost to an initial prime provided by BCG. There may be additional boosts provided by repeat inoculation of either DNA, polypeptide or viral vector or (less commonly) recombinant BCG. Groups of six to eight animals are immunised and then rested for 6 weeks prior to challenge. A group of positive control animals are inoculated sub-cutaneously with 5.times.10.sup.4 colony forming units (CFU) of BCG Danish (1331), and a group of negative control animals remain unvaccinated. Six weeks following the final vaccination, fine particle aerosols of M. tuberculosis (2 .mu.m mean diameter; generated in a Collison nebuliser), are delivered directly to the animal snout using a contained Henderson apparatus. 24 weeks post-aerosol challenge, the animals are euthanised and the lungs and spleen removed for CFU determination. Homogenised samples are serially diluted and plated on Middlebrook 7H11 selective agar and the mean CFU for each treatment group is determined. Vaccine efficacy is assessed in terms of reduction in bacterial counts in lungs or spleens compared to the unvaccinated control group. The reduction in bacterial load of test groups can be expressed as a proportion of the reduction achieved by BCG alone, or as the additional reduction in bacterial burden achieved. Protective efficacy in animal models is indicative of the ability of the mycobacterial antigen to protect humans and animals from pathogenic mycobacterial infection.

Sequence CWU 1

1

6511515DNAMycobacterium tuberculosis 1atgtggtggt tccgccgccg agaccgggcg ccgctgcgcg ccaccagctc attatccctg 60cggtggcggg tcatgctgct ggcgatgtcc atggtcgcga tggtggttgt gctgatgtcg 120ttcgccgtct atgcggtgat ctcggccgcg ctctacagcg acatcgacaa ccaactgcag 180agccgggcgc aactgctcat cgccagtggc tcgctggcag ctgatccggg taaggcaatc 240gagggtaccg cctattcgga tgtcaacgcg atgctggtca accccggcca gtccatctac 300accgctcaac agccgggcca gacgctgccg gtcggtgctg ccgagaaggc ggtgatccgt 360ggcgagttgt tcatgtcgcg gcgcaccacc gccgaccaac gggtgcttgc catccgtctg 420accaacggta gttcgctgct gatctccaaa agtctcaagc ccaccgaagc agtcatgaac 480aagctgcgtt gggtgctatt gatcgtgggt gggatcgggg tggcggtcgc cgcggtggcc 540ggggggatgg tcacccgggc cgggctgagg ccggtgggcc gcctcaccga agcggccgag 600cgggtggcgc gaaccgacga cctgcggccc atccccgtct tcggcagcga cgaattggcc 660aggctgacag aggcattcaa tttaatgctg cgggcgctgg ccgagtcacg ggaacggcag 720gcaaggctgg ttaccgacgc cggacatgaa ttgcgtaccc cgctaacgtc gctgcgcacc 780aatgtcgaac tcttgatggc ctcgatggcc ccgggggctc cgcggctacc caagcaggag 840atggtcgacc tgcgtgccga tgtgctggct caaatcgagg aattgtccac actggtaggc 900gatttggtgg acctgtcccg aggcgacgcc ggagaagtgg tgcacgagcc ggtcgacatg 960gctgacgtcg tcgaccgcag cctggagcgg gtcaggcggc ggcgcaacga tatccttttc 1020gacgtcgagg tgattgggtg gcaggtttat ggcgataccg ctggattgtc gcggatggcg 1080cttaacctga tggacaacgc cgcgaagtgg agcccgccgg gcggccacgt gggtgtcagg 1140ctgagccagc tcgacgcgtc gcacgctgag ctggtggttt ccgaccgcgg cccgggcatt 1200cccgtgcagg agcgccgtct ggtgtttgaa cggttttacc ggtcggcatc ggcacgggcg 1260ttgccgggtt cgggcctcgg gttggcgatc gtcaaacagg tggtgctcaa ccacggcgga 1320ttgctgcgca tcgaagacac cgacccaggc ggccagcccc ctggaacgtc gatttacgtg 1380ctgctccccg gccgtcggat gccgattccg cagcttcccg gtgcgacggc tggcgctcgg 1440agcacggaca tcgagaactc tcggggttcg gcgaacgtta tctcagtgga atctcagtcc 1500acgcgcgcaa cctag 15152504PRTMycobacterium tuberculosis 2Met Trp Trp Phe Arg Arg Arg Asp Arg Ala Pro Leu Arg Ala Thr Ser 1 5 10 15 Ser Leu Ser Leu Arg Trp Arg Val Met Leu Leu Ala Met Ser Met Val 20 25 30 Ala Met Val Val Val Leu Met Ser Phe Ala Val Tyr Ala Val Ile Ser 35 40 45 Ala Ala Leu Tyr Ser Asp Ile Asp Asn Gln Leu Gln Ser Arg Ala Gln 50 55 60 Leu Leu Ile Ala Ser Gly Ser Leu Ala Ala Asp Pro Gly Lys Ala Ile 65 70 75 80 Glu Gly Thr Ala Tyr Ser Asp Val Asn Ala Met Leu Val Asn Pro Gly 85 90 95 Gln Ser Ile Tyr Thr Ala Gln Gln Pro Gly Gln Thr Leu Pro Val Gly 100 105 110 Ala Ala Glu Lys Ala Val Ile Arg Gly Glu Leu Phe Met Ser Arg Arg 115 120 125 Thr Thr Ala Asp Gln Arg Val Leu Ala Ile Arg Leu Thr Asn Gly Ser 130 135 140 Ser Leu Leu Ile Ser Lys Ser Leu Lys Pro Thr Glu Ala Val Met Asn 145 150 155 160 Lys Leu Arg Trp Val Leu Leu Ile Val Gly Gly Ile Gly Val Ala Val 165 170 175 Ala Ala Val Ala Gly Gly Met Val Thr Arg Ala Gly Leu Arg Pro Val 180 185 190 Gly Arg Leu Thr Glu Ala Ala Glu Arg Val Ala Arg Thr Asp Asp Leu 195 200 205 Arg Pro Ile Pro Val Phe Gly Ser Asp Glu Leu Ala Arg Leu Thr Glu 210 215 220 Ala Phe Asn Leu Met Leu Arg Ala Leu Ala Glu Ser Arg Glu Arg Gln 225 230 235 240 Ala Arg Leu Val Thr Asp Ala Gly His Glu Leu Arg Thr Pro Leu Thr 245 250 255 Ser Leu Arg Thr Asn Val Glu Leu Leu Met Ala Ser Met Ala Pro Gly 260 265 270 Ala Pro Arg Leu Pro Lys Gln Glu Met Val Asp Leu Arg Ala Asp Val 275 280 285 Leu Ala Gln Ile Glu Glu Leu Ser Thr Leu Val Gly Asp Leu Val Asp 290 295 300 Leu Ser Arg Gly Asp Ala Gly Glu Val Val His Glu Pro Val Asp Met 305 310 315 320 Ala Asp Val Val Asp Arg Ser Leu Glu Arg Val Arg Arg Arg Arg Asn 325 330 335 Asp Ile Leu Phe Asp Val Glu Val Ile Gly Trp Gln Val Tyr Gly Asp 340 345 350 Thr Ala Gly Leu Ser Arg Met Ala Leu Asn Leu Met Asp Asn Ala Ala 355 360 365 Lys Trp Ser Pro Pro Gly Gly His Val Gly Val Arg Leu Ser Gln Leu 370 375 380 Asp Ala Ser His Ala Glu Leu Val Val Ser Asp Arg Gly Pro Gly Ile 385 390 395 400 Pro Val Gln Glu Arg Arg Leu Val Phe Glu Arg Phe Tyr Arg Ser Ala 405 410 415 Ser Ala Arg Ala Leu Pro Gly Ser Gly Leu Gly Leu Ala Ile Val Lys 420 425 430 Gln Val Val Leu Asn His Gly Gly Leu Leu Arg Ile Glu Asp Thr Asp 435 440 445 Pro Gly Gly Gln Pro Pro Gly Thr Ser Ile Tyr Val Leu Leu Pro Gly 450 455 460 Arg Arg Met Pro Ile Pro Gln Leu Pro Gly Ala Thr Ala Gly Ala Arg 465 470 475 480 Ser Thr Asp Ile Glu Asn Ser Arg Gly Ser Ala Asn Val Ile Ser Val 485 490 495 Glu Ser Gln Ser Thr Arg Ala Thr 500 31194DNAMycobacterium tuberculosis 3atgacggctc cgggactgac agcagccgtc gaggggatcg cacacaacaa gggcgagctg 60ttcgcctcct ttgacgtgga cgcgttcgag gttccgcacg gccgcgacga gatctggcgg 120ttcaccccgt tgcggcggct gcgtggcctg cacgacggct ccgcgcgggc caccggtagc 180gccacgatca cggtcagcga gcggccgggc gtatacaccc agaccgtgcg ccgcggcgat 240ccacgactgg gcgagggcgg cgtacccacc gaccgcgttg ccgcccaagc gttttcgtcg 300ttcaactccg cgactctggt caccgtcgag cgcgacaccc aggtcgtcga gccggtaggc 360atcaccgtga ccgggccggg ggagggcgcg gtggcctatg ggcacctgca ggtgcgtatc 420gaggagcttg gcgaggcggt cgtggtcatc gaccaccggg gcggcggaac ctacgccgac 480aacgtcgagt tcgttgtcga cgacgccgct cggctgaccg ccgtgtggat cgccgactgg 540gccgacaaca ccgttcacct cagcgcgcac catgctcgga tcggcaagga cgcggtgctg 600cgccacgtca ccgtcatgtt gggcggcgac gtggtgcgaa tgtcggcggg cgtgcggttc 660tgcggtgcgg gtggggacgc ggaactgctg gggctgtatt tcgccgacga cggccagcac 720ctggagtcgc ggctgctggt ggaccacgcc caccccgact gcaagtcgaa cgtgctgtat 780aagggtgcac tgcaaggtga tccggcgtcg tcgttgcccg acgcacacac ggtctgggtg 840ggtgacgtgc tgatccgtgc gcaggccacc ggcaccgaca ccttcgaggt gaaccggaac 900ctggtgctca ccgacggcgc gcgtgccgac tcggtgccca acctggagat cgagaccggc 960gagatcgtcg gcgccggaca cgccagcgcc accggtcgct tcgacgatga gcaattgttc 1020tacctgcgtt cgcgcggtat tcccgaagca caggcccgcc ggctggtggt ccgcggcttc 1080ttcggtgaga tcatcgccaa gatcgcggtg cccgaggtac gcgagcgcct gaccgcagcc 1140atcgaacacg agctggaaat cacggaatca acggaaaaga caacagtctc atga 11944397PRTMycobacterium tuberculosis 4Met Thr Ala Pro Gly Leu Thr Ala Ala Val Glu Gly Ile Ala His Asn 1 5 10 15 Lys Gly Glu Leu Phe Ala Ser Phe Asp Val Asp Ala Phe Glu Val Pro 20 25 30 His Gly Arg Asp Glu Ile Trp Arg Phe Thr Pro Leu Arg Arg Leu Arg 35 40 45 Gly Leu His Asp Gly Ser Ala Arg Ala Thr Gly Ser Ala Thr Ile Thr 50 55 60 Val Ser Glu Arg Pro Gly Val Tyr Thr Gln Thr Val Arg Arg Gly Asp 65 70 75 80 Pro Arg Leu Gly Glu Gly Gly Val Pro Thr Asp Arg Val Ala Ala Gln 85 90 95 Ala Phe Ser Ser Phe Asn Ser Ala Thr Leu Val Thr Val Glu Arg Asp 100 105 110 Thr Gln Val Val Glu Pro Val Gly Ile Thr Val Thr Gly Pro Gly Glu 115 120 125 Gly Ala Val Ala Tyr Gly His Leu Gln Val Arg Ile Glu Glu Leu Gly 130 135 140 Glu Ala Val Val Val Ile Asp His Arg Gly Gly Gly Thr Tyr Ala Asp 145 150 155 160 Asn Val Glu Phe Val Val Asp Asp Ala Ala Arg Leu Thr Ala Val Trp 165 170 175 Ile Ala Asp Trp Ala Asp Asn Thr Val His Leu Ser Ala His His Ala 180 185 190 Arg Ile Gly Lys Asp Ala Val Leu Arg His Val Thr Val Met Leu Gly 195 200 205 Gly Asp Val Val Arg Met Ser Ala Gly Val Arg Phe Cys Gly Ala Gly 210 215 220 Gly Asp Ala Glu Leu Leu Gly Leu Tyr Phe Ala Asp Asp Gly Gln His 225 230 235 240 Leu Glu Ser Arg Leu Leu Val Asp His Ala His Pro Asp Cys Lys Ser 245 250 255 Asn Val Leu Tyr Lys Gly Ala Leu Gln Gly Asp Pro Ala Ser Ser Leu 260 265 270 Pro Asp Ala His Thr Val Trp Val Gly Asp Val Leu Ile Arg Ala Gln 275 280 285 Ala Thr Gly Thr Asp Thr Phe Glu Val Asn Arg Asn Leu Val Leu Thr 290 295 300 Asp Gly Ala Arg Ala Asp Ser Val Pro Asn Leu Glu Ile Glu Thr Gly 305 310 315 320 Glu Ile Val Gly Ala Gly His Ala Ser Ala Thr Gly Arg Phe Asp Asp 325 330 335 Glu Gln Leu Phe Tyr Leu Arg Ser Arg Gly Ile Pro Glu Ala Gln Ala 340 345 350 Arg Arg Leu Val Val Arg Gly Phe Phe Gly Glu Ile Ile Ala Lys Ile 355 360 365 Ala Val Pro Glu Val Arg Glu Arg Leu Thr Ala Ala Ile Glu His Glu 370 375 380 Leu Glu Ile Thr Glu Ser Thr Glu Lys Thr Thr Val Ser 385 390 395 52520DNAMycobacterium tuberculosis 5atggcggttc gtcaggtcac cgtcggctat tcggacggca cgcacaagac gatgccggtg 60cggtgcgacc agacggtcct ggatgccgcc gaggaacacg gcgtggccat cgtcaacgaa 120tgccaaagcg ggatatgtgg cacctgcgtg gccacctgca ccgccggccg ctaccagatg 180ggacgcaccg agggactgtc cgatgtcgag cgggcggcgc gaaagatcct cacctgccag 240acgtttgtta cctccgattg ccggatcgag ctgcagtatc cggtcgacga caacgccgcc 300ctgctggtca ccggtgacgg tgtggtgacc gcggtcgagt tggtgtcgcc cagcaccgcc 360atcctgcggg tggacacctc tggcatggcc ggcgcgctga gataccgggc cggccagttc 420gcccaattgc aggttcccgg taccaacgta tggcgcaact actcctacgc ccatccggcc 480gacggccgcg gtgagtgcga gttcatcatc aggttgctgc cggacggcgt gatgtcgaat 540tatcttcgcg accgcgccca gcccggtgac catatcgcgc tgcgctgcag caagggcagc 600ttttatctgc gcccgatcgt gcgaccggtg atcctggtcg ccggaggaac cggcctgtca 660gcgatcctgg cgatggccca gagcctggat gccgatgtcg ctcacccggt ctacctgctc 720tacggggtcg agcgcaccga agacctgtgc aagctcgacg aactcaccga gctgcgccgc 780cgcgttggcc gcctggaggt gcacgtcgtc gtcgctcgcc cggaccccga ctgggatggg 840cgcaccgggc tggtcaccga cctgctcgac gagcggatgc tggcgagcgg tgacgccgac 900gtgtatctgt gcggtccggt cgccatggtc gacgcagccc gaacctggct ggaccacaat 960ggctttcacc gtgtcgggtt gtactacgag aagttcgtgg ccagcggggc ggcgcgccgc 1020cgcaccccgg ctcggctgga ttacgcgggc gtggacattg ccgaggtgtg ccgccgcggc 1080cgcggcaccg cggtggtcat cggcggcagc atcgcgggca tcgcggcggc gaaaatgctc 1140agcgagacct tcgatcgcgt catcgtgctg gagaaggacg gcccgcaccg tcgccgcgag 1200ggcaggccgg gcgcggcaca gggttggcac ctgcaccacc tgctgaccgc cgggcagatc 1260gagctggagc gcatcttccc tggcatcgtc gacgacatgg tgcgcgaggg agcgttcaag 1320gtcgacatgg ccgcgcagta ccgtatccgg ctgggcggca cctggaagaa gcccggcact 1380agtgacatcg agatcgtctg cgcgggaagg ccgctgctcg aatggtgtgt gcgccgccgg 1440ctcgacgacg aaccgcgcat cgacttccgc tacgaatcgg aggtggccga tctcgccttc 1500gaccgcgcca acaatgccat cgtcggcgtc gccgtggaca atggcgacgc cgacggaggc 1560gacggtttgc aggtggtgcc cgccgagttc gtcgtggacg cgtcgggcaa gaacacccgc 1620gtgccggagt tcttggagcg tctcggtgtt ggcgctcccg aggccgagca ggacatcatc 1680aactgcttct actccacgat gcagcaccgg gttccgccgg agcggcggtg gcaggacaag 1740gtgatggtga tctgctatgc gtaccgccct ttcgaggata cctacgccgc gcagtactac 1800accgacagct cccgcaccat cctgtccacc tcactggtgg cctacaactg ctattcgccg 1860ccgcgtaccg cccgagaatt ccgcgcgttc gccgacctga tgccgtcccc ggtcatcggg 1920gagaacatcg acgggctgga gccggcatcg cccatctaca atttccgcta tcccaacatg 1980ctgcggctgc gctacgagaa gaagcgcaac ctgccgcggg ctttgctggc ggtgggcgat 2040gcctacacca gcgccgaccc ggtgtcgggt ctgggtatga gcctggcgct caaggaagtt 2100cgggagatgc aggcgctgct ggctaaatac ggcgccggtc accgggatct gccgcgccgg 2160tactaccggg cgatcgccaa gatggccgac acggcctggt tcgtgatccg cgagcagaac 2220ctgcgcttcg actggatgaa ggacgtcgac aagaagcgcc cgttctattt cggtgtgctg 2280acctggtaca tggaccgcgt gctggagctg gtgcatgacg atctcgacgc gtaccgggaa 2340ttcttggccg tcgtccatct ggtcaagccg ccgtcggcgc tgatgcgacc caggatcgcc 2400agccgcgtcc tcggcaaatg ggcacgaacc cgattgtcgg gccagaagac gttgattgcc 2460cgcaactacg aaaatcatcc gataccagcc gaacccgcgg accaacttgt aaacgcttag 25206839PRTMycobacterium tuberculosis 6Met Ala Val Arg Gln Val Thr Val Gly Tyr Ser Asp Gly Thr His Lys 1 5 10 15 Thr Met Pro Val Arg Cys Asp Gln Thr Val Leu Asp Ala Ala Glu Glu 20 25 30 His Gly Val Ala Ile Val Asn Glu Cys Gln Ser Gly Ile Cys Gly Thr 35 40 45 Cys Val Ala Thr Cys Thr Ala Gly Arg Tyr Gln Met Gly Arg Thr Glu 50 55 60 Gly Leu Ser Asp Val Glu Arg Ala Ala Arg Lys Ile Leu Thr Cys Gln 65 70 75 80 Thr Phe Val Thr Ser Asp Cys Arg Ile Glu Leu Gln Tyr Pro Val Asp 85 90 95 Asp Asn Ala Ala Leu Leu Val Thr Gly Asp Gly Val Val Thr Ala Val 100 105 110 Glu Leu Val Ser Pro Ser Thr Ala Ile Leu Arg Val Asp Thr Ser Gly 115 120 125 Met Ala Gly Ala Leu Arg Tyr Arg Ala Gly Gln Phe Ala Gln Leu Gln 130 135 140 Val Pro Gly Thr Asn Val Trp Arg Asn Tyr Ser Tyr Ala His Pro Ala 145 150 155 160 Asp Gly Arg Gly Glu Cys Glu Phe Ile Ile Arg Leu Leu Pro Asp Gly 165 170 175 Val Met Ser Asn Tyr Leu Arg Asp Arg Ala Gln Pro Gly Asp His Ile 180 185 190 Ala Leu Arg Cys Ser Lys Gly Ser Phe Tyr Leu Arg Pro Ile Val Arg 195 200 205 Pro Val Ile Leu Val Ala Gly Gly Thr Gly Leu Ser Ala Ile Leu Ala 210 215 220 Met Ala Gln Ser Leu Asp Ala Asp Val Ala His Pro Val Tyr Leu Leu 225 230 235 240 Tyr Gly Val Glu Arg Thr Glu Asp Leu Cys Lys Leu Asp Glu Leu Thr 245 250 255 Glu Leu Arg Arg Arg Val Gly Arg Leu Glu Val His Val Val Val Ala 260 265 270 Arg Pro Asp Pro Asp Trp Asp Gly Arg Thr Gly Leu Val Thr Asp Leu 275 280 285 Leu Asp Glu Arg Met Leu Ala Ser Gly Asp Ala Asp Val Tyr Leu Cys 290 295 300 Gly Pro Val Ala Met Val Asp Ala Ala Arg Thr Trp Leu Asp His Asn 305 310 315 320 Gly Phe His Arg Val Gly Leu Tyr Tyr Glu Lys Phe Val Ala Ser Gly 325 330 335 Ala Ala Arg Arg Arg Thr Pro Ala Arg Leu Asp Tyr Ala Gly Val Asp 340 345 350 Ile Ala Glu Val Cys Arg Arg Gly Arg Gly Thr Ala Val Val Ile Gly 355 360 365 Gly Ser Ile Ala Gly Ile Ala Ala Ala Lys Met Leu Ser Glu Thr Phe 370 375 380 Asp Arg Val Ile Val Leu Glu Lys Asp Gly Pro His Arg Arg Arg Glu 385 390 395 400 Gly Arg Pro Gly Ala Ala Gln Gly Trp His Leu His His Leu Leu Thr 405 410 415 Ala Gly Gln Ile Glu Leu Glu Arg Ile Phe Pro Gly Ile Val Asp Asp 420 425 430 Met Val Arg Glu Gly Ala Phe Lys Val Asp Met Ala Ala Gln Tyr Arg 435 440 445 Ile Arg Leu Gly Gly Thr Trp Lys Lys Pro Gly Thr Ser Asp Ile Glu 450 455 460 Ile Val Cys Ala Gly Arg Pro Leu Leu Glu Trp Cys Val Arg Arg Arg 465 470 475 480 Leu Asp Asp Glu Pro Arg Ile Asp Phe Arg Tyr Glu Ser Glu Val Ala 485 490 495 Asp Leu Ala Phe Asp Arg Ala Asn Asn Ala Ile Val Gly Val Ala Val 500 505 510 Asp Asn Gly Asp Ala Asp Gly Gly Asp Gly Leu Gln Val Val Pro Ala 515 520 525 Glu Phe Val Val Asp Ala Ser Gly Lys Asn Thr Arg Val Pro Glu Phe 530 535 540 Leu Glu Arg Leu Gly Val Gly Ala Pro Glu Ala Glu Gln Asp Ile Ile 545 550 555 560 Asn Cys Phe Tyr Ser Thr Met Gln His Arg Val Pro Pro Glu Arg Arg

565 570 575 Trp Gln Asp Lys Val Met Val Ile Cys Tyr Ala Tyr Arg Pro Phe Glu 580 585 590 Asp Thr Tyr Ala Ala Gln Tyr Tyr Thr Asp Ser Ser Arg Thr Ile Leu 595 600 605 Ser Thr Ser Leu Val Ala Tyr Asn Cys Tyr Ser Pro Pro Arg Thr Ala 610 615 620 Arg Glu Phe Arg Ala Phe Ala Asp Leu Met Pro Ser Pro Val Ile Gly 625 630 635 640 Glu Asn Ile Asp Gly Leu Glu Pro Ala Ser Pro Ile Tyr Asn Phe Arg 645 650 655 Tyr Pro Asn Met Leu Arg Leu Arg Tyr Glu Lys Lys Arg Asn Leu Pro 660 665 670 Arg Ala Leu Leu Ala Val Gly Asp Ala Tyr Thr Ser Ala Asp Pro Val 675 680 685 Ser Gly Leu Gly Met Ser Leu Ala Leu Lys Glu Val Arg Glu Met Gln 690 695 700 Ala Leu Leu Ala Lys Tyr Gly Ala Gly His Arg Asp Leu Pro Arg Arg 705 710 715 720 Tyr Tyr Arg Ala Ile Ala Lys Met Ala Asp Thr Ala Trp Phe Val Ile 725 730 735 Arg Glu Gln Asn Leu Arg Phe Asp Trp Met Lys Asp Val Asp Lys Lys 740 745 750 Arg Pro Phe Tyr Phe Gly Val Leu Thr Trp Tyr Met Asp Arg Val Leu 755 760 765 Glu Leu Val His Asp Asp Leu Asp Ala Tyr Arg Glu Phe Leu Ala Val 770 775 780 Val His Leu Val Lys Pro Pro Ser Ala Leu Met Arg Pro Arg Ile Ala 785 790 795 800 Ser Arg Val Leu Gly Lys Trp Ala Arg Thr Arg Leu Ser Gly Gln Lys 805 810 815 Thr Leu Ile Ala Arg Asn Tyr Glu Asn His Pro Ile Pro Ala Glu Pro 820 825 830 Ala Asp Gln Leu Val Asn Ala 835 71185DNAMycobacterium tuberculosis 7atgaccacaa cgactacaac gatttctggg gggatattac ccaaggaata ccaagatctt 60cgggatacgg tggccgattt tgcgcgcacc gtggtcgcgc cggtatcggc caaacacgat 120gcggaacaca gcttcccata cgaaattgtc gccaagatgg gagagatggg cctgttcggg 180ctgccgtttc cggaggagta cggcggcatg ggcggcgact acttcgcgct gtcgctggta 240cttgaggagc tgggcaaggt tgaccaatcg gtagcgatca cgctggaggc cgcggtgggc 300ctgggtgcga tgccgatcta ccggttcggt accgaggagc agaaacagaa gtggttgccc 360gacttgacgt ctggccgtgc gctcgccggt tttggtctca ccgagccggg agcgggatcg 420gacgcgggca gcacccgcac cacggcgcgt ctcgaaggtg acgagtggat catcaacggc 480tccaagcaat ttatcaccaa ctcgggcacc gacatcacat cgctggtcac cgtcactgcg 540gttaccggga ccaccggaac cgctgcggat gccaagaaag agatttcgac gatcatcgtg 600cccagcggca caccgggatt caccgtggaa ccggtctata acaaggtcgg ctggaacgcc 660tcggacaccc acccactgac atttgccgat gcgcgggtcc cgagggagaa cctgctggga 720gcccggggga gcggctatgc caacttcttg tccatcctgg acgagggccg gattgcgatt 780gcagcgctgg ccaccggcgc ggcgcagggc tgtgttgacg agagcgtcaa gtacgccaac 840cagcgtcagt cgtttggcca gccgatcggc gcttatcagg cgatcggctt caagatcgcg 900cggatggagg cacgcgccca tgttgcccgc acagcgtact atgatgccgc cgcaaagatg 960ttggcgggca agcccttcaa gaaggaggcg gcgatcgcga agatgatctc ctcggaggcg 1020gcgatggaca actcccgcga tgccacccag atacacggcg gatacggctt tatgaacgaa 1080tatccggtgg cgcgtcatta ccgcgacagc aaggtgctcg agattggtga gggcaccacg 1140gaagtgcagc tgatgcttat cgcgcgatcg ttgggactgc agtga 11858394PRTMycobacterium tuberculosis 8Met Thr Thr Thr Thr Thr Thr Ile Ser Gly Gly Ile Leu Pro Lys Glu 1 5 10 15 Tyr Gln Asp Leu Arg Asp Thr Val Ala Asp Phe Ala Arg Thr Val Val 20 25 30 Ala Pro Val Ser Ala Lys His Asp Ala Glu His Ser Phe Pro Tyr Glu 35 40 45 Ile Val Ala Lys Met Gly Glu Met Gly Leu Phe Gly Leu Pro Phe Pro 50 55 60 Glu Glu Tyr Gly Gly Met Gly Gly Asp Tyr Phe Ala Leu Ser Leu Val 65 70 75 80 Leu Glu Glu Leu Gly Lys Val Asp Gln Ser Val Ala Ile Thr Leu Glu 85 90 95 Ala Ala Val Gly Leu Gly Ala Met Pro Ile Tyr Arg Phe Gly Thr Glu 100 105 110 Glu Gln Lys Gln Lys Trp Leu Pro Asp Leu Thr Ser Gly Arg Ala Leu 115 120 125 Ala Gly Phe Gly Leu Thr Glu Pro Gly Ala Gly Ser Asp Ala Gly Ser 130 135 140 Thr Arg Thr Thr Ala Arg Leu Glu Gly Asp Glu Trp Ile Ile Asn Gly 145 150 155 160 Ser Lys Gln Phe Ile Thr Asn Ser Gly Thr Asp Ile Thr Ser Leu Val 165 170 175 Thr Val Thr Ala Val Thr Gly Thr Thr Gly Thr Ala Ala Asp Ala Lys 180 185 190 Lys Glu Ile Ser Thr Ile Ile Val Pro Ser Gly Thr Pro Gly Phe Thr 195 200 205 Val Glu Pro Val Tyr Asn Lys Val Gly Trp Asn Ala Ser Asp Thr His 210 215 220 Pro Leu Thr Phe Ala Asp Ala Arg Val Pro Arg Glu Asn Leu Leu Gly 225 230 235 240 Ala Arg Gly Ser Gly Tyr Ala Asn Phe Leu Ser Ile Leu Asp Glu Gly 245 250 255 Arg Ile Ala Ile Ala Ala Leu Ala Thr Gly Ala Ala Gln Gly Cys Val 260 265 270 Asp Glu Ser Val Lys Tyr Ala Asn Gln Arg Gln Ser Phe Gly Gln Pro 275 280 285 Ile Gly Ala Tyr Gln Ala Ile Gly Phe Lys Ile Ala Arg Met Glu Ala 290 295 300 Arg Ala His Val Ala Arg Thr Ala Tyr Tyr Asp Ala Ala Ala Lys Met 305 310 315 320 Leu Ala Gly Lys Pro Phe Lys Lys Glu Ala Ala Ile Ala Lys Met Ile 325 330 335 Ser Ser Glu Ala Ala Met Asp Asn Ser Arg Asp Ala Thr Gln Ile His 340 345 350 Gly Gly Tyr Gly Phe Met Asn Glu Tyr Pro Val Ala Arg His Tyr Arg 355 360 365 Asp Ser Lys Val Leu Glu Ile Gly Glu Gly Thr Thr Glu Val Gln Leu 370 375 380 Met Leu Ile Ala Arg Ser Leu Gly Leu Gln 385 390 9747DNAMycobacterium tuberculosis 9atggacaagg tggtggccac cgccgcggag gcggtcgcag acatagccaa cgggtcgtcg 60cttgcggttg gtggattcgg gctttgcggc atccccgaag cactgatcgc agcgttggtg 120gatagcggtg tcaccgacct ggaaacagtc tcgaacaact gcggaatcga cggtgttggt 180ctgggactat tgttgcaaca caagcgaatt cgccggacag tctcctccta cgtgggggag 240aacaaggagt tcgcccgcca gttcctcgcg ggcgagctcg aggtggaact gaccccgcag 300ggcacgctgg ccgagcggtt gcgggccgga gggatgggca taccggcctt ctatacaccg 360gcaggggtcg gtacccaggt cgccgacggc gggttgccgt ggcgctacga cgcctcgggc 420ggggtggcgg tggtgtcgcc ggccaaggag actcgggagt tcgatggtgt cacctatgtc 480ctcgagcggg ggatccggac cgacttcgca ctggtgcatg cctggcaggg ggaccggcac 540ggcaacctga tgtaccgcca cgccgcggcc aacttcaacc cggagtgcgc atccgcaggc 600aggatcacga tcgccgaggt cgagcacttg gtcgagccgg gtgagatcga ccctgccacc 660gtacacaccc cgggcgtgtt tgtgcaccgg gtggttcatg tgcccaaccc cgccaagaag 720atcgagaggg agacggtgcg gcaatga 74710248PRTMycobacterium tuberculosis 10Met Asp Lys Val Val Ala Thr Ala Ala Glu Ala Val Ala Asp Ile Ala 1 5 10 15 Asn Gly Ser Ser Leu Ala Val Gly Gly Phe Gly Leu Cys Gly Ile Pro 20 25 30 Glu Ala Leu Ile Ala Ala Leu Val Asp Ser Gly Val Thr Asp Leu Glu 35 40 45 Thr Val Ser Asn Asn Cys Gly Ile Asp Gly Val Gly Leu Gly Leu Leu 50 55 60 Leu Gln His Lys Arg Ile Arg Arg Thr Val Ser Ser Tyr Val Gly Glu 65 70 75 80 Asn Lys Glu Phe Ala Arg Gln Phe Leu Ala Gly Glu Leu Glu Val Glu 85 90 95 Leu Thr Pro Gln Gly Thr Leu Ala Glu Arg Leu Arg Ala Gly Gly Met 100 105 110 Gly Ile Pro Ala Phe Tyr Thr Pro Ala Gly Val Gly Thr Gln Val Ala 115 120 125 Asp Gly Gly Leu Pro Trp Arg Tyr Asp Ala Ser Gly Gly Val Ala Val 130 135 140 Val Ser Pro Ala Lys Glu Thr Arg Glu Phe Asp Gly Val Thr Tyr Val 145 150 155 160 Leu Glu Arg Gly Ile Arg Thr Asp Phe Ala Leu Val His Ala Trp Gln 165 170 175 Gly Asp Arg His Gly Asn Leu Met Tyr Arg His Ala Ala Ala Asn Phe 180 185 190 Asn Pro Glu Cys Ala Ser Ala Gly Arg Ile Thr Ile Ala Glu Val Glu 195 200 205 His Leu Val Glu Pro Gly Glu Ile Asp Pro Ala Thr Val His Thr Pro 210 215 220 Gly Val Phe Val His Arg Val Val His Val Pro Asn Pro Ala Lys Lys 225 230 235 240 Ile Glu Arg Glu Thr Val Arg Gln 245 112157DNAMycobacterium tuberculosis 11atgaccctgg aagtggtatc ggacgcggcc ggacgcatgc gggtcaaagt cgactgggtc 60cgttgcgatt cccggcgcgc ggtcgcggtc gaagaggccg ttgccaagca gaacggtgtg 120cgcgtcgtgc acgcctaccc gcgcaccggg tccgtggtcg tgtggtattc acccagacgc 180gccgaccgcg cggcggtgct ggcggcgatc aagggcgccg cgcacgtcgc cgccgaactg 240atccccgcgc gtgcgccgca ctcggccgag atccgcaaca ccgacgtgct ccggatggtc 300atcggcgggg tggcactggc cttgctcggg gtgcgccgct acgtgttcgc gcggccaccg 360ctgctcggaa ccaccgggcg gacggtggcc accggtgtca ccattttcac cgggtatccg 420ttcctgcgtg gcgcgctgcg ctcgctgcgc tccggaaagg ccggcaccga tgccctggtc 480tccgcggcga cggtggcaag cctcatcctg cgcgagaacg tggtcgcact caccgtcctg 540tggttgctca acatcggtga gtacctgcag gatctgacgc tgcggcggac ccggcgggcc 600atctcggagc tgctgcgcgg caaccaggac acggcctggg tgcgcctcac cgatccttct 660gcaggctccg acgcggccac cgaaatccag gtcccgatcg acaccgtgca gatcggtgac 720gaggtggtgg tccacgagca cgtcgcgata ccggtcgacg gtgaggtggt cgacggcgaa 780gcgatcgtca atcagtccgc gatcaccggg gaaaacctgc cggtcagcgt cgtggtcgga 840acgcgcgtgc acgccggttc ggtcgtggtg cgcggacgcg tggtggtgcg cgcccacgcg 900gtaggcaacc aaaccaccat cggtcgcatc attagcaggg tcgaagaggc tcagctcgac 960cgggcaccca tccagacggt gggcgagaac ttctcccgcc gcttcgttcc cacctcgttc 1020atcgtctcgg ccatcgcgtt gctgatcacc ggcgacgtgc ggcgcgcgat gaccatgttg 1080ttgatcgcat gcccgtgcgc ggtgggactg tccaccccga ccgcgatcag cgcagcgatc 1140ggcaacggcg cgcgccgtgg catcctgatc aagggcggat cccacctcga gcaggcgggc 1200cgcgtcgacg ccatcgtgtt cgacaagacc gggacgttga ccgtgggccg ccccgtggtc 1260accaatatcg ttgccatgca taaagattgg gagcccgagc aagtgctggc ctatgccgcc 1320agctcggaga tccactcacg tcatccgctg gccgaggcgg tgatccgctc gacggaggaa 1380cgccgcatca gcatcccacc acacgaggag tgcgaggtgc tggtcggcct gggcatgcgg 1440acctgggccg acggtcggac cctgctgctg ggcagtccgt cgttgctgcg cgccgaaaaa 1500gttcgggtgt ccaagaaggc gtcggagtgg gtcgacaagc tgcgccgcca ggcggagacc 1560ccgctgctgc tcgcggtgga cggcacgctg gtcggcctga tcagcctgcg cgacgaggtg 1620cgtccggagg cggcccaggt gctgacgaag ctgcgggcca atgggattcg ccggatcgtc 1680atgctcaccg gcgaccaccc ggagatcgcc caggttgtcg ccgacgaact ggggattgat 1740gagtggcgcg ccgaggtcat gccggaggac aagctcgcgg cggtgcgcga gctgcaggac 1800gacggctacg tcgtcgggat ggtcggcgac ggcatcaacg acgccccggc gctggccgcc 1860gccgatatcg ggatcgccat gggccttgcc ggaaccgacg tcgccgtcga gaccgccgat 1920gtcgcgctgg ccaacgacga cctgcaccgc ctgctcgacg ttggggacct gggcgagcgg 1980gcagtggatg taatccggca gaactacggc atgtccatcg ccgtcaacgc ggccgggctg 2040ctgatcggcg cgggcggtgc gctctcgccg gtgctggcgg cgatcctgca caacgcgtcg 2100tcggtggcgg tggtggccaa cagttcccgg ttgatccgct accgcctgga ccgctag 215712718PRTMycobacterium tuberculosis 12Met Thr Leu Glu Val Val Ser Asp Ala Ala Gly Arg Met Arg Val Lys 1 5 10 15 Val Asp Trp Val Arg Cys Asp Ser Arg Arg Ala Val Ala Val Glu Glu 20 25 30 Ala Val Ala Lys Gln Asn Gly Val Arg Val Val His Ala Tyr Pro Arg 35 40 45 Thr Gly Ser Val Val Val Trp Tyr Ser Pro Arg Arg Ala Asp Arg Ala 50 55 60 Ala Val Leu Ala Ala Ile Lys Gly Ala Ala His Val Ala Ala Glu Leu 65 70 75 80 Ile Pro Ala Arg Ala Pro His Ser Ala Glu Ile Arg Asn Thr Asp Val 85 90 95 Leu Arg Met Val Ile Gly Gly Val Ala Leu Ala Leu Leu Gly Val Arg 100 105 110 Arg Tyr Val Phe Ala Arg Pro Pro Leu Leu Gly Thr Thr Gly Arg Thr 115 120 125 Val Ala Thr Gly Val Thr Ile Phe Thr Gly Tyr Pro Phe Leu Arg Gly 130 135 140 Ala Leu Arg Ser Leu Arg Ser Gly Lys Ala Gly Thr Asp Ala Leu Val 145 150 155 160 Ser Ala Ala Thr Val Ala Ser Leu Ile Leu Arg Glu Asn Val Val Ala 165 170 175 Leu Thr Val Leu Trp Leu Leu Asn Ile Gly Glu Tyr Leu Gln Asp Leu 180 185 190 Thr Leu Arg Arg Thr Arg Arg Ala Ile Ser Glu Leu Leu Arg Gly Asn 195 200 205 Gln Asp Thr Ala Trp Val Arg Leu Thr Asp Pro Ser Ala Gly Ser Asp 210 215 220 Ala Ala Thr Glu Ile Gln Val Pro Ile Asp Thr Val Gln Ile Gly Asp 225 230 235 240 Glu Val Val Val His Glu His Val Ala Ile Pro Val Asp Gly Glu Val 245 250 255 Val Asp Gly Glu Ala Ile Val Asn Gln Ser Ala Ile Thr Gly Glu Asn 260 265 270 Leu Pro Val Ser Val Val Val Gly Thr Arg Val His Ala Gly Ser Val 275 280 285 Val Val Arg Gly Arg Val Val Val Arg Ala His Ala Val Gly Asn Gln 290 295 300 Thr Thr Ile Gly Arg Ile Ile Ser Arg Val Glu Glu Ala Gln Leu Asp 305 310 315 320 Arg Ala Pro Ile Gln Thr Val Gly Glu Asn Phe Ser Arg Arg Phe Val 325 330 335 Pro Thr Ser Phe Ile Val Ser Ala Ile Ala Leu Leu Ile Thr Gly Asp 340 345 350 Val Arg Arg Ala Met Thr Met Leu Leu Ile Ala Cys Pro Cys Ala Val 355 360 365 Gly Leu Ser Thr Pro Thr Ala Ile Ser Ala Ala Ile Gly Asn Gly Ala 370 375 380 Arg Arg Gly Ile Leu Ile Lys Gly Gly Ser His Leu Glu Gln Ala Gly 385 390 395 400 Arg Val Asp Ala Ile Val Phe Asp Lys Thr Gly Thr Leu Thr Val Gly 405 410 415 Arg Pro Val Val Thr Asn Ile Val Ala Met His Lys Asp Trp Glu Pro 420 425 430 Glu Gln Val Leu Ala Tyr Ala Ala Ser Ser Glu Ile His Ser Arg His 435 440 445 Pro Leu Ala Glu Ala Val Ile Arg Ser Thr Glu Glu Arg Arg Ile Ser 450 455 460 Ile Pro Pro His Glu Glu Cys Glu Val Leu Val Gly Leu Gly Met Arg 465 470 475 480 Thr Trp Ala Asp Gly Arg Thr Leu Leu Leu Gly Ser Pro Ser Leu Leu 485 490 495 Arg Ala Glu Lys Val Arg Val Ser Lys Lys Ala Ser Glu Trp Val Asp 500 505 510 Lys Leu Arg Arg Gln Ala Glu Thr Pro Leu Leu Leu Ala Val Asp Gly 515 520 525 Thr Leu Val Gly Leu Ile Ser Leu Arg Asp Glu Val Arg Pro Glu Ala 530 535 540 Ala Gln Val Leu Thr Lys Leu Arg Ala Asn Gly Ile Arg Arg Ile Val 545 550 555 560 Met Leu Thr Gly Asp His Pro Glu Ile Ala Gln Val Val Ala Asp Glu 565 570 575 Leu Gly Ile Asp Glu Trp Arg Ala Glu Val Met Pro Glu Asp Lys Leu 580 585 590 Ala Ala Val Arg Glu Leu Gln Asp Asp Gly Tyr Val Val Gly Met Val 595 600 605 Gly Asp Gly Ile Asn Asp Ala Pro Ala Leu Ala Ala Ala Asp Ile Gly 610 615 620 Ile Ala Met Gly Leu Ala Gly Thr Asp Val Ala Val Glu Thr Ala Asp 625 630 635 640 Val Ala Leu Ala Asn Asp Asp Leu His Arg Leu Leu Asp Val Gly Asp 645 650 655 Leu Gly Glu Arg Ala Val Asp Val Ile Arg Gln Asn Tyr Gly Met Ser 660 665 670 Ile Ala Val Asn Ala Ala Gly Leu Leu Ile Gly Ala Gly Gly Ala Leu 675 680 685 Ser Pro Val Leu Ala Ala Ile Leu His Asn Ala Ser Ser Val Ala Val 690 695 700 Val Ala Asn Ser Ser Arg Leu Ile Arg Tyr Arg Leu Asp Arg 705 710 715 131692DNAMycobacterium tuberculosis 13atgactgtgc aggagttcga cgtcgtggtg

gtcggcagcg gcgccgccgg catggttgct 60gcgctggtcg ccgctcaccg aggtctctcg acggtagtcg tcgagaaggc cccgcactac 120ggcggctcca ccgcacgctc gggcggcggc gtctggatcc ccaacaacga ggtcctcaag 180cgccgcggcg ttcgagatac accggaggcg gcacgcacct atctgcacgg catcgtcggc 240gaaatcgtcg agccggaacg catcgatgct tacctcgacc gcgggcccga gatgctgtcg 300ttcgtgctga agcacacgcc gctgaagatg tgctgggtac ccggctactc cgactactac 360cccgaggctc cgggcggccg cccgggcgga cgttcgatcg agccgaaacc gttcaacgcg 420cgcaagcttg gtgccgacat ggccgggctg gagcccgcgt atggcaaggt tccgctcaat 480gtggttgtga tgcagcagga ctacgttcgc ctcaatcagc tcaaacgtca cccccgtggc 540gtgctgcgca gcatgaaggt cggcgcccgc acgatgtggg cgaaggcaac aggtaagaac 600ctggtcggca tgggtcgagc cctcattggg ccgttgcgga tcgggttgca gcgcgccgga 660gtgccggtcg aactcaacac cgccttcacc gatcttttcg tcgaaaatgg cgtcgtgtcc 720ggggtatacg tccgcgattc ccacgaggcg gaatccgctg agccgcagct gatccgggct 780cgccgcggcg tgatcctggc ctgtggtggt ttcgagcata acgagcagat gcgaatcaag 840taccagcggg cacccatcac caccgagtgg accgtgggcg ccagcgccaa taccggtgac 900ggcattctcg ccgccgaaaa gctcggcgca gcactggatc tgatggatga cgcttggtgg 960ggcccgacgg taccgctggt cggcaaacca tggttcgcgc tctcggagcg caactctccc 1020ggttcgatca tcgtcaacat gtcaggcaag cgattcatga acgaatcgat gccatacgtc 1080gaagcctgtc atcatatgta cggcggcgaa cacggccagg ggcccggacc gggcgagaac 1140attccggcgt ggctggtgtt cgaccagcga taccgggacc gctacatctt cgcgggacta 1200caaccagggc aacgcattcc gagcaggtgg ctggattccg gcgtcatcgt ccaggccgat 1260acccttgcgg agctggccgg caaggccggt ctacccgcgg acgaactcac tgccaccgtc 1320cagcgtttca acgcattcgc ccggtccggt gtcgacgagg actaccaccg cggggaaagt 1380gcctacgatc gctactacgg cgacccgagc aacaagccca atccgaacct cggcgaggtc 1440ggccacccgc cctattatgg cgccaagatg gttccgggcg acctggggac caagggcggt 1500atccgcaccg atgtcaacgg acgtgctctg cgggacgacg gcagcatcat cgacggcctt 1560tacgctgcag gcaatgtcag tgccccagtg atgggacaca cctaccccgg tccgggcggc 1620acgataggcc cggcgatgac gttcgggtac ctggcggcgc tgcacattgc cgatcaggcg 1680ggaaagcgct ga 169214563PRTMycobacterium tuberculosis 14Met Thr Val Gln Glu Phe Asp Val Val Val Val Gly Ser Gly Ala Ala 1 5 10 15 Gly Met Val Ala Ala Leu Val Ala Ala His Arg Gly Leu Ser Thr Val 20 25 30 Val Val Glu Lys Ala Pro His Tyr Gly Gly Ser Thr Ala Arg Ser Gly 35 40 45 Gly Gly Val Trp Ile Pro Asn Asn Glu Val Leu Lys Arg Arg Gly Val 50 55 60 Arg Asp Thr Pro Glu Ala Ala Arg Thr Tyr Leu His Gly Ile Val Gly 65 70 75 80 Glu Ile Val Glu Pro Glu Arg Ile Asp Ala Tyr Leu Asp Arg Gly Pro 85 90 95 Glu Met Leu Ser Phe Val Leu Lys His Thr Pro Leu Lys Met Cys Trp 100 105 110 Val Pro Gly Tyr Ser Asp Tyr Tyr Pro Glu Ala Pro Gly Gly Arg Pro 115 120 125 Gly Gly Arg Ser Ile Glu Pro Lys Pro Phe Asn Ala Arg Lys Leu Gly 130 135 140 Ala Asp Met Ala Gly Leu Glu Pro Ala Tyr Gly Lys Val Pro Leu Asn 145 150 155 160 Val Val Val Met Gln Gln Asp Tyr Val Arg Leu Asn Gln Leu Lys Arg 165 170 175 His Pro Arg Gly Val Leu Arg Ser Met Lys Val Gly Ala Arg Thr Met 180 185 190 Trp Ala Lys Ala Thr Gly Lys Asn Leu Val Gly Met Gly Arg Ala Leu 195 200 205 Ile Gly Pro Leu Arg Ile Gly Leu Gln Arg Ala Gly Val Pro Val Glu 210 215 220 Leu Asn Thr Ala Phe Thr Asp Leu Phe Val Glu Asn Gly Val Val Ser 225 230 235 240 Gly Val Tyr Val Arg Asp Ser His Glu Ala Glu Ser Ala Glu Pro Gln 245 250 255 Leu Ile Arg Ala Arg Arg Gly Val Ile Leu Ala Cys Gly Gly Phe Glu 260 265 270 His Asn Glu Gln Met Arg Ile Lys Tyr Gln Arg Ala Pro Ile Thr Thr 275 280 285 Glu Trp Thr Val Gly Ala Ser Ala Asn Thr Gly Asp Gly Ile Leu Ala 290 295 300 Ala Glu Lys Leu Gly Ala Ala Leu Asp Leu Met Asp Asp Ala Trp Trp 305 310 315 320 Gly Pro Thr Val Pro Leu Val Gly Lys Pro Trp Phe Ala Leu Ser Glu 325 330 335 Arg Asn Ser Pro Gly Ser Ile Ile Val Asn Met Ser Gly Lys Arg Phe 340 345 350 Met Asn Glu Ser Met Pro Tyr Val Glu Ala Cys His His Met Tyr Gly 355 360 365 Gly Glu His Gly Gln Gly Pro Gly Pro Gly Glu Asn Ile Pro Ala Trp 370 375 380 Leu Val Phe Asp Gln Arg Tyr Arg Asp Arg Tyr Ile Phe Ala Gly Leu 385 390 395 400 Gln Pro Gly Gln Arg Ile Pro Ser Arg Trp Leu Asp Ser Gly Val Ile 405 410 415 Val Gln Ala Asp Thr Leu Ala Glu Leu Ala Gly Lys Ala Gly Leu Pro 420 425 430 Ala Asp Glu Leu Thr Ala Thr Val Gln Arg Phe Asn Ala Phe Ala Arg 435 440 445 Ser Gly Val Asp Glu Asp Tyr His Arg Gly Glu Ser Ala Tyr Asp Arg 450 455 460 Tyr Tyr Gly Asp Pro Ser Asn Lys Pro Asn Pro Asn Leu Gly Glu Val 465 470 475 480 Gly His Pro Pro Tyr Tyr Gly Ala Lys Met Val Pro Gly Asp Leu Gly 485 490 495 Thr Lys Gly Gly Ile Arg Thr Asp Val Asn Gly Arg Ala Leu Arg Asp 500 505 510 Asp Gly Ser Ile Ile Asp Gly Leu Tyr Ala Ala Gly Asn Val Ser Ala 515 520 525 Pro Val Met Gly His Thr Tyr Pro Gly Pro Gly Gly Thr Ile Gly Pro 530 535 540 Ala Met Thr Phe Gly Tyr Leu Ala Ala Leu His Ile Ala Asp Gln Ala 545 550 555 560 Gly Lys Arg 15843DNAMycobacterium tuberculosis 15gtgagtccgg cgcccgtgca ggtgatgggg gttctaaacg tcacggacga ctctttctcg 60gacggcgggt gttatctcga tctcgacgat gcggtgaagc acggtctggc gatggcagcc 120gcaggtgcgg gcatcgtcga cgtcggtggt gagtcgagcc ggcccggtgc cactcgggtt 180gacccggcgg tggagacgtc tcgtgtcata cccgtcgtca aagagcttgc agcacaaggc 240atcaccgtca gcatcgatac catgcgcgcg gatgtcgctc gggcggcgtt gcagaacggt 300gcccagatgg tcaacgacgt gtcgggtggg cgggccgatc cggcgatggg gccgctgttg 360gccgaggccg atgtgccgtg ggtgttgatg cactggcggg cggtatcggc cgataccccg 420catgtgcctg tgcgctacgg caacgtggtg gccgaggtcc gtgccgacct gctggccagc 480gtcgccgacg cggtggccgc aggcgtcgac ccggcaaggc tggtgctcga tcccgggctt 540ggattcgcca agacggcgca acataattgg gcgatcttgc atgcccttcc ggaactggtc 600gcgaccggaa tcccagtgct ggtgggtgct tcgcgcaagc gcttcctcgg tgcgttgttg 660gccgggcccg acggcgtgat gcggccaacc gatgggcgtg acaccgcgac ggcggtgatt 720tccgcgctgg ccgcactgca cggggcctgg ggtgtgcggg tgcatgatgt gcgggcctcg 780gtcgatgcca tcaaggtggt cgaagcgtgg atgggagcgg aaaggataga acgcgatggc 840tga 84316280PRTMycobacterium tuberculosis 16Val Ser Pro Ala Pro Val Gln Val Met Gly Val Leu Asn Val Thr Asp 1 5 10 15 Asp Ser Phe Ser Asp Gly Gly Cys Tyr Leu Asp Leu Asp Asp Ala Val 20 25 30 Lys His Gly Leu Ala Met Ala Ala Ala Gly Ala Gly Ile Val Asp Val 35 40 45 Gly Gly Glu Ser Ser Arg Pro Gly Ala Thr Arg Val Asp Pro Ala Val 50 55 60 Glu Thr Ser Arg Val Ile Pro Val Val Lys Glu Leu Ala Ala Gln Gly 65 70 75 80 Ile Thr Val Ser Ile Asp Thr Met Arg Ala Asp Val Ala Arg Ala Ala 85 90 95 Leu Gln Asn Gly Ala Gln Met Val Asn Asp Val Ser Gly Gly Arg Ala 100 105 110 Asp Pro Ala Met Gly Pro Leu Leu Ala Glu Ala Asp Val Pro Trp Val 115 120 125 Leu Met His Trp Arg Ala Val Ser Ala Asp Thr Pro His Val Pro Val 130 135 140 Arg Tyr Gly Asn Val Val Ala Glu Val Arg Ala Asp Leu Leu Ala Ser 145 150 155 160 Val Ala Asp Ala Val Ala Ala Gly Val Asp Pro Ala Arg Leu Val Leu 165 170 175 Asp Pro Gly Leu Gly Phe Ala Lys Thr Ala Gln His Asn Trp Ala Ile 180 185 190 Leu His Ala Leu Pro Glu Leu Val Ala Thr Gly Ile Pro Val Leu Val 195 200 205 Gly Ala Ser Arg Lys Arg Phe Leu Gly Ala Leu Leu Ala Gly Pro Asp 210 215 220 Gly Val Met Arg Pro Thr Asp Gly Arg Asp Thr Ala Thr Ala Val Ile 225 230 235 240 Ser Ala Leu Ala Ala Leu His Gly Ala Trp Gly Val Arg Val His Asp 245 250 255 Val Arg Ala Ser Val Asp Ala Ile Lys Val Val Glu Ala Trp Met Gly 260 265 270 Ala Glu Arg Ile Glu Arg Asp Gly 275 280 172190DNAMycobacterium tuberculosis 17atgagtatta ccaggccgac gggcagctat gccagacaga tgctggatcc gggcggctgg 60gtggaagccg atgaagacac tttctatgac cgggcccagg aatatagcca ggttttgcaa 120agggtcaccg atgtattgga cacctgccgc cagcagaaag gccacgtctt cgaaggcggc 180ctatggtccg gcggcgccgc caatgctgcc aacggcgccc tgggtgcaaa catcaatcaa 240ttgatgacgc tgcaggatta tctcgccacg gtgattacct ggcacaggca tattgccggg 300ttgattgagc aagctaaatc cgatatcggc aataatgtgg atggcgctca acgggagatc 360gatatcctgg agaatgaccc tagcctggat gctgatgagc gccataccgc catcaattca 420ttggtcacgg cgacgcatgg ggccaatgtc agtctggtcg ccgagaccgc tgagcgggtg 480ctggaatcca agaattggaa acctccgaag aacgcactcg aggatttgct tcagcagaag 540tcgccgccac ccccagacgt gcctaccctg gtcgtgccat ccccgggcac accgggcaca 600ccgggaaccc cgatcacccc gggaaccccg atcaccccgg gaaccccaat cacacccatc 660ccgggagcgc cggtaactcc gatcacacca acgcccggca ctcccgtcac gccggtgacc 720ccgggcaagc cggtcacccc ggtgaccccg gtcaaaccgg gcacaccagg cgagccaacc 780ccgatcacgc cggtcacccc cccggtcgcc ccggccacac cggcaacccc ggccacgccc 840gttaccccag ctcccgctcc acacccgcag ccggctccgg caccggcgcc atcgcctggg 900ccccagccgg ttacaccggc cactcccggt ccgtctggtc cagcaacacc gggcacccca 960gggggcgagc cggcgccgca cgtcaaaccc gcggcgttgg cggagcaacc tggtgtgccg 1020ggccagcatg cgggcggggg gacgcagtcg gggcctgccc atgcggacga atccgccgcg 1080tcggtgacgc cggctgcggc gtccggtgtc ccgggcgcac gggcggcggc cgccgcgccg 1140agcggtaccg ccgtgggagc gggcgcgcgt tcgagcgtgg gtacggccgc ggcctcgggc 1200gcggggtcgc atgctgccac tgggcgggcg ccggtggcta cctcggacaa ggcggcggca 1260ccgagcacgc gggcggcctc ggcgcggacg gcacctcctg cccgcccgcc gtcgaccgat 1320cacatcgaca aacccgatcg cagcgagtct gcagatgacg gtacgccggt gtcgatgatc 1380ccggtgtcgg cggctcgggc ggcacgcgac gccgccactg cagctgccag cgcccgccag 1440cgtggccgcg gtgatgcgct gcggttggcg cgacgcatcg cggcggcgct caacgcgtcc 1500gacaacaacg cgggcgacta cgggttcttc tggatcaccg cggtgaccac cgacggttcc 1560atcgtcgtgg ccaacagcta tgggctggcc tacatacccg acgggatgga attgccgaat 1620aaggtgtact tggccagcgc ggatcacgca atcccggttg acgaaattgc acgctgtgcc 1680acctacccgg ttttggccgt gcaagcctgg gcggctttcc acgacatgac gctgcgggcg 1740gtgatcggta ccgcggagca gttggccagt tcggatcccg gtgtggccaa gattgtgctg 1800gagccagatg acattccgga gagcggcaaa atgacgggcc ggtcgcggct ggaggtcgtc 1860gacccctcgg cggcggctca gctggccgac actaccgatc agcgtttgct cgacttgttg 1920ccgccggcgc cggtggatgt caatccaccg ggcgatgagc ggcacatgct gtggttcgag 1980ctgatgaagc ccatgaccag caccgctacc ggccgcgagg ccgctcatct gcgggcgttc 2040cgggcctacg ctgcccactc acaggagatt gccctgcacc aagcgcacac tgcgactgac 2100gcggccgtcc agcgtgtggc cgtcgcggac tggctgtact ggcaatacgt caccgggttg 2160ctcgaccggg ccctggccgc cgcatgctga 219018729PRTMycobacterium tuberculosis 18Met Ser Ile Thr Arg Pro Thr Gly Ser Tyr Ala Arg Gln Met Leu Asp 1 5 10 15 Pro Gly Gly Trp Val Glu Ala Asp Glu Asp Thr Phe Tyr Asp Arg Ala 20 25 30 Gln Glu Tyr Ser Gln Val Leu Gln Arg Val Thr Asp Val Leu Asp Thr 35 40 45 Cys Arg Gln Gln Lys Gly His Val Phe Glu Gly Gly Leu Trp Ser Gly 50 55 60 Gly Ala Ala Asn Ala Ala Asn Gly Ala Leu Gly Ala Asn Ile Asn Gln 65 70 75 80 Leu Met Thr Leu Gln Asp Tyr Leu Ala Thr Val Ile Thr Trp His Arg 85 90 95 His Ile Ala Gly Leu Ile Glu Gln Ala Lys Ser Asp Ile Gly Asn Asn 100 105 110 Val Asp Gly Ala Gln Arg Glu Ile Asp Ile Leu Glu Asn Asp Pro Ser 115 120 125 Leu Asp Ala Asp Glu Arg His Thr Ala Ile Asn Ser Leu Val Thr Ala 130 135 140 Thr His Gly Ala Asn Val Ser Leu Val Ala Glu Thr Ala Glu Arg Val 145 150 155 160 Leu Glu Ser Lys Asn Trp Lys Pro Pro Lys Asn Ala Leu Glu Asp Leu 165 170 175 Leu Gln Gln Lys Ser Pro Pro Pro Pro Asp Val Pro Thr Leu Val Val 180 185 190 Pro Ser Pro Gly Thr Pro Gly Thr Pro Gly Thr Pro Ile Thr Pro Gly 195 200 205 Thr Pro Ile Thr Pro Gly Thr Pro Ile Thr Pro Ile Pro Gly Ala Pro 210 215 220 Val Thr Pro Ile Thr Pro Thr Pro Gly Thr Pro Val Thr Pro Val Thr 225 230 235 240 Pro Gly Lys Pro Val Thr Pro Val Thr Pro Val Lys Pro Gly Thr Pro 245 250 255 Gly Glu Pro Thr Pro Ile Thr Pro Val Thr Pro Pro Val Ala Pro Ala 260 265 270 Thr Pro Ala Thr Pro Ala Thr Pro Val Thr Pro Ala Pro Ala Pro His 275 280 285 Pro Gln Pro Ala Pro Ala Pro Ala Pro Ser Pro Gly Pro Gln Pro Val 290 295 300 Thr Pro Ala Thr Pro Gly Pro Ser Gly Pro Ala Thr Pro Gly Thr Pro 305 310 315 320 Gly Gly Glu Pro Ala Pro His Val Lys Pro Ala Ala Leu Ala Glu Gln 325 330 335 Pro Gly Val Pro Gly Gln His Ala Gly Gly Gly Thr Gln Ser Gly Pro 340 345 350 Ala His Ala Asp Glu Ser Ala Ala Ser Val Thr Pro Ala Ala Ala Ser 355 360 365 Gly Val Pro Gly Ala Arg Ala Ala Ala Ala Ala Pro Ser Gly Thr Ala 370 375 380 Val Gly Ala Gly Ala Arg Ser Ser Val Gly Thr Ala Ala Ala Ser Gly 385 390 395 400 Ala Gly Ser His Ala Ala Thr Gly Arg Ala Pro Val Ala Thr Ser Asp 405 410 415 Lys Ala Ala Ala Pro Ser Thr Arg Ala Ala Ser Ala Arg Thr Ala Pro 420 425 430 Pro Ala Arg Pro Pro Ser Thr Asp His Ile Asp Lys Pro Asp Arg Ser 435 440 445 Glu Ser Ala Asp Asp Gly Thr Pro Val Ser Met Ile Pro Val Ser Ala 450 455 460 Ala Arg Ala Ala Arg Asp Ala Ala Thr Ala Ala Ala Ser Ala Arg Gln 465 470 475 480 Arg Gly Arg Gly Asp Ala Leu Arg Leu Ala Arg Arg Ile Ala Ala Ala 485 490 495 Leu Asn Ala Ser Asp Asn Asn Ala Gly Asp Tyr Gly Phe Phe Trp Ile 500 505 510 Thr Ala Val Thr Thr Asp Gly Ser Ile Val Val Ala Asn Ser Tyr Gly 515 520 525 Leu Ala Tyr Ile Pro Asp Gly Met Glu Leu Pro Asn Lys Val Tyr Leu 530 535 540 Ala Ser Ala Asp His Ala Ile Pro Val Asp Glu Ile Ala Arg Cys Ala 545 550 555 560 Thr Tyr Pro Val Leu Ala Val Gln Ala Trp Ala Ala Phe His Asp Met 565 570 575 Thr Leu Arg Ala Val Ile Gly Thr Ala Glu Gln Leu Ala Ser Ser Asp 580 585 590 Pro Gly Val Ala Lys Ile Val Leu Glu Pro Asp Asp Ile Pro Glu Ser 595 600 605 Gly Lys Met Thr Gly Arg Ser Arg Leu Glu Val Val Asp Pro Ser Ala 610 615 620 Ala Ala Gln Leu Ala Asp Thr Thr Asp Gln Arg Leu Leu Asp Leu Leu 625 630 635 640 Pro Pro Ala Pro Val Asp Val Asn Pro Pro Gly Asp Glu Arg His Met 645 650 655 Leu Trp Phe Glu Leu Met Lys Pro Met Thr Ser Thr Ala Thr Gly Arg 660 665 670 Glu Ala Ala His Leu Arg Ala Phe Arg Ala Tyr Ala Ala His Ser Gln 675 680 685 Glu Ile Ala Leu His Gln Ala His Thr Ala Thr Asp Ala Ala Val Gln 690 695 700

Arg Val Ala Val Ala Asp Trp Leu Tyr Trp Gln Tyr Val Thr Gly Leu 705 710 715 720 Leu Asp Arg Ala Leu Ala Ala Ala Cys 725 19338PRTMycobacterium tuberculosis 19Met 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 20325PRTMycobacterium tuberculosis 20Met 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 Ser Leu Gly Ala Gly 325 2195PRTMycobacterium tuberculosis 21Met 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 2296PRTMycobacterium tuberculosis 22Met 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 23355PRTMycobacterium tuberculosis 23Met Ser Asn Ser Arg Arg Arg Ser Leu Arg Trp Ser Trp Leu Leu Ser 1 5 10 15 Val Leu Ala Ala Val Gly Leu Gly Leu Ala Thr Ala Pro Ala Gln Ala 20 25 30 Ala Pro Pro Ala Leu Ser Gln Asp Arg Phe Ala Asp Phe Pro Ala Leu 35 40 45 Pro Leu Asp Pro Ser Ala Met Val Ala Gln Val Gly Pro Gln Val Val 50 55 60 Asn Ile Asn Thr Lys Leu Gly Tyr Asn Asn Ala Val Gly Ala Gly Thr 65 70 75 80 Gly Ile Val Ile Asp Pro Asn Gly Val Val Leu Thr Asn Asn His Val 85 90 95 Ile Ala Gly Ala Thr Asp Ile Asn Ala Phe Ser Val Gly Ser Gly Gln 100 105 110 Thr Tyr Gly Val Asp Val Val Gly Tyr Asp Arg Thr Gln Asp Val Ala 115 120 125 Val Leu Gln Leu Arg Gly Ala Gly Gly Leu Pro Ser Ala Ala Ile Gly 130 135 140 Gly Gly Val Ala Val Gly Glu Pro Val Val Ala Met Gly Asn Ser Gly 145 150 155 160 Gly Gln Gly Gly Thr Pro Arg Ala Val Pro Gly Arg Val Val Ala Leu 165 170 175 Gly Gln Thr Val Gln Ala Ser Asp Ser Leu Thr Gly Ala Glu Glu Thr 180 185 190 Leu Asn Gly Leu Ile Gln Phe Asp Ala Ala Ile Gln Pro Gly Asp Ser 195 200 205 Gly Gly Pro Val Val Asn Gly Leu Gly Gln Val Val Gly Met Asn Thr 210 215 220 Ala Ala Ser Asp Asn Phe Gln Leu Ser Gln Gly Gly Gln Gly Phe Ala 225 230 235 240 Ile Pro Ile Gly Gln Ala Met Ala Ile Ala Gly Gln Ile Arg Ser Gly 245 250 255 Gly Gly Ser Pro Thr Val His Ile Gly Pro Thr Ala Phe Leu Gly Leu 260 265 270 Gly Val Val Asp Asn Asn Gly Asn Gly Ala Arg Val Gln Arg Val Val 275 280 285 Gly Ser Ala Pro Ala Ala Ser Leu Gly Ile Ser Thr Gly Asp Val Ile 290 295 300 Thr Ala Val Asp Gly Ala Pro Ile Asn Ser Ala Thr Ala Met Ala Asp 305 310 315 320 Ala Leu Asn Gly His His Pro Gly Asp Val Ile Ser Val Thr Trp Gln 325 330 335 Thr Lys Ser Gly Gly Thr Arg Thr Gly Asn Val Thr Leu Ala Glu Gly 340 345 350 Pro Pro Ala 355 24391PRTMycobacterium tuberculosis 24Met 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 25236PRTMycobacterium tuberculosis 25Met Arg Thr Pro Arg Arg His Cys Arg Arg Ile Ala Val Leu Ala Ala 1 5 10 15 Val Ser Ile Ala Ala Thr Val Val Ala Gly Cys Ser Ser Gly Ser Lys 20 25 30 Pro Ser Gly Gly Pro Leu Pro Asp Ala Lys Pro Leu Val Glu Glu Ala 35 40 45 Thr Ala Gln Thr Lys Ala Leu Lys Ser Ala His Met Val Leu Thr Val 50 55 60 Asn Gly Lys Ile Pro Gly Leu Ser Leu Lys Thr Leu Ser Gly Asp Leu 65 70 75 80 Thr Thr Asn Pro Thr Ala Ala Thr Gly Asn Val Lys Leu Thr Leu Gly 85 90 95 Gly Ser Asp Ile Asp Ala Asp Phe Val Val Phe Asp Gly Ile Leu Tyr 100 105 110 Ala Thr Leu Thr Pro Asn Gln Trp Ser Asp Phe Gly Pro Ala Ala Asp 115 120 125 Ile Tyr Asp Pro Ala Gln Val Leu Asn Pro Asp Thr Gly Leu Ala Asn 130 135 140 Val Leu Ala Asn Phe Ala Asp Ala Lys Ala Glu Gly Arg Asp Thr Ile 145 150 155 160 Asn Gly Gln Asn Thr Ile Arg Ile Ser Gly Lys Val Ser Ala Gln Ala 165 170 175 Val Asn Gln Ile Ala Pro Pro Phe Asn Ala Thr Gln Pro Val Pro Ala 180 185 190 Thr Val Trp Ile Gln Glu Thr Gly Asp His Gln Leu Ala Gln Ala Gln 195 200 205 Leu Asp Arg Gly Ser Gly Asn Ser Val Gln Met Thr Leu Ser Lys Trp 210 215 220 Gly Glu Lys Val Gln Val Thr Lys Pro Pro Val Ser 225 230 235 26540PRTMycobacterium tuberculosis 26Met Ala Lys Thr Ile Ala Tyr Asp Glu Glu Ala Arg Arg Gly Leu Glu 1 5 10 15 Arg Gly Leu Asn Ala Leu Ala Asp Ala Val Lys Val Thr Leu Gly Pro 20 25 30 Lys Gly Arg Asn Val Val Leu Glu Lys Lys Trp Gly Ala Pro Thr Ile 35 40 45 Thr Asn Asp Gly Val Ser Ile Ala Lys Glu Ile Glu Leu Glu Asp Pro 50 55 60 Tyr Glu Lys Ile Gly Ala Glu Leu Val Lys Glu Val Ala Lys Lys Thr 65 70 75 80 Asp Asp Val Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala Gln 85 90 95 Ala Leu Val Arg Glu Gly Leu Arg Asn Val Ala Ala Gly Ala Asn Pro 100 105 110 Leu Gly Leu Lys Arg Gly Ile Glu Lys Ala Val Glu Lys Val Thr Glu 115 120 125 Thr Leu Leu Lys Gly Ala Lys Glu Val Glu Thr Lys Glu Gln Ile Ala 130 135 140 Ala Thr Ala Ala Ile Ser Ala Gly Asp Gln Ser Ile Gly Asp Leu Ile 145 150 155 160 Ala Glu Ala Met Asp Lys Val Gly Asn Glu Gly Val Ile Thr Val Glu 165 170 175 Glu Ser Asn Thr Phe Gly Leu Gln Leu Glu Leu Thr Glu Gly Met Arg 180 185 190 Phe Asp Lys Gly Tyr Ile Ser Gly Tyr Phe Val Thr Asp Pro Glu Arg 195 200 205 Gln Glu Ala Val Leu Glu Asp Pro Tyr Ile Leu Leu Val Ser Ser Lys 210 215 220 Val Ser Thr Val Lys Asp Leu Leu Pro Leu Leu Glu Lys Val Ile Gly 225 230 235 240 Ala Gly Lys Pro Leu Leu Ile Ile Ala Glu Asp Val Glu Gly Glu Ala 245 250 255 Leu Ser Thr Leu Val Val Asn Lys Ile Arg Gly Thr Phe Lys Ser Val 260 265 270 Ala Val Lys Ala Pro Gly Phe Gly Asp Arg Arg Lys Ala Met Leu Gln 275 280 285 Asp Met Ala Ile Leu Thr Gly Gly Gln Val Ile Ser Glu Glu Val Gly 290 295 300 Leu Thr Leu Glu Asn Ala Asp Leu Ser Leu Leu Gly Lys Ala Arg Lys 305 310 315 320 Val Val Val Thr Lys Asp Glu Thr Thr Ile Val Glu Gly Ala Gly Asp 325 330 335 Thr Asp Ala Ile Ala Gly Arg Val Ala Gln Ile Arg Gln Glu Ile Glu 340 345 350 Asn Ser Asp Ser Asp Tyr Asp Arg Glu Lys Leu Gln Glu Arg Leu Ala 355 360 365 Lys Leu Ala Gly Gly Val Ala Val Ile Lys Ala Gly Ala Ala Thr Glu 370 375 380 Val Glu Leu Lys Glu Arg Lys His Arg Ile Glu Asp Ala Val Arg Asn 385 390 395 400 Ala Lys Ala Ala

Val Glu Glu Gly Ile Val Ala Gly Gly Gly Val Thr 405 410 415 Leu Leu Gln Ala Ala Pro Thr Leu Asp Glu Leu Lys Leu Glu Gly Asp 420 425 430 Glu Ala Thr Gly Ala Asn Ile Val Lys Val Ala Leu Glu Ala Pro Leu 435 440 445 Lys Gln Ile Ala Phe Asn Ser Gly Leu Glu Pro Gly Val Val Ala Glu 450 455 460 Lys Val Arg Asn Leu Pro Ala Gly His Gly Leu Asn Ala Gln Thr Gly 465 470 475 480 Val Tyr Glu Asp Leu Leu Ala Ala Gly Val Ala Asp Pro Val Lys Val 485 490 495 Thr Arg Ser Ala Leu Gln Asn Ala Ala Ser Ile Ala Gly Leu Phe Leu 500 505 510 Thr Thr Glu Ala Val Val Ala Asp Lys Pro Glu Lys Glu Lys Ala Ser 515 520 525 Val Pro Gly Gly Gly Asp Met Gly Gly Met Asp Phe 530 535 540 27199PRTMycobacterium tuberculosis 27Met 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 28375PRTMycobacterium tuberculosis 28Val Thr Gln Thr Gly Lys Arg Gln Arg Arg Lys Phe Gly Arg Ile Arg 1 5 10 15 Gln Phe Asn Ser Gly Arg Trp Gln Ala Ser Tyr Thr Gly Pro Asp Gly 20 25 30 Arg Val Tyr Ile Ala Pro Lys Thr Phe Asn Ala Lys Ile Asp Ala Glu 35 40 45 Ala Trp Leu Thr Asp Arg Arg Arg Glu Ile Asp Arg Gln Leu Trp Ser 50 55 60 Pro Ala Ser Gly Gln Glu Asp Arg Pro Gly Ala Pro Phe Gly Glu Tyr 65 70 75 80 Ala Glu Gly Trp Leu Lys Gln Arg Gly Ile Lys Asp Arg Thr Arg Ala 85 90 95 His Tyr Arg Lys Leu Leu Asp Asn His Ile Leu Ala Thr Phe Ala Asp 100 105 110 Thr Asp Leu Arg Asp Ile Thr Pro Ala Ala Val Arg Arg Trp Tyr Ala 115 120 125 Thr Thr Ala Val Gly Thr Pro Thr Met Arg Ala His Ser Tyr Ser Leu 130 135 140 Leu Arg Ala Ile Met Gln Thr Ala Leu Ala Asp Asp Leu Ile Asp Ser 145 150 155 160 Asn Pro Cys Arg Ile Ser Gly Ala Ser Thr Ala Arg Arg Val His Lys 165 170 175 Ile Arg Pro Ala Thr Leu Asp Glu Leu Glu Thr Ile Thr Lys Ala Met 180 185 190 Pro Asp Pro Tyr Gln Ala Phe Val Leu Met Ala Ala Trp Leu Ala Met 195 200 205 Arg Tyr Gly Glu Leu Thr Glu Leu Arg Arg Lys Asp Ile Asp Leu His 210 215 220 Gly Glu Val Ala Arg Val Arg Arg Ala Val Val Arg Val Gly Glu Gly 225 230 235 240 Phe Lys Val Thr Thr Pro Lys Ser Asp Ala Gly Val Arg Asp Ile Ser 245 250 255 Ile Pro Pro His Leu Ile Pro Ala Ile Glu Asp His Leu His Lys His 260 265 270 Val Asn Pro Gly Arg Glu Ser Leu Leu Phe Pro Ser Val Asn Asp Pro 275 280 285 Asn Arg His Leu Ala Pro Ser Ala Leu Tyr Arg Met Phe Tyr Lys Ala 290 295 300 Arg Lys Ala Ala Gly Arg Pro Asp Leu Arg Val His Asp Leu Arg His 305 310 315 320 Ser Gly Ala Val Leu Ala Ala Ser Thr Gly Ala Thr Leu Ala Glu Leu 325 330 335 Met Gln Arg Leu Gly His Ser Thr Ala Gly Ala Ala Leu Arg Tyr Gln 340 345 350 His Ala Ala Lys Gly Arg Asp Arg Glu Ile Ala Ala Leu Leu Ser Lys 355 360 365 Leu Ala Glu Asn Gln Glu Met 370 375 2975PRTMycobacterium tuberculosis 29Val 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 30144PRTMycobacterium tuberculosis 30Met 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 80 Arg Thr Glu Gln Lys Asp Phe Asp Gly Arg Ser Glu Phe Ala Tyr Gly 85 90 95 Ser Phe Val Arg Thr Val Ser Leu Pro Val Gly Ala Asp Glu Asp Asp 100 105 110 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 31407PRTMycobacterium tuberculosis 31Met Ser Gly Arg His Arg Lys Pro Thr Thr Ser Asn Val Ser Val Ala 1 5 10 15 Lys Ile Ala Phe Thr Gly Ala Val Leu Gly Gly Gly Gly Ile Ala Met 20 25 30 Ala Ala Gln Ala Thr Ala Ala Thr Asp Gly Glu Trp Asp Gln Val Ala 35 40 45 Arg Cys Glu Ser Gly Gly Asn Trp Ser Ile Asn Thr Gly Asn Gly Tyr 50 55 60 Leu Gly Gly Leu Gln Phe Thr Gln Ser Thr Trp Ala Ala His Gly Gly 65 70 75 80 Gly Glu Phe Ala Pro Ser Ala Gln Leu Ala Ser Arg Glu Gln Gln Ile 85 90 95 Ala Val Gly Glu Arg Val Leu Ala Thr Gln Gly Arg Gly Ala Trp Pro 100 105 110 Val Cys Gly Arg Gly Leu Ser Asn Ala Thr Pro Arg Glu Val Leu Pro 115 120 125 Ala Ser Ala Ala Met Asp Ala Pro Leu Asp Ala Ala Ala Val Asn Gly 130 135 140 Glu Pro Ala Pro Leu Ala Pro Pro Pro Ala Asp Pro Ala Pro Pro Val 145 150 155 160 Glu Leu Ala Ala Asn Asp Leu Pro Ala Pro Leu Gly Glu Pro Leu Pro 165 170 175 Ala Ala Pro Ala Asp Pro Ala Pro Pro Ala Asp Leu Ala Pro Pro Ala 180 185 190 Pro Ala Asp Val Ala Pro Pro Val Glu Leu Ala Val Asn Asp Leu Pro 195 200 205 Ala Pro Leu Gly Glu Pro Leu Pro Ala Ala Pro Ala Asp Pro Ala Pro 210 215 220 Pro Ala Asp Leu Ala Pro Pro Ala Pro Ala Asp Leu Ala Pro Pro Ala 225 230 235 240 Pro Ala Asp Leu Ala Pro Pro Ala Pro Ala Asp Leu Ala Pro Pro Val 245 250 255 Glu Leu Ala Val Asn Asp Leu Pro Ala Pro Leu Gly Glu Pro Leu Pro 260 265 270 Ala Ala Pro Ala Glu Leu Ala Pro Pro Ala Asp Leu Ala Pro Ala Ser 275 280 285 Ala Asp Leu Ala Pro Pro Ala Pro Ala Asp Leu Ala Pro Pro Ala Pro 290 295 300 Ala Glu Leu Ala Pro Pro Ala Pro Ala Asp Leu Ala Pro Pro Ala Ala 305 310 315 320 Val Asn Glu Gln Thr Ala Pro Gly Asp Gln Pro Ala Thr Ala Pro Gly 325 330 335 Gly Pro Val Gly Leu Ala Thr Asp Leu Glu Leu Pro Glu Pro Asp Pro 340 345 350 Gln Pro Ala Asp Ala Pro Pro Pro Gly Asp Val Thr Glu Ala Pro Ala 355 360 365 Glu Thr Pro Gln Val Ser Asn Ile Ala Tyr Thr Lys Lys Leu Trp Gln 370 375 380 Ala Ile Arg Ala Gln Asp Val Cys Gly Asn Asp Ala Leu Asp Ser Leu 385 390 395 400 Ala Gln Pro Tyr Val Ile Gly 405 32362PRTMycobacterium tuberculosis 32Met Leu Arg Leu Val Val Gly Ala Leu Leu Leu Val Leu Ala Phe Ala 1 5 10 15 Gly Gly Tyr Ala Val Ala Ala Cys Lys Thr Val Thr Leu Thr Val Asp 20 25 30 Gly Thr Ala Met Arg Val Thr Thr Met Lys Ser Arg Val Ile Asp Ile 35 40 45 Val Glu Glu Asn Gly Phe Ser Val Asp Asp Arg Asp Asp Leu Tyr Pro 50 55 60 Ala Ala Gly Val Gln Val His Asp Ala Asp Thr Ile Val Leu Arg Arg 65 70 75 80 Ser Arg Pro Leu Gln Ile Ser Leu Asp Gly His Asp Ala Lys Gln Val 85 90 95 Trp Thr Thr Ala Ser Thr Val Asp Glu Ala Leu Ala Gln Leu Ala Met 100 105 110 Thr Asp Thr Ala Pro Ala Ala Ala Ser Arg Ala Ser Arg Val Pro Leu 115 120 125 Ser Gly Met Ala Leu Pro Val Val Ser Ala Lys Thr Val Gln Leu Asn 130 135 140 Asp Gly Gly Leu Val Arg Thr Val His Leu Pro Ala Pro Asn Val Ala 145 150 155 160 Gly Leu Leu Ser Ala Ala Gly Val Pro Leu Leu Gln Ser Asp His Val 165 170 175 Val Pro Ala Ala Thr Ala Pro Ile Val Glu Gly Met Gln Ile Gln Val 180 185 190 Thr Arg Asn Arg Ile Lys Lys Val Thr Glu Arg Leu Pro Leu Pro Pro 195 200 205 Asn Ala Arg Arg Val Glu Asp Pro Glu Met Asn Met Ser Arg Glu Val 210 215 220 Val Glu Asp Pro Gly Val Pro Gly Thr Gln Asp Val Thr Phe Ala Val 225 230 235 240 Ala Glu Val Asn Gly Val Glu Thr Gly Arg Leu Pro Val Ala Asn Val 245 250 255 Val Val Thr Pro Ala His Glu Ala Val Val Arg Val Gly Thr Lys Pro 260 265 270 Gly Thr Glu Val Pro Pro Val Ile Asp Gly Ser Ile Trp Asp Ala Ile 275 280 285 Ala Gly Cys Glu Ala Gly Gly Asn Trp Ala Ile Asn Thr Gly Asn Gly 290 295 300 Tyr Tyr Gly Gly Val Gln Phe Asp Gln Gly Thr Trp Glu Ala Asn Gly 305 310 315 320 Gly Leu Arg Tyr Ala Pro Arg Ala Asp Leu Ala Thr Arg Glu Glu Gln 325 330 335 Ile Ala Val Ala Glu Val Thr Arg Leu Arg Gln Gly Trp Gly Ala Trp 340 345 350 Pro Val Cys Ala Ala Arg Ala Gly Ala Arg 355 360 33176PRTMycobacterium tuberculosis 33Val His Pro Leu Pro Ala Asp His Gly Arg Ser Arg Cys Asn Arg His 1 5 10 15 Pro Ile Ser Pro Leu Ser Leu Ile Gly Asn Ala Ser Ala Thr Ser Gly 20 25 30 Asp Met Ser Ser Met Thr Arg Ile Ala Lys Pro Leu Ile Lys Ser Ala 35 40 45 Met Ala Ala Gly Leu Val Thr Ala Ser Met Ser Leu Ser Thr Ala Val 50 55 60 Ala His Ala Gly Pro Ser Pro Asn Trp Asp Ala Val Ala Gln Cys Glu 65 70 75 80 Ser Gly Gly Asn Trp Ala Ala Asn Thr Gly Asn Gly Lys Tyr Gly Gly 85 90 95 Leu Gln Phe Lys Pro Ala Thr Trp Ala Ala Phe Gly Gly Val Gly Asn 100 105 110 Pro Ala Ala Ala Ser Arg Glu Gln Gln Ile Ala Val Ala Asn Arg Val 115 120 125 Leu Ala Glu Gln Gly Leu Asp Ala Trp Pro Thr Cys Gly Ala Ala Ser 130 135 140 Gly Leu Pro Ile Ala Leu Trp Ser Lys Pro Ala Gln Gly Ile Lys Gln 145 150 155 160 Ile Ile Asn Glu Ile Ile Trp Ala Gly Ile Gln Ala Ser Ile Pro Arg 165 170 175 34154PRTMycobacterium tuberculosis 34Met Thr Pro Gly Leu Leu Thr Thr Ala Gly Ala Gly Arg Pro Arg Asp 1 5 10 15 Arg Cys Ala Arg Ile Val Cys Thr Val Phe Ile Glu Thr Ala Val Val 20 25 30 Ala Thr Met Phe Val Ala Leu Leu Gly Leu Ser Thr Ile Ser Ser Lys 35 40 45 Ala Asp Asp Ile Asp Trp Asp Ala Ile Ala Gln Cys Glu Ser Gly Gly 50 55 60 Asn Trp Ala Ala Asn Thr Gly Asn Gly Leu Tyr Gly Gly Leu Gln Ile 65 70 75 80 Ser Gln Ala Thr Trp Asp Ser Asn Gly Gly Val Gly Ser Pro Ala Ala 85 90 95 Ala Ser Pro Gln Gln Gln Ile Glu Val Ala Asp Asn Ile Met Lys Thr 100 105 110 Gln Gly Pro Gly Ala Trp Pro Lys Cys Ser Ser Cys Ser Gln Gly Asp 115 120 125 Ala Pro Leu Gly Ser Leu Thr His Ile Leu Thr Phe Leu Ala Ala Glu 130 135 140 Thr Gly Gly Cys Ser Gly Ser Arg Asp Asp 145 150 35172PRTMycobacterium tuberculosis 35Leu Lys Asn Ala Arg Thr Thr Leu Ile Ala Ala Ala Ile Ala Gly Thr 1 5 10 15 Leu Val Thr Thr Ser Pro Ala Gly Ile Ala Asn Ala Asp Asp Ala Gly 20 25 30 Leu Asp Pro Asn Ala Ala Ala Gly Pro Asp Ala Val Gly Phe Asp Pro 35 40 45 Asn Leu Pro Pro Ala Pro Asp Ala Ala Pro Val Asp Thr Pro Pro Ala 50 55 60 Pro Glu Asp Ala Gly Phe Asp Pro Asn Leu Pro Pro Pro Leu Ala Pro 65 70 75 80 Asp Phe Leu Ser Pro Pro Ala Glu Glu Ala Pro Pro Val Pro Val Ala 85 90 95 Tyr Ser Val Asn Trp Asp Ala Ile Ala Gln Cys Glu Ser Gly Gly Asn 100 105 110 Trp Ser Ile Asn Thr Gly Asn Gly Tyr Tyr Gly Gly Leu Arg Phe Thr 115 120 125 Ala Gly Thr Trp Arg Ala Asn Gly Gly Ser Gly Ser Ala Ala Asn Ala 130 135 140 Ser Arg Glu Glu Gln Ile Arg Val Ala Glu Asn Val Leu Arg Ser Gln 145 150 155 160 Gly Ile Arg Ala Trp Pro Val Cys Gly Arg Arg Gly 165 170 36210PRTMycobacterium tuberculosis 36Met Ile Ala Thr Thr Arg Asp Arg Glu Gly Ala Thr Met Ile Thr Phe 1 5 10 15 Arg Leu Arg Leu Pro Cys Arg Thr Ile Leu Arg Val Phe Ser Arg Asn 20 25 30 Pro Leu Val Arg Gly Thr Asp Arg Leu Glu Ala Val Val Met Leu Leu 35 40 45 Ala Val Thr Val Ser Leu Leu Thr Ile

Pro Phe Ala Ala Ala Ala Gly 50 55 60 Thr Ala Val Gln Asp Ser Arg Ser His Val Tyr Ala His Gln Ala Gln 65 70 75 80 Thr Arg His Pro Ala Thr Ala Thr Val Ile Asp His Glu Gly Val Ile 85 90 95 Asp Ser Asn Thr Thr Ala Thr Ser Ala Pro Pro Arg Thr Lys Ile Thr 100 105 110 Val Pro Ala Arg Trp Val Val Asn Gly Ile Glu Arg Ser Gly Glu Val 115 120 125 Asn Ala Lys Pro Gly Thr Lys Ser Gly Asp Arg Val Gly Ile Trp Val 130 135 140 Asp Ser Ala Gly Gln Leu Val Asp Glu Pro Ala Pro Pro Ala Arg Ala 145 150 155 160 Ile Ala Asp Ala Ala Leu Ala Ala Leu Gly Leu Trp Leu Ser Val Ala 165 170 175 Ala Val Ala Gly Ala Leu Leu Ala Leu Thr Arg Ala Ile Leu Ile Arg 180 185 190 Val Arg Asn Ala Ser Trp Gln His Asp Ile Asp Ser Leu Phe Cys Thr 195 200 205 Gln Arg 210 37339PRTMycobacterium tuberculosis 37Met Thr Glu Pro Ala Ala Trp Asp Glu Gly Lys Pro Arg Ile Ile Thr 1 5 10 15 Leu Thr Met Asn Pro Ala Leu Asp Ile Thr Thr Ser Val Asp Val Val 20 25 30 Arg Pro Thr Glu Lys Met Arg Cys Gly Ala Pro Arg Tyr Asp Pro Gly 35 40 45 Gly Gly Gly Ile Asn Val Ala Arg Ile Val His Val Leu Gly Gly Cys 50 55 60 Ser Thr Ala Leu Phe Pro Ala Gly Gly Ser Thr Gly Ser Leu Leu Met 65 70 75 80 Ala Leu Leu Gly Asp Ala Gly Val Pro Phe Arg Val Ile Pro Ile Ala 85 90 95 Ala Ser Thr Arg Glu Ser Phe Thr Val Asn Glu Ser Arg Thr Ala Lys 100 105 110 Gln Tyr Arg Phe Val Leu Pro Gly Pro Ser Leu Thr Val Ala Glu Gln 115 120 125 Glu Gln Cys Leu Asp Glu Leu Arg Gly Ala Ala Ala Ser Ala Ala Phe 130 135 140 Val Val Ala Ser Gly Ser Leu Pro Pro Gly Val Ala Ala Asp Tyr Tyr 145 150 155 160 Gln Arg Val Ala Asp Ile Cys Arg Arg Ser Ser Thr Pro Leu Ile Leu 165 170 175 Asp Thr Ser Gly Gly Gly Leu Gln His Ile Ser Ser Gly Val Phe Leu 180 185 190 Leu Lys Ala Ser Val Arg Glu Leu Arg Glu Cys Val Gly Ser Glu Leu 195 200 205 Leu Thr Glu Pro Glu Gln Leu Ala Ala Ala His Glu Leu Ile Asp Arg 210 215 220 Gly Arg Ala Glu Val Val Val Val Ser Leu Gly Ser Gln Gly Ala Leu 225 230 235 240 Leu Ala Thr Arg His Ala Ser His Arg Phe Ser Ser Ile Pro Met Thr 245 250 255 Ala Val Ser Gly Val Gly Ala Gly Asp Ala Met Val Ala Ala Ile Thr 260 265 270 Val Gly Leu Ser Arg Gly Trp Ser Leu Ile Lys Ser Val Arg Leu Gly 275 280 285 Asn Ala Ala Gly Ala Ala Met Leu Leu Thr Pro Gly Thr Ala Ala Cys 290 295 300 Asn Arg Asp Asp Val Glu Arg Phe Phe Glu Leu Ala Ala Glu Pro Thr 305 310 315 320 Glu Val Gly Gln Asp Gln Tyr Val Trp His Pro Ile Val Asn Pro Glu 325 330 335 Ala Ser Pro 38331PRTMycobacterium tuberculosis 38Met Pro Asp Thr Met Val Thr Thr Asp Val Ile Lys Ser Ala Val Gln 1 5 10 15 Leu Ala Cys Arg Ala Pro Ser Leu His Asn Ser Gln Pro Trp Arg Trp 20 25 30 Ile Ala Glu Asp His Thr Val Ala Leu Phe Leu Asp Lys Asp Arg Val 35 40 45 Leu Tyr Ala Thr Asp His Ser Gly Arg Glu Ala Leu Leu Gly Cys Gly 50 55 60 Ala Val Leu Asp His Phe Arg Val Ala Met Ala Ala Ala Gly Thr Thr 65 70 75 80 Ala Asn Val Glu Arg Phe Pro Asn Pro Asn Asp Pro Leu His Leu Ala 85 90 95 Ser Ile Asp Phe Ser Pro Ala Asp Phe Val Thr Glu Gly His Arg Leu 100 105 110 Arg Ala Asp Ala Ile Leu Leu Arg Arg Thr Asp Arg Leu Pro Phe Ala 115 120 125 Glu Pro Pro Asp Trp Asp Leu Val Glu Ser Gln Leu Arg Thr Thr Val 130 135 140 Thr Ala Asp Thr Val Arg Ile Asp Val Ile Ala Asp Asp Met Arg Pro 145 150 155 160 Glu Leu Ala Ala Ala Ser Lys Leu Thr Glu Ser Leu Arg Leu Tyr Asp 165 170 175 Ser Ser Tyr His Ala Glu Leu Phe Trp Trp Thr Gly Ala Phe Glu Thr 180 185 190 Ser Glu Gly Ile Pro His Ser Ser Leu Val Ser Ala Ala Glu Ser Asp 195 200 205 Arg Val Thr Phe Gly Arg Asp Phe Pro Val Val Ala Asn Thr Asp Arg 210 215 220 Arg Pro Glu Phe Gly His Asp Arg Ser Lys Val Leu Val Leu Ser Thr 225 230 235 240 Tyr Asp Asn Glu Arg Ala Ser Leu Leu Arg Cys Gly Glu Met Leu Ser 245 250 255 Ala Val Leu Leu Asp Ala Thr Met Ala Gly Leu Ala Thr Cys Thr Leu 260 265 270 Thr His Ile Thr Glu Leu His Ala Ser Arg Asp Leu Val Ala Ala Leu 275 280 285 Ile Gly Gln Pro Ala Thr Pro Gln Ala Leu Val Arg Val Gly Leu Ala 290 295 300 Pro Glu Met Glu Glu Pro Pro Pro Ala Thr Pro Arg Arg Pro Ile Asp 305 310 315 320 Glu Val Phe His Val Arg Ala Lys Asp His Arg 325 330 39143PRTMycobacterium tuberculosis 39Met Thr Thr Ala Arg Asp Ile Met Asn Ala Gly Val Thr Cys Val Gly 1 5 10 15 Glu His Glu Thr Leu Thr Ala Ala Ala Gln Tyr Met Arg Glu His Asp 20 25 30 Ile Gly Ala Leu Pro Ile Cys Gly Asp Asp Asp Arg Leu His Gly Met 35 40 45 Leu Thr Asp Arg Asp Ile Val Ile Lys Gly Leu Ala Ala Gly Leu Asp 50 55 60 Pro Asn Thr Ala Thr Ala Gly Glu Leu Ala Arg Asp Ser Ile Tyr Tyr 65 70 75 80 Val Asp Ala Asn Ala Ser Ile Gln Glu Met Leu Asn Val Met Glu Glu 85 90 95 His Gln Val Arg Arg Val Pro Val Ile Ser Glu His Arg Leu Val Gly 100 105 110 Ile Val Thr Glu Ala Asp Ile Ala Arg His Leu Pro Glu His Ala Ile 115 120 125 Val Gln Phe Val Lys Ala Ile Cys Ser Pro Met Ala Leu Ala Ser 130 135 140 40413PRTMycobacterium tuberculosis 40Met Ala Ser Ser Ala Ser Asp Gly Thr His Glu Arg Ser Ala Phe Arg 1 5 10 15 Leu Ser Pro Pro Val Leu Ser Gly Ala Met Gly Pro Phe Met His Thr 20 25 30 Gly Leu Tyr Val Ala Gln Ser Trp Arg Asp Tyr Leu Gly Gln Gln Pro 35 40 45 Asp Lys Leu Pro Ile Ala Arg Pro Thr Ile Ala Leu Ala Ala Gln Ala 50 55 60 Phe Arg Asp Glu Ile Val Leu Leu Gly Leu Lys Ala Arg Arg Pro Val 65 70 75 80 Ser Asn His Arg Val Phe Glu Arg Ile Ser Gln Glu Val Ala Ala Gly 85 90 95 Leu Glu Phe Tyr Gly Asn Arg Arg Trp Leu Glu Lys Pro Ser Gly Phe 100 105 110 Phe Ala Gln Pro Pro Pro Leu Thr Glu Val Ala Val Arg Lys Val Lys 115 120 125 Asp Arg Arg Arg Ser Phe Tyr Arg Ile Phe Phe Asp Ser Gly Phe Thr 130 135 140 Pro His Pro Gly Glu Pro Gly Ser Gln Arg Trp Leu Ser Tyr Thr Ala 145 150 155 160 Asn Asn Arg Glu Tyr Ala Leu Leu Leu Arg His Pro Glu Pro Arg Pro 165 170 175 Trp Leu Val Cys Val His Gly Thr Glu Met Gly Arg Ala Pro Leu Asp 180 185 190 Leu Ala Val Phe Arg Ala Trp Lys Leu His Asp Glu Leu Gly Leu Asn 195 200 205 Ile Val Met Pro Val Leu Pro Met His Gly Pro Arg Gly Gln Gly Leu 210 215 220 Pro Lys Gly Ala Val Phe Pro Gly Glu Asp Val Leu Asp Asp Val His 225 230 235 240 Gly Thr Ala Gln Ala Val Trp Asp Ile Arg Arg Leu Leu Ser Trp Ile 245 250 255 Arg Ser Gln Glu Glu Glu Ser Leu Ile Gly Leu Asn Gly Leu Ser Leu 260 265 270 Gly Gly Tyr Ile Ala Ser Leu Val Ala Ser Leu Glu Glu Gly Leu Ala 275 280 285 Cys Ala Ile Leu Gly Val Pro Val Ala Asp Leu Ile Glu Leu Leu Gly 290 295 300 Arg His Cys Gly Leu Arg His Lys Asp Pro Arg Arg His Thr Val Lys 305 310 315 320 Met Ala Glu Pro Ile Gly Arg Met Ile Ser Pro Leu Ser Leu Thr Pro 325 330 335 Leu Val Pro Met Pro Gly Arg Phe Ile Tyr Ala Gly Ile Ala Asp Arg 340 345 350 Leu Val His Pro Arg Glu Gln Val Thr Arg Leu Trp Glu His Trp Gly 355 360 365 Lys Pro Glu Ile Val Trp Tyr Pro Gly Gly His Thr Gly Phe Phe Gln 370 375 380 Ser Arg Pro Val Arg Arg Phe Val Gln Ala Ala Leu Glu Gln Ser Gly 385 390 395 400 Leu Leu Asp Ala Pro Arg Thr Gln Arg Asp Arg Ser Ala 405 410 41120PRTMycobacterium tuberculosis 41Met Ser Thr Gln Arg Pro Arg His Ser Gly Ile Arg Ala Val Gly Pro 1 5 10 15 Tyr Ala Trp Ala Gly Arg Cys Gly Arg Ile Gly Arg Trp Gly Val His 20 25 30 Gln Glu Ala Met Met Asn Leu Ala Ile Trp His Pro Arg Lys Val Gln 35 40 45 Ser Ala Thr Ile Tyr Gln Val Thr Asp Arg Ser His Asp Gly Arg Thr 50 55 60 Ala Arg Val Pro Gly Asp Glu Ile Thr Ser Thr Val Ser Gly Trp Leu 65 70 75 80 Ser Glu Leu Gly Thr Gln Ser Pro Leu Ala Asp Glu Leu Ala Arg Ala 85 90 95 Val Arg Ile Gly Asp Trp Pro Ala Ala Tyr Ala Ile Gly Glu His Leu 100 105 110 Ser Val Glu Ile Ala Val Ala Val 115 120 421017DNAMycobacterium tuberculosis 42atgcagcttg ttgacagggt tcgtggcgcc gtcacgggta tgtcgcgtcg actcgtggtc 60ggggccgtcg gcgcggccct agtgtcgggt ctggtcggcg ccgtcggtgg cacggcgacc 120gcgggggcat tttcccggcc gggcttgccg gtggagtacc tgcaggtgcc gtcgccgtcg 180atgggccgtg acatcaaggt ccaattccaa agtggtggtg ccaactcgcc cgccctgtac 240ctgctcgacg gcctgcgcgc gcaggacgac ttcagcggct gggacatcaa caccccggcg 300ttcgagtggt acgaccagtc gggcctgtcg gtggtcatgc cggtgggtgg ccagtcaagc 360ttctactccg actggtacca gcccgcctgc ggcaaggccg gttgccagac ttacaagtgg 420gagaccttcc tgaccagcga gctgccgggg tggctgcagg ccaacaggca cgtcaagccc 480accggaagcg ccgtcgtcgg tctttcgatg gctgcttctt cggcgctgac gctggcgatc 540tatcaccccc agcagttcgt ctacgcggga gcgatgtcgg gcctgttgga cccctcccag 600gcgatgggtc ccaccctgat cggcctggcg atgggtgacg ctggcggcta caaggcctcc 660gacatgtggg gcccgaagga ggacccggcg tggcagcgca acgacccgct gttgaacgtc 720gggaagctga tcgccaacaa cacccgcgtc tgggtgtact gcggcaacgg caagccgtcg 780gatctgggtg gcaacaacct gccggccaag ttcctcgagg gcttcgtgcg gaccagcaac 840atcaagttcc aagacgccta caacgccggt ggcggccaca acggcgtgtt cgacttcccg 900gacagcggta cgcacagctg ggagtactgg ggcgcgcagc tcaacgctat gaagcccgac 960ctgcaacggg cactgggtgc cacgcccaac accgggcccg cgccccaggg cgcctag 101743978DNAMycobacterium tuberculosis 43atgacagacg tgagccgaaa gattcgagct tggggacgcc gattgatgat cggcacggca 60gcggctgtag tccttccggg cctggtgggg cttgccggcg gagcggcaac cgcgggcgcg 120ttctcccggc cggggctgcc ggtcgagtac ctgcaggtgc cgtcgccgtc gatgggccgc 180gacatcaagg ttcagttcca gagcggtggg aacaactcac ctgcggttta tctgctcgac 240ggcctgcgcg cccaagacga ctacaacggc tgggatatca acaccccggc gttcgagtgg 300tactaccagt cgggactgtc gatagtcatg ccggtcggcg ggcagtccag cttctacagc 360gactggtaca gcccggcctg cggtaaggct ggctgccaga cttacaagtg ggaaaccttc 420ctgaccagcg agctgccgca atggttgtcc gccaacaggg ccgtgaagcc caccggcagc 480gctgcaatcg gcttgtcgat ggccggctcg tcggcaatga tcttggccgc ctaccacccc 540cagcagttca tctacgccgg ctcgctgtcg gccctgctgg acccctctca ggggatgggg 600cctagcctga tcggcctcgc gatgggtgac gccggcggtt acaaggccgc agacatgtgg 660ggtccctcga gtgacccggc atgggagcgc aacgacccta cgcagcagat ccccaagctg 720gtcgcaaaca acacccggct atgggtttat tgcgggaacg gcaccccgaa cgagttgggc 780ggtgccaaca tacccgccga gttcttggag aacttcgttc gtagcagcaa cctgaagttc 840caggatgcgt acaacgccgc gggcgggcac aacgccgtgt tcaacttccc gcccaacggc 900acgcacagct gggagtactg gggcgctcag ctcaacgcca tgaagggtga cctgcagagt 960tcgttaggcg ccggctga 97844288DNAMycobacterium tuberculosis 44atgacagagc agcagtggaa tttcgcgggt atcgaggccg cggcaagcgc aatccaggga 60aatgtcacgt ccattcattc cctccttgac gaggggaagc agtccctgac caagctcgca 120gcggcctggg gcggtagcgg ttcggaggcg taccagggtg tccagcaaaa atgggacgcc 180acggctaccg agctgaacaa cgcgctgcag aacctggcgc ggacgatcag cgaagccggt 240caggcaatgg cttcgaccga aggcaacgtc actgggatgt tcgcatag 28845291DNAMycobacterium tuberculosis 45atgtcgcaaa tcatgtacaa ctaccccgcg atgttgggtc acgccgggga tatggccgga 60tatgccggca cgctgcagag cttgggtgcc gagatcgccg tggagcaggc cgcgttgcag 120agtgcgtggc agggcgatac cgggatcacg tatcaggcgt ggcaggcaca gtggaaccag 180gccatggaag atttggtgcg ggcctatcat gcgatgtcca gcacccatga agccaacacc 240atggcgatga tggcccgcga cacggccgaa gccgccaaat ggggcggcta g 291461068DNAMycobacterium tuberculosis 46atgagcaatt cgcgccgccg ctcactcagg tggtcatggt tgctgagcgt gctggctgcc 60gtcgggctgg gcctggccac ggcgccggcc caggcggccc cgccggcctt gtcgcaggac 120cggttcgccg acttccccgc gctgcccctc gacccgtccg cgatggtcgc ccaagtgggg 180ccacaggtgg tcaacatcaa caccaaactg ggctacaaca acgccgtggg cgccgggacc 240ggcatcgtca tcgatcccaa cggtgtcgtg ctgaccaaca accacgtgat cgcgggcgcc 300accgacatca atgcgttcag cgtcggctcc ggccaaacct acggcgtcga tgtggtcggg 360tatgaccgca cccaggatgt cgcggtgctg cagctgcgcg gtgccggtgg cctgccgtcg 420gcggcgatcg gtggcggcgt cgcggttggt gagcccgtcg tcgcgatggg caacagcggt 480gggcagggcg gaacgccccg tgcggtgcct ggcagggtgg tcgcgctcgg ccaaaccgtg 540caggcgtcgg attcgctgac cggtgccgaa gagacattga acgggttgat ccagttcgat 600gccgcgatcc agcccggtga ttcgggcggg cccgtcgtca acggcctagg acaggtggtc 660ggtatgaaca cggccgcgtc cgataacttc cagctgtccc agggtgggca gggattcgcc 720attccgatcg ggcaggcgat ggcgatcgcg ggccagatcc gatcgggtgg ggggtcaccc 780accgttcata tcgggcctac cgccttcctc ggcttgggtg ttgtcgacaa caacggcaac 840ggcgcacgag tccaacgcgt ggtcgggagc gctccggcgg caagtctcgg catctccacc 900ggcgacgtga tcaccgcggt cgacggcgct ccgatcaact cggccaccgc gatggcggac 960gcgcttaacg ggcatcatcc cggtgacgtc atctcggtga cctggcaaac caagtcgggc 1020ggcacgcgta cagggaacgt gacattggcc gagggacccc cggcctga 1068471176DNAMycobacterium tuberculosis 47atggtggatt tcggggcgtt accaccggag atcaactccg cgaggatgta cgccggcccg 60ggttcggcct cgctggtggc cgcggctcag atgtgggaca gcgtggcgag tgacctgttt 120tcggccgcgt cggcgtttca gtcggtggtc tggggtctga cggtggggtc gtggataggt 180tcgtcggcgg gtctgatggt ggcggcggcc tcgccgtatg tggcgtggat gagcgtcacc 240gcggggcagg ccgagctgac cgccgcccag gtccgggttg ctgcggcggc ctacgagacg 300gcgtatgggc tgacggtgcc cccgccggtg atcgccgaga accgtgctga actgatgatt 360ctgatagcga ccaacctctt ggggcaaaac accccggcga tcgcggtcaa cgaggccgaa 420tacggcgaga tgtgggccca agacgccgcc gcgatgtttg gctacgccgc ggcgacggcg 480acggcgacgg cgacgttgct gccgttcgag gaggcgccgg agatgaccag cgcgggtggg 540ctcctcgagc aggccgccgc ggtcgaggag gcctccgaca ccgccgcggc gaaccagttg 600atgaacaatg tgccccaggc gctgcaacag ctggcccagc ccacgcaggg caccacgcct 660tcttccaagc tgggtggcct gtggaagacg gtctcgccgc atcggtcgcc gatcagcaac 720atggtgtcga tggccaacaa ccacatgtcg atgaccaact cgggtgtgtc gatgaccaac 780accttgagct cgatgttgaa gggctttgct ccggcggcgg ccgcccaggc cgtgcaaacc 840gcggcgcaaa acggggtccg ggcgatgagc tcgctgggca gctcgctggg ttcttcgggt 900ctgggcggtg gggtggccgc caacttgggt cgggcggcct cggtcggttc gttgtcggtg 960ccgcaggcct gggccgcggc caaccaggca gtcaccccgg cggcgcgggc gctgccgctg 1020accagcctga ccagcgccgc ggaaagaggg cccgggcaga tgctgggcgg gctgccggtg 1080gggcagatgg gcgccagggc cggtggtggg ctcagtggtg tgctgcgtgt tccgccgcga 1140ccctatgtga

tgccgcattc tccggcggcc ggctag 117648711DNAMycobacterium tuberculosis 48atgcggaccc ccagacgcca ctgccgtcgc atcgccgtcc tcgccgccgt tagcatcgcc 60gccactgtcg ttgccggctg ctcgtcgggc tcgaagccaa gcggcggacc acttccggac 120gcgaagccgc tggtcgagga ggccaccgcg cagaccaagg ctctcaagag cgcgcacatg 180gtgctgacgg tcaacggcaa gatcccggga ctgtctctga agacgctgag cggcgatctc 240accaccaacc ccaccgccgc gacgggaaac gtcaagctca cgctgggtgg gtctgatatc 300gatgccgact tcgtggtgtt cgacgggatc ctgtacgcca ccctgacgcc caaccagtgg 360agcgatttcg gtcccgccgc cgacatctac gaccccgccc aggtgctgaa tccggatacc 420ggcctggcca acgtgctggc gaatttcgcc gacgcaaaag ccgaagggcg ggataccatc 480aacggccaga acaccatccg catcagcggg aaggtatcgg cacaggcggt gaaccagata 540gcgccgccgt tcaacgcgac gcagccggtg ccggcgaccg tctggattca ggagaccggc 600gatcatcaac tggcacaggc ccagttggac cgcggctcgg gcaattccgt ccagatgacc 660ttgtcgaaat ggggcgagaa ggtccaggtc acgaagcccc cggtgagctg a 711491623DNAMycobacterium tuberculosis 49atggccaaga caattgcgta cgacgaagag gcccgtcgcg gcctcgagcg gggcttgaac 60gccctcgccg atgcggtaaa ggtgacattg ggccccaagg gccgcaacgt cgtcctggaa 120aagaagtggg gtgcccccac gatcaccaac gatggtgtgt ccatcgccaa ggagatcgag 180ctggaggatc cgtacgagaa gatcggcgcc gagctggtca aagaggtagc caagaagacc 240gatgacgtcg ccggtgacgg caccacgacg gccaccgtgc tggcccaggc gttggttcgc 300gagggcctgc gcaacgtcgc ggccggcgcc aacccgctcg gtctcaaacg cggcatcgaa 360aaggccgtgg agaaggtcac cgagaccctg ctcaagggcg ccaaggaggt cgagaccaag 420gagcagattg cggccaccgc agcgatttcg gcgggtgacc agtccatcgg tgacctgatc 480gccgaggcga tggacaaggt gggcaacgag ggcgtcatca ccgtcgagga gtccaacacc 540tttgggctgc agctcgagct caccgagggt atgcggttcg acaagggcta catctcgggg 600tacttcgtga ccgacccgga gcgtcaggag gcggtcctgg aggaccccta catcctgctg 660gtcagctcca aggtgtccac tgtcaaggat ctgctgccgc tgctcgagaa ggtcatcgga 720gccggtaagc cgctgctgat catcgccgag gacgtcgagg gcgaggcgct gtccaccctg 780gtcgtcaaca agatccgcgg caccttcaag tcggtggcgg tcaaggctcc cggcttcggc 840gaccgccgca aggcgatgct gcaggatatg gccattctca ccggtggtca ggtgatcagc 900gaagaggtcg gcctgacgct ggagaacgcc gacctgtcgc tgctaggcaa ggcccgcaag 960gtcgtggtca ccaaggacga gaccaccatc gtcgagggcg ccggtgacac cgacgccatc 1020gccggacgag tggcccagat ccgccaggag atcgagaaca gcgactccga ctacgaccgt 1080gagaagctgc aggagcggct ggccaagctg gccggtggtg tcgcggtgat caaggccggt 1140gccgccaccg aggtcgaact caaggagcgc aagcaccgca tcgaggatgc ggttcgcaat 1200gccaaggccg ccgtcgagga gggcatcgtc gccggtgggg gtgtgacgct gttgcaagcg 1260gccccgaccc tggacgagct gaagctcgaa ggcgacgagg cgaccggcgc caacatcgtg 1320aaggtggcgc tggaggcccc gctgaagcag atcgccttca actccgggct ggagccgggc 1380gtggtggccg agaaggtgcg caacctgccg gctggccacg gactgaacgc tcagaccggt 1440gtctacgagg atctgctcgc tgccggcgtt gctgacccgg tcaaggtgac ccgttcggcg 1500ctgcagaatg cggcgtccat cgcggggctg ttcctgacca ccgaggccgt cgttgccgac 1560aagccggaaa aggagaaggc ttccgttccc ggtggcggcg acatgggtgg catggatttc 1620tga 162350600DNAMycobacterium tuberculosis 50atggctgaaa actcgaacat tgatgacatc aaggctccgt tgcttgccgc gcttggagcg 60gccgacctgg ccttggccac tgtcaacgag ttgatcacga acctgcgtga gcgtgcggag 120gagactcgta cggacacccg cagccgggtc gaggagagcc gtgctcgcct gaccaagctg 180caggaagatc tgcccgagca gctcaccgag ctgcgtgaga agttcaccgc cgaggagctg 240cgtaaggccg ccgagggcta cctcgaggcc gcgactagcc ggtacaacga gctggtcgag 300cgcggtgagg ccgctctaga gcggctgcgc agccagcaga gcttcgagga agtgtcggcg 360cgcgccgaag gctacgtgga ccaggcggtg gagttgaccc aggaggcgtt gggtacggtc 420gcatcgcaga cccgcgcggt cggtgagcgt gccgccaagc tggtcggcat cgagctgcct 480aagaaggctg ctccggccaa gaaggccgct ccggccaaga aggccgctcc ggccaagaag 540gcggcggcca agaaggcgcc cgcgaagaag gcggcggcca agaaggtcac ccagaagtag 600511128DNAMycobacterium tuberculosis 51gtgacgcaaa ccggcaagcg tcagagacgc aaattcggtc gcatccgaca gttcaactcc 60ggccgctggc aagccagcta caccggcccc gacggccgcg tgtacatcgc ccccaaaacc 120ttcaacgcca agatcgacgc cgaagcatgg ctcaccgacc gccgccgcga aatcgaccga 180caactatggt ccccggcatc gggtcaggaa gaccgccccg gagccccatt cggtgagtac 240gccgaaggat ggctgaagca gcgtggaatc aaggaccgca cccgcgccca ctatcgcaaa 300ctgctggaca accacatcct ggccaccttc gctgacaccg acctacgcga catcaccccg 360gccgccgtgc gccgctggta cgccaccacc gccgtgggca caccgaccat gcgggcacac 420tcctacagct tgctgcgcgc aatcatgcag accgccttgg ccgacgacct gatcgactcc 480aacccctgcc gcatctcagg cgcgtccacc gcccgccgcg tccacaagat caggcccgcc 540accctcgacg agctggaaac catcaccaaa gccatgcccg acccctacca ggcgttcgtg 600ctgatggcgg catggctggc catgcgctac ggcgagctga ccgaattacg ccgcaaagac 660atcgacctgc acggcgaggt tgcgcgggtg cggcgggctg tcgttcgggt gggcgaaggc 720ttcaaggtga cgacaccgaa aagcgatgcg ggagtgcgcg acataagtat cccgccacat 780ctgatacccg ccatcgaaga ccaccttcac aaacacgtca accccggccg ggagtccctg 840ctgttcccat cggtcaacga ccccaaccgt cacctagcac cctcggcgct gtaccgcatg 900ttctacaagg cccgaaaagc cgccggccga ccagacttac gggtgcacga ccttcgacac 960tccggcgccg tgttggctgc atccaccggc gccacactgg ccgaactgat gcagcggcta 1020ggacacagca cagccggcgc cgcactccgc taccagcacg ccgccaaggg ccgggaccgc 1080gaaatcgccg cactgttaag caaactggcc gagaaccagg agatgtga 112852228DNAMycobacterium tuberculosis 52gtgatagcgg gcgtcgacca ggcgcttgca gcaacaggcc aggctagcca gcgggcggca 60ggcgcatctg gtggggtcac cgtcggtgtc ggcgtgggca cggaacagag gaacctttcg 120gtggttgcac cgagtcagtt cacatttagt tcacgcagcc cagattttgt ggatgaaacc 180gcaggtcaat cgtggtgcgc gatactggga ttgaaccagt ttcactag 22853435DNAMycobacterium tuberculosis 53atggccacca cccttcccgt tcagcgccac ccgcggtccc tcttccccga gttttctgag 60ctgttcgcgg ccttcccgtc attcgccgga ctccggccca ccttcgacac ccggttgatg 120cggctggaag acgagatgaa agaggggcgc tacgaggtac gcgcggagct tcccggggtc 180gaccccgaca aggacgtcga cattatggtc cgcgatggtc agctgaccat caaggccgag 240cgcaccgagc agaaggactt cgacggtcgc tcggaattcg cgtacggttc cttcgttcgc 300acggtgtcgc tgccggtagg tgctgacgag gacgacatta aggccaccta cgacaagggc 360attcttactg tgtcggtggc ggtttcggaa gggaagccaa ccgaaaagca cattcagatc 420cggtccacca actga 435541224DNAMycobacterium tuberculosis 54atgagtggac gccaccgtaa gcccaccaca tccaacgtca gcgtcgccaa gatcgccttt 60accggcgcag tactcggtgg cggcggcatc gccatggccg ctcaggcgac cgcggccacc 120gacggggaat gggatcaggt ggcccgctgc gagtcgggcg gcaactggtc gatcaacacc 180ggcaacggtt acctcggtgg cttgcagttc actcaaagca cctgggccgc acatggtggc 240ggcgagttcg ccccgtcggc tcagctggcc agccgggagc agcagattgc cgtcggtgag 300cgggtgctgg ccacccaggg tcgcggcgcc tggccggtgt gcggccgcgg gttatcgaac 360gcaacacccc gcgaagtgct tcccgcttcg gcagcgatgg acgctccgtt ggacgcggcc 420gcggtcaacg gcgaaccagc accgctggcc ccgccgcccg ccgacccggc gccacccgtg 480gaacttgccg ctaacgacct gcccgcaccg ctgggtgaac ccctcccggc agctcccgcc 540gacccggcac cacccgccga cctggcacca cccgcgcccg ccgacgtcgc gccacccgtg 600gaacttgccg taaacgacct gcccgcaccg ctgggtgaac ccctcccggc agctcccgcc 660gacccggcac cacccgccga cctggcacca cccgcgcccg ccgacctggc gccacccgcg 720cccgccgacc tggcgccacc cgcgcccgcc gacctggcac cacccgtgga acttgccgta 780aacgacctgc ccgcgccgct gggtgaaccc ctcccggcag ctcccgccga actggcgcca 840cccgccgatc tggcacccgc gtccgccgac ctggcgccac ccgcgcccgc cgacctggcg 900ccacccgcgc ccgccgaact ggcgccaccc gcgcccgccg acctggcacc acccgctgcg 960gtgaacgagc aaaccgcgcc gggcgatcag cccgccacag ctccaggcgg cccggttggc 1020cttgccaccg atttggaact ccccgagccc gacccccaac cagctgacgc accgccgccc 1080ggcgacgtca ccgaggcgcc cgccgaaacg ccccaagtct cgaacatcgc ctatacgaag 1140aagctgtggc aggcgattcg ggcccaggac gtctgcggca acgatgcgct ggactcgctc 1200gcacagccgt acgtcatcgg ctga 1224551089DNAMycobacterium tuberculosis 55atgttgcgcc tggtagtcgg tgcgctgctg ctggtgttgg cgttcgccgg tggctatgcg 60gtcgccgcat gcaaaacggt gacgttgacc gtcgacggaa ccgcgatgcg ggtgaccacg 120atgaaatcgc gggtgatcga catcgtcgaa gagaacgggt tctcagtcga cgaccgcgac 180gacctgtatc ccgcggccgg cgtgcaggtc catgacgccg acaccatcgt gctgcggcgt 240agccgtccgc tgcagatctc gctggatggt cacgacgcta agcaggtgtg gacgaccgcg 300tcgacggtgg acgaggcgct ggcccaactc gcgatgaccg acacggcgcc ggccgcggct 360tctcgcgcca gccgcgtccc gctgtccggg atggcgctac cggtcgtcag cgccaagacg 420gtgcagctca acgacggcgg gttggtgcgc acggtgcact tgccggcccc caatgtcgcg 480gggctgctga gtgcggccgg cgtgccgctg ttgcaaagcg accacgtggt gcccgccgcg 540acggccccga tcgtcgaagg catgcagatc caggtgaccc gcaatcggat caagaaggtc 600accgagcggc tgccgctgcc gccgaacgcg cgtcgtgtcg aggacccgga gatgaacatg 660agccgggagg tcgtcgaaga cccgggggtt ccggggaccc aggatgtgac gttcgcggta 720gctgaggtca acggcgtcga gaccggccgt ttgcccgtcg ccaacgtcgt ggtgaccccg 780gcccacgaag ccgtggtgcg ggtgggcacc aagcccggta ccgaggtgcc cccggtgatc 840gacggaagca tctgggacgc gatcgccggc tgtgaggccg gtggcaactg ggcgatcaac 900accggcaacg ggtattacgg tggtgtgcag tttgaccagg gcacctggga ggccaacggc 960gggctgcggt atgcaccccg cgctgacctc gccacccgcg aagagcagat cgccgttgcc 1020gaggtgaccc gactgcgtca aggttggggc gcctggccgg tatgtgctgc acgagcgggt 1080gcgcgctga 108956531DNAMycobacterium tuberculosis 56gtgcatcctt tgccggccga ccacggccgg tcgcggtgca atagacaccc gatctcacca 60ctctctctaa tcggtaacgc ttcggccact tccggcgata tgtcgagcat gacaagaatc 120gccaagccgc tcatcaagtc cgccatggcc gcaggactcg tcacggcatc catgtcgctc 180tccaccgccg ttgcccacgc cggtcccagc ccgaactggg acgccgtcgc gcagtgcgaa 240tccgggggca actgggcggc caacaccgga aacggcaaat acggcggact gcagttcaag 300ccggccacct gggccgcatt cggcggtgtc ggcaacccag cagctgcctc tcgggaacaa 360caaatcgcag ttgccaatcg ggttctcgcc gaacagggat tggacgcgtg gccgacgtgc 420ggcgccgcct ctggccttcc gatcgcactg tggtcgaaac ccgcgcaggg catcaagcaa 480atcatcaacg agatcatttg ggcaggcatt caggcaagta ttccgcgctg a 53157465DNAMycobacterium tuberculosis 57atgacaccgg gtttgcttac tactgcgggt gctggccgac cacgtgacag gtgcgccagg 60atcgtatgca cggtgttcat cgaaaccgcc gttgtcgcga ccatgtttgt cgcgttgttg 120ggtctgtcca ccatcagctc gaaagccgac gacatcgatt gggacgccat cgcgcaatgc 180gaatccggcg gcaattgggc ggccaacacc ggtaacgggt tatacggtgg tctgcagatc 240agccaggcga cgtgggattc caacggtggt gtcgggtcgc cggcggccgc gagtccccag 300caacagatcg aggtcgcaga caacattatg aaaacccaag gcccgggtgc gtggccgaaa 360tgtagttctt gtagtcaggg agacgcaccg ctgggctcgc tcacccacat cctgacgttc 420ctcgcggccg agactggagg ttgttcgggg agcagggacg attga 46558519DNAMycobacterium tuberculosis 58ttgaagaacg cccgtacgac gctcatcgcc gccgcgattg ccgggacgtt ggtgaccacg 60tcaccagccg gtatcgccaa tgccgacgac gcgggcttgg acccaaacgc cgcagccggc 120ccggatgccg tgggctttga cccgaacctg ccgccggccc cggacgctgc acccgtcgat 180actccgccgg ctccggagga cgcgggcttt gatcccaacc tccccccgcc gctggccccg 240gacttcctgt ccccgcctgc ggaggaagcg cctcccgtgc ccgtggccta cagcgtgaac 300tgggacgcga tcgcgcagtg cgagtccggt ggaaactggt cgatcaacac cggtaacggt 360tactacggcg gcctgcggtt caccgccggc acctggcgtg ccaacggtgg ctcggggtcc 420gcggccaacg cgagccggga ggagcagatc cgggtggctg agaacgtgct gcgttcgcag 480ggtatccgcg cctggccggt ctgcggccgc cgcggctga 51959633DNAMycobacterium tuberculosis 59atgatcgcca caacccgcga tcgtgaagga gccaccatga tcacgtttag gctgcgcttg 60ccgtgccgga cgatactgcg ggtgttcagc cgcaatccgc tggtgcgtgg gacggatcga 120ctcgaggcgg tcgtcatgct gctggccgtc acggtctcgc tgctgactat cccgttcgcc 180gccgcggccg gcaccgcagt ccaggattcc cgcagccacg tctatgccca ccaggcccag 240acccgccatc ccgcaaccgc gaccgtgatc gatcacgagg gggtgatcga cagcaacacg 300accgccacgt cagcgccgcc gcgcacgaag atcaccgtgc ctgcccgatg ggtcgtgaac 360ggaatagaac gcagcggtga ggtcaacgcg aagccgggaa ccaaatccgg tgaccgcgtc 420ggcatttggg tcgacagtgc cggtcagctg gtcgatgaac cagctccgcc ggcccgtgcc 480attgcggatg cggccctggc cgccttggga ctctggttga gcgtcgccgc ggttgcgggc 540gccctgctgg cgctcactcg ggcgattctg atccgcgttc gcaacgccag ttggcaacac 600gacatcgaca gcctgttctg cacgcagcgg tga 633601020DNAMycobacterium tuberculosis 60atgacggagc cagcggcgtg ggacgaaggc aagccgcgaa tcatcacttt gaccatgaac 60cccgccttgg acatcacgac gagcgtcgac gtggtgcgcc cgaccgagaa aatgcgttgt 120ggcgcacctc gctacgatcc cggcggcggc ggtatcaatg tcgcccgcat tgtgcatgtc 180ctcggcggtt gctcgacagc actgttcccg gccggcgggt cgaccgggag cctgctgatg 240gcgctgctcg gtgatgcggg agtgccattt cgcgtcattc cgatcgcggc ctcgacgcgg 300gagagcttca cggtcaacga gtccaggacc gccaagcagt atcgtttcgt gcttccgggg 360ccgtcgctga ccgtcgcgga gcaggagcaa tgcctcgacg aactgcgcgg tgcggcggct 420tcggccgcct ttgtggtggc cagtggcagc ctgccgccag gtgtggctgc cgactactat 480cagcgggttg ccgacatctg ccgccgatcg agcactccgc tgatcctgga tacatctggt 540ggcgggttgc agcacatttc gtccggggtg tttcttctca aggcgagcgt gcgggaactg 600cgcgagtgcg tcggatccga actgctgacc gagcccgaac aactggccgc cgcacacgaa 660ctcattgacc gtgggcgcgc cgaggtcgtg gtggtctcgc ttggatctca gggcgcgcta 720ttggccacac gacatgcgag ccatcgattt tcgtcgattc cgatgaccgc ggttagcggt 780gtcggcgccg gcgacgcgat ggtggccgcg attaccgtgg gcctcagccg tggctggtcg 840ctcatcaagt ccgttcgctt gggaaacgcg gcaggtgcag ccatgctgct gacgccaggc 900accgcggcct gcaatcgcga cgatgtggag aggttcttcg agctggcggc cgaacccacc 960gaagtcgggc aggatcaata cgtttggcac ccgatcgtta acccggaagc ctcgccatga 102061996DNAMycobacterium tuberculosis 61atgccggaca ccatggtgac caccgatgtc atcaagagcg cggtgcagtt ggcctgccgc 60gcaccgtcgc tccacaacag ccagccctgg cgctggatag ccgaggacca cacggttgcg 120ctgttcctcg acaaggatcg ggtgctttac gcgaccgacc actccggccg ggaagcgctg 180ctggggtgcg gcgccgtact cgaccacttt cgggtggcga tggcggccgc gggtaccacc 240gccaatgtgg aacggtttcc caaccccaac gatcctttgc atctggcgtc aattgacttc 300agcccggccg atttcgtcac cgagggccac cgtctaaggg cggatgcgat cctactgcgc 360cgtaccgacc ggctgccttt cgccgagccg ccggattggg acttggtgga gtcgcagttg 420cgcacgaccg tcaccgccga cacggtgcgc atcgacgtca tcgccgacga tatgcgtccc 480gaactggcgg cggcgtccaa actcaccgaa tcgctgcggc tctacgattc gtcgtatcat 540gccgaactct tttggtggac aggggctttt gagacttctg agggcatacc gcacagttca 600ttggtatcgg cggccgaaag tgaccgggtc accttcggac gcgacttccc ggtcgtcgcc 660aacaccgata ggcgcccgga gtttggccac gaccgctcta aggtcctggt gctctccacc 720tacgacaacg aacgcgccag cctactgcgc tgcggcgaga tgctttccgc cgtattgctt 780gacgccacca tggctgggct tgccacctgc acgctgaccc acatcaccga actgcacgcc 840agccgagacc tggtcgcagc gctgattggg cagcccgcaa ctccgcaagc cttggttcgc 900gtcggtctgg ccccggagat ggaagagccg ccaccggcaa cgcctcggcg accaatcgat 960gaagtgtttc acgttcgggc taaggatcac cggtag 99662432DNAMycobacterium tuberculosis 62atgaccaccg cacgcgacat catgaacgca ggtgtgacct gtgttggcga acacgagacg 60ctaaccgctg ccgctcaata catgcgtgag cacgacatcg gcgcgttgcc gatctgcggg 120gacgacgacc ggctgcacgg catgctcacc gaccgcgaca ttgtgatcaa aggcctggct 180gcgggcctag acccgaatac cgccacggct ggcgagttgg cccgggacag catctactac 240gtcgatgcga acgcaagcat ccaggagatg ctcaacgtca tggaagaaca tcaggtccgc 300cgtgttccgg tcatctcaga gcaccgcttg gtcggaatcg tcaccgaagc cgacatcgcc 360cgacacctgc ccgagcacgc cattgtgcag ttcgtcaagg caatctgctc gcccatggcc 420ctcgccagct ag 432631242DNAMycobacterium tuberculosis 63atggcaagtt ctgcgagcga cggcacccac gaacgctcgg cttttcgcct gagtccaccg 60gtcttgagcg gcgccatggg accgttcatg cacaccggtc tgtacgtcgc tcaatcgtgg 120cgcgactatc tgggtcaaca gcccgataaa ctgccgatcg cacggcccac tattgcctta 180gcggcgcaag cctttcgaga cgaaatcgtc ctgctgggcc tcaaggcacg acgtccggtc 240agcaatcatc gagtgttcga gcgcatcagc caagaagtgg ccgctggact ggagttctat 300gggaatcgca gatggctgga gaagcctagc ggattttttg cccagccccc accgctcacc 360gaggtcgcgg tccgaaaggt caaggaccgc agacgctcct tttatcgcat cttcttcgac 420agtgggttta cgccgcatcc gggtgaaccg ggcagccaac ggtggctctc atacactgcg 480aacaatcgcg agtacgccct gttactgcgg cacccagagc cgcgtccctg gctggtttgt 540gtacacggca ccgagatggg cagggccccg ttggatctcg cggtgttccg cgcctggaag 600ctgcatgacg aactcggcct gaacattgtc atgccggttc ttccgatgca tggtccccgc 660gggcaaggtc tgccgaaggg cgccgttttt cccggagaag atgttctcga cgatgtgcat 720gggacggctc aagcggtgtg ggatatccgg cggctgttgt cctggatacg atcgcaggag 780gaggagtcgc tgatcgggtt gaacggtctc tcgctgggcg gctacatcgc gtcattggtc 840gccagcctcg aagaaggtct cgcctgcgcg attctcggtg tcccagtggc tgatctgatc 900gagttgttgg gccgccactg cggtcttcgg cacaaagacc cccgccgcca caccgtcaag 960atggccgaac cgatcggccg aatgatctcg ccgctctcac ttacgccact ggtgcccatg 1020ccgggccgct ttatctacgc gggcattgcc gaccgactcg tgcatccacg cgaacaggtg 1080actcgcctct gggagcactg gggcaaaccc gaaatcgtgt ggtatccagg cggtcacact 1140ggcttcttcc agtcgcggcc ggtacgacgg tttgtccagg ctgcgctgga gcagtcgggc 1200ctgttggacg cgccacggac acagcgcgac cgttccgcct aa 124264363DNAMycobacterium tuberculosis 64atgtccacgc aacgaccgag gcactccggt attcgggctg ttggccccta cgcatgggcc 60ggccgatgtg gtcggatagg caggtggggg gtgcaccagg aggcgatgat gaatctagcg 120atatggcacc cgcgcaaggt gcaatccgcc accatctatc aggtgaccga tcgctcgcac 180gacgggcgca cagcacgggt gcctggtgac gagatcacta gcaccgtgtc cggttggttg 240tcggagttgg gcacccaaag cccgttggcc gatgagcttg cgcgtgcggt gcggatcggc 300gactggcccg ctgcgtacgc aatcggtgag cacctgtccg ttgagattgc cgttgcggtc 360taa 36365399PRTMycobacterium tuberculosis 65Leu Asp Phe Ala Thr Leu Pro Pro Glu Ile Asn Ser Ala Arg Met Tyr 1 5 10 15 Ser Gly Ala Gly Ser Ala Pro Met Leu Ala Ala Ala Ser Ala Trp His 20 25 30 Gly Leu Ser Ala Glu Leu Arg Ala Ser Ala Leu Ser Tyr Ser Ser Val 35 40 45 Leu Ser Thr Leu Thr Gly Glu Glu Trp His Gly Pro Ala Ser Ala Ser 50 55 60 Met Thr Ala Ala Ala Ala Pro Tyr Val Ala Trp Met Ser Val Thr Ala 65 70 75 80 Val Arg Ala Glu Gln Ala Gly Ala Gln Ala Glu Ala Ala Ala Ala Ala 85 90 95 Tyr Glu Ala Ala Phe Ala Ala Thr Val Pro Pro Pro Val Ile Glu Ala 100 105

110 Asn Arg Ala Gln Leu Met Ala Leu Ile Ala Thr Asn Val Leu Gly Gln 115 120 125 Asn Ala Pro Ala Ile Ala Ala Thr Glu Ala Gln Tyr Ala Glu Met Trp 130 135 140 Ser Gln Asp Ala Met Ala Met Tyr Gly Tyr Ala Gly Ala Ser Ala Ala 145 150 155 160 Ala Thr Gln Leu Thr Pro Phe Thr Glu Pro Val Gln Thr Thr Asn Ala 165 170 175 Ser Gly Leu Ala Ala Gln Ser Ala Ala Ile Ala His Ala Thr Gly Ala 180 185 190 Ser Ala Gly Ala Gln Gln Thr Thr Leu Ser Gln Leu Ile Ala Ala Ile 195 200 205 Pro Ser Val Leu Gln Gly Leu Ser Ser Ser Thr Ala Ala Thr Phe Ala 210 215 220 Ser Gly Pro Ser Gly Leu Leu Gly Ile Val Gly Ser Gly Ser Ser Trp 225 230 235 240 Leu Asp Lys Leu Trp Ala Leu Leu Asp Pro Asn Ser Asn Phe Trp Asn 245 250 255 Thr Ile Ala Ser Ser Gly Leu Phe Leu Pro Ser Asn Thr Ile Ala Pro 260 265 270 Phe Leu Gly Leu Leu Gly Gly Val Ala Ala Ala Asp Ala Ala Gly Asp 275 280 285 Val Leu Gly Glu Ala Thr Ser Gly Gly Leu Gly Gly Ala Leu Val Ala 290 295 300 Pro Leu Gly Ser Ala Gly Gly Leu Gly Gly Thr Val Ala Ala Gly Leu 305 310 315 320 Gly Asn Ala Ala Thr Val Gly Thr Leu Ser Val Pro Pro Ser Trp Thr 325 330 335 Ala Ala Ala Pro Leu Ala Ser Pro Leu Gly Ser Ala Leu Gly Gly Thr 340 345 350 Pro Met Val Ala Pro Pro Pro Ala Val Ala Ala Gly Met Pro Gly Met 355 360 365 Pro Phe Gly Thr Met Gly Gly Gln Gly Phe Gly Arg Ala Val Pro Gln 370 375 380 Tyr Gly Phe Arg Pro Asn Phe Val Ala Arg Pro Pro Ala Ala Gly 385 390 395

Claims

1. An antigenic composition comprising either: (i) a first mycobacterial antigenic polypeptide, wherein said first mycobacterial antigenic polypeptide comprises a polypeptide sequence having at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 6, 12, 2, 18, 8, 10, 16, 4, or a fragment thereof having at least 150 consecutive amino acids thereof, wherein said fragment has a common antigenic cross-reactivity with a polypeptide selected from SEQ ID NOs: 6, 12, 2, 18, 8, 10, 16, 4; or (ii) a first mycobacterial polynucleotide, wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence encoding said first mycobacterial antigenic polypeptide, or wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence selected from SEQ ID NO: 5, 11, 1, 17, 7, 9, 15, 3; wherein said antigenic composition is for use in treating, suppressing or preventing a mycobacterial infection in a subject.

2. The antigenic composition according to claim 1, wherein said first mycobacterial polynucleotide comprises a polynucleotide sequence having at least 90% nucleotide sequence identity to the nucleic acid sequence of SEQ ID NO: 5, 11, 1, 17, 7, 9, 15, 3, or a fragment thereof having at least 450 consecutive nucleotides thereof, wherein said fragment encodes a polypeptide that has a common cross-reactivity with a polypeptide selected from SEQ ID NOs: 6, 12, 2, 18, 8, 10, 16, 4.

3. The antigenic composition according to claim 1, further comprising at least one additional (second) mycobacterial polypeptide, which is different from said first mycobacterial antigenic polypeptide; or at least one additional (second) mycobacterial polynucleotide encoding a second mycobacterial antigenic polypeptide, wherein said second mycobacterial polypeptide is different from said first mycobacterial polypeptide.

4. The antigenic composition according to claim 1, wherein said antigenic composition comprises at least one vector and wherein said mycobacterial polynucleotide(s) is/are incorporated into said at least one vector.

5. The antigenic composition according to claim 4, wherein said vector is an expression vector or a viral vector.

6. The antigenic composition according to claim 1, wherein said antigenic composition comprises at least one cell, and wherein said cell comprises at least one of said mycobacterial antigenic polypeptides and/or mycobacterial polynucleotides.

7. A method for producing a therapeutic or prophylactic formulation, the method comprising combining a pharmaceutically acceptable carrier with either: (i) mycobacterial antigenic polypeptide according to claim 1; or (ii) a first mycobacterial polynucleotide according to claim 1.

8. (canceled)

9. An in vitro method of diagnosing a mycobacterial infection, comprising incubating a test sample containing an immune cell such as a T-lymphocyte from a subject with: (a) antigenic composition according to claim 1; or (b) a first mycobacterial antigenic polypeptide or first mycobacterial polynucleotide according to claim 1; and detecting for activation of said immune cell, wherein activation of said immune cell is indicative of a mycobacterial infection in the subject.

10. An in vitro method of diagnosing a mycobacterial infection, comprising incubating a test sample from a subject with: (a) antigenic composition according to any of claim 1; or (b) a first mycobacterial antigenic polypeptide or first mycobacterial polynucleotide according to claim 1; wherein said incubating is performed under conditions that allow binding of said first mycobacterial antigen with antibodies in the sample to form antigen-antibody complexes; and then detecting for the formation of such complexes, wherein the presence of antigen-antibody complexes is indicative of a mycobacterial infection in the subject.

11. An in vitro method of diagnosing a mycobacterial infection, comprising incubating a test sample from a subject with: (a) an antigenic/immunogenic composition according to claim 1; or (b) a first antibody, wherein said first antibody binds a first mycobacterial antigenic polypeptide according to claim 1; wherein said incubating is performed under conditions that allow binding of said first and second antibodies with antigens in the sample to form antigen-antibody complexes; and then detecting for the formation of such complexes, wherein the presence of antigen-antibody complexes is indicative of a mycobacterial infection in the subject.

12. A method for treating, suppressing or preventing a mycobacterial infection in a subject, said method comprising administering an antigenic composition according to any of claim 1 to said subject.

13. The antigenic composition of claim 4, wherein said vector is a plasmid.

14. The antigenic composition of claim 5, wherein said viral vector is an attenuated vaccinia virus vector or an adenoviral vector.

15. The antigenic composition of claim 6, wherein said cell is an attenuated microbial carrier.

16. The antigenic composition of claim 15, wherein said attenuated microbial carrier is attenuated salmonella, attenuated M. bovis or attenuated M. tuberculosis.

17. The antigenic composition of claim 16, wherein said attenuated M. bovis is a BCG strain of M. bovis.

18. The method of claim 7, wherein said formulation is a vaccine.

19. The method of claim 9, wherein said mycobacterial infection is an early stage mycobacterial infection.

20. The method of claim 10, wherein said mycobacterial infection is an early stage mycobacterial infection.

21. The method of claim 11, wherein said mycobacterial infection is an early stage mycobacterial infection.

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