THERAPEUTIC TB VACCINE

Abstract

Therapeutic vaccines comprising polypeptides expressed during the latent stage of mycobacteria infection are provided, as are multiphase vaccines, and methods for treating and preventing tuberculosis.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a division of U.S. patent application Ser. No. 12/785,053, filed May 21, 2010, which is a continuation of U.S. patent application Ser. No. 10/617,038, filed Jul. 11, 2003, now abandoned, which claims the benefit of the priority of U.S. Provisional Patent Application No. 60/401,725, filed Aug. 7, 2002, now expired, and the priority of Danish Patent Patent Application No. PA 2002 01098, filed Jul. 13, 2002, which applications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] The present invention discloses a therapeutic vaccine against latent or active tuberculosis infection caused by the tuberculosis complex microorganisms (Mycobacterium tuberculosis, M. bovis, M. africanum). The invention furthermore discloses a multi-phase vaccine that can be administered either prophylactically or therapeutically as well as a diagnostic reagent for the detection of latent stages of tuberculosis.

[0003] Human tuberculosis caused by Mycobacterium tuberculosis (M. tuberculosis) is a severe global health problem, responsible for approx. 3 million deaths annually, according to the WHO. The worldwide incidence of new tuberculosis (TB) cases had been falling during the 1960s and 1970s but during recent decades this trend has markedly changed in part due to the advent of AIDS and the appearance of multidrug resistant strains of M. tuberculosis.

[0004] Organisms of the tuberculosis complex can cause a variety of diseases, but the commonest route of invasion is by inhalation of bacteria. This initiates an infection in the lung, which can ultimately spread to other parts of the body. Normally, this infection is restricted in growth by the immune system, so that the majority of infected individuals show few signs apart from cough and fever, which eventually abates. Approximately 30% of individuals are unable to contain the infection and they will develop primary disease, which in many cases will eventually prove fatal. However, it is believed that even those individuals who apparently control the infection remain infected, probably for the rest of their life. Certainly, individuals who have been healthy for years or even decades can suddenly develop tuberculosis, which has proven to be caused by the same organism they were infected with many years previously. M. tuberculosis and other organisms of the TB complex are unique in that the mycobacteria can evade the immune response and survive for long periods in a refractory non-replicating or slowly-replicating stage. This is referred to as latent TB and is at present a very significant global health problem that is estimated to affect approximately 1/3 of the world's population (Anon., 2001).

[0005] The course of a M. tuberculosis infection runs essentially through 3 phases, as illustrated in FIG. 1. During the acute phase, the bacteria proliferate in the organs, until the immune response increases to the point at which it can control the infection, whereupon the bacterial load peaks and starts declining. After this, a latent phase is established where the bacterial load is kept stable at a low level. In this phase M. tuberculosis goes from active multiplication to dormancy, essentially becoming non-replicating and remaining inside the granuloma. In some cases, the infection goes to the reactivation phase, where the dormant bacteria start replicating again. The full nature of the immune response that controls latent infection and the factors that lead to reactivation are largely unknown. However, there is some evidence for a shift in the dominant cell types responsible. While CD4 T cells are essential and sufficient for control of infection during the acute phase, studies suggest that CD8 T cell responses are more important in the latent phase. It is also likely that changes in the antigen-specificity of the response occur, as the bacterium modulates gene expression during its transition from active replication to dormancy.

[0006] The only vaccine presently available for clinical use is BCG, a vaccine whose efficacy remains a matter of controversy. Although BCG consistently performs well in animal models of primary infection, it has clearly failed to control the TB epidemic. Consistent with that, BCG vaccination appears to provide protection against pediatric TB (which is due to primary infection), while offering little or no protection against adult disease (which is often reactivation of latent infection acquired in childhood). It has also been shown that vaccination of individuals who are currently sensitized to mycobacteria or latently infected is ineffective. Thus, current vaccination strategies, while effective against primary disease, fail to activate immune responses that efficiently control surviving dormant bacteria.

[0007] At this point no vaccine has been developed that confers protection against reactivation whether given as a prophylactic vaccine prior to infection or as a therapeutic vaccine given to already latently infected individuals.

[0008] This makes the development of a new and improved vaccine against TB an urgent matter, which has been given a very high priority by the WHO. Many attempts to define protective mycobacterial substances have been made, and different investigators have reported increased resistance after experimental vaccination. However, these efforts have almost exclusively focused on the development of prophylactic vaccines for the prevention of disease (Doherty, 2002), and such vaccines have not been demonstrated to work if given in an immunotherapeutic fashion (J. Turner et al., Infect and Immunity, 2000, pp. 1706-1709).

[0009] It has been suggested that the transition of M. tuberculosis from primary infection to latency is accompanied by changes in gene expression (see, for example, Honer zu Bentrup, 2001, which is incorporated herein by reference). In vitro hypoxic culture conditions, which mimic the conditions of low oxygen tension and restricted nutrients found in the granuloma (the location of the latent infection), have been used to analyze changes in gene expression and a number of antigens have been found that are induced or markedly upregulated under these conditions e.g. the 16 kDa antigen α-crystalline (Boon, 2001, Monahan, 2001, Florczyk 2001, Sherman 2001, Manganelli, 2001, all of which are incorporated herein by reference) and Rv0569 as described in Rosenkrands, 2002, and which is described in WO0179274.

[0010] As noted in the references cited above, it is already known that some genes are upregulated under conditions that mimic latency. However, these are a limited subset of the total gene expression during latent infection. Moreover, as one skilled in the art will readily appreciate, expression of a gene is not sufficient to make it a good vaccine candidate. The only way to determine if a protein is recognized by the immune system during latent infection with M. tuberculosis is to produce the given protein and test it in an appropriate assay as described herein. Of the more than 200 hundred antigens known to be expressed during primary infection, and tested as vaccines, less than a half dozen have demonstrated significant potential. So far only one antigen has been shown to have any potential as a therapeutic vaccine (Lowrie, 1999). However this vaccine only worked if given as a DNA vaccine, an experimental technique so far not approved for use in humans. Moreover, the technique has proved controversial, with other groups claiming that vaccination using this protocol induces either non-specific protection or even worsens disease (J. Turner et al., Infect and Immunity, 2000, pp. 1706-1709).

[0011] What are needed are therapeutic vaccines that treat latent TB infection.

SUMMARY OF THE INVENTION

[0012] The present invention provides therapeutic vaccines based on molecules that are induced or upregulated under the conditions of low oxygen transmission and restricted nutrients found in the granuloma (i.e., the location of latent TB infection). These vaccines are therapeutic and contrast with prior art vaccines which are designed to elicit protective immune responses prior to infection (prophylactic vaccination) that are only effective against primary infection. The immune responses elicited are powerless against the latent stage of the disease, because the bacteria have changed the antigens that they produce so that in essence they have altered their appearance and the immune system can no longer recognize them. However, latency is a dynamic process, maintained by the immune response, as indicated by the dramatic increase in the risk of reactivation of TB after HIV infection or other events that compromise immunity. Therefore, an effective vaccination strategy to protect infected individuals (therapeutic vaccination) is possible, but only if it is directed against those antigens expressed in the latent stage.

[0013] Further, the present invention provides a multiphase vaccine that combines components with prophylactic and therapeutic activity. In contrast, existing TB vaccines do not result in sterilizing immunity but rather control the infection at a subclinical level (thereby resulting in the subsequent establishment of latent infection. After conventional prophylactic vaccination, the evasion of the primary immune response and the subsequent development of latent disease are probably at least in part due to the change in the antigenic profile of the invading bacteria. Thus, vaccinating with antigens associated with latent TB prevents or reduces the establishment of latent infection and therefore, a vaccine incorporating antigens expressed by the bacteria both in the first logarithmic growth phase and during latent disease improve long-term immunity when used as a prophylactic vaccine. A multiphase vaccine of the invention will also be efficient as a therapeutic vaccine thereby addressing the problem that the majority of the population in the third world who would receive a future TB vaccine could be already latently infected.

[0014] For a number of years, a major effort has been put into the identification of protective antigens for the development of novel prophylactic vaccines against TB and today a few antigens with demonstrated protective activity in prophylactic vaccines have been identified (e.g. ESAT-6, the 38 kDa antigen, Ag85A and Ag85B). Such molecules are useful components, which in combination with latency associated antigens such as α-crystalline, form a multiphase vaccine of the invention. Advantageously and in contrast to antigens in the art, the antigens described in the invention are incorporated in vaccines through the use of well-recognized vaccination technology, as demonstrated in provided examples.

[0015] Finally, the immunodominant antigens identified in this invention may be used as diagnostic reagents. Our group has abundantly demonstrated that antigens expressed by mycobacteria during the early stages of the infection, such as ESAT-6 (Early Secretory Antigen Target-6) are recognized in individuals who are in the process of developing primary TB, even though they are healthy at the time of diagnosis (Doherty 2002). However, the large numbers of contacts who are exposed, and almost certainly infected, remain negative to this antigen (Doherty 2002). Since those individuals latently infected remain healthy by making an immune response against the latent bacteria, they must be making an immune response to those antigens expressed by the latent bacteria. Thus, the antigens of the invention may also be used to diagnose latent infection and differentiate it from primary acute TB.

[0016] Other aspects and advantages of the invention will be readily apparent to one of skill in the art.

BRIEF DESCRIPTION OF THE DRAWINGS

[0017] FIGS. 1A and 1B illustrate the results of testing in TB vaccination models. A schematic time schedule of the models for FIG. 1A, prophylactic vaccination and FIG. 1B, therapeutic vaccination. Each square on the time axis represents one week. Three prophylactic vaccinations two weeks apart are given 6 weeks prior to an aerosol infection. The protective effect of the vaccines is measured 6 weeks after infection, in the acute phase of the infection. For analysis of therapeutic vaccinations a reactivation model is established, where aerosol infected mice are treated with anti-M tuberculosis drugs for 8 weeks from the peak of infection (6 weeks after infection). This induces a latent infection phase with a low bacterial load. Four to five weeks into the latency phase three therapeutic vaccinations are given two weeks apart and the protective effect of the vaccines is measured as bacterial load in the reactivation phase, seven weeks after the last immunization.

[0018] FIGS. 2A and 2B illustrate prophylactic and therapeutic vaccine induced protection. C57Bl/6j mice were immunized 3 times with a 2-week interval with recombinant ESAT6, BCG or recombinant Rv2031c. In FIG. 2A, the immunization was given as a prophylactic vaccine 6 weeks before the mice were given a M. tuberculosis infection (approx. 250 bacilli) through the aerosol route with. Bacterial numbers in the lung was enumerated 6 weeks post infection. In FIG. 2B, the immunization was given as a therapeutic vaccine after a latent infection had been established. Bacterial numbers in the lung was enumerated 8 weeks after the last immunization. The data represents the mean of 5 individual mice.

[0019] FIG. 3 illustrates Rv2031c specific IFN-γ responses. Latent infected C57Bl/6j mice were either not immunized or immunized with 3 μg recombinant Rv2031 3 times with a two-week interval. One and two weeks post immunization mice were bleed and PBMCs isolated. The frequency of IFN-γ producing cells specific for either ESAT6 or Rv2031c was determined for both the rRv2031c immunized and the unimmunized group. In an ELIspot plate precoated with anti-IFN-γ antibodies graded numbers of PBMCs were incubated with either 2 μg/ml rRv2031c or 2 μg/ml rESAT6. After 32 h the plate was washed and incubated with biotinylated anti-INF-γ antibodies followed by a streptavidin-alkalinephosphatase incubation. The INFγ spots, representing individual IFN-γ producing cells were visualized using BCIP/NBT substrate. The results are shown as number Rv2031c specific IFN-γ producing cell (black bars) and number of ESAT6 specific IFN-γ producing cell (hatch bars) per 10⁶ PBMCs.

[0020] FIGS. 4A and 4B illustrate the results of epitope screening of Rv2031c. PBMCs from rRv2031c immunized latently infected C57Bl/6j mice were analyzed for recognition of 20' mer overlapping peptides scanning through Rv2031c. In FIG. 4A, the peptides were analyzed in pools of 3-4 peptides. PBMCs (2×10⁵) were incubated for 72 h with the peptide pools at 5 μg/ml per peptide. Supernatant was harvested and secreted IFN-γ was quantitated by ELISA. In FIG. 4B, individual peptides of positive pools were reanalyzed. PBMCs (2×10⁵) were incubated for 72 h with 1 μg/ml of each peptide. Secreted IFN-γ in the supernatant was quantitated.

[0021] FIGS. 5A and 5B illustrate protection against reactivation conferred by therapeutic vaccine given during latent infection. Latent infected C57Bl/6j mice were immunized 3 times with or without rRv2031c. Bacterial numbers in lung (FIG. 5A) and spleen (FIG. 5B) was enumerated 8 weeks after the last immunization. The data represents the mean of 8 individual mice.

[0022] FIGS. 6A and 6B illustrate Rv0569 specific IFN-γ responses. Latent infected C57Bl/6j mice were vaccinated with 3 μg of either recombinant Rv0569 or recombinant ESAT6 in a DDA/MPL adjuvant. The vaccines were given as 3 s.c. injections with a two-week interval and the induced immune response were evaluated 7 weeks after the last vaccination. Isolated splenocytes (2×10⁵) were incubated for 72 h with antigen at 1 μg/ml. Supernatant was harvested and secreted IFN-γ was quantitated by ELISA using paired anti-murine IFN-γ antibodies (PharMingen) and recombinant IFN-γ (PharMingen) as standard. In FIG. 6A, Rv0569 specific response is measured in Rv0569-vaccinated and un-vaccinated latently infected mice; in FIG. 6B, the ESAT6 specific response is measured in ESAT6-vaccinated and un-vaccinated latently infected mice

[0023] FIGS. 7A and 7B illustrate therapeutic vaccine induced protection against reactivation. Latently infected C57Bl/6j mice were vaccinated once with BCG or 3 times with a 2-week interval with either recombinant Rv0569 or recombinant ESAT6. Seven weeks after the last vaccination the bacterial numbers was enumerated in FIG. 7A, the lung and in FIG. 7B, the spleen of vaccinated and un-vaccinated mice. The data represents the mean of Log CFU per organ of 6-8 individual mice.

DETAILED DESCRIPTION OF THE INVENTION

[0024] The invention is related to preventing, treating and detecting infections caused by species of the tuberculosis complex (Mycobacterium tuberculosis, M. bovis, M. africanum) by the use of a polypeptide comprising a M. tuberculosis antigen or an immunogenic portion or other variant thereof, or by the use of a DNA sequence encoding a M. tuberculosis antigen or an immunogenic portion or other variant thereof. The invention discloses a new therapeutic vaccine against tuberculosis comprising antigens induced during the latent stage of TB-infection. It also discloses a multiphase vaccine incorporating a combination of prophylactic and therapeutic antigens as well as diagnostic reagents for the detection of the latent stage of M. tuberculosis infection.

[0025] The present invention discloses the use of one or more polypeptides, nucleic acids encoding these polypeptides or fragments hereof, which polypeptides are expressed during the latent stage of the mycobacteria infection, which stage is characterized by low-oxygen tension in the microenvironment of the mycobacteria, for a therapeutic vaccine against tuberculosis.

[0026] The polypeptides comprises one or more amino acid sequences selected from

[0027] (a) The sequences presented in Table 1.

[0028] (b) an immunogenic portion, e.g. a T-cell epitope, of any one of the sequences in (a); and/or

[0029] (c) an amino acid sequence analogue having at least 70% sequence identity to any one of the sequences in (a) or (b) and at the same time being immunogenic.

TABLE-US-00001

[0029] TABLE 1 Amino acid sequences of selected low oxygen induced antigens Rv no. SEQ ID NO: Sequence Rv0569 91 MKAKVGDWLVIKGATIDQPDHRGLIIEVRSSDGSPPYVVRWLETDHVATV IPGPDAVVVTAEEQNAADERAQHRFGAVQSAILHARGT Rv0079 1 VEPKRSRLVVCAPEPSHAREFPDVAVFSGGRANASQAERLARAVGRVLAD RGVTGGARVRLTMANCADGPTLVQINLQVGDTPLRAQAATAGIDDLRPAL IRLDRQIVRASAQWCPRPWPDRPRRRLTTPAEALVTRRKPVVLRRATPLQ AIAAMDAMDYDVHLFTDAETGEDAVVYRAGPSGLRLARQHHVFPPGWSRC RAPAGPPVPLIVNSRPTPVLTEAAAVDRAREHGLPFLETTDQATGRGQLL YSRYDGNLGLITPTGDGVADGLA Rv0080 2 MSPGSRRASPQSAREVVELDRDEAMRLLASVDHGRVVFTRAALPAIRPVN HLVVDGRVIGRTRLTAKVSVAVRSSADAGVVVAYEADDLDPRRRTGWSVV VTGLATEVSDPEQVARYQRLLHPWVNMAMDTVVAIEPEIVTGIRIVADSR TP Rv0081 3 VESEPLYKLKAEFEKTLAHPARIRILELLVERDRSVGELLSSDVGLESSN LSQQLGVLRRAGVVAARRDGNAMIYSIAAPDIAELLAVARKVLARVLSDR VAVLEDLRAGGSAT Rv0363c 4 MPIATPEVYAEMLGQAKQNSYAFPAINCTSSETVNAAIKGFADAGSDGII QFSTGGAEFGSGLGVKDMVTGAVALAEFTHVIAAKYPVNVALHTDHCPKD KLDSYVRPLLAISAQRVSKGGNPLFQSHMWDGSAVPIDENLAIAQELLKA AAAAKIILEIEIGVVGGEEDGVANEINEKLYTSPEDFEKTIEALGAGEHG KYLLAATFGNVHGVYKPGNVKLRPDILAQGQQVAAAKLGLPADAKPFDFV FHGGSGSLKSEIEEALRYGVVKMNVDTDTQYAFTRPIAGHMFTNYDGVLK VDGEVGVKKVYDPRSYLKKAEASMSQRVVQACNDLHCAGKSLTH Rv0572c 5 MGEHAIKRHMRQRKPTKHPLAQKRGARILVFTDDPRRSVLIVPGCHLDSM RREKNAYYFQDGNALVGMVVSGGTVEYDADDRTYVVQLTDGRHTTESSFE HSSPSRSPQSDDL Rv0574c 6 VAGNPDVVTVLLGGDVMLGRGVDQILPHPGKPQLRERYMRDATGYVRLAE RVNGRIPLPVDWRWPWGEALAVLENTATDVCLINLETTITADGEFADRKP VCYRMHPDNVPALTALRPHVCALANNHILDFGYQGLTDTVAALAGAGIQS VGAGADLLAARRSALVTVGHERRVIVGSVAAESSGVPESWAARRDRPGVW LIRDPAQRDVADDVAAQVLADKRPGDIAIVSMHWGSNWGYATAPGDVAFA HRLIDAGIDMVHGHSSHHPRPIEIYRGKPILYGCGDVVDDYEGIGGHESF RSELRLLYLTVTDPASGNLISLQMLPLRVSRMRLQRASQTDTEWLRNTIE RISRRFGIRVVTRPDNLLEVVPAANLTSKE Rv1264 7 VTDHVREADDANIDDLLGDLGGTARAERAKLVEWLLEQGITPDEIRATNP PLLLATRHLVGDDGTYVSAREISENYGVDLELLQRVQRAVGLARVDDPDA VVHMRADGEAAARAQRFVELGLNPDQVVLVVRVLAEGLSHAAEAMRYTAL EAIMRPGATELDIAKGSQALVSQIVPLLGPMIQDMLFMQLRHMMETEAVN AGERAAGKPLPGARQVTVAFADLVGFTQLGEVVSAEELGHLAGRLAGLAR DLTAPPVWFIKTIGDAVMLVCPDPAPLLDTVLKLVEVVDTDNNFPRLRAG VASGMAVSRAGDWFGSPVNVASRVTGVARPGAVLVADSVREALGDAPEAD GFQWSFAGPRRLRGIRGDVRLFRVRRGATRTGSGGAAQDDDLAGSSP Rv1592c 8 MVEPGNLAGATGAEWIGRPPHEELQRKVRPLLPSDDPFYFPPAGYQHAVP GTVLRSRDVELAFMGLIPQPVTATQLLYRTTNMYGNPEATVTTVIVPAEL APGQTCPLLSYQCAIDAMSSRCFPSYALRRRAKALGSLTQMELLMISAAL AEGWAVSVPDHEGPKGLWGSPYEPGYRVLDGIRAALNSERVGLSPATPIG LWGYSGGGLASAWAAEACGEYAPDLDIVGAVLGSPVGDLGHTFRRLNGTL LAGLPALVVAALQHSYPGLARVIKEHANDEGRQLLEQLTEMTTVDAVIRM AGRDMGDFLDEPLEDILSTPEISHVFGDTKLGSAVPTPPVLIVQAVHDYL IDVSDIDALADSYTAGGANVTYHRDLFSEHVSLHPLSAPMTLRWLTDRFA GKPLTDHRVRTTWPTIFNPMTYAGMARLAVIAAKVITGRKLSRRPL Rv1733c 9 MIATTRDREGATMITFRLRLPCRTILRVFSRNPLVRGTDRLEAVVMLLAV TVSLLTIPFAAAAGTAVQDSRSHVYAHQAQTRHPATATVIDHEGVIDSNT TATSAPPRTKITVPARWVVNGIERSGEVNAKPGTKSGDRVGIWVDSAGQL VDEPAPPARAIADAALAALGLWLSVAAVAGALLALTRAILIRVRNASWQH DIDSLFCTQR Rv1734c 10 MTNVGDQGVDAVFGVIYPPQVALVSFGKPAQRVCAVDGAIHVMTTVLATL PADHGCSDDHRGALFFLSINELTRCAAVTG Rv1736c 11 VTVTPRTGSRIEELLARSGRFFIPGEISADLRTVTRRGGRDGDVFYRDRW SHDKVVRSTHGVNCTGSCSWKIYVKDDIITWETQETDYPSVGPDRPEYEP RGCPRGAAFSWYTYSPTRVRHPYARGVLVEMYREAKARLGDPVAAWADIQ ADPRRRRRYQRARGKGGLVRVSWAEATEMIAAAHVHTISTYGPDRVAGFS PIPAMSMVSHAAGSREVELIGGVMTSFYDWYADLPVASPQVFGDQTDVPE SGDWWDVVWQCASVLLTYPNSRQLGTAEELLAHIDGPAADLLGRTVSELR RADPLTAATRYVDTFDLRGRATLYLTYWTAGDTRNRGREMLAFAQTYRST DVAPPRGETPDFLPVVLEFAATVDPEAGRRLLSGYRVPTAALCNALTEAA LPYAHTVAAVCRTGDMMGELFWTVVPYVTMTIVAVGSWWRYRYDKFGWTT RSSQLYESRLLRIASPMFHFGILVVIVGHGIGLVIPQSWTQAAGLSEGAY HVQAVVLGSIAGITTLAGVTLLIYRRRTRGPVFMATTVNDKVMYLVLVAA IVAGLGATALGSGVVGEAYNYRETVSVWFRSVWVLQPRGDLMAEAPLYYQ IHVLIGLALFALWPFTRLVHAFSAPIGYLFRPYIIYRSREELVLTRPRRR GW Rv1737c 12 MRGQAANLVLATWISVVNFWAWNLIGPLSTSYARDMSLSSAEASLLVATP ILVGALGRIVTGPLTDRFGGRANLIAVTLASILPVLAVGVAATMGSYALL VFFGLFLGVAGTIFAVGIPFANNWYQPARRGESTGVEGMGMVGTALSAFE TPREVRWEGLFTTHAIVAAALASTAVVAMVVLRDAPYFRPNADPVLPRLK AAARLPVTWEMSFLYAIVEGGEVAFSNYLPTYITTIYGESTVDAGARTAG FALAAVLARPVGGWLSDRIAPRHVVLASLAGTALLAFAAALQPPPEVWSA ATFITLAVCLGVGTGGVFAWVARRAPAASVGSVTGIVAAAGGLGGYFPPL VMGATYDPVDNDYTVGLLLLVATALVACTYTALHAREPVSEEASR Rv1738c 13 MCGDQSDHVLQHWTVDISIDEHEGLTRAKARLRWREKELVGVGLARLNPA DRNVPEIGDELSVARALSDLGKRMLKVSTHDIEAVTHQPARLLY Rv1739c 14 MIPTMTSAGWAPGVVQFREYQRRWLRGDVLAGLTVAAYLIPQAMAYATVA GLPPAAGLWASIAPLAIYALLGSSRQLSIGPESATALMTAAVLAPMAAGD LRRYAVLAATLGLLVGLICLLAGTARLGFLASLRSRPVLVGYMAGIALVM ISSQLGTITGTSVEGNEFFSEVHSFATSVTRVHWPTFVLAMSVLALLTML TRWAPRAPGPIIAVLAATMLVAVMSLDAKGIAIVGRIPSGLPTPGVPPVS VEDLRALIIPAAGIAIVTFTDGVLTARAFAARRGQEVNANAELRAVGACN IAAGLTHGFPVSSSSSRTALADVVGGRTQLYSLIALGLVVIVMVFASGLL AMFPTAALGALVVYAALRLIDLSEFRRLARFRRSELMLALATTAAVLGLG VFYGVLAAVALSILELLRRVAHPHDSVLGFVPGIAGMHDIDDYPQAKRVP GLVVYRYDAPLCFANAEDFRRRALTVVDQDPGQVEWFVLNAESNVEVDLT ALDALDQLRTELLRRGIVFAMARVKQDLRESLRAASLLDKIGEDHIFMTL PTAVQAFRRR Rv1813c 15 MITNLRRRTAMAAAGLGAALGLGILLVPTVDAHLANGSMSEVMMSE IAGLPIPPIIHYGAIAYAPSGASGKAWHQRTPARAEQVALEKCGDK TCKVVSRFTRCGAVAYNGSKYQGGTGLTRRAAEDDAVNRLEGGRIV NWACN Rv1997c 16 LSASVSATTAHHGLPAHEVVLLLESDPYHGLSDGEAAQRLERFGPNTLAV VTRASLLARILRQFHHPLIYVLLVAGTITAGLKEFVDAAVIFGVVVINAI VGFIQESKAEAALQGLRSMVHTHAKVVREGHEHTMPSEELVPGDLVLLAA GDKVPADLRLVRQTGLSVNESALTGESTPVHKDEVALPEGTPVADRRNIA YSGTLVTAGHGAGIVVATGAETELGEIHRLVGAAEVVATPLTAKLAWFSK FLTTATLGLAALTFGVGLLRRQDAVETFTAAIALAVGAIPEGLPTAVTIT LAIGMARMAKRRAVIRRLPAVETLGSTTVICADKTGTLTENQMTVQSIWT PHGEIRATGTGYAPDVLLCDTDDAPVPVNANAALRWSLLAGACSNDAALV RDGTRWQIVGDPTEGAMLVVAAKAGFNPERLATTLPQVAAIPFSSERQYM ATLHRDGTDHVVLAKGAVERMLDLCGTEMGADGALRPLDRATVLRATEML TSRGLRVLATGMGAGAGTPDDFDENVIPGSLALTGLQAMSDPPRAAAASA VAACHSAGIAVKMITGDHAGTATAIATEVGLLDNTEPAAGSVLTGAELAA LSADQYPEAVDTASVFARVSPEQKLRLVQALQARGHVVAMTGDGVNDAPA LRQANIGVAMGRGGTEVAKDAADMVLTDDDFATIEAAVEEGRGVFDNLTK FITWTLPTNLGEGLVILAAIAVGVALPILPTQILWINMTTAIALGLMLAF EPKEAGIMTRPPRDPDQPLLTGWLVRRTLLVSTLLVASAWWLFAWELDNG AGLHEARTAALNLFVVVEAFYLFSCRSLTRSAWRLGMFANRWIILGVSAQ AIAQFATTYLPAMNMVFDTAPIDIGVWVRIFAVATAITIVVATDTLLPRI RAQPP Rv1998c 17 MSFHDLHHQGVPFVLPNAWDVPSALAYLAEGFTAIGTTSFGVSSSGGHPD GHRATRGANIALAAALAPLQCYVSVDIEDGYSDEPDAIADYVAQLSTAGI NIEDSSAEKLIDPALAAAKIVAIKQRNPEVFVNARVDTYWLRQHADTTST IQRALRYVDAGADGVFVPLANDPDELAELTRNIPCPVNTLPVPGLTIADL GELGVARVSTGSVPYSAGLYAAAHAARAVSDGEQLPRSVPYAELQARLVD YENRTSTT Rv2003c 18 VVKRSRATRLSPSIWSGWESPQCRSIRARLLLPRGRSRPPNADCCWNQLA VTPDTRMPASSAAGRDAAAYDAWYDSPTGRPILATEVAALRPLIEVFAQP RLEIGVGTGRFADLLGVREGLDPSRDALMFARRRGVLVANAVGEAVPFVS RHFGAVLMAFTLCFVTDPAAIFRETRRLLADGGGLVIGELPRGTPWADLY ALRAARGQPGYRDARFYTAAELEQLLADSGFRVIARRCTLHQPPGLARYD IEAAHDGIQAGAGFVAISAVDQAHEPKDDHPLESE Rv2005c 19 MSKPRKQHGVVVGVDGSLESDAAACWGATDAAMRNIPLTVVHVVNADVAT WPPMPYPETWGVWQEDEGRQIVANAVKLAKEAVGADRKLSVKSELVFSTP VPTMVEISNEAEMVVLGSSGRGALARGLLGSVSSSLVRRAGCPVAVIHSD DAVIPDPQHAPVLVGIDGSPVSELATAVAFDEASRRGVELIAVHAWSDVE VVELPGLDFSAVQQEAELSLAERLAGWQERYPDVPVSRVVVCDRPARKLV QKSASAQLVVVGSHGRGGLTGMLLGSVSNAVLHAARVPVIVARQS Rv2007c 20 VTYVIGSECVDVMDKSCVQECPVDCIYEGARMLYINPDECVDCGACKPAC RVEAIYWEGDLPDDQHQHLGDNAAEFHQVLPGRVAPLGSPGGAAAVGPIG VDTPLVAAIPVECP Rv2028c 21 MNQSHKPPSIVVGIDGSKPAVQAALWAVDEAASRDIPLRLLYAIEPDDPG YAAHGAAARKLAAAENAVRYAFTAVEAADRPVKVEVEITQERPVTSLIRA SAAAALVCVGAIGVHHFRPERVGSTAAALALSAQCPVAIVRPHRVPIGRD AAWIVVEADGSSDIGVLLGAVMAEARLRDSPVRVVTCRQSGVGDTGDDVR ASLDRWLARWQPRYPDVRVQSAAVHGELLDYLAGLGRSVHMVVLSASDQE HVEQLVGAPGNAVLQEAGCTLLVVGQQYL Rv2029c 22 MTEPAAWDEGKPRIITLTMNPALDITTSVDVVRPTEKMRCGAPRYDPGGG GINVARIVHVLGGCSTALFPAGGSTGSLLMALLGDAGVPFRVIPIAASTR ESFTVNESRTAKQYRFVLPGPSLTVAEQEQCLDELRGAAASAAFVVASGS LPPGVAADYYQRVADICRRSSTPLILDTSGGGLQHISSGVFLLKASVREL RECVGSELLTEPEQLAAAHELIDRGRAEVVVVSLGSQGALLATRHASHRF SSIPMTAVSGVGAGDAMVAAITVGLSRGWSLIKSVRLGNAAGAAMLLTPG TAACNRDDVERFFELAAEPTEVGQDQYVWHPIVNPEASP Rv2030c 23 VLMTAAADVTRRSPRRVFRDRREAGRVLAELLAAYRDQPDVIVLGLARGG LPVAWEVAAALHAPLDAFVVRKLGAPGHDEFAVGALASGGRVVVNDDVVR GLRITPQQLRDIAEREGRELLRRESAYRGERPPTDITGKTVIVVDDGLAT GASMFAAVQALRDAQPAQIVIAVPAAPESTCREFAGLVDDVVCATMPTPF LAVGESFWDFRQVTDEEVRRLLATPTAGPSLRRPAASTAADVLRRVAIDA PGGVPTHEVLAELVGDARIVLIGESSHGTHEFYQARAAMTQWLIEEKGFG AVAAEADWPDAYRVNRYVRGLGEDTNADEALSGFEREPAWMWRNTVVRDF VEWLRTRNQRYESGALRQAGFYGLDLYSLHRSIQEVISYLDKVDPRAAAR ARARYACFDHACADDGQAYGFAAAFGAGPSCEREAVEQLVDVQRNALAYA RQDGLLAEDELFYAQQNAQTVRDAEVYYRAMFSGRVTSWNLRDQHMAQTL GSLLTHLDRHLDAPPARIVVWAHNSHVGDARATEVWADGQLTLGQIVRER YGDESRSIGESTYTGTVTAASEWGGIAQRKAVRPALHGSVEELFHQTADS FLVSARLSRDAEAPLDVVRLGRAIGVVYLPATERQSHYLHVRPADQFDAM IHIDQTRALEPLEVTSRWIAGENPETYPTGL Rv2031c 24 MATTLPVQRHPRSLFPEESELFAAFPSFAGLRPTFDTRLMRLEDEMKEGR YEVRAELPGVDPDKDVDIMVRDGQLTIKAERTEQKDFDGRSEFAYGSFVR TVSLPVGADEDDIKATYDKGILTVSVAVSEGKPTEKHIQIRSTN Rv2032 25 MPDTMVTTDVIKSAVQLACRAPSLHNSQPWRWIAEDHTVALFLDKDRVLY ATDHSGREALLGCGAVLDHFRVAMAAAGTTANVERFPNPNDPLHLASIDF SPADFVTEGHRLRADAILLRRTDRLPFAEPPDWDLVESQLRTTVTADTVR IDVIADDMRPELAAASKLTESLRLYDSSYHAELFWWTGAFETSEGIPHSS LVSAAESDRVTFGRDEPVVANTDRRPEFGHDRSKVLVLSTYDNERASLLR CGEMLSAVLLDATMAGLATCTLTHITELHASRDLVAALIGQPATPQALVR VGLAPEMEEPPPATPRRPIDEVFHVRAKDHR Rv2428 26 MPLLTIGDQFPAYQLTALIGGDLSKVDAKQPGDYFTTITSDEHPGKWRVV FEWPKDFTEVCPTEIAAFSKLNDEFEDRDAQILGVSIDSEFAHFQWRAQH NDLKTLPFPMLSDIKRELSQAAGVLNADGVADRVTFIVDPNNEIQFVSAT AGSVGRNVDEVLRVLDALQSDELCACNWRKGDPTLDAGELLKASA Rv2624c 27 MSGRGEPTMKTIIVGIDGSHAAITAALWGVDEAISRAVPLRLVSVIKPTH PSPDDYDRDLAHAERSLREAQSAVEAAGKLVKIETDIPRGPAGPVLVEAS RDAEMICVGSVGIGRYASSILGSTATELAEKAHCPVAVMRSKVDQPASDI NWIVVRMTDAPDNEAVLEYAAREAKLRQAPILALGGRPEELREIPDGEFE RRVQDWHHRHPDVRVYPITTHTGIARFLADHDERVQLAVIGGGEAGQLAR LVGPSGHPVFRHAECSVLVVRR Rv2625c 28 MRDAIPLGRIAGFVVNVHWSVLVILWLFTWSLATMLPGTVGGYPAVVYWL LGAGGAVMLLASLLAHELAHAVVARRAGVSVESVTLWLFGGVTALGGEAK TPKAAFRIAFAGPATSLALSATFGALAITLAGVRTPAIVISVAWWLATVN LLLGLFNLLPGAPLDGGRLVRAYLWRRHGDSVRAGIGAARAGRVVALVLI ALGLAEFVAGGLVGGVWLAFIGWFIFAAAREEETRISTQQLFAGVRVADA MTAQPHTAPGWINVEDFIQRYVLGERHSAYPVADRDGSITGLVALRQLRD VAPSRRSTTSVGDIALPLHSVPTARPQEPLTALLERMAPLGPRSRALVTE GSAVVGIVTPSDVARLIDVYRLAQPEPTFTTSPQDADRFSDAG Rv2727c 29 MASSASGDTHERSAFRLSPPVLSGAMGPFMHTGLYVAQSWRDYLGQQPDK LPTARPTIALAAQAFRDEIVLLGLKARRPVSNHRVFERISQEVAAGLEFY GNRRWLEKPSGFFAQPPPLTEVAVRKVKDRRRSFYRIFEDSGFTPHPGEP GSQRWLSYTANNREYALLLRHPEPRPWLVCVHGTEMGRAPLDLAVFRAWK LHDELGLNIVMPVLPMHGPRGQGLPKGAVFPGEDVLDDVHGTAQAVWDIR RLLSWIRSQEEESLIGLNGLSLGGYIASLVASLEEGLACAILGVPVADLI ELLGRHCGLRHKDPRRHTVKMAEPIGRMISPLSLTPLVPMPGRFIYAGIA DRLVHPREQVTRLWEHWGKPEIVWYPGGHTGFFQSRPVRRFVQAALEQSG LLDAPRTQRDRSA Rv2628 30 MSTQRPRHSGIRAVGPYAWAGRCGRIGRWGVHQEAMMNLAIWHPRKVQSA TIYQVTDRSHDGRTARVPGDEITSTVSGWLSELGTQSPLADELARAVRIG DWPAAYAIGEHLSVEIAVAV Rv2629 31 MRSERLRWLVAAEGPFASVYFDDSHDTLDAVERREATWRDVRKHLESRDA KQELIDSLEEAVRDSRPAVGQRGRALIATGEQVLVNEHLIGPPPATVIRL SDYPYVVPLIDLEMRRPTYVFAAVDHTGADVKLYQGATISSTKIDGVGYP VHKPVTAGWNGYGDFQHTTEEAIRMNCRAVADHLTRLVDAADPEVVFVSG EVRSRTDLLSTLPQRVAVRVSQLHAGPRKSALDEEEIWDLTSAEFTRRRY AEITNVAQQFEAEIGRGSGLAAQGLAEVCAALRDGDVDTLIVGELGEATV VTGKARTTVARDADMLSELGEPVDRVARADEALPFAAIAVGAALVRDDNR IAPLDGVGALLRYAATNRLGSHRS Rv2630 32 MLHRDDHINPPRPRGLDVPCARLRATNPLRALARCVQAGKPGTSSGHRSV

PHTADLRIEAWAPTRDGCIRQAVLGTVESFLDLESAHAVHTRLRRLTADR DDDLLVAVLEEVIYLLDTVGETPVDLRLRDVDGGVDVTFATTDASTLVQV GAVPKAVSLNELRFSQGRHGWRCAVTLDV Rv2659c 33 VTQTGKRQRRKFGRIRQFNSGRWQASYTGPDGRVYIAPKTFNAKIDAEAW LTDRRREIDRQLWSPASGQEDRPGAPFGEYAEGWLKQRGIKDRTRAHYRK LLDNHILATFADTDLRDITPAAVRRWYATTAVGTPTMRAHSYSLLRAIMQ TALADDLIDSNPCRISGASTARRVHKIRPATLDELETITKAMPDPYQAFV LMAAWLAMRYGELTELRRKDIDLHGEVARVRRAVVRVGEGFKVTTPKSDA GVRDISIPPHLIPAIEDHLHKHVNPGRESLLFPSVNDPNRHLAPSALYRM FYKARKAAGRPDLRVHDLRHSGAVLAASTGATLAELMQRLGHSTAGAALR YQHAAKGRDREIAALLSKLAENQEM Rv2780 34 MRVGIPTETKNNEFRVAITPAGVAELTRRGHEVLIQAGAGEGSAITDADF KAAGAQLVGTADQVWADADLLLKVKEPIAAEYGRLRHGQILFTFLHLAAS RACTDALLDSGTTSIAYETVQTADGALPLLAPMSEVAGRLAAQVGAYHLM RTQGGRGVLMGGVPGVEPADVVVIGAGTAGYNAARIANGMGATVTVLDIN IDKLRQLDAEFCGRIHTRYSSAYELEGAVKRADLVIGAVLVPGAKAPKLV SNSLVAHMKPGAVLVDIAIDQGGCFEGSRPTTYDHPTFAVHDTLFYCVAN MPASVPKTSTYALTNATMPYVLELADHGWRAACRSNPALAKGLSTHEGAL LSERVATDLGVPFTEPASVLA Rv3126c 35 MVIRFDQIGSLVLSMKSLASLSFQRCLRENSSLVAALDRLDAAVDELSAL SFDALTTPERDRARRDRDHHPWSRSRSQLSPRMAHGAVHQCQWPKAVWAV IDNP Rv3127 36 VLKNAVLLACRAPSVHNSQPWRWVAESGSEHTTVHLFVNRHRTVPATDHS GRQAIISCGAVLDHLRIAMTAAHWQANITRFPQPNQPDQLATVEFSPIDH VTAGQRNRAQAILQRRTDRLPFDSPMYWHLFEPALRDAVDKDVAMLDVVS DDQRTRLVVASQLSEVLRRDDPYYHAELEWWTSPFVLAHGVPPDTLASDA ERLRVDLGRDFPVRSYQNRRAELADDRSKVLVLSTPSDTRADALRCGEVL STILLECTMAGMATCTLTHLIESSDSRDIVRGLTRQRGEPQALIRVGIAP PLAAVPAPTPRRPLDSVLQIRQTPEKGRNASDRNARETGWFSPP Rv3128c 37 VWSASGGQCGKYLAASMVLQLDGLERHGVLEFGRDRYGPEVREELLAMSA ASIDRYLKTAKAKDQISGVSTTKPSPLLRNSIKVRRAGDEVEAEPGFFEG DTVAHCGPTLKGEFAHTLNLTDVHIGWVFTRTVRNNARTHILAGLKASVT EIPHGITGLDFDNGTVELNKPVISWAGDNGIYFTRFRPYKKNH*ATIESK NNHLVRKYAFYYRYDTAEERAVLNRMWKLVNDRLNYLTPTIKPIGYASSA DGRRRRLYDAPQTPLDRPLAARVLSAAQQADLITYRDSLNPAQIGRKIAD LQNRLLILAKEKTEQLYLANIPTALPDIHKGILIKAG Rv3129 38 VVQGRTVLFRTAEGAKLFSAVAKCAVAFEADDHNVAEGWSVIVKVRAQVL TTDAGVREAERAQLLPWTATLKRHCVRVIPWEITGRHFREGPEPDRSQTF ACEASSHNQR Rv3130c 39 MNHLTTLDAGFLKAEDVDRHVSLAIGALAVIEGPAPDQEAFLSSLAQRLR PCTRFGQRLRLRPFDLGAPKWVDDPDFDLGRHVWRIALPRPGNEDQLFEL IADLMARRLDRGRPLWEVWVIEGLADSKWAILTKLHHCMADGIAATHLLA GLSDESMSDSFASNIHTTMQSQSASVRRGGFRVNPSEALTASTAVMAGIV RAAKGASEIAAGVLSPAASSLNGPISDLRRYSAAKVPLADVEQVCRKFDV TINDVALAAITESYRNVLIQRGERPREDSLRTLVPVSTRSNSALSKTDNR VSLMLPNLPVDQENPLQRLRIVHSRLTRAKAGGQRQFGNTLMAIANRLPF PMTAWAVGLLMRLPQRGVVTVATNVPGPRRPLQIMGRRVLDLYPVSPIAM QLRTSVAMLSYADDLYFGILADYDVVADAGQLARGIEDAVARLVAISKRR KVTRRRGALSLVV Rv3131 40 MNTHFPDAETVRTVLTLAVRAPSIHNTQPWRWRVCPTSLELFSRRDMQLR STDPDGRELILSCGVALHHCVVALASLGWQAKVNRFPDPKDRCHLATIGV QPLVPDQADVALAAAIPRRRTDRRAYSCWPVPGGDIALMAARAARGGVML RQVSALDRMKAIVAQAVLDHVTDEEYLRELTIWSGRYGSVAGVPARNEPP SDPSAPIPGRLFAGPGLSQPSDVLPADDGAAILALGTETDDRLARLRAGE AASIVLLTATAMGLACCPITEPLETAKTRDAVRAEVEGAGGYPQMLLRVG WAPINADPLPPTPRRELSQVVEWPEELLRQRC Rv3132 41 MTTGGLVDENDGAAMRPLRHTLSQLRLHELLVEVQDRVEQIVEGRDRLDG LVEAMLVVTAGLDLEATLRAIVHSATSLVDARYGAMEVHDRQHRVLHEVY EGIDEETVRRIGHLPKGLGVIGLLIEDPKPLRLDDVSAHPASIGFPPYHP PMRTFLGVPVRVRDESEGTLYLTDKTNGQPFSDDDEVLVQALAAAAGIAV ANARLYQQAKARQSWIEATRDTATELLSGTEPATVFRLVAAEALKLTAAD AALVAVPVDEDMPAADVGELLVIETVGSAVASIVGRTIPVAGAVLREVFV NGIPRRVDRVDLEGLDELADAGPALLLPLRARGTVAGVVVVLSQGGPGAF TDEQLEMMAAFADQAALAWQLATSQRRMRELDVLTDRDRIARDLHDHVIQ RLFATGLALQGAVPHERNPEVQQRLSDVVDDLQDVIQEIRTTIYDLHGAS QGITRLRQRIDAAVAQFADSGLRTSVQFVGPLSVVDSALADQAEAVVREA VSNAVRHAKASTLTVRVKVDDDLCIEVTDNGRGLPDEFTGSGLTNLRQRA EQAGGEFTLASVPGASGTVLRWSAPLSQ Rv3134c 42 MSDPRPARAVVVGIDGSRAATHAALWAVDEAVNRDIPLRLVYVIDPSQLS AAGEGGGQSAARAALHDASRKVEATGQPVKIETEVLCGRPLTKLMQESRS AAMLCVGSVGLDHVRGRRGSVAATLAGSALCPVAVIHPSPAEPATTSQVS AVVAEVDNGVVLRHAFEEARLRGVPLRAVAVHAAETPDDVEQGSRLAHVH LSRRLAHWTRLYPEVRVDRAIAGGSACRHLAANAKPGQLFVADSHSAHEL CGAYQPGCAVLTVRSANL Rv3841 43 MTEYEGPKTKFHALMQEQIHNEFTAAQQYVAIAVYFDSEDLPQLAKHFYS QAVEERNHAMMLVQHLLDRDLRVEIPGVDTVRNQFDRPREALALALDQER TVTDQVGRLTAVARDEGDFLGEQFMQWFLQEQIEEVALMATLVRVADRAG ANLFELENFVAREVDVAPAASGAPHAAGGRL Rv3842c 44 MTWADEVLAGHPFVVAHRGASAARPEHTLAAYDLALKEGADGVECDVRLT RDGHLVCVHDRRLDRTSTGAGLVSTMTLAQLRELEYGAWHDSWRPDGSHG DTSLLTLDALVSLVLDWHRPVKIFVETKHPVRYGSLVENKLLALLHRFGI AAPASADRSRAVVMSFSAAAVWRIRRAAPLLPTVLLGKTPRYLTSSAATA VGATAVGPSLPALKEYPQLVDRSAAQGRAVYCWNVDEYEDIDFCREVGVA WIGTHHPGRTKAWLEDGRANGTTR Rv3908 45 VSDGEQAKSRRRRGRRRGRRAAATAENHMDAQPAGDATPTPATAKRSRSR SPRRGSTRMRTVHETSAGGLVIDGIDGPRDAQVAALIGRVDRRGRLLWSL PKGHIELGETAEQTAIREVAEETGIRGSVLAALGRIDYWFVTDGRRVHKT VHHYLMRFLGGELSDEDLEVAEVAWVPIRELPSRLAYADERRLAEVADEL IDKLQSDGPAALPPLPPSSPRRRPQTHSRARHADDSAPGQHNGPGPGP

TABLE-US-00002 TABLE 2 DNA sequences of selected low oxygen induced antigens Rv no. SEQ ID NO: Sequence Rv0079c 46 gtggaaccgaaacgcagtcgcctcgtcgtatgtgcacccgagccatcgca cgcgcgggaattcccggatgtcgccgtattctccggcggccgggctaacg catcccaggccgaacggttggctcgtgccgtgggtcgcgtgttggccgat cggggcgtcaccgggggtgctcgggtgcggctgaccatggcgaactgcgc cgatgggccgacgctggtgcagataaacctgcaggtaggtgacaccccat taagggcgcaggccgccaccgcgggcatcgatgatctgcgacccgcactg atcagactggatcgacagatcgtgcgggcgtcggcacagtggtgcccccg gccttggccggatcggccccgccggcgattgaccacgccggccgaggcgc tagtcacccgccgcaaaccggtcgtgctaaggcgcgcaaccccgttgcag gcgattgccgctatggacgccatggactacgacgtgcatttgttcaccga cgccgagacgggggaggacgctgtggtctatcgggctggaccgtcggggc tgcggctggcccgccagcaccacgtatttcccccaggatggtcacgttgt cgcgccccagccgggccgccggtgccgctgattgtgaattcgcgtccgac accggttctcacggaggccgccgcggtggaccgggcgcgcgaacatggac tgccattcctgtttttcaccgaccaggccaccggccgcggccagctgctc tactcccgctacgacggcaacctcgggttgatcaccccgaccggtgacgg cgttgccgacggtctggca Rv0080 47 atgagcccgggctcgcggcgcgccagcccgcaaagcgcccgggaggtggt cgagctcgaccgtgacgaggcgatgcggttgctggccagcgttgaccatg ggcgtgtggtgttcacccgcgcggcgctgccggcgatccgtccagtcaat cacctcgtggtcgacggtcgggtgatcgggcgcacccgcctgacggccaa ggtgtccgttgcggtgcgatcgagcgccgatgccggtgtcgtggtcgcct acgaagccgacgaccttgatccgcggcgtcggacggggtggagtgtggtg gtgacgggactggcgaccgaggtcagcgatcccgagcaggttgcccgcta ccagcggctgctacacccgtgggtgaacatggcgatggacaccgtggtcg cgatcgaacccgagatcgtcaccggcatccgcatcgttgctgactcgcgt acgccg Rv0081 48 gtggagtccgaaccgctgtacaagctcaaggcggagttcttcaaaaccct tgcgcatccggcgcggatcaggattttggagctgctggtcgagcgggacc gttcggtcggtgagttgctgtcctcggacgtcggcctggagtcgtcgaac ctgtcccagcagctgggtgtgctacgccgggcgggtgttgtcgcggcacg tcgtgacggcaacgcgatgatctattcgattgccgcacccgatatcgccg agctgctggcggtggcacgcaaggtgctggccagggtgctcagcgaccgg gtggcggtgctagaggacctccgcgccggcggctcggccacg Rv0363c 49 atgcctatcgcaacgcccgaggtctacgcggagatgctcggtcaggccaa acaaaactcgtacgctttcccggctatcaactgcacctcctcggaaaccg tcaacgccgcgatcaaaggtttcgccgacgccggcagtgacggaatcatc cagttctcgaccggtggcgcagaattcggctccggcctcggggtcaaaga catggtgaccggtgcggtcgccttggcggagttcacccacgttatcgcgg ccaagtacccggtcaacgtggcgctgcacaccgaccactgccccaaggac aagttggacagctatgtccggcccttgctggcgatctcggcgcaacgcgt gagcaaaggtggcaatcctttgttccagtcgcacatgtgggacggctcgg cagtgccaatcgatgagaacctggccatcgcccaggagctgctcaaggcg gcggcggccgccaagatcattctggagatcgagatcggcgtcgtcggcgg cgaagaggacggcgtggcgaacgagatcaacgagaagctgtacaccagcc cggaggacttcgagaaaaccatcgaggcgctgggcgccggtgagcacggc aaatacctgctggccgcgacgttcggcaacgtgcatggcgtctacaagcc cggcaacgtcaagcttcgccccgacatccttgcgcaagggcaacaggtgg cggcggccaagctcggactgccggccgacgccaagccgttcgacttcgtg ttccacggcggctcgggttcgcttaagtcggagatcgaggaggcgctgcg ctacggcgtggtgaagatgaacgtcgacaccgacacccagtacgcgttca cccgcccgatcgccggtcacatgttcaccaactacgacggagtgctcaag gtcgatggcgaggtgggtgtcaagaaggtctacgacccgcgcagctacct caagaaggccgaagcttcgatgagccagcgggtcgttcaggcgtgcaatg acctgcactgcgccggaaagtccctaacccac Rv0572c 50 atgggtgagcacgccatcaagcggcacatgcggcaacggaagcctacgaa gcatcccctagcccagaaacggggcgcgcggattctggtcttcaccgacg atccccgcaggagcgtcctcatagtgcccggttgccacctggattccatg cgccgagaaaagaacgcgtactacttccaggacggcaatgcgttggttgg gatggttgtctcgggcggcacggttgagtacgacgccgacgaccgcacat atgtcgtgcagctcaccgacggaaggcacaccactgagtcatctttcgaa cactcatcgccgagtcgatcacctcaatccgatgaccta Rv0574c 51 gtggctggcaatcctgatgtggtgacggtgctgctgggcggtgacgtcat gctcggccgtggcgtcgatcagatcctgcctcatcccggcaaaccgcaat tgcgcgaacggtatatgcgggatgcgaccggctatgttcgcctggccgag cgggtgaacgggcgcattccgctccccgtggattggcgctggccctgggg cgaggcgttggcggtccttgagaacaccgcgaccgacgtctgtttgatca atctggagacgacgatcaccgccgacggtgaattcgccgaccgcaaaccg gtctgctaccggatgcacccggataacgtgccggcgctgacggcattgcg gccgcacgtgtgcgcgctggccaacaaccacattctcgatttcggctacc aggggctgaccgatacggtcgcggctctcgccggtgcggggatccagagt gtcggggcgggagccgatttgctcgccgctcgccgctcggcgctagtcac ggttggccatgaacgccgggtgatcgtcggctcggtagcggcggaatcca gcggcgtccccgaatcctgggccgcccgccgcgaccggcccggagtgtgg ttgatccgggatccggcgcaacgcgacgtcgccgacgatgtggcggcaca ggtgctggcggacaaacgccccggcgatatcgccatagtctcgatgcatt ggggatccaattggggctatgcgaccgcacccggcgacgtcgcgttcgcg caccgactgatcgacgccggcatcgacatggtccacggacattcctcgca ccatccgcggccaatcgagatatatcgcggtaaaccgatcctgtacggat gcggtgacgtcgttgacgactacgaaggcatcggcgggcacgagtcgttc cgcagtgaactgcgactgctgtatctgaccgtcaccgatcccgccagcgg gaacctgatctcgctgcagatgcttccactgcgagtgtcgcggatgcgcc tacagcgtgcctcccagaccgacaccgaatggctccgcaacaccattgag cgcatcagccgccggttcgggattcgagtcgtgactcgacccgacaacct gctggaggtcgttcccgctgccaacctaacgagcaaggag Rv1264 52 gtgacagaccacgtgcgcgaggcggacgacgcgaacatcgacgatctgtt gggcgacctgggcggtaccgcgcgcgccgagcgtgcgaagcttgtcgagt ggttgctcgagcagggcatcacccccgacgagattcgggcgaccaacccg ccgttgctgctggccacccgccacctcgtcggcgacgacggcacctacgt atccgcaagggagattagcgagaactatggcgttgacctcgagctgctgc agcgggtgcagcgcgctgtcggtctggccagagtggatgatcctgacgcg gtggtgcacatgcgtgccgacggtgaggcggccgcacgcgcacagcggtt cgttgagctggggctgaatcccgaccaagtcgtgctggtcgtgcgtgtgc tcgccgagggcttgtcacacgccgccgaggccatgcgctacaccgcgctg gaggccattatgcggccgggggctaccgagttggacatcgcgaaggggtc gcaggcgctggtgagccagatcgtgccgctgctggggccgatgatccagg acatgctgttcatgcagctgcggcacatgatggagacggaggccgtcaac gccggagagcgtgcggccggcaagccgctaccgggagcgcgacaggtcac cgttgccttcgccgacctggtcggtttcacccagctaggcgaagtggtgt cggccgaagagctagggcacctcgccgggcggctggccggcctcgcgcgt gacctgaccgctccgccggtgtggttcattaagacgatcggcgacgcggt catgttggtctgtcctgatccggcgccattgctggacaccgtgctgaagc tggtcgaggtcgtcgacaccgacaacaactttccccggctgcgagccggc gtcgcctccgggatggcggttagccgggccggcgactggttcggcagccc ggtcaacgtggcaagccgggtgaccggggtggcgcgcccgggtgccgtgc tggtcgcggattcggtgcgggaggcccttggtgatgcccccgaagccgac ggatttcagtggtccttcgccggcccccgtcgcctcaggggaatccgggg tgacgtcaggctttttcgagtccggcgaggggccactcgcaccggctccg gcggcgcggcccaagacgacgatttggccggctcgtcaccg Rv1592c 53 atggtagagcccggcaatttggcaggcgcgaccggcgccgaatggatcgg ccggccaccgcacgaggaattgcagcgcaaagtgcgcccgctgctgccat ccgacgatccgttctacttcccacctgccggctaccagcatgccgtgccc ggaacggtgttgcgctcgcgcgatgtcgaactggcgtttatgggcttgat tccgcagcccgtcaccgctacccagctgctgtaccggaccacgaacatgt acggcaaccccgaggcgacggtgaccacggtgatcgtcccagcggagctt gccccgggtcagacctgccccttgctgtcgtaccagtgtgcgatcgatgc catgtcgtcgcgctgttttccgtcatatgccctgcgacgacgggccaagg ccctggggtcactgacccaaatggagctgttgatgatcagcgccgcactt gccgaaggatgggcggtatcagtacccgaccatgaagggccgaaagggct gtgggggtcgccgtatgaacccggttaccgagtcctcgacggaatccggg ctgccttgaattccgagcgtgtcgggttgtccccggcaacgccgatcggg ctgtggggctactccggcggcgggctggccagcgcgtgggccgccgaagc atgcggcgagtacgcaccggacctagacatcgtcggcgccgtgctgggat cacccgtcggtgaccttggtcacacgttccgccggctcaatggcactctt cttgccggtctgcccgcgttggtggtggccgcgctgcaacacagctaccc cggcctggcccgggtgatcaaggagcacgccaacgacgaaggacgtcagc tgctggagcaactgacggagatgacaacggtagacgcagtgatccggatg gccggcagggacatgggtgacttcctcgacgaaccccttgaggacattct gtcgacgccggaaatttcccatgtcttcggcgacaccaagctgggtagcg cggtgcccaccccgccggtattgatcgtgcaggccgtgcatgactacctc atcgacgtctctgacatcgacgcgctcgctgacagctatacagccggcgg cgccaacgtcacctaccaccgcgacctgttcagcgaacatgtgtccctgc acccgctgtcggccccaatgacgcttcgctggctcaccgaccggttcgcc ggcaagccactgaccgaccaccgcgtccggaccacgtggccgaccatctt caacccgatgacctacgccggcatggcgagactggccgtgatcgcggcca aggtgatcaccggcaggaagttgagccgccgtccgctc Rv1733c 54 atgatcgccacaacccgcgatcgtgaaggagccaccatgatcacgtttag gctgcgcttgccgtgccggacgatactgcgggtgttcagccgcaatccgc tggtgcgtgggacggatcgactcgaggcggtcgtcatgctgctggccgtc acggtctcgctgctgactatcccgttcgccgccgcggccggcaccgcagt ccaggattcccgcagccacgtctatgcccaccaggcccagacccgccatc ccgcaaccgcgaccgtgatcgatcacgagggggtgatcgacagcaacacg accgccacgtcagcgccgccgcgcacgaagatcaccgtgcctgcccgatg ggtcgtgaacggaatagaacgcagcggtgaggtcaacgcgaagccgggaa ccaaatccggtgaccgcgtcggcatttgggtcgacagtgccggtcagctg gtcgatgaaccagctccgccggcccgtgccattgcggatgcggccctggc cgccttgggactctggttgagcgtcgccgcggttgcgggcgccctgctgg cgctcactcgggcgattctgatccgcgttcgcaacgccagttggcaacac gacatcgacagcctgttctgcacgcagcgg Rv1734c 55 atgaccaacgtcggtgaccagggggttgacgcggtcttcggggtgatcta cccacctcaggtcgcgctggtcagtttcggcaagccggcacaacgagttt gcgccgtcgacggcgcgatccacgtcatgacgaccgtgctggctacgctg cccgctgaccacggctgcagcgatgaccatcgcggcgcgctgttcttcct gtcgatcaacgagctgacgcggtgcgccgcagtaacagga Rv1736c 56 gtgacggtgacaccacggaccggcagccgcatcgaggagctgcttgcacg cagcggccggttcttcatcccgggtgagatctcggcggatctgcgtaccg tgacccgccgcggcggccgcgacggcgacgtgttctatcgagaccggtgg agccacgacaaggtggtccgctccacacacggggtgaattgcaccgggtc gtgttcttggaagatctacgtcaaagacgacatcatcacctgggagacgc aggagaccgactatccgtcggtgggcccggaccggcccgagtatgagccc cgcggctgcccgcgcggcgcggcgttttcctggtacacgtattcgccgac gcgggtgcgccatccgtacgcccgcggcgtgcttgtcgagatgtatcggg aggcgaaggcacgtttgggtgatccggtggcggcctgggccgacatccag gccgacccgcggcggcgccgccgctaccagcgcgcccgcggcaagggcgg gctggtccgggtcagctgggccgaggccaccgagatgatcgccgccgccc acgtgcacaccatctccacatacggcccggaccgggttgccggcttctcc cccatcccggcgatgtccatggtgagccacgccgcggggtcgcggttcgt ggagctaatcggcggggtgatgacgtcgttctacgactggtacgccgacc tgccggtggcctccccgcaggtgttcggcgaccagaccgacgtgccggag tccggagattggtgggacgtggtgtggcaatgcgcctcggtgctgctgac ctacccgaactcacggcaactcggcaccgcagaggaattgctggcccaca tcgacggtccggccgcggatctgttggggcgcacggtctctgagctgcgc cgtgccgatccgctgaccgcggcgacccgctacgtcgacaccttcgacct gcgaggccgcgccaccctgtacctgacctactggaccgccggcgacaccc gcaaccgcggccgggagatgctggccttcgcccagacctaccgcagcacc gacgtcgcaccaccgcgcggcgagaccccggacttcctgccggtggtgct cgaattcgccgcgaccgtcgaccccgaggcggggcgacggttgctgagcg ggtaccgggtgcccatcgccgcgctgtgcaatgccctgaccgaggccgca ttgccatacgcacacacggtggccgcggtatgccggacgggtgacatgat gggcgaactcttctggaccgtcgtgccgtatgtgacgatgacgatcgtcg cggtcggctcctggtggcgctaccgctatgacaaattcggctggaccacc cgctcgtcccagctgtacgagtcgcggctgctgcggatcgccagcccgat gtttcatttcggcatcctggtggtcatcgtcggccacggtatcgggctcg tgatcccgcagtcgtggactcaggccgccggtttgagcgagggcgcatat cacgtgcaggccgtcgtgctggggtcgatcgccggcatcaccaccttggc cggcgttaccctgctgatctaccggcggcgcacccgcgggccggtgttca tggctaccaccgtcaacgacaaggtgatgtacctcgtgctggtggcggcg atcgtcgcgggactgggtgcgacggcgttgggctccggcgttgtcggcga ggcgtacaactaccgcgagacggtgtcggtgtggttccgctcggtgtggg tactgcaaccgcgcggggacctgatggccgaggctccgctgtattaccag atccatgtgctgatcgggttggcgttgttcgcgttgtggccgttcacccg gctggtacacgcgttcagcgccccgatcggctatctgttccgcccgtaca tcatctaccgcagccgcgaggagctggtgctaacgcggccgcggcggcgc gggtgg Rv1737c 57 atgagagggcaagcggccaatctcgtgctggccacctggatctcggtggt caacttctgggcgtggaacctgatcggcccgctgtcgaccagctacgcgc gtgacatgtcactgtccagcgccgaggcgtcgctgctcgtcgccaccccg atcctggtgggtgcccttggccgcatcgtcaccgggccgctcaccgaccg cttcggcgggcgcgccatgctcatcgcggtgacgctggcgtcgatcctcc cggtgctcgcggtcggggtcgcggcaaccatgggctcctacgcgttgctg gtgtttttcgggctcttcctgggcgttgccggcacgatcttcgccgtcgg catcccgttcgccaacaactggtaccagccggcgcggcgcggtttctcca ccggcgtgttcggtatgggcatggtcggcaccgcgctctcggcgttcttc accccgcggtttgtacggtggttcggcctgttcaccacccacgccatcgt cgcggccgcgctcgcgtcgaccgccgtggtggccatggtcgtgcttcgtg atgcaccctactttcggcccaacgccgacccggtgctgcccaggctcaag gccgcggcacggttgccggtgacctgggagatgtcgtttctgtacgcgat cgtgttcggcgggttcgtggcgttcagcaactacctgcccacctacatca ccacgatctacgggttctccacggtcgacgcgggcgctcgcaccgccggg ttcgccctggcggcggtgctggcccggccggtgggcgggtggctctccga ccggatcgcaccgaggcacgtggtgctggcctcgctcgccgggaccgcgc tgctggcgttcgccgcggcgttgcagccgccgccggaggtgtggtcggcg gccaccttcatcaccctggcggtctgcctcggcgtgggcaccggcggcgt gttcgcgtgggtggcccgccgcgccccggccgcatcggtcggctcggtca ccggaatcgtcgccgcggcaggcggattgggcggttacttcccgccgctg gtgatgggcgcgacctacgacccggtcgacaacgactacacggtcgggtt gctgctgctggtggcgaccgcgctggtcgcgtgtacctacaccgcgctgc acgcgcgggagccggtgagtgaggaggcgtccagg Rv1738 58 atgtgcggcgaccagtcggatcacgtgctgcagcactggaccgtcgacat atcgatcgacgaacacgaaggattgactcgggcgaaggcacggctgcgtt ggcgggaaaaggaattggtgggtgttggcctggcaaggctcaatccggcc gaccgcaacgtccccgagatcggcgatgaactctcggtcgcccgagcctt gtccgacttggggaagcgaatgttgaaggtgtcgacccacgacatcgaag ctgttacccatcagccggcgcgattgttgtat Rv1739c 59 atgattcccacgatgacatcggccggctgggcaccaggggtggtgcagtt ccgcgaataccaacggcgttggctgcgcggcgatgtcctcgccggcctga ccgtggccgcctatctgatcccgcaagcgatggcgtatgcgaccgtggcg ggcctaccgccggcagccgggctgtgggcgtcgatcgcgccgcttgccat ttacgcactgctcggatcgtcccggcagctttcaatcggcccggaatccg ccaccgccttgatgacggcggccgtgctcgctccgatggccgccggggat cttcgacgctatgccgttctggcggcaaccctcggattgctagtcggcct tatctgcctactcgctggcacggcgcgactaggtttcctcgccagcctgc

gatcgcggccggtgctcgtcggatacatggccggcatcgcgcttgtcatg atctccagccaactcggcactatcaccggcacctcggtcgaaggcaacga attcttcagcgaagtacactctttcgcgactagcgtcacgcgagttcact ggccgacttttgtgttagccatgtctgtcctagcgctgctaactatgctc acgcggtgggcgccgcgcgcccccggaccgatcatcgcggttcttgcggc cacgatgctagtggccgttatgtccttggatgccaaaggtattgcgattg tgggtcggataccttccggtctgccgacgccgggtgtgccgcccgtttcg gtggaagacttgcgggcactgatcattccggctgccgggatcgcgattgt taccttcaccgacggtgtgttgaccgcacgcgccttcgccgctcgtcgag gtcaggaagtcaatgccaacgccgagctgcgcgcggtcggggcctgcaac atcgccgccgggctgacacacggttttccggtgagttccagcagcagccg taccgccctcgccgacgtcgtcggtggccgcacccagctgtactcgctga tcgcgttggggcttgttgtcatcgtgatggttttcgcgagtgggctgctg gccatgtttccgatcgccgctctgggcgctttggtggtatatgccgcgct acgcttgatcgacttgtcagaattccggcgactggcgcggtttcggcgca gcgaactcatgctggcactagccaccacagcagccgtgttaggcctagga gtgttctatggagtcctcgccgcggttgccctgtccatcctcgaactgct tcgtcgggtcgcacatccgcatgacagcgttctcgggttcgtgccgggca ttgccggcatgcacgacatcgatgactatccgcaggccaagcgcgtgccc gggctggtggtgtatcgctatgacgcgccgttgtgcttcgccaatgccga agacttccgcaggcgagcactgaccgtggtcgatcaggatccggggcaag tcgagtggttcgtactcaacgccgaatccaatgtggaggtcgacctgact gcgctggatgcgctcgaccaactccgcaccgagctgctgcgtcggggaat agtgttcgccatggcccgggtcaaacaagacttgcgtgaatcactcaggg ccgccagtcttctcgataagattggcgaagaccatatctttatgacattg cctaccgcagtgcaggcgttccgtcggcgc Rv1813c 60 atgatcacaaacctccgacgccgaaccgcgatggcagccgccggcctagg ggctgctctcgggctgggcatcctgctggttccgacggtggacgcccatc tcgccaacggttcgatgtcggaagtcatgatgtcggaaattgccgggttg cctatccctccgattatccattacggggcgattgcctatgcccccagcgg cgcgtcgggcaaagcgtggcaccagcgcacaccggcgcgagcagagcaag tcgcactagaaaagtgcggtgacaagacttgcaaagtggttagtcgcttc accaggtgcggcgcggtcgcctacaacggctcgaaataccaaggcggaac cggactcacgcgccgcgcggcagaagacgacgccgtgaaccgactcgaag gcgggcggatcgtcaactgggcgtgcaac Rv1997 61 ttgtcggcgtcagtgtctgccacgacggctcatcatggcttgccagcaca tgaagtggtgctgctgctggagagcgatccatatcacgggctgtccgacg gcgaggccgcccaacgactagaacgcttcgggcccaacaccttggcggtg gtaacgcgcgctagcttgctggcccgcatcctgcggcagtttcatcaccc gctgatctacgttctgctcgttgccgggacgatcaccgccggtcttaagg aattcgttgacgccgcagtgatcttcggtgtggtggtgatcaatgcgatc gtgggtttcattcaagaatccaaggcagaggccgcactgcagggcctgcg ctccatggtgcacacccacgccaaggtggtgcgcgagggtcacgagcaca caatgccatccgaagagctggttcccggtgaccttgtgctgttagcggcc ggtgacaaggttcccgccgatttgcggctggtgcgacagaccggattgag cgtgaacgagtcagcacttaccggcgagtcgacgccggttcacaaggacg aggtggcgttgccggagggcacaccggtcgctgatcgtcgcaatatcgcg tattccggcacattggtaaccgcgggccatggcgccgggatcgtcgtcgc gaccggcgccgaaaccgaactcggtgagattcatcggctcgttggggccg ccgaggttgtcgccacaccgctgaccgcgaagctggcgtggttcagcaag tttctgaccatcgccatcctgggtctggcagcgctcacgttcggcgtggg tttgctgcgccggcaagatgccgtcgaaacgttcaccgctgcgatcgcgc tggcggtcggggcaattcccgaaggtctgcccaccgccgtgaccatcacc ttggccatcggcatggcccggatggccaagcgccgcgcggtcattcgacg tctacccgcggtggaaacgctgggcagcaccacggtcatctgcgccgaca agaccggaacgctgaccgagaatcagatgacggtccagtcgatctggaca ccccacggtgagatccgggcgaccggaacgggctatgcacccgacgtcct cctgtgcgacaccgacgacgcgccggttccggtgaatgccaatgcggccc ttcgctggtcgctgctggccggtgcctgcagcaacgacgccgcactggtt cgcgacggcacacgctggcagatcgtcggcgatcccaccgagggcgcgat gctcgtcgtggccgccaaggccggcttcaacccggagcggctggcgacaa ctctgccgcaagtggcagccataccgttcagttccgagcggcaatacatg gccaccctgcatcgcgacgggacggatcatgtggtgctggccaagggtgc tgtggagcgcatgctcgacctgtgcggcaccgagatgggcgccgacggcg cattgcggccgctggaccgcgccaccgtgttgcgtgccaccgaaatgttg acttcccgggggttgcgggtgctggcaaccgggatgggtgccggcgccgg cactcccgacgacttcgacgaaaacgtgataccaggttcgctggcgctga ccggcctgcaagcgatgagcgatccaccacgagcggccgcggcatcggcg gtggcggcctgccacagtgccggcattgcggtaaaaatgattaccggtga ccacgcgggcaccgccacggcgatcgcaaccgaggtggggttgctcgaca acactgaaccggcggcaggctcggtcctgacgggtgccgagctggccgcg ctgagcgcagaccagtacccggaggccgtggatacagccagcgtgtttgc cagggtctctcccgagcagaagctgcggttggtgcaagcattgcaggcca gggggcacgtcgtcgcgatgaccggcgacggcgtcaacgacgccccggcc ttgcgtcaggccaacattggcgtcgcgatgggccgcggtggcaccgaggt cgccaaggatgccgccgacatggtgttgaccgacgacgacttcgccacca tcgaagccgcggtcgaggaaggccgcggcgtattcgacaatctgaccaag ttcatcacctggacgctgcccaccaacctcggtgagggcctagtgatctt ggccgccatcgctgttggcgtcgccttgccgattctgcccacccaaattc tgtggatcaacatgaccacagcgatcgcgctcggactcatgctcgcgttc gagcccaaggaggccggaatcatgacccggccaccgcgcgaccccgacca accgctgctgaccggctggcttgtcaggcggactcttctggtttccacct tgctcgtcgccagcgcgtggtggctgtttgcatgggagctcgacaatggc gcgggcctgcatgaggcgcgcacggcggcgctgaacctgttcgtcgtcgt cgaggcgttctatctgttcagctgccggtcgctgacccgatcggcctggc ggctcggcatgttcgccaaccgctggatcatcctcggcgtcagtgcgcag gccatcgcgcaattcgcgatcacatatctacccgcgatgaatatggtgtt cgacaccgcgccaatcgatatcggggtgtgggtgcgcatattcgctgtcg cgaccgcaatcacgattgtggtggccaccgacacgctgctgccgagaata cgggcgcaaccgcca Rv1998c 62 atgagtttccacgatcttcatcaccaaggtgttccgttcgtgttgcccaa cgcctgggatgtgccgtcggccctggcctacctcgcggagggcttcacgg ctatcggcacaaccagtttcggggtctcgtccagcggcgggcacccggac gggcaccgcgccactcgcggcgccaacatcgcactggcggccgccctggc accgctgcaatgctacgtcagcgtcgacatcgaggacggatacagcgacg aacccgacgccattgctgactacgtcgcacaactgtcgacagccggaatc aatatcgaggacagtagcgccgaaaagctcatcgaccccgccctggcagc cgctaaaatcgttgcgatcaaacaacgtaaccccgaggtgttcgtcaacg cccgcgtcgacacctattggttgcgccagcacgccgataccaccagcacg atccagcgcgcacttcgctacgtcgatgccggcgccgacggcgtctttgt cccactggccaacgatcccgacgaacttgctgagctcactcgcaacattc cgtgcccggttaacacgttgcccgtgcccggcttgacgatcgccgacctt ggtgagctcggggtggcccgggtgtcaaccggttcagtgccctacagcgc ggggttgtatgcagcggcccacgcggctcgggccgtgagcgacggagagc agctgccacggtccgtaccgtacgccgaactgcaggcacgcttggttgac tacgagaaccgcacgagtacaacg Rv2003c 63 gtggtcaagcgctctcgggcaacccgactttcgccgagcatctggtccgg atgggaatcacctcagtgtcggtccattcgggcgcgattgctgctacccc ggggtcggtcgcggccgccgaacgccgattgttgctggaatcagctcgcg gtgacgcctgacacccggatgccggcatcgtcggccgccgggcgcgacgc ggcggcctacgacgcctggtatgactcacccaccgggcggccgatcctgg cgaccgaggtcgccgcgttgcggccgctcatcgaggtctttgcccagcca cgcttggaaatcggtgtcggtacaggacgtttcgccgacctgctcggcgt gcggttcggactcgatccatcccgtgatgcgctgatgttcgcacgccggc gcggcgtcctggtcgccaatgccgtcggcgaggcggtccctttcgtcagc cggcacttcggggcggtcctcatggcattcacgctctgtttcgtcaccga cccggccgccatattccgggaaacgcggcgtctgctcgccgacggcggcg gccttgttatcgggttcttgcctcgcgggacaccgtgggccgacctgtac gctctgcgcgcggcccgcggacagccaggctaccgcgacgcccgcttcta caccgcggccgaactcgaacaactgctcgcagactcgggattccgggtca tcgcccgccgctgcacgctgcaccaaccgccgggactcgcccggtacgac atcgaagccgcccatgacggtatccaagccggcgccggcttcgttgctat ctcggcggtcgaccaagcgcacgagcctaaggatgatcacccactcgagt cggaa Rv2005c 64 atgtctaaaccccgcaagcagcacggagttgtcgtcggggtagatggttc gctcgaatcggatgccgccgcctgttggggtgccaccgatgcggcgatga ggaacattccgctgaccgtggtccacgtggtgaacgccgatgtagcgacg tggccgccgatgccgtatccggagacctggggggtttggcaggaggacga gggtcgccagatcgtcgccaacgccgtcaagctcgccaaagaggcggttg gagcggatcgaaagctcagcgtaaagagcgagctcgtattttccacgccg gtacctaccatggttgaaatctccaacgaggcagagatggtggtgttggg cagctcgggccggggagcgctggcccgaggcttgctcggttcggtcagct cgagcctggtgcgacgcgccgggtgcccggtcgcggtcatccacagcgat gatgcggtgatccctgatccgcagcacgctcccgtgctggtgggaatcga cggttcgccggtttcggagcttgcgacggcggtggcatttgacgaggcgt cgcgccgcggcgtcgaactgatcgccgtgcacgcgtggagtgacgtcgaa gtggtggaacttccgggtttggacttctcggctgtacagcaggaagcgga gcttagtctcgccgaacgcttggcaggttggcaagaacgctatcccgatg tgccggtgagccgggttgtcgtttgcgatcgcccggcgcggaagctggtg caaaagtcggcgtccgcccagcttgtcgtcgttggcagtcatggccgagg tggcttgaccggcatgcttctggggtcggtcagtaacgcggtcttacacg ccgcgcgggtgccagtgatcgtggcacggcagtcg Rv2007c 65 gtgacctatgtgatcggtagtgagtgcgtggatgtgatggacaagtcctg tgtgcaggagtgtccggtcgactgtatctatgagggcgcccgaatgctct acatcaaccccgacgagtgcgtggattgtggtgcgtgcaaaccggcctgc cgcgtcgaggcgatctactgggaaggcgatctacccgacgatcaacacca gcatctgggggacaacgccgcctttttccaccaagtcctgccgggccgag tggctccgctgggttcgccgggtggtgccgcagcggtgggcccgatcgga gtcgacacgcctctggtcgcggctatcccggtggagtgccct Rv2028c 66 atgaaccaatcacacaaacccccatcgatcgtcgtcggtattgatggctc gaagccggccgtgcaagccgcactgtgggcggtcgacgaggcagccagcc gtgacatcccgctgcgtctgctgtacgcgatcgaacccgacgatcccggg tacgccgcacacggcgcggcggctcgcaaactcgccgccgccgagaacgc ggtgcgctacgcgttcacagcggtcgaggcggcggaccggccggtcaagg tcgaggtggagatcacccaggagcggccggtcacctcgttgatccgcgct tcggcggctgctgccctggtgtgcgttggcgctatcggcgtgcaccactt ccgaccggagcgggtgggatctaccgcagcggccctggcgttatcggcgc agtgcccagtggcgatcgtgcgaccccaccgggtccccatcggacgcgac gccgcatggatcgtcgtcgaggcggacgggtcgtccgatatcggtgtttt gctgggggcggtgatggccgaagcacggctgcgcgactcgccggttcggg tggtcacctgccggcaatccggagtgggcgataccggggacgacgtccgt gccagcctggaccgctggcttgcccgttggcaaccacggtatcccgatgt gcgggtgcaatcggcggcagtgcacggcgagctgctggattatctggctg ggctgggtcgatcggtacacatggtggtgctcagcgcgagcgaccaggag catgtggagcaacttgtgggagcgccgggcaacgccgtgttgcaggaggc cggctgcaccctgctggtcgtcggtcagcagtatctg Rv2029c 67 atgacggagccagcggcgtgggacgaaggcaagccgcgaatcatcacttt gaccatgaaccccgccttggacatcacgacgagcgtcgacgtggtgcgcc cgaccgagaaaatgcgttgtggcgcacctcgctacgatcccggcggcggc ggtatcaatgtcgcccgcattgtgcatgtcctcggcggttgctcgacagc actgttcccggccggcgggtcgaccgggagcctgctgatggcgctgctcg gtgatgcgggagtgccatttcgcgtcattccgatcgcggcctcgacgcgg gagagcttcacggtcaacgagtccaggaccgccaagcagtatcgtttcgt gcttccggggccgtcgctgaccgtcgcggagcaggagcaatgcctcgacg aactgcgcggtgcggcggcttcggccgcctttgtggtggccagtggcagc ctgccgccaggtgtggctgccgactactatcagcgggttgccgacatctg ccgccgatcgagcactccgctgatcctggatacatctggtggcgggttgc agcacatttcgtccggggtgtttcttctcaaggcgagcgtgcgggaactg cgcgagtgcgtcggatccgaactgctgaccgagcccgaacaactggccgc cgcacacgaactcattgaccgtgggcgcgccgaggtcgtggtggtctcgc ttggatctcagggcgcgctattggccacacgacatgcgagccatcgattt tcgtcgattccgatgaccgcggttagcggtgtcggcgccggcgacgcgat ggtggccgcgattaccgtgggcctcagccgtggctggtcgctcatcaagt ccgttcgcttgggaaacgcggcaggtgcagccatgctgctgacgccaggc accgcggcctgcaatcgcgacgatgtggagaggttcttcgagctggcggc cgaacccaccgaagtcgggcaggatcaatacgtttggcacccgatcgtta acccggaagcctcggca Rv2030c 68 gtgctgatgaccgcagcggctgatgtcacccggcgctcgccgcggcgcgt gttccgtgaccgccgcgaggccggccgggtgctggcggaattactcgccg cctatcgggaccagccggacgtgattgtgctcggcttggcccggggtggc ctcccggtcgcatgggaggttgccgcggcactgcatgccccgctagacgc cttcgtcgtgcgcaaacttggtgccccggggcatgacgagttcgccgttg gtgcactggccagcggcggccgcgtcgtggtcaatgacgacgtcgtgcgg ggcctgcggatcacaccgcagcaactgcgcgacatcgccgaacgtgaggg tcgggaactgcttcggcgcgagtccgcctaccgcggcgagcgcccgccca ccgatatcaccggcaagacggtcattgtcgtcgatgacggtttggccacc ggcgcaagcatgttcgcggcggtacaggcattgcgcgatgcgcaaccagc gcagatcgtgattgccgtgccggcggcgccggagtccacgtgccgggagt tcgccggcctcgtcgacgacgttgtgtgcgcgaccatgccgaccccgttc ctggccgtcggtgagtcgttttgggacttccggcaggtcaccgacgagga ggtccgccggctcctggccaccccgaccgctgggccgtcgctgcgccggc ccgcggcgtcaacggcggccgatgttctgcgcagagtcgcgatcgacgcc cccgggggtgttccgacgcacgaggtgttggcggagctggtcggcgatgc acgaatcgtgttgatcggcgaaagctcgcacggcacacacgagttctacc aggcccgggccgccatgacacagtggctgatcgaggagaagggctttggt gcggtagccgccgaggcggactggcccgacgcctaccgggtcaatcggta cgttcgcggcctcggcgaggacaccaacgctgacgaggcgcttagcggat tcgagcggtttcccgcctggatgtggcgcaacaccgtggtccgagatttt gtggaatggctgcgcacacgcaaccagcgctacgagtcgggcgcgctgcg gcaagccggcttctacggtctggatctttacagcctgcatcggtcgatcc aagaggtgatcagctatctcgacaaggtcgacccgcgtgcggcggcacgg gcgcgggcccggtatgcgtgcttcgaccatgcctgcgccgatgacggtca ggcgtacggattcgcggccgcattcggcgccggtccgtcgtgcgaacgtg aagccgtcgagcaactggtcgacgttcagcgcaatgccctggcgtatgcg cgccaagacgggctgcttgccgaggacgaactgttctacgcccagcaaaa cgcgcagacggtgcgcgacgcagaggtgtattaccgggccatgttcagtg gacgcgttacctcgtggaacctgcgcgaccagcacatggcgcagaccctt ggcagtttgctgacgcatttggaccgacacctcgatgcgccgccggcgcg aatagtggtgtgggctcataactcccacgtgggtgacgcacgcgctaccg aggtgtgggccgacgggcagctcaccctcggccagatagtccgtgagcga tacggtgacgagtcgcgcagcatcggattcagcacgtacacgggcaccgt caccgcggccagcgaatggggtggtatcgcccaacgcaaagcggttcggc cggcactgcacggcagtgtcgaggagctcttccaccagactgcagacagt ttcctggtgtcagcgcggctaagccgcgacgccgaagccccgctggacgt tgtccggttgggacgtgccatcggcgtcgtttatctaccggcaacggaac ggcaaagtcactacttgcacgtgcggcccgccgaccagttcgacgccatg atccacatcgatcagacccgtgccctggaacctctcgaggtgacgagccg gtggatcgccggcgagaacccggaaacctacccgaccggtctg Rv2031c 69 Atggccaccacccttcccgttcagcgccacccgcggtccctcttccccga gttttctgagctgttcgcggccttcccgtcattcgccggactccggccca ccttcgacacccggttgatgcggctggaagacgagatgaaagaggggcgc tacgaggtacgcgcggagcttcccggggtcgaccccgacaaggacgtcga cattatggtccgcgatggtcagctgaccatcaaggccgagcgcaccgagc agaaggacttcgacggtcgctcggaattcgcgtacggttccttcgttcgc acggtgtcgctgccggtaggtgctgacgaggacgacattaaggccaccta cgacaagggcattcttactgtgtcggtggcggtttcggaagggaagccaa ccgaaaagcacattcagatccggtccaccaac Rv2032 70 atgccggacaccatggtgaccaccgatgtcatcaagagcgcggtgcagtt ggcctgccgcgcaccgtcgctccacaacagccagccctggcgctggatag ccgaggaccacacggttgcgctgttcctcgacaaggatcgggtgctttac

gcgaccgaccactccggccgggaagcgctgctggggtgcggcgccgtact cgaccactttcgggtggcgatggcggccgcgggtaccaccgccaatgtgg aacggtttcccaaccccaacgatcctttgcatctggcgtcaattgacttc agcccggccgatttcgtcaccgagggccaccgtctaagggcggatgcgat cctactgcgccgtaccgaccggctgcctttcgccgagccgccggattggg acttggtggagtcgcagttgcgcacgaccgtcaccgccgacacggtgcgc atcgacgtcatcgccgacgatatgcgtcccgaactggcggcggcgtccaa actcaccgaatcgctgcggctctacgattcgtcgtatcatgccgaactct tttggtggacaggggcttttgagacttctgagggcataccgcacagttca ttggtatcggcggccgaaagtgaccgggtcaccttcggacgcgacttccc ggtcgtcgccaacaccgataggcgcccggagtttggccacgaccgctcta aggtcctggtgctctccacctacgacaacgaacgcgccagcctactgcgc tgcggcgagatgctttccgccgtattgcttgacgccaccatggctgggct tgccacctgcacgctgacccacatcaccgaactgcacgccagccgagacc tggtcgcagcgctgattgggcagcccgcaactccgcaagccttggttcgc gtcggtctggccccggagatggaagagccgccaccggcaacgcctcggcg accaatcgatgaagtgtttcacgttcgggctaaggatcaccgg Rv2428 71 atgccactgctaaccattggcgatcaattccccgcctaccagctcaccgc tctcatcggcggtgacctgtccaaggtcgacgccaagcagcccggcgact acttcaccactatcaccagtgacgaacacccaggcaagtggcgggtggtg ttcttttggccgaaagacttcacgttcgtgtgccctaccgagatcgcggc gttcagcaagctcaatgacgagttcgaggaccgcgacgcccagatcctgg gggtttcgattgacagcgaattcgcgcatttccagtggcgtgcacagcac aacgacctcaaaacgttacccttcccgatgctctccgacatcaagcgcga actcagccaagccgcaggtgtcctcaacgccgacggtgtggccgaccgcg tgacctttatcgtcgaccccaacaacgagatccagttcgtctcggccacc gccggttcggtgggacgcaacgtcgatgaggtactgcgagtgctcgacgc cctccagtccgacgagctgtgcgcatgcaactggcgcaagggcgacccga cgctagacgctggcgaactcctcaaggcttcggcc Rv2624c 72 atgtctgggagaggagagccgacgatgaaaacaatcattgttggtatcga tggttcgcacgcggcgattacggccgcattgtggggggttgacgaggcca tcagccgagcggtgccgctgcgactggtctcagtgatcaagccgacacat ccgtccccggacgactacgaccgcgaccttgcgcatgctgaaagatcgct tcgggaagcgcagtccgctgttgaggccgcgggcaagctcgtcaagatcg aaaccgacatcccccgcgggccagccggcccggtgcttgtggaggcatcg cgcgacgccgagatgatctgcgtcggctccgtgggaatcgggcgctacgc cagctcgatcttgggttcgacggcaaccgagctggccgaaaaggcgcatt gcccggtcgccgtcatgcgctcaaaagtggaccagccagcgtctgacatc aactggatcgtggtgcgcatgaccgacgcaccggataacgaggccgtgct ggaatacgctgcccgggaagcgaagttgcggcaagcgcccatactggcac tcggcgggcgaccggaggagctccgggagattccggacggcgaattcgaa cgtcgcgtgcaggattggcaccaccgtcatcccgatgtgcgcgtctaccc gatcaccactcacacgggtattgcccggttcctggccgaccacgacgagc gcgtacagctggcagtgatcggcggtggtgaggccggtcagctagcgcgg ctggtcgggccatccggacatccggtgttccgtcacgccgagtgttcggt gcttgtcgttcgccgc Rv2625c 73 atgcgtgatgcgatcccgcttgggcggatcgccgggtttgtggtgaacgt ccactggagcgtgttggtgatcctgtggttgttcacctggagtctggcga ccatgttgccgggtaccgtcggaggctacccggccgtggtctattggctt ctcggcgcaggtggcgcggtcatgttgctggcgtcgctgttggctcatga gctcgcgcacgccgtcgtcgctcgtcgcgccggggtatccgttgagagcg tgacgttgtggctgttcggcggggtgaccgcgcttggcggcgaggcaaag acgcccaaagccgctttccggatcgcgttcgcgggtccggctaccagcct ggcgctgtcggcgacattcggtgcgttggccatcacgctcgccggcgtgc ggaccccggccatcgtgatcagcgttgcttggtggttggctactgtcaac ctgctgctggggctgttcaatctgctgcctggcgcgccgttggacggtgg gcggttggtccgggcctatctgtggcgccgccacggcgatagtgtgcgcg ccgggatcggtgcggcgcgggccggacgggtggttgcgctggtcttgatc gcgttgggattggccgagtttgtggctggtggcctcgtcggtggggtctg gttagccttcattggctggtttatcttcgctgccgctcgcgaggaggaga cccggatttcgacccagcagctgtttgccggggtgcgtgtggccgatgcg atgaccgcccaaccgcatacggctcccggatggatcaatgtcgaggattt catccagcgttacgtgcttggtgaacggcactcggcatatccggttgccg atcgggacggatcgatcacgggcctggtggcattgcggcagctgcgcgat gttgcgcctagccggcgcagcactaccagcgtaggtgacattgcgctgcc gctgcacagcgtgccgaccgcccgaccacaagagccgctgaccgcgctcc tagagcggatggcaccgctcggcccgcgcagccgtgcgctggtcaccgaa gggagcgcggtggtcggcatcgtcactcccagcgatgtcgcgcggctgat tgacgtctaccggttggcccagccggaaccgacctttaccacgagtcccc aagatgcggacaggttttccgatgcgggg Rv2627c 74 atggcaagttctgcgagcgacggcacccacgaacgctcggcttttcgcct gagtccaccggtcttgagcggcgccatgggaccgttcatgcacaccggtc tgtacgtcgctcaatcgtggcgcgactatctgggtcaacagcccgataaa ctgccgatcgcacggcccactattgccttagcggcgcaagcctttcgaga cgaaatcgtcctgctgggcctcaaggcacgacgtccggtcagcaatcatc gagtgttcgagcgcatcagccaagaagtggccgctggactggagttctat gggaatcgcagatggctggagaagcctagcggattttttgcccagccccc accgctcaccgaggtcgcggtccgaaaggtcaaggaccgcagacgctcct tttatcgcatcttcttcgacagtgggtttacgccgcatccgggtgaaccg ggcagccaacggtggctctcatacactgcgaacaatcgcgagtacgccct gttactgcggcacccagagccgcgtccctggctggtttgtgtacacggca ccgagatgggcagggccccgttggatctcgcggtgttccgcgcctggaag ctgcatgacgaactcggcctgaacattgtcatgccggttcttccgatgca tggtccccgcgggcaaggtctgccgaagggcgccgtttttcccggagaag atgttctcgacgatgtgcatgggacggctcaagcggtgtgggatatccgg cggctgttgtcctggatacgatcgcaggaggaggagtcgctgatcgggtt gaacggtctctcgctgggcggctacatcgcgtcattggtcgccagcctcg aagaaggtctcgcctgcgcgattctcggtgtcccagtggctgatctgatc gagttgttgggccgccactgcggtcttcggcacaaagacccccgccgcca caccgtcaagatggccgaaccgatcggccgaatgatctcgccgctctcac ttacgccactggtgcccatgccgggccgctttatctacgcgggcattgcc gaccgactcgtgcatccacgcgaacaggtgactcgcctctgggagcactg gggcaaacccgaaatcgtgtggtatccaggcggtcacactggcttcttcc agtcgcggccggtacgacggtttgtccaggctgcgctggagcagtcgggc ctgttggacgcgccacggacacagcgcgaccgttccgcc Rv2628 75 Atgtccacgcaacgaccgaggcactccggtattcgggctgttggccccta cgcatgggccggccgatgtggtcggataggcaggtggggggtgcaccagg aggcgatgatgaatctagcgatatggcacccgcgcaaggtgcaatccgcc accatctatcaggtgaccgatcgctcgcacgacgggcgcacagcacgggt gcctggtgacgagatcactagcaccgtgtccggttggttgtcggagttgg gcacccaaagcccgttggccgatgagcttgcgcgtgcggtgcggatcggc gactggcccgctgcgtacgcaatcggtgagcacctgtccgttgagattgc cgttgcggtc Rv2629 76 atgcgatcagaacgtctccggtggctggtagccgcagaaggtccgttcgc ctcggtgtatttcgacgactcgcacgacactcttgatgccgtcgagcgcc gggaagcgacgtggcgcgatgtccggaagcatctcgaaagccgcgacgcg aagcaggagctcatcgacagcctcgaagaggcggtgcgggattctcgacc ggccgtcggccagcgtggccgcgcgctgatcgcgaccggcgagcaagtac tggtcaacgagcatctgatcggcccaccaccggctacggtgattcggctg tcggattatccgtacgtcgtgccattgatagaccttgagatgcggcgacc gacgtatgtatttgccgcggttgatcacaccggcgccgacgtcaagctgt atcagggggccaccatcagttccacgaaaatcgatggggtcggctacccg gtgcacaagccggtcaccgccggctggaacggctacggcgacttccagca caccaccgaagaagccatccgaatgaactgccgcgcggtcgccgaccatc tcacccgactggtagacgctgccgaccccgaggtggtgttcgtgtccggc gaggtgcggtcacgcacagacctgctttccacattgccgcagcgggtggc ggtccgggtgtcgcagctgcatgccggaccgcgcaaaagcgccttagacg aggaagagatctgggacctgacatccgcggagttcacccggcggcggtac gccgaaatcaccaatgtcgcacaacaatttgaggcggagatcggacgcgg atcggggctggcggcccaagggttggcggaggtgtgtgcggctctgcgtg acggcgacgtcgacacgctgatcgtcggagagctaggcgaggccaccgtg gtcaccggtaaagcgcgtactacggtcgcgcgggatgccgacatgttgtc cgaactcggcgaaccggtagatcgcgtggcaagggccgatgaggcgttgc cattcgccgcgatcgcggtaggtgccgcattggtccgtgacgacaaccgg atcgcgccactagatggggtgggcgcattgctgcgttatgccgccaccaa ccgactcggcagccatagatcc Rv2630 77 Atgctgcaccgcgacgatcacatcaatccgccgcggccccgcgggttgga tgttccttgcgcccgcctacgagcgacaaatcccctgcgcgccttggcgc gttgcgttcaggcgggcaagccgggcaccagttcagggcatcggtccgtg ccgcatacggcggacttgcgaatcgaagcctgggcaccgacccgtgacgg ctgtatccggcaggcggtgctgggtaccgtcgagagcttcctcgacctgg aatccgcgcacgcggtccatacccggctgcgccggctgaccgcggatcgc gacgacgatctactggtcgcggtgctcgaggaggtcatttatttgctgga caccgtcggtgaaacgcctgtcgatctcaggctgcgcgacgttgacgggg gtgtcgacgtcacattcgcaacgaccgatgcgagtacgctagttcaggtg ggtgccgtgccgaaggcggtgtcactcaacgaacttcggttctcgcaggg tcgccacggctggcgatgtgcggtaacgctcgatgtg Rv2659c 78 Gtgacgcaaaccggcaagcgtcagagacgcaaattcggtcgcatccgaca gttcaactccggccgctggcaagccagctacaccggccccgacggccgcg tgtacatcgcccccaaaaccttcaacgccaagatcgacgccgaagcatgg ctcaccgaccgccgccgcgaaatcgaccgacaactatggtccccggcatc gggtcaggaagaccgccccggagccccattcggtgagtacgccgaaggat ggctgaagcagcgtggaatcaaggaccgcacccgcgcccactatcgcaaa ctgctggacaaccacatcctggccaccttcgctgacaccgacctacgcga catcaccccggccgccgtgcgccgctggtacgccaccaccgccgtgggca caccgaccatgcgggcacactcctacagcttgctgcgcgcaatcatgcag accgccttggccgacgacctgatcgactccaacccctgccgcatctcagg cgcgtccaccgcccgccgcgtccacaagatcaggcccgccaccctcgacg agctggaaaccatcaccaaagccatgcccgacccctaccaggcgttcgtg ctgatggcggcatggctggccatgcgctacggcgagctgaccgaattacg ccgcaaagacatcgacctgcacggcgaggttgcgcgggtgcggcgggctg tcgttcgggtgggcgaaggcttcaaggtgacgacaccgaaaagcgatgcg ggagtgcgcgacataagtatcccgccacatctgatacccgccatcgaaga ccaccttcacaaacacgtcaaccccggccgggagtccctgctgttcccat cggtcaacgaccccaaccgtcacctagcaccctcggcgctgtaccgcatg ttctacaaggcccgaaaagccgccggccgaccagacttacgggtgcacga ccttcgacactccggcgccgtgttggctgcatccaccggcgccacactgg ccgaactgatgcagcggctaggacacagcacagccggcgccgcactccgc taccagcacgccgccaagggccgggaccgcgaaatcgccgcactgttaag caaactggccgagaaccaggagatg Rv2780 79 Atgcgcgtcggtattccgaccgagaccaaaaacaacgaattccgggtggc catcaccccggccggcgtcgcggaactaacccgtcgtggccatgaggtgc tcatccaggcaggtgccggagagggctcggctatcaccgacgcggatttc aaggcggcaggcgcgcaactggtcggcaccgccgaccaggtgtgggccga cgctgatttattgctcaaggtcaaagaaccgatagcggcggaatacggcc gcctgcgacacgggcagatcttgttcacgttcttgcatttggccgcgtca cgtgcttgcaccgatgcgttgttggattccggcaccacgtcaattgccta cgagaccgtccagaccgccgacggcgcactacccctgcttgccccgatga gcgaagtcgccggtcgactcgccgcccaggttggcgcttaccacctgatg cgaacccaagggggccgcggtgtgctgatgggcggggtgcccggcgtcga accggccgacgtcgtggtgatcggcgccggcaccgccggctacaacgcag cccgcatcgccaacggcatgggcgcgaccgttacggttctagacatcaac atcgacaaacttcggcaactcgacgccgagttctgcggccggatccacac tcgctactcatcggcctacgagctcgagggtgccgtcaaacgtgccgacc tggtgattggggccgtcctggtgccaggcgccaaggcacccaaattagtc tcgaattcacttgtcgcgcatatgaaaccaggtgcggtactggtggatat agccatcgaccagggcggctgtttcgaaggctcacgaccgaccacctacg accacccgacgttcgccgtgcacgacacgctgttttactgcgtggcgaac atgcccgcctcggtgccgaagacgtcgacctacgcgctgaccaacgcgac gatgccgtatgtgctcgagcttgccgaccatggctggcgggcggcgtgcc ggtcgaatccggcactagccaaaggtctttcgacgcacgaaggggcgtta ctgtccgaacgggtggccaccgacctgggggtgccgttcaccgagcccgc cagcgtgctggcc Rv3126c 80 Atggtcatccggtttgatcaaatagggtcattggtcctctcaatgaaatc ccttgcgtcactgtcgtttcagcggtgtctgcgcgagaattctagtttgg tcgcggcgctggaccggctcgatgctgcggtcgatgagctgagcgctttg tcgtttgatgcgttgaccactccggagcgggatcgcgcccgtcgcgaccg ggaccatcatccttggtcccgctcccgctcgcagttgtcgccacgaatgg cgcacggtgcagtgcaccaatgccagtggccgaaggcggtttgggctgtc attgacaatcca Rv3127 81 Gtgctcaagaacgcagtcttgctggcatgccgggcgccgtcggtgcacaa cagccagccctggcgttgggtggccgaaagcggctccgagcacactactg tgcacctgttcgtcaaccgccaccgaacggtgccggccaccgaccattcc ggccggcaagcgatcatcagttgcggtgccgtactcgatcaccttcgcat cgccatgacggccgcgcactggcaggcgaatatcactcgctttccccagc cgaaccaacctgaccagttggccaccgtcgaattcagtcccatcgatcac gtcacggcgggacagcgaaaccgcgcccaggcgattctgcagcgccgaac cgatcggcttccgtttgacagcccgatgtactggcacctgtttgagcccg cgctgcgcgacgccgtcgacaaagacgttgcgatgcttgatgtggtatcc gacgaccagcgaacacgactggtggtagcgtcacaactcagcgaagtcct gcggcgggacgatccgtactatcacgccgaactcgaatggtggacttcac cgttcgtgctggcccatggtgtgccgccggatacgctggcatcagacgcc gaacgcttgcgggttgacctgggccgtgacttcccggtccggagctacca gaatcgccgtgccgagctagctgatgaccgatcgaaagtccttgtgctgt cgacccctagcgacacgcgagccgacgcactgaggtgtggcgaagtgctg tcgaccatcctactcgagtgcaccatggccggcatggctacctgcacgtt gacccatctgatcgaatccagtgacagtcgtgacatcgtgcggggcctga cgaggcagcgaggcgagccgcaagccttgatccgggtagggatagccccg ccgttggcagcagttcccgcccccacaccacggcggccgctggacagcgt cttgcagattcgccagacgcccgagaaagggcgtaatgcctcagatagaa atgcccgtgaaacgggttggttcagcccgcct Rv3128c 82 gtgtggtccgcctcgggtgggcagtgcgggaagtatcttgccgcctcgat ggtgctgcagcttgatgggttggaacgtcacggtgtgttggagtttgggc gtgaccgctatggccccgaggtgcgtgaggagctgttggcgatgagtgcg gccagcatcgatcgttatctgaagaccgcgaaggccaaagaccagatatc gggtgtgtcgacgacgaaaccctcaccactgctgcgtaattcgatcaagg ttcgcagggccggcgatgaggtcgaggcggagccggggttcttcgagggc gacaccgtcgcccattgcggtccgacgctcaaaggcgagttcgcccacac cctgaacttgaccgacgtgcacatcggatgggtgttcacccgcaccgtcc gcaacaacgcccgtacccacatcctcgccgggctcaaagcttctgtcacc gagatcccgcatgggataacgggtttagatttcgacaacggcaccgtgtt tctcaacaagccggtcatcagctgggccggcgacaacggtatctacttca cccgctttcgcccgtacaagaaaaaccactaggccaccatcgagtccaag aacaaccacctggtccgcaagtacgcgttctactaccgctatgacaccgc cgaggaacgcgccgtgctcaaccggatgtggaagctggtcaacgaccgcc tcaactacctcaccccgaccatcaaaccgatcgggtatgccagcagcgcc gacggccgccgccgacgcctctacgatgccccacagacgccgctggaccg gccactggccgcaagggtgctctccgcggcccagcaggccgacctgatca cctaccgagacagcctcaaccccgcccagatcggccgcaaaatcgccgac ctgcagaaccgactcctcatcttggccaaggagaaaaccgagcagctcta cctcgctaacatccccaccgccctacccgacatccacaaaggcatcctga tcaaggcgggc Rv3129 83 Gtggtgcaaggccgcaccgtgctgtttcgtaccgcggagggcgccaaatt attttcagccgtcgcgaagtgcgcggtggctttcgaggcggacgaccaca acgttgccgagggctggagcgtgatcgtcaaggttcgcgcccaggtgctg acgaccgacgcgggggtccgcgaagccgaacgcgcccagttactaccgtg gaccgcgacgctgaaacgtcactgtgtgcgggtgatcccgtgggagatca ccggccgccacttcaggttcggtccggaaccggaccgcagccagaccttt

gcctgcgaggcctcgtcacacaaccagcga Rv3130c 84 atgaatcacctaacgacacttgacgccgggtttctcaaggcagaagacgt ggatcggcacgtgagtctggcaatcggcgctctggcggtcatcgaggggc cggctcccgatcaggaagccttcttatcgtcgctcgctcaacgcctacgt ccctgtacccggttcgggcagcggttacgcctgcgcccgttcgacctcgg tgcacccaaatgggtggacgatcccgacttcgatcttggccgtcatgtgt ggcgcatcgccttgccgcggcctggcaacgaagaccagttattcgagctg atcgccgatctgatggcgcgtcgtttggaccggggtcgaccgctgtggga ggtctgggtcatcgaaggcctggcggacagcaagtgggcgatcctgacca aactgcaccactgcatggccgacggaatcgcggcgactcacctgctagct gggctctccgatgaaagtatgagcgacagcttcgcgagcaacatccacac gaccatgcagtcgcaatccgcatctgtgcggcggggtggattccgtgtca atccaagcgaggcgttgaccgcgtcgaccgccgtgatggcaggcatcgtt cgcgcggccaagggtgccagtgagatcgcggccggcgtgctaagtcccgc cgcgtcgtcgttgaacgggccgatcagtgatttgcgtcgctacagcgcag caaaggtccctctcgccgacgtcgaacaggtgtgccggaaattcgacgtc accatcaatgatgttgcgcttgccgcgattacggaaagctaccgcaacgt cctcatccagcggggtgagcggcctaggtttgattcgctgcgtacgctag tgccggtctcgacgcgttccaacagcgctttgagcaagaccgataaccgt gtttcgttaatgctgcccaacctgccggtggatcaagagaacccgctgca gcggctgcggatcgtgcactcgcggctgactcgggccaaggcggggggac agagacaattcggaaatactttgatggcgattgccaaccgccttccgttc cccatgaccgcatgggcggtcgggctgttgatgcggctgccgcagcgtgg tgttgtcaccgtggcgacaaatgtgccgggtccacgacggccgctgcaga ttatgggcagacgggtgcttgacctatacccggtttcgccgatcgcgatg caactgcgcaccagtgtcgcgatgctcagctacgccgacgacctgtactt cgggatcctggccgactacgacgtggtagcagatgccggccagctggcgc gaggaattgaagacgccgtcgcacggctggtggcgatcagtaagcggcgc aaggtgactcgcaggcgcggagcgctatcgctggttgtg Rv3131 85 atgaacacccatttcccggacgccgaaaccgtgcgaacggttctcaccct ggccgtccgggccccctccatccacaacacgcagccgtggcggtggcggg tatgcccgacgagtctggagctgttctctagacccgatatgcagctgcgt agcaccgatccggacgggcgtgagttgatcctcagctgtggtgtggcatt gcaccactgcgtcgtcgctttggcgtcgctgggctggcaggccaaggtaa accgtttccccgatcccaaggaccgctgccatctggccaccatcggggta caaccgcttgttcccgatcaggccgatgtcgccttggcggcggccatacc gcggcgacgcaccgatcggcgcgcctacagttgctggccggtgccaggag gtgacatcgcgttgatggccgcaagagcagcccgtggcggggtcatgctg cggcaggtcagtgccctagaccgaatgaaagccattgtggcgcaggctgt cttggaccacgtgaccgacgaggaatatctgcgcgagctcaccatttgga gtgggcgctacggttcagtggccggggttcccgcccgcaacgagccgcca tcagaccccagtgccccgatccccggtcgcctgttcgccgggcccggtct gtctcagccgtccgacgtcttacccgctgacgacggcgccgcgatcctgg cactaggcaccgagacagacgaccggttggcccggctgcgcgccggcgag gccgccagcatcgtcttgttgaccgcgacggcaatggggctggcgtgctg cccgatcaccgaaccgctggagatcgccaagacccgcgacgcggtccgtg ccgaggtgttcggcgccggcggctacccccagatgctgctgcgagtgggt tgggcaccgatcaatgccgacccgttgccaccgacgccacggcgcgaact gtcccaggtcgttgagtggccggaagagctactgcgacaacggtgc Rv3132c 86 atgacaacagggggcctcgtcgacgaaaacgacggcgccgcaatgcgtcc actgcgtcacacgctctcccaactacgcctgcacgagctgctggtcgagg tgcaggaccgggtcgagcagatcgtcgagggccgggaccgcctcgatggt ctggtggaggccatgctcgtggtcacagcgggcctggacctggaggcaac cctacgcgctatcgtgcattcagcgaccagccttgtcgatgcgcgctatg gcgctatggaggtgcacgaccggcagcatcgggtattgcactttgtctat gaaggcatcgacgaggagaccgttcggcggatcggccacctaccgaaagg cctaggcgtcatcgggctgctcatcgaagatcccaaaccgttacggctgg acgatgtttctgcgcacccggcctcgattggttttccgccgtatcatccg ccgatgcgtaccttcctcggggtaccggttcgggtgcgcgatgaatcgtt cggcactctgtacctgactgacaagaccaacgggcaaccgttcagcgacg acgacgaggttctggtccaggcgctggcggccgccgcgggtatcgcagtc gcgaatgcccggctctaccagcaggctaaggcgcgtcagtcgtggatcga ggccacccgtgacatcgccaccgagttgttgtccggcaccgaacccgcga cggtgttccggcttgtcgccgcggaggcgctcaagctgacggcggctgac gctgccctggtagccgttcccgtcgacgaggacatgcctgccgctgacgt gggggagctgctggtgattgaaacagtcggcagcgctgtggcttccattg ttgggcgaacgattccggtggcgggcgcggtgctgcgggaggtcttcgtc aacggcattccgcgacgggtcgaccgggtcgatttggaaggcctggacga actggccgacgcaggtccggcgctgctgttgccgctgcgggccagaggta ccgtagcgggtgtcgttgttgtgctgagtcaaggcggtccaggggctttc accgacgaacaactcgagatgatggccgcgttcgccgaccaggccgcgct ggcttggcaattggccacttcgcaacgtcggatgcgcgaactcgacgtac tgaccgaccgggatcgtatcgcccgtgacctccatgaccatgtcatccat cggctcttcgcgattggcctggctttgcagggtgctgtcccgcacgaacg taatcctgaagtgcagcaacgactctcggacgtggtagacgatctgcaag acgttatacaggaaatccggaccaccatttatgacctgcacggagcatcg cagggtatcactcggctccggcagcgaatcgatgcggccgtagcccaatt tgccgactcggggttgcgcaccagcgttcaattcgtgggtccattgtcgg tggtcgacagcgcgctcgccgatcaggccgaggcggtggttcgggaagcg gtcagcaacgcggttcgccatgcgaaggccagcacgttgaccgtccgggt caaagtcgacgacgacttgtgcatcgaggtgaccgacaacggccgcgggc tgcccgacgagttcaccggaagcggcttaacgaacctgcggcagcgggca gagcaggccggcggcgaattcaccctcgcgagcgtaccgggcgcgagcgg aacagtgctgcgatggtcagcaccgttgtcgcag Rv3134c 87 atgagcgatcctcggccagctcgggcagtggtcgttggtatcgacgggtc aagggcggcaacgcatgcggcgttgtgggcggtcgatgaggcggtgaacc gagacattccgctgcgactggtgtacgtcatcgatccgtcccaactgtcc gccgccggcgagggcggtgggcaatcagcggcccgagcggcgctgcacga cgcctctcggaaggtcgaggccaccgggcaaccggtcaagatcgaaacgg aggttctgtgcggcaggccgctcaccaagctgatgcaggagtccaggtcc gcggcgatgctgtgcgtcggttcggtggggcttgatcatgtccgcggtcg ccggggttcggtcgcggcgaccctggctgggtcggccttatgccccgtgg cggtgattcacccgtcgccggccgagccagcgacaacctcccaggtcagc gcggttgtcgcggaggtggacaatggtgtggtgctgcggcacgcattcga ggaggccaggctgcgcggagttccgctgcgggccgtggctgtccacgctg ctgaaacacccgatgacgtcgaacagggcagccggttggcgcatgtacac ctgagccgtcggctcgcccactggacccggctctaccccgaggtgcgggt ggatcgggccatcgccggcggcagtgcgtgccgtcatctggccgccaacg caaagccgggtcagctgttcgtcgcggactcacactccgcgcacgaattg tgcggtgcataccagcccggatgcgccgtacttacggtacgcagtgccaa cttg Rv3841 88 atgacagaatacgaagggcctaagacaaaattccacgcgttaatgcagga acagattcataacgaattcacagcggcacaacaatatgtcgcgatcgcgg tttatttcgacagcgaagacctgccgcagttggcgaagcatttttacagc caagcggtcgaggaacgaaaccatgcaatgatgctcgtgcaacacctgct cgaccgcgaccttcgtgtcgaaattcccggcgtagacacggtgcgaaacc agttcgacagaccccgcgaggcactggcgctggcgctcgatcaggaacgc acagtcaccgaccaggtcggtcggctgacagcggtggcccgcgacgaggg cgatttcctcggcgagcagttcatgcagtggttcttgcaggaacagatcg aagaggtggccttgatggcaaccctggtgcgggttgccgatcgggccggg gccaacctgttcgagctagagaacttcgtcgcacgtgaagtggatgtggc gccggccgcatcaggcgccccgcacgctgccgggggccgcctc Rv3842c 89 atgacatgggccgacgaggtgctcgccggacatccctttgtggttgctca ccgtggtgcgtcggcggctcggccggagcatacccttgccgcctacgacc tggcgctcaaagagggcgccgacggcgtggaatgtgatgtgcggttgacc cgggacgggcatctggtctgtgtgcatgaccgccgcctggaccgaacctc gacgggagccggcttggtcagcacgatgacgctggcccagctacgcgagc tggagtacggcgcgtggcacgacagctggcgccccgacggttcgcacggc gacaccagtctgctgaccctggacgcgcttgtttcgctggttttggactg gcaccggccggtgaagatcttcgtcgagaccaagcatcccgtccgatacg gctcgctggtggaaaacaagctgctggcgctgctacaccggttcggtatt gccgcacccgcctccgcagatcgatcccgtgcggtggtgatgtcgttttc ggccgccgcggtctggcggatccggcgggctgcaccgctgctgccgacgg tgttgctcggcaagaccccccgatacctgaccagcagtgcggccacggcg gtcggggcaaccgccgtgggaccctcactgcctgcgttaaaggaatatcc gcaactcgttgaccgctcggcagctcagggccgggcggtgtactgctgga acgtcgatgagtacgaggacatcgacttttgccgggaggtcggggtggcc tggattggtactcaccaccccggccgcaccaaggcctggctggaagacgg gcgggcgaacgggaccactcgc Rv3908 90 gtgtccgacggcgaacaagccaaatcacgtcgacgccgggggcggcgccg cgggcggcgcgctgcggctacagccgagaatcacatggacgcccaaccgg ccggcgacgccaccccgaccccggcaacggcgaagcggtcccggtcccgc tcacctcgtcgcgggtcgactcggatgcgcaccgtgcacgaaacatcggc tggagggttggtcattgacggtatcgacggtccacgagacgcgcaggtcg cggctctgatcggccgcgtcgaccggcgcggccggctgctgtggtcgcta cccaaggggcacatcgagttgggcgagaccgccgagcagaccgccatccg cgaggtcgccgaggagaccggcatccgcggcagtgtgctcgccgcgctgg ggcgcatcgactactggttcgtcaccgacggccggcgggtgcacaagacc gtccaccattatttgatgcggtttttaggcggagagctgtccgacgaaga cctcgaggtagccgaggtagcctgggtgccgatccgggaactgccgtctc gactggcctacgccgacgaacgtcgactagccgaggtggccgacgaactg atcgacaagctgcagagcgacggccccgccgcgcttccgccgctaccacc cagctcgcctcgtcgacggccgcaaacgcattcacgcgctcgtcatgccg atgactcagcaccgggtcagcacaacggtcccgggccggggccg

[0030] Preferably the immunogenic portions are selected from the group consisting of the sequences presented in Table 1 and the nucleic acid sequences are selected from the sequences presented in Table 2.

[0031] In another embodiment, the vaccine is a multiphase vaccine, where the polypeptides or fragments hereof are fused to other antigens with efficacy as prophylactic vaccines, where the fusion partner is selected from e.g. the group consisting of ESAT-6, TB10.4, CFP10, RD1-ORF5, RD1-ORF2, Rv1036, MPB64, MPT64, Ag85A, Ag85B (MPT59), MPB59, Ag85C, 19 kDa lipoprotein, MPT32.

[0032] The invention further discloses a therapeutic vaccine against tuberculosis comprising one or more polypeptides or fragments hereof, which polypeptides are expressed during the latent stage of the mycobacteria infection, which stage is characterized by low-oxygen tension in the microenvironment of the mycobacteria, or nucleic acids encoding these polypeptides.

[0033] Preferably, the therapeutic and multiphase vaccine comprises an additional delivery system selected from among, live recombinant vaccines, that is gene-modified organisms such as bacteria or viruses expressing mycobacteria genes, or immunogenic delivery systems such as, DNA vaccines, that is plasmids expressing genes or gene fragments for the proteins described above, or protein vaccines, that is the proteins themselves or synthetic peptides derived from the proteins themselves delivered in a delivery system such as an adjuvant.

[0034] The invention further discloses a therapeutic vaccine in which the amino acid sequence is lipidated so as to allow a self-adjuvanting effect of the polypeptide.

[0035] The invention also discloses a method for treating an animal, including a human being, with tuberculosis caused by virulent mycobacteria, e.g., by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, comprising administering to the animal the above-mentioned vaccine.

[0036] The invention also discloses a method for immunizing an animal, including a human being, against tuberculosis caused by virulent mycobacteria, e.g., by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, comprising administering to the animal the above mentioned vaccine.

[0037] In a still further embodiment, the invention discloses an immunogenic composition comprising a polypeptide as defined above, preferably in the form of a vaccine or in the form of a diagnostic reagent. The diagnostic reagent can be in the form of a skin test reagent (administered by the transcutaneous, subcutaneous or intradermal routes), a serological reagent or a reagent for stimulating a cell-mediated reaction.

[0038] In another embodiment, the invention discloses a nucleic acid fragment in isolated form which

[0039] (a) comprises a nucleic acid sequence which encodes a polypeptide as defined above, or comprises a nucleic acid sequence complementary thereto; or

[0040] (b) has a length of at least 10 nucleotides and hybridizes readily under stringent hybridization conditions with a nucleotide sequence selected from the nucleotide sequences presented in Table 2 or a sequence complementary thereto, or with a nucleotide sequence selected from a sequence in (a)

[0041] The nucleic acid fragment is preferably a DNA fragment. The fragment can be used as a pharmaceutical.

[0042] In another embodiment, the invention discloses a vaccine comprising a nucleic acid fragment according to the invention, optionally inserted in a vector, the vaccine effecting in vivo expression of antigen by a human being or other mammal or animal, to whom the vaccine has been administered, the amount of expressed antigen being effective to confer substantially increased resistance to tuberculosis caused by virulent mycobacteria, e.g. by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, in an animal, including a human being.

[0043] In a further embodiment, the invention discloses the use of a nucleic acid fragment according to the invention for the preparation of a composition for the diagnosis of tuberculosis caused by virulent mycobacteria, e.g., by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, and the use of a nucleic acid fragment according to the invention for the preparation of a pharmaceutical composition for the vaccination against tuberculosis caused by virulent mycobacteria, e.g., by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis.

[0044] In a still further embodiment, the invention discloses a vaccine for immunizing an human being or other mammal or animal, against tuberculosis caused by virulent mycobacteria, e.g. by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, comprising as the effective component a non-pathogenic microorganism, wherein at least one copy of a DNA fragment comprising a DNA sequence encoding a polypeptide as defined above has been incorporated into the microorganism (e.g., placed on a plasmid or in the genome) in a manner allowing the microorganism to express and optionally secrete the polypeptide.

[0045] In another embodiment, the invention discloses a replicable expression vector, which comprises a nucleic acid fragment according to the invention, and a transformed cell harboring at least one such vector.

[0046] In another embodiment, the invention discloses a method for producing a polypeptide as defined above, comprising

[0047] (a) inserting a nucleic acid fragment according to the invention into a vector that is able to replicate in a host cell, introducing the resulting recombinant vector into the host cell, culturing the host cell in a culture medium under conditions sufficient to effect expression of the polypeptide, and recovering the polypeptide from the host cell or culture medium;

[0048] (b) isolating the polypeptide from a whole mycobacterium, e.g. Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, from culture filtrate or from lysates or fractions thereof; or

[0049] (c) synthesizing the polypeptide e.g. by solid or liquid phase peptide synthesis.

[0050] The invention also discloses a method of diagnosing tuberculosis caused by virulent mycobacteria, e.g. by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, in an animal, including a human being, comprising intradermally injecting, in the animal, a polypeptide as defined above or an immunogenic composition as defined above, a positive skin response at the location of injection being indicative of the animal having tuberculosis, and a negative skin response at the location of injection being indicative of the animal not having tuberculosis.

[0051] In another embodiment, the invention discloses a method for immunizing an animal, including a human being, against tuberculosis caused by virulent mycobacteria, e.g. by Mycobacterium tuberculosis, Mycobacterium africanum or Mycobacterium bovis, comprising administering to the animal the polypeptide as defined above, the immunogenic composition according to the invention, or the vaccine according to the invention.

[0052] Another embodiment of the invention discloses a monoclonal or polyclonal antibody, which is specifically reacting with a polypeptide as defined above in an immuno assay, or a specific binding fragment of said antibody. Preferably, said antibody is for use as a diagnostic reagent, e.g. for detection of mycobacteria antigens in sputum, urine or other body fluids of an infected animal, including a human being.

[0053] In a further embodiment the invention discloses a pharmaceutical composition that comprises an immunologically responsive amount of at least one member selected from the group consisting of:

[0054] (a) a polypeptide selected from the sequences presented in Table 1, or an immunogenic portion thereof;

[0055] (b) an amino acid sequence which has a sequence identity of at least 70% to any one of said polypeptides in (a) and is immunogenic;

[0056] (c) a fusion polypeptide comprising at least one polypeptide or amino acid sequence according to (a) or (b) and at least one fusion partner;

[0057] (d) a nucleic acid sequence which encodes a polypeptide or amino acid sequence according to (a), (b) or (c);

[0058] (e) a nucleic acid sequence which is complementary to a sequence according to (d);

[0059] (f) a nucleic acid sequence which has a length of at least 10 nucleotides and which hybridizes under stringent conditions with a nucleic acid sequence according to (d) or (e); and

[0060] (g) a non-pathogenic micro-organism which has incorporated (e.g. placed on a plasmid or in the genome) therein a nucleic acid sequence according to (d), (e) or (f) in a manner to permit expression of a polypeptide encoded thereby.

[0061] In a still further embodiment the invention discloses a method for stimulating an immunogenic response in an animal which comprises administering to said animal an immunologically stimulating amount of at least one member selected from the group consisting of:

[0062] (a) a polypeptide selected from the sequences presented in Table 1, or an immunogenic portion thereof;

[0063] (b) an amino acid sequence which has a sequence identity of at least 70% to any one of said polypeptides in (a) and is immunogenic;

[0064] (c) a fusion polypeptide comprising at least one polypeptide or amino acid sequence according to (a) or (b) and at least one fusion partner;

[0065] (d) a nucleic acid sequence which encodes a polypeptide or amino acid sequence according to (a), (b) or (c);

[0066] (e) a nucleic acid sequence that is complementary to a sequence according to (d);

[0067] (f) a nucleic acid sequence which has a length of at least 10 nucleotides and which hybridizes under stringent conditions with a nucleic acid sequence according to (d) or (e); and

[0068] (g) a non-pathogenic micro-organism which has incorporated therein (e.g. placed on a plasmid or in the genome) a nucleic acid sequence according to (d), (e) or (f) in a manner to permit expression of a polypeptide encoded thereby.

[0069] The vaccine, immunogenic composition and pharmaceutical composition according to the invention can be used therapeutically in a subject infected with a virulent mycobacterium combined with a prophylactic composition in a subject to prevent further infection with a virulent mycobacterium.

[0070] The invention also discloses a method for diagnosing previous or ongoing infection with a virulent mycobacterium, said method comprising

[0071] (a) contacting a sample, e.g. a blood sample, with a composition comprising an antibody according to the invention, a nucleic acid fragment according to the invention and/or a polypeptide as defined above, or

[0072] (b) contacting a sample, e.g. a blood sample comprising mononuclear cells (e.g. T-lymphocytes), with a composition comprising one or more polypeptides as defined above in order to detect a positive reaction, e.g. proliferation of the cells or release of cytokines such as IFN-γ.

[0073] Finally, the invention discloses a method of diagnosing Mycobacterium tuberculosis infection in a subject comprising:

[0074] (a) contacting a polypeptide as defined above with a bodily fluid of the subject;

[0075] (b) detecting binding of a antibody to said polypeptide, said binding being an indication that said subject is infected by Mycobacterium tuberculosis or is susceptible to Mycobacterium tuberculosis infection.

DEFINITIONS

Polypeptides

[0076] The word "polypeptide" in the present invention should have its usual meaning. That is an amino acid chain of any length, including a full-length protein, oligopeptides, short peptides and fragments thereof, wherein the amino acid residues are linked by covalent peptide bonds.

[0077] The polypeptide may be chemically modified by being glycosylated, by being lipidated (e.g. by chemical lipidation with palmitoyloxy succinimide as described by Mowat et al. 1991 or with dodecanoyl chloride as described by Lustig et al. 1976), by comprising prosthetic groups, or by containing additional amino acids such as e.g. a his-tag or a signal peptide.

[0078] Each polypeptide may thus be characterized by specific amino acids and be encoded by specific nucleic acid sequences. It will be understood that such sequences include analogues and variants produced by recombinant or synthetic methods wherein such polypeptide sequences have been modified by substitution, insertion, addition or deletion of one or more amino acid residues in the recombinant polypeptide and still be immunogenic in any of the biological assays described herein. Substitutions are preferably "conservative". These are defined according to the following table. Amino acids in the same block in the second column and preferably in the same line in the third column may be substituted for each other. The amino acids in the third column are indicated in one-letter code.

TABLE-US-00003 ALIPHATIC Non-polar GAP ILV Polar-uncharged CSTM NQ Polar-charged DE KR AROMATIC HFWY

[0079] A preferred polypeptide within the present invention is an immunogenic antigen from M. tuberculosis produced when the organism is subjected to the stresses associated with latent infection. Such antigen can for example also be derived from the M. tuberculosis cell and/or M. tuberculosis culture filtrate. Thus, a polypeptide comprising an immunogenic portion of one of the above antigens may consist entirely of the immunogenic portion, or may contain additional sequences. The additional sequences may be derived from the native M. tuberculosis antigen or be heterologous and such sequences may, but need not, be immunogenic.

[0080] Each polypeptide is encoded by a specific nucleic acid sequence. It will be understood that such sequences include analogues and variants hereof wherein such nucleic acid sequences have been modified by substitution, insertion, addition or deletion of one or more nucleic acids. Substitutions are preferably silent substitutions in the codon usage that will not lead to any change in the amino acid sequence, but may be introduced to enhance the expression of the protein.

[0081] In the present context the term "substantially pure polypeptide fragment" means a polypeptide preparation which contains at most 5% by weight of other polypeptide material with which it is natively associated (lower percentages of other polypeptide material are preferred, e.g. at most 4%, at most 3%, at most 2%, at most 1%, and at most 1/2%). It is preferred that the substantially pure polypeptide is at least 96% pure, i.e. that the polypeptide constitutes at least 96% by weight of total polypeptide material present in the preparation, and higher percentages are preferred, such as at least 97%, at least 98%, at least 99%, at least 99.25%, at least 99.5%, and at least 99.75%. It is especially preferred that the polypeptide fragment is in "essentially pure form", i.e. that the polypeptide fragment is essentially free of any other antigen with which it is natively associated, i.e. free of any other antigen from bacteria belonging to the tuberculosis complex or a virulent mycobacterium. This can be accomplished by preparing the polypeptide fragment by means of recombinant methods in a non-mycobacterial host cell as will be described in detail below, or by synthesizing the polypeptide fragment by the well-known methods of solid or liquid phase peptide synthesis, e.g. by the method described by Merrifield, 1963, or variations thereof.

[0082] By the term "virulent mycobacterium" is understood a bacterium capable of causing the tuberculosis disease in an animal or in a human being. Examples of virulent mycobacteria include but are not limited to M. tuberculosis, M. africanum, and M. bovis. Examples of relevant animals are cattle, possums, badgers and kangaroos.

[0083] By "a TB patient" is understood an individual with culture or microscopically proven infection with virulent mycobacteria, and/or an individual clinically diagnosed with TB and who is responsive to anti-TB chemotherapy. Culture, microscopy and clinical diagnosis of TB are well known by any person skilled in the art.

[0084] By the term "PPD-positive individual" is understood an individual with a positive Mantoux test or an individual where PPD induces a positive in vitro recall response determined by release of IFN-γ.

[0085] By "a latently infected individual" is understood an individual, who has been infected by a virulent mycobacterium, e.g. M. tuberculosis, but shows no sign of active tuberculosis. It is likely that individuals who have been vaccinated, e.g. by BCG, or treated for TB may still retain the mycobacteria within their bodies, although this is currently impossible to prove since such individuals would be expected to be positive if tested for PPD reactivity. Nonetheless, in its most accurate sense, "latently-infected" may be used to describe any individual who has M. tuberculosis residing in their tissues but who is not clinically ill.

[0086] By the term "delayed type hypersensitivity reaction" (DTH) is understood a T-cell mediated inflammatory response elicited after the injection of a polypeptide into, or application to, the skin, said inflammatory response appearing 72-96 hours after the polypeptide injection or application.

[0087] By the term "IFN-γ" is understood interferon-gamma. The measurement of IFN-γ is used as an indication of an immunological response.

[0088] By the terms "nucleic acid fragment" and "nucleic acid sequence" are understood any nucleic acid molecule including DNA, RNA, LNA (locked nucleic acids), PNA, RNA, dsRNA and RNA-DNA-hybrids. Also included are nucleic acid molecules comprising non-naturally occurring nucleosides. The term includes nucleic acid molecules of any length e.g. from 10 to 10000 nucleotides, depending on the use. When the nucleic acid molecule is for use as a pharmaceutical, e.g. in DNA therapy, or for use in a method for producing a polypeptide according to the invention, a molecule encoding at least one epitope is preferably used, having a length from about 18 to about 1000 nucleotides, the molecule being optionally inserted into a vector. When the nucleic acid molecule is used as a probe, as a primer or in antisense therapy, a molecule having a length of 10-100 is preferably used. According to the invention, other molecule lengths can be used, for instance a molecule having at least 12, 15, 21, 24, 27, 30, 33, 36, 39, 42, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500 or 1000 nucleotides (or nucleotide derivatives), or a molecule having at most 10000, 5000, 4000, 3000, 2000, 1000, 700, 500, 400, 300, 200, 100, 50, 40, 30 or 20 nucleotides (or nucleotide derivatives).

[0089] The term "stringent" when used in conjunction with hybridization conditions is as defined in the art, i.e. the hybridization is performed at a temperature not more than 15-20° C. under the melting point T_m, cf. Sambrook et al, 1989, pages 11.45-11.49. Preferably, the conditions are "highly stringent", i.e. 5-10° C. under the melting point T_m.

[0090] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations thereof such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.

Sequence Identity

[0091] The term "sequence identity" indicates a quantitative measure of the degree of homology between two amino acid sequences of equal length or between two nucleotide sequences of equal length. The two sequences to be compared must be aligned to best possible fit allowing the insertion of gaps or alternatively, truncation at the ends of the protein sequences. The sequence identity can be calculated as

( N ref - N dif ) 100 N ref , ##EQU00001##

wherein N_dif is the total number of non-identical residues in the two sequences when aligned and wherein N_ref is the number of residues in one of the sequences. Hence, the DNA sequence AGTCAGTC [SEQ ID NO: 184] will have a sequence identity of 75% with the sequence AATCAATC, SEQ ID NO: 185 (N_dif=2 and N_ref=8). A gap is counted as non-identity of the specific residue(s), i.e. the DNA sequence AGTGTC [SEQ ID NO: 186] will have a sequence identity of 75% with the DNA sequence AGTCAGTC, SEQ ID NO: 187, (N_dif=2 and N_ref=8). Sequence identity can alternatively be calculated by the BLAST program e.g. the BLASTP program (Pearson, 1988, or online through the NIH website). In one aspect of the invention, alignment is performed with the sequence alignment method ClustalW with default parameters as described by Thompson J., et al. 1994 and as available through online sources.

[0092] A preferred minimum percentage of sequence identity is at least 80%, such as at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, and at least 99.5%.

Immunogenic Portion

[0093] In a preferred embodiment of the invention, the polypeptide comprises an immunogenic portion of the polypeptide, such as an epitope for a B-cell or T-cell. The immunogenic portion of a polypeptide is a part of the polypeptide, which elicits an immune response in an animal or a human being, and/or in a biological sample determined by any of the biological assays described herein. The immunogenic portion of a polypeptide may be a T-cell epitope or a B-cell epitope. Immunogenic portions can be related to one or a few relatively small parts of the polypeptide, they can be scattered throughout the polypeptide sequence or be situated in specific parts of the polypeptide. For a few polypeptides, epitopes have even been demonstrated to be scattered throughout the polypeptide covering the full sequence (Ravn et al 1999). In order to identify relevant T-cell epitopes which are recognized during an immune response, it is possible to use overlapping oligopeptides for the detection of MHC class II epitopes, preferably synthetic, having a length of e.g. 20 amino acid residues derived from the polypeptide. These peptides can be tested in biological assays (e.g. the IFN-γ assay as described herein) and some of these will give a positive response (and thereby be immunogenic) as evidence for the presence of a T cell epitope in the peptide. For the detection of MHC class I epitopes it is possible to predict peptides that will bind (Stryhn et al. 1996) and hereafter produce these peptides synthetic and test them in relevant biological assays, e.g. the IFN-γ assay as described herein. The peptides preferably having a length of, e.g., 8 to 11 amino acid residues derived from the polypeptide. B-cell epitopes can be determined by analyzing the B cell recognition to overlapping peptides covering the polypeptide of interest as, e.g., described in Harboe et al 1998.

[0094] Although the minimum length of a T-cell epitope has been shown to be at least 6 amino acids, it is normal that such epitopes are constituted of longer stretches of amino acids. Hence, it is preferred that the polypeptide fragment of the invention has a length of at least 7 amino acid residues, such as at least 8, at least 9, at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 22, at least 24, and at least 30 amino acid residues. Hence, in important embodiments of the inventive method, it is preferred that the polypeptide fragment has a length of at most 50 amino acid residues, such as at most 40, 35, 30, 25, and 20 amino acid residues. It is expected that the peptides having a length of from 10 to 20 amino acid residues will prove to be most efficient as MHC class II epitopes and therefore especially preferred lengths of the polypeptide fragment used in the inventive method are 18, such as 15, 14, 13, 12 and even 11 amino acid residues. It is expected that the peptides having a length of from 7 to 12 amino acid residues will prove to be most efficient as MHC class I epitopes and therefore especially preferred lengths of the polypeptide fragment used in the inventive method are 11, such as 10, 9, 8 and even 7 amino acid residues.

[0095] Immunogenic portions of polypeptides may be recognized by a broad part (high frequency) or by a minor part (low frequency) of the genetically heterogeneous human population. In addition some immunogenic portions induce high immunological responses (dominant), whereas others induce lower, but still significant, responses (subdominant). High frequency or low frequency can be related to the immunogenic portion binding to widely distributed MHC molecules (HLA type) or even by multiple MHC molecules (Sinigaglia, 1988, Kilgus, 1991).

[0096] In the context of providing candidate molecules for a new vaccine against tuberculosis, the subdominant epitopes are however as relevant as are the dominant epitopes since it has been shown (Olsen, 2000) that such epitopes can induce protection regardless of the fact that they are not as strongly or broadly recognized.

Variants

[0097] A common feature of the polypeptides of the invention is their capability to induce an immunological response as illustrated in the examples. It is understood that a variant of a polypeptide of the invention produced by substitution, insertion, addition or deletion may also be immunogenic as determined by any of the assays described herein.

Immune Individual

[0098] An immune individual is defined as a person or an animal, which has cleared or controlled an infection with virulent mycobacteria or has received a vaccination with M. bovis BCG.

Immune Response

[0099] The immune response may be monitored by one of the following methods:

[0100] An in vitro cellular response is determined by induction of the release of a relevant cytokine such as IFN-γ from, or the induction of proliferation in lymphocytes withdrawn from an animal or human being currently or previously infected with virulent mycobacteria or immunized with the relevant polypeptide. The induction being performed by the addition of the polypeptide or the immunogenic portion of the polypeptide to a suspension comprising from 2×10⁵ cells to 4×10⁵ cells per well. The cells being isolated from either the blood, the spleen, the liver or the lung and the addition of the polypeptide or the immunogenic portion resulting in a concentration of not more than 20 μg per ml suspension and the stimulation being performed from two to five days. For monitoring cell proliferation the cells are pulsed with radioactive labeled Thymidine and after 16-22 hours of incubation detecting the proliferation by liquid scintillation counting. A positive response is defined as being a response more than background plus two standard deviations. The release of IFN-γ can be determined by the ELISA method, which is well known to a person skilled in the art. A positive response being a response more than background plus two standard deviations. Other cytokines than IFN-γ could be relevant when monitoring the immunological response to the polypeptide, such as IL-12, TNF-α, IL-4, IL-5, IL-10, IL-6, TGF-β. Another and more sensitive method for detecting the immune response is the ELISpot method, in which the frequency of IFN-γ producing cells is determined. In an ELIspot plate (MAHA, Millipore) precoated with anti-murine IFN-γ antibodies (PharMingen) graded numbers of cells isolated from either blood, spleen, or lung (typically between 1 to 4×10⁵ cells/well) are incubated for 24-32 hrs in the presence of the polypeptide or the immunogenic portion resulting in a concentration of not more than 20 μg per ml. The plates are subsequently incubated with biotinylated anti-IFN-γ antibodies followed by a streptavidin-alkaline phosphatase incubation. The IFN-γ producing cells are identified by adding BCIP/NBT (Sigma), the relevant substrate giving rise to spots. These spots can be enumerated using a dissection microscope. It is also a possibility to determine the presence of mRNA coding for the relevant cytokine by the use of the PCR technique. Usually one or more cytokines will be measured utilizing for example PCR, ELISPOT or ELISA. It will be appreciated by a person skilled in the art that a significant increase or decrease in the amount of any of these cytokines induced by a specific polypeptide can be used in evaluation of the immunological activity of the polypeptide.

[0101] An in vitro cellular response may also be determined by the use of T cell lines derived from an immune individual or an M. tuberculosis-infected person where the T cell lines have been driven with either live mycobacteria, extracts from the bacterial cell or culture filtrate for 10 to 20 days with the addition of IL-2. The induction being performed by addition of not more than 20 μg polypeptide per ml suspension to the T cell lines containing from 1×10⁵ cells to 3×10⁵ cells per well and incubation being performed from two to six days. The induction of IFN-γ or release of another relevant cytokine is detected by ELISA. The stimulation of T cells can also be monitored by detecting cell proliferation using radioactively labeled Thymidine as described above. For both assays a positive response being a response more than background plus two standard deviations.

[0102] An in vivo cellular response may be determined as a positive DTH response after intradermal injection or local application patch of at most 100 μg of the polypeptide or the immunogenic portion to an individual who is clinically or subclinically infected with a virulent mycobacterium, a positive response having a diameter of at least 5 mm 72-96 hours after the injection or application.

[0103] An in vitro humoral response is determined by a specific antibody response in an immune or infected individual. The presence of antibodies may be determined by an ELISA technique or a Western blot where the polypeptide or the immunogenic portion is absorbed to either a nitrocellulose membrane or a polystyrene surface. The serum is preferably diluted in PBS from 1:10 to 1:100 and added to the absorbed polypeptide and the incubation being performed from 1 to 12 hours. By the use of labeled secondary antibodies the presence of specific antibodies can be determined by measuring the OD e.g. by ELISA where a positive response is a response of more than background plus two standard deviations or alternatively a visual response in a Western blot.

[0104] Another relevant parameter is measurement of the protection in animal models induced after vaccination with the polypeptide in an adjuvant or after DNA vaccination. Suitable animal models include primates, guinea pigs or mice, which are challenged with an infection of a virulent Mycobacterium. Readout for induced protection could be decrease of the bacterial load in target organs compared to non-vaccinated animals, prolonged survival times compared to non-vaccinated animals and diminished weight loss compared to non-vaccinated animals.

Preparation Methods

[0105] In general, M. tuberculosis antigens, and DNA sequences encoding such antigens, may be prepared using any one of a variety of procedures.

[0106] They may be purified as native proteins from the M. tuberculosis cell or culture filtrate by procedures such as those described above. Immunogenic antigens may also be produced recombinantly using a DNA sequence encoding the antigen, which has been inserted into an expression vector and expressed in an appropriate host. Examples of host cells are E. coli. The polypeptides or immunogenic portion hereof can also be produced synthetically having fewer than about 100 amino acids, and generally fewer than 50 amino acids and may be generated using techniques well known to those ordinarily skilled in the art, such as commercially available solid-phase techniques where amino acids are sequentially added to a growing amino acid chain.

[0107] In the construction and preparation of plasmid DNA encoding the polypeptide as defined for DNA vaccination a host strain such as E. coli can be used. Plasmid DNA can then be prepared from cultures of the host strain carrying the plasmid of interest, and purified using e.g. the Qiagen Giga-Plasmid column kit (Qiagen, Santa Clarita, Calif., USA) including an endotoxin removal step. It is preferred that plasmid DNA used for DNA vaccination is endotoxin free.

Fusion Proteins

[0108] The immunogenic polypeptides may also be produced as fusion proteins, by which methods superior characteristics of the polypeptide of the invention can be achieved. For instance, fusion partners that facilitate export of the polypeptide when produced recombinantly, fusion partners that facilitate purification of the polypeptide, and fusion partners which enhance the immunogenicity of the polypeptide fragment of the invention are all interesting possibilities. Therefore, the invention also pertains to a fusion polypeptide comprising at least one polypeptide or immunogenic portion defined above and at least one fusion partner. The fusion partner can, in order to enhance immunogenicity, be another polypeptide derived from M. tuberculosis, such as of a polypeptide fragment derived from a bacterium belonging to the tuberculosis complex, such as ESAT-6, TB10.4, CFP10, RD1-ORF5, RD1-ORF2, Rv1036, MPB64, MPT64, Ag85A, Ag85B (MPT59), MPB59, Ag85C, 19 kDa lipoprotein, MPT32 and alpha-crystalline, or at least one T-cell epitope of any of the above mentioned antigens (Skjot et al 2000; Danish Patent application PA 2000 00666; Danish Patent application PA 1999 01020; U.S. patent application Ser. No. 09/505,739; Rosenkrands et al 1998; Nagai et al 1991). The invention also pertains to a fusion polypeptide comprising mutual fusions of two or more of the polypeptides (or immunogenic portions thereof) of the invention.

[0109] Other fusion partners, which could enhance the immunogenicity of the product, are lymphokines such as IFN-γ, IL-2 and IL-12. In order to facilitate expression and/or purification, the fusion partner can e.g. be a bacterial fimbrial protein, e.g. the pilus components pilin and papA; protein A; the ZZ-peptide (ZZ-fusions are marketed by Pharmacia in Sweden); the maltose binding protein; glutathione S-transferase; β-galactosidase; or poly-histidine. Fusion proteins can be produced recombinantly in a host cell, which could be E. coli, and it is a possibility to induce a linker region between the different fusion partners.

[0110] Other interesting fusion partners are polypeptides, which are lipidated so that the immunogenic polypeptide is presented in a suitable manner to the immune system. This effect is e.g. known from vaccines based on the Borrelia burgdorferi OspA polypeptide as described in e.g. WO 96/40718 A or vaccines based on the Pseudomonas aeruginosa OprI lipoprotein (Cote-Sierra J 1998). Another possibility is N-terminal fusion of a known signal sequence and an N-terminal cystein to the immunogenic polypeptide. Such a fusion results in lipidation of the immunogenic polypeptide at the N-terminal cystein, when produced in a suitable production host.

Uses

Protein Vaccine

[0111] Another part of the invention pertains to a vaccine composition comprising a polypeptide (or at least one immunogenic portion thereof) or fusion polypeptide according to the invention. In order to ensure optimum performance of such a vaccine composition it is preferred that it comprises an immunologically and pharmaceutically acceptable carrier, vehicle or adjuvant.

[0112] An effective vaccine, wherein a polypeptide of the invention is recognized by the animal, will in an animal model be able to decrease bacterial load in target organs, prolong survival times and/or diminish weight loss after challenge with a virulent Mycobacterium, compared to non-vaccinated animals

[0113] Suitable carriers are selected from the group consisting of a polymer to which the polypeptide(s) is/are bound by hydrophobic non-covalent interaction, such as a plastic, e.g. polystyrene, or a polymer to which the polypeptide(s) is/are covalently bound, such as a polysaccharide, or a polypeptide, e.g. bovine serum albumin, ovalbumin or keyhole limpet haemocyanin. Suitable vehicles are selected from the group consisting of a diluent and a suspending agent. The adjuvant is preferably selected from the group consisting of dimethyldioctadecylammonium bromide (DDA), Quil A, poly I:C, aluminum hydroxide, Freund's incomplete adjuvant, IFN-γ, IL-2, IL-12, monophosphoryl lipid A (MPL), Trehalose Dimycolate (TDM), Trehalose Dibehenate and muramyl dipeptide (MDP).

[0114] Preparation of vaccines which contain peptide sequences as active ingredients is generally well understood in the art, as exemplified by U.S. Pat. Nos. 4,608,251; 4,601,903; 4,599,231 and 4,599,230, all incorporated herein by reference.

[0115] Other methods of achieving adjuvant effect for the vaccine include use of agents such as aluminum hydroxide or phosphate (alum), synthetic polymers of sugars (Carbopol), aggregation of the protein in the vaccine by heat treatment, aggregation by reactivating with pepsin treated (Fab) antibodies to albumin, mixture with bacterial cells such as C. parvum or endotoxins or lipopolysaccharide components of gram-negative bacteria, emulsion in physiologically acceptable oil vehicles such as mannide mono-oleate (Aracel A) or emulsion with 20 percent solution of a perfluorocarbon (Fluosol-DA) used as a block substitute may also be employed. Other possibilities involve the use of immune modulating substances such as cytokines or synthetic IFN-γ inducers such as poly I:C in combination with the above-mentioned adjuvants.

[0116] Another interesting possibility for achieving adjuvant effect is to employ the technique described in Gosselin et al., 1992 (which is hereby incorporated by reference herein). In brief, a relevant antigen such as an antigen of the present invention can be conjugated to an antibody (or antigen binding antibody fragment) against the Fcγ receptors on monocytes/macrophages.

[0117] The vaccines are administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutically effective and immunogenic. The quantity to be administered depends on the subject to be treated, including, e.g., the capacity of the individual's immune system to mount an immune response, and the degree of protection desired. Suitable dosage ranges are of the order of several hundred micrograms active ingredient per vaccination with a preferred range from about 0.1 μg to 1000 μg, such as in the range from about 1 μg to 300 μg, and especially in the range from about 10 μg to 50 μg. Suitable regimens for initial administration and booster shots are also variable but are typified by an initial administration followed by subsequent inoculations or other administrations.

[0118] The manner of application may be varied widely. Any of the conventional methods for administration of a vaccine are applicable. These are believed to include oral application on a solid physiologically acceptable base or in a physiologically acceptable dispersion, parenterally, by injection or the like. The dosage of the vaccine will depend on the route of administration and will vary according to the age of the person to be vaccinated and, to a lesser degree, the size of the person to be vaccinated.

[0119] The vaccines are conventionally administered parenterally, by injection, for example, either subcutaneously or intramuscularly. Additional formulations which are suitable for other modes of administration include suppositories and, in some cases, oral formulations. 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 and advantageously contain 10-95% of active ingredient, preferably 25-70%.

[0120] In many instances, it will be necessary to have multiple administrations of the vaccine. Especially, vaccines can be administered to prevent an infection with virulent mycobacteria, a prophylactic vaccine, and/or to treat established mycobacterial infection, a therapeutic vaccine. When administered to prevent an infection, the vaccine is given prophylactically, before definitive clinical signs, diagnosis or identification of an infection TB are present. Since the current vaccine BCG appears to induce an effective, but short-lived immune response, prophylactic vaccines may also be designed to be used as booster vaccines. Such booster vaccines are given to individuals who have previously received a vaccination, with the intention of prolonging the period of protection. In instances where the individual has already become infected or is suspected to have become infected, the previous vaccination may have provided sufficient immunity to prevent primary disease, but as discussed previously, boosting this immune response will not help against the latent infection. In such a situation, the vaccine will necessarily have to be a therapeutic vaccine designed for efficacy against the latent stage of infection. A combination of a prophylactic vaccine and a therapeutic vaccine, which is active against both primary and latent infection, constitutes a multiphase vaccine.

[0121] Due to genetic variation, different individuals may react with immune responses of varying strength to the same polypeptide. Therefore, the vaccine according to the invention may comprise several different polypeptides in order to increase the immune response. The vaccine may comprise two or more polypeptides or immunogenic portions, where all of the polypeptides are as defined above, or some but not all of the peptides may be derived from virulent mycobacteria. In the latter example, the polypeptides not necessarily fulfilling the criteria set forth above for polypeptides may either act due to their own immunogenicity or merely act as adjuvants.

[0122] The vaccine may comprise 1-20, such as 2-20 or even 3-20 different polypeptides or fusion polypeptides, such as 3-10 different polypeptides or fusion polypeptides.

[0123] The invention also pertains to a method for immunizing an animal, including a human being, against TB caused by virulent mycobacteria, comprising administering to the animal the polypeptide of the invention, or a vaccine composition of the invention as described above, or a living vaccine described above.

[0124] The invention also pertains to a method for producing an immunologic composition according to the invention, the method comprising preparing, synthesizing or isolating a polypeptide according to the invention, and solubilizing or dispersing the polypeptide in a medium for a vaccine, and optionally adding other M. tuberculosis antigens and/or a carrier, vehicle and/or adjuvant substance.

DNA Vaccine.

[0125] The nucleic acid fragments of the invention may be used for effecting in vivo expression of antigens, i.e. the nucleic acid fragments may be used in so-called DNA vaccines as reviewed in Ulmer et al 1993, which is included by reference.

[0126] Hence, the invention also relates to a vaccine comprising a nucleic acid fragment according to the invention, the vaccine effecting in vivo expression of antigen by an animal, including a human being, to whom the vaccine has been administered, the amount of expressed antigen being effective to confer substantially increased resistance to infections caused by virulent mycobacteria in an animal, including a human being.

[0127] The above mentioned definitions and distinctions of prophylactic-, booster-, therapeutic- and multiphase vaccines also applies for DNA vaccines

[0128] The efficacy of such a DNA vaccine can possibly be enhanced by administering the gene encoding the expression product together with a DNA fragment encoding a polypeptide that has the capability of modulating an immune response.

Live Recombinant Vaccines

[0129] One possibility for effectively activating a cellular immune response for a vaccine can be achieved by expressing the relevant antigen in a vaccine in a non-pathogenic microorganism or virus. Well-known examples of such microorganisms are Mycobacterium bovis BCG, Salmonella and Pseudomona and examples of viruses are Vaccinia Virus and Adenovirus.

[0130] Therefore, another important aspect of the present invention is an improvement of the living BCG vaccine presently available, wherein one or more copies of a DNA sequence encoding one or more polypeptide as defined above has been incorporated into the genome of the micro-organism in a manner allowing the micro-organism to express and secrete the polypeptide. The incorporation of more than one copy of a nucleotide sequence of the invention is contemplated to enhance the immune response.

[0131] Another possibility is to integrate the DNA encoding the polypeptide according to the invention in an attenuated virus such as the vaccinia virus or Adenovirus (Rolph et al 1997). The recombinant vaccinia virus is able to replicate within the cytoplasma of the infected host cell and the polypeptide of interest can therefore induce an immune response, which is envisioned to induce protection against TB.

Therapeutic Vaccine.

[0132] The invention also relates to the use of a polypeptide or nucleic acid of the invention for use as therapeutic vaccines as have been described by D. Lowrie (Lowrie, 1999) using DNA vaccine encoding HSP65 from M. leprae. Antigens with therapeutic properties may be identified based on their ability to diminish the severity of M. tuberculosis infection in experimental animals or prevent reactivation of previous infection, when administered as a vaccine. The composition used for therapeutic vaccines can be prepared as described above for vaccines.

Diagnostic Protein

[0133] The invention also relates to a method of diagnosing latent TB caused by a virulent mycobacterium in an animal, including a human being, comprising intradermally injecting, in the animal, a polypeptide according to the invention, a positive skin response at the location of injection being indicative of the animal having TB, and a negative skin response at the location of injection being indicative of the animal not having TB.

[0134] When diagnosis of latent infection with virulent mycobacteria is the aim, a blood sample comprising mononuclear cells (i.e. T-lymphocytes) from a patient is contacted with a sample of one or more polypeptides of the invention. This contacting can be performed in vitro and a positive reaction could e.g. be proliferation of the T-cells or release of cytokines such as IFN-γ into the extracellular phase. It is also conceivable to contact a serum sample from a subject with a polypeptide of the invention, the demonstration of a binding between antibodies in the serum sample and the polypeptide being indicative of previous or ongoing infection.

[0135] The invention therefore also relates to an in vitro method for diagnosing latent infection in an animal or a human being with a virulent mycobacterium, the method comprising providing a blood sample from the animal or human being, and contacting the sample from the animal with the polypeptide of the invention, a significant release into the extracellular phase of at least one cytokine by mononuclear cells in the blood sample being indicative of the animal being sensitized. A positive response being a response more than release from a blood sample derived from a patient without the TB diagnosis plus two standard deviations. The invention also relates to the in vitro method for diagnosing ongoing or previous sensitization in an animal or a human being with a virulent mycobacterium, the method comprising providing a blood sample from the animal or human being, and by contacting the sample from the animal with the polypeptide of the invention demonstrating the presence of antibodies recognizing the polypeptide of the invention in the serum sample.

[0136] The immunogenic composition used for diagnosing may comprise 1-20, such as 2-20 or even 3-20 different polypeptides or fusion polypeptides, such as 3-10 different polypeptides or fusion polypeptides.

Diagnostic DNA

[0137] The nucleic acid probes encoding the polypeptide of the invention can be used in a variety of diagnostic assays for detecting the presence of pathogenic organisms in a given sample.

[0138] A method of determining the presence of mycobacterial nucleic acids in an animal, including a human being, or in a sample, comprising administering a nucleic acid fragment of the invention to the animal or incubating the sample with the nucleic acid fragment of the invention or a nucleic acid fragment complementary thereto, and detecting the presence of hybridized nucleic acids resulting from the incubation (by using the hybridization assays which are well-known in the art), is also included in the invention. Such a method of diagnosing TB might involve the use of a composition comprising at least a part of a nucleotide sequence as defined above and detecting the presence of nucleotide sequences in a sample from the animal or human being to be tested which hybridize with the nucleic acid fragment (or a complementary fragment) by the use of PCR technique.

Antibodies

[0139] A monoclonal or polyclonal antibody, which is specifically reacting with a polypeptide of the invention in an immunoassay, or a specific binding fragment of said antibody, is also a part of the invention. The antibodies can be produced by methods known to a person skilled in the art. Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of a polypeptide according to the present invention and, if desired, an adjuvant. The monoclonal antibodies according to the present invention may, for example, be produced by the hybridoma method first described by Kohler and Milstein (Kohler and Milstein, 1975), or may be produced by recombinant DNA methods such as described in U.S. Pat. No. 4,816,567. The monoclonal antibodies may also be isolated from phage libraries generated using the techniques described by McCafferty et al (McCafferty, 1990), for example. Methods for producing antibodies are described in the literature, e.g. in U.S. Pat. No. 6,136,958.

[0140] A sample of a potentially infected organ or body fluid from an infected individual may be contacted with such an antibody recognizing a polypeptide of the invention. The demonstration of the reaction by means of methods well known in the art between the sample and the antibody will be indicative of an ongoing infection. It is of course also a possibility to demonstrate the presence of anti-mycobacterial antibodies in serum or other body fluids by contacting a serum sample from a subject with at least one of the polypeptide fragments of the invention and using well-known methods for visualizing the reaction between the antibody and antigen.

[0141] In diagnostics, an antibody, a nucleic acid fragment and/or a polypeptide of the invention can be used either alone, or as a constituent in a composition. Such compositions are known in the art, and comprise compositions in which the antibody, the nucleic acid fragment or the polypeptide of the invention is coupled, preferably covalently, to at least one other molecule, e.g. a label (e.g. radioactive or fluorescent) or a carrier molecule.

[0142] It will be understood that the following examples are illustrative of the present invention and are not a limitation thereof. A number of variations on the techniques, reagents, and conditions described in the following examples will be readily apparent to one of skill in the art.

EXAMPLES

Example 1

Cloning and Expression of Low Oxygen Induced M. Tuberculosis Antigens in E. coli

[0143] A number of M. tuberculosis genes are induced under low oxygen conditions. The up-regulation of the genes listed in table 2 has been determined at either the mRNA (Sherman, 2001) or protein (Boon, 2001, Rosenkrands, 2002) level. The coding region of these selected antigens is amplified by PCR using the primer sets listed in Table 3.

TABLE-US-00004 TABLE 3 Primer sequences for PCR amplification of selected low oxygen induced antigens SEQ Rv no. ID NO: Primer sequence Rv0079 Fwd 92 CACCGTGGAACCGAAACGCAGTCG Rvs 93 TTATGCCAGACCGTCGGCA Rv0080 Fwd 94 CACCATGAGCCCGGGCTCG Rvs 95 TTACGGCGTACGCGAGTCAG Rv0081 Fwd 96 CACCGTGGAGTCCGAACCGCTGTA Rvs 97 TTACGTGGCCGAGCCGC Rv0363c Fwd 98 CACCATGCCTATCGCAACGCCC (fba) Rvs 99 TTAGTGGGTTAGGGACTTTCCGG Rv0569 Fwd 100 GGGGACAAGTTTGTACAAAAAAGCAGGCTTAAAGGCAAAGGTCGGGGAC Rvs 101 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTACGTTCCCCTGGCATGGA Rv0572c Fwd 102 CACCATGGGTGAGCACGCCATC Rvs 103 TTATAGGTCATCGGATTGAGGTGATC Rv0574c Fwd 104 CACCGTGGCTGGCAATCCTGATGT Rvs 105 TTACTCCTTGCTCGTTAGGTTGGC Rv1264 Fwd 106 CACCGTGACAGACCACGTGCGC Rvs 107 TTACGGTGACGAGCCGGC Rv1592c Fwd 108 CACCATGGTAGAGCCCGGCAATTTG Rvs 109 TTAGAGCGGACGGCGGCT Rv1733c Fwd 110 CACCATGATCGCCACAACCCGC Rvs 111 TTACCGCTGCGTGCAGAACA Rv1734c Fwd 112 CACCATGACCAACGTCGGTGACCA Rvs 113 TTATCCTGTTACTGCGGCGCA Rv1736c Fwd 114 CACCGTGACGGTGACACCACGGAC (narX) Rvs 115 TTACCACCCGCGCCGC Rv1737c Fwd 116 CACCATGAGAGGGCAAGCGGC (narK2) Rvs 117 TTACCTGGACGCCTCCTCACTC Rv1738 Fwd 118 CACCATGTGCGGCGACCAGTC Rvs 119 TTAATACAACAATCGCGCCGG Rv1739c Fwd 120 CACCATGATTCCCACGATGACATCG Rvs 121 TTAGCGCCGACGGAACG Rv1813c Fwd 122 GGGGACAAGTTTGTACAAAAAAGCAGGCTTAATCACAAACCTCCGACGC Rvs 123 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTAGTTGCACGCCCAGTTGA C Rv1997 Fwd 124 CACCTTGTCGGCGTCAGTGTCTGC (ctpF) Rvs 125 TTATGGCGGTTGCGCCC Rv1998c Fwd 126 CACCATGAGTTTCCACGATCTTCATCACC Rvs 127 TTACGTTGTACTCGTGCGGTTCTC Rv2003c Fwd 128 CACCGTGGTCAAGCGCTCTCGG Rvs 129 TTATTCCGACTCGAGTGGGTGA Rv2005c Fwd 130 CACCATGTCTAAACCCCGCAAGCA Rvs 131 TTACGACTGCCGTGCCACG Rv2007c Fwd 132 CACCGTGACCTATGTGATCGGTAGTGAGTG (fdxA) Rvs 133 TTAAGGGCACTCCACCGGGA Rv2028c Fwd 134 CACCATGAACCAATCACACAAACCCC Rvs 135 TTACAGATACTGCTGACCGACGACC Rv2029c Fwd 136 CACCATGACGGAGCCAGCGG (PfkB) Rvs 137 TTATGGCGAGGCTTCCGG Rv2030c Fwd 138 GGGGACAAGTTTGTACAAAAAAGCAGGCTTACTGATGACCGCAGCGGCT Rvs 139 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTACAGACCGGTCGGGTAGG TTT Rv2031c Fwd 140 GGGGACAAGTTTGTACAAAAAAGCAGGCTTAGCCACCACCCTTCCCGT (hspX) Rvs 141 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTAGTTGGTGGACCGGATCT GAAT Rv2032 Fwd 142 CACCATGCCGGACACCATGGTG Rvs 143 TTAGTGATCCTTAGCCCGAACGTG Rv2428 Fwd 144 GGGGACAAGTTTGTACAAAAAAGCAGGCTTAATGCCACTGCTAACCATTG GC (ahpC) Rvs 145 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTAGGCCGAAGCCTTGAGGA GT Rv2624c Fwd 146 CACCATGTCTGGGAGAGGAGAGCCG Rvs 147 TTAGCGAACGACAAGCACCGA Rv2625c Fwd 148 GGGGACAAGTTTGTACAAAAAAGCAGGCTTACGTGATGCGATCCCGCT Rvs 149 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTACCCCGCATCGGAAAACC Rv2627c Fwd 150 GGGGACAAGTTTGTACAAAAAAGCAGGCTTAATGGCAAGTTCTGCGAGCG A Rvs 151 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTAGGAACGGTCGCGCTGTG T Rv2628 Fwd 152 CACCATGTCCACGCAACGACCG Rvs 153 TTAACCGCAACGGCAATCTCA Rv2629 Fwd 154 CACCATGCGATCAGAACGTCTCCG Rvs 155 TTAGGATCTATGGCTGCCGAGTC Rv2630 Fwd 156 CACCATGCTGCACCGCGACGA Rvs 157 TTACACATCGAGCGTTACCGCAC Rv2659c Fwd 158 GGGGACAAGTTTGTACAAAAAAGCAGGCTTAGTGACGCAAACCGGCAA Rvs 159 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTACATCTCCTGGTTCTCGG CC Rv2780 Fwd 160 GGGGACAAGTTTGTACAAAAAAGCAGGCTTACGCGTCGGTATTCCGACC Rvs 161 GGGGACCACTTTGTACAAGAAAGCTGGGTCCTACACGCTGGCGGGCTC Rv3126c Fwd 162 CACCATGGTCATCCGGTTTGATCAAATA Rvs 163 TTATGGATTGTCAATGACAGCCCA Rv3127 Fwd 164 CACCGTGCTCAAGAACGCAGTCTTGC Rvs 165 TTAAGGCGGGCTGAACCAACC Rv3128c Fwd 166 CACCGTGTGGTCCGCCTCGG Rvs 167 TTAGCCCGCCTTGATCAGGA Rv3129 Fwd 168 CACCGTGGTGCAAGGCCGCA Rvs 169 TTATCGCTGGTTGTGTGACGAG Rv3130c Fwd 170 CACCATGAATCACCTAACGACACTTGACG Rvs 171 TTACACAACCAGCGATAGCGCTC Rv3131 Fwd 172 CACCATGAACACCCATTTCCCGG Rvs 173 TTAGCACCGTTGTCGCAGTAGCT Rv3132c Fwd 174 CACCATGACAACAGGGGGCCTCG Rvs 175 TTACTGCGACAACGGTGCTGAC Rv3134c Fwd 176 CACCATGAGCGATCCTCGGCCA Rvs 177 TTACAAGTTGGCACTGCGTACCG Rv3841 Fwd 178 CCGGCTGAGATCTATGACAGAATACGAAGGGC (bfrB) Rvs 179 CCCCGCCAGGGAACTAGAGGCGGC Rv3842c Fwd 180 CACCATGACATGGGCCGACGAG (glpQ1) Rvs 181 TTAGCGAGTGGTCCCGTTCG Rv3908 Fwd 182 CACCGTGTCCGACGGCGAACAA Rvs 183 TTACGGCCCCGGCCC

[0144] PCR reactions were carried out using Platinum Tag DNA Polymerase (GIBCO BRL) in a 50 μl reaction volume containing 60 mM Tris-SO₄ (pH 8.9), 18 mM Ammonium Sulfate, 0.2 mM of each of the four nucleotides, 0.2 μM of each primer and 10 ng of M. tuberculosis H37Rv chromosomal DNA. The reaction mixtures were initially heated to 95° C. for 5 min., followed by 35 cycles of: 95° C. for 45 sec, 60° C. for 45 sec and 72° C. for 2 min. The amplification products were precipitated by PEG/MgCl₂, and dissolved in 50 μL TE buffer.

[0145] DNA fragments were cloned and expressed in Gateway Cloning system (Life Technology). First, to create Entry Clones, 5 μL of DNA fragment was mixed with 1 μL of pDONR201, 2 μL of BP CLONASE enzyme mix and 2 μL of BP reaction buffer. The recombination reactions were carried out at 25° C. for 60 min. After Proteinase K treatment at 37° C. for 10 min., 5 μL of each sample was used to transform E. coli DH5α competent cells. Transformants were selected on LB plates containing 50 μg/mL kanamycin. One bacterial clone from each transformation was grown in 3 mL LB medium containing 50 μg/mL kanamycin and plasmid DNA was isolated (Qiagen).

[0146] Second, to create expression clones, 2 μL of each entry clone DNA was mixed with μL of His-tagged expression vector (pDest17), 2 μL LR reaction buffer, 2 μL LR CLONASE enzyme mix and 3 μL TE. After recombination at 25° C. for 60 min. and Proteinase K treatment at 37° C. for 10 min., 5 μL of each sample was used to transform E. coli BL21-SI competent cells. Transformants were selected on LBON (LB without NaCl) plates containing 100 μg/mL ampicillin. The resulting E. coli clones express recombinant proteins carrying a 6-histine tag at the N-terminal. All clones were confirmed by DNA sequencing.

[0147] Recombinant proteins were purified from transformed E. coli BL21-SI cells cultured in 900 mL LBON medium containing 100 μg/mL at 30° C. until OD₆₀₀=0.4-0.6. At this point 100 mL 3 M NaCl was added and 3 hours later bacteria were harvested by centrifugation. Bacteria pellets were resuspended in 20 mL bacterial protein extraction reagent (Pierce) incubated for 10 min. at room temperature and pelleted by centrifugation. Bacteria were lysed and their DNA digested by treating with lysozyme (0.1 mg/mL) and DNase I (2.5 μg/mL) at room temperature for 30 minutes, with gentle agitation. The recombinant protein forms inclusion bodies and can be pelleted by centrifugation at 27.000×g for 15 min. Protein pellets were solubilized by adding 20 ml of sonication buffer (8 M urea, 50 mM Na₂HPO₄, 100 mM Tris-HCl, pH 8.0) and sonicating 5×30 sec pulses interrupted by a 30 sec pause. After another centrifugation at 27.000×g for 15 min., supernatants were applied to 10 mL TALON columns (Clontech). The columns were washed with 50 mL sonication buffer. Bound proteins were eluted by lowering pH (8 M urea, 50 mM Na₂HPO₄, 100 mM Tris-HCl, pH 4.5). 5 mL fractions were collected and analyzed by SDS-PAGE. Fractions containing recombinant protein were pooled. Further purification was achieved by anion- or cation-exchange chromatography on Hitrap columns (Pharmacia). Bound protein was eluted using a NaCl gradient from 0-500 mM in 3 M urea, 10 mM Tris-HCl, pH 8.0. All fractions were collected and analyzed on SDS-PAGE using Coomassie staining. Fractions containing recombinant protein were pooled. Final protein concentrations were determined by micro BCA (Pierce).

Example 2

Prophylactic Versus Therapeutic Vaccine

Murine Vaccination Models.

[0148] A prophylactic vaccine given prior to infection should induce an immune response sufficiently strong to prevent or dampen the initial proliferation of the bacteria in the acute phase and thereby reduce the ensuing disease. In the murine prophylactic vaccine model outlined in FIG. 1A, nai{umlaut over (v)}e mice are immunized 3 times, 2 weeks apart with recombinant antigens. Six weeks after the last immunization, the mice are given an aerosol infection with approximately 250 M. tuberculosis bacilli. The protective capacity of the vaccine is evaluated by enumeration of the bacteria in spleen and lung 6 weeks post-infection.

[0149] To define the optimal components for a therapeutic vaccine, a murine reactivation model of latent TB has been established (van Pinxteren, 2000) (FIG. 1B). An aerosol infection with approximately 250 M. tuberculosis bacilli is given and at the peak of infection 6 weeks later, the mice receive an 8-week course of anti-mycobacterial drug treatment of isoniazid and rifabutin given in the drinking water. This reduces the bacterial load in spleen and lung to a low level (about 500 bacteria per organ). This latent phase of low chronic infection is stable for 9-10 weeks after which a slow spontaneous reactivation occurs. The therapeutic vaccine is given as 3 subcutaneous (s.c.) immunizations about 5 weeks after cessation of drug treatment. The effect of the therapeutic vaccine is evaluated as protection against reactivation determined by enumeration of bacteria in spleen and lung 7 weeks after the last immunization.

[0150] The effect of the antigens in a prophylactic or a therapeutic vaccine.

[0151] BCG, ESAT6, and Rv2031c, one of the most prominent proteins induced under low oxygen conditions (Rosenkrands, 2002), were analyzed for their prophylactic and therapeutic vaccine potential. Naive or latently infected C57Bl mice were immunized with one s.c. injection of 2.5×10⁵ BCG, or 3 s.c. immunizations of 10 μg of either recombinant ESAT6 or recombinant Rv2031c in a DDA/MPL adjuvant. The vaccinations were done in groups of 5 mice and protective capacity of the vaccines was evaluated as described above. FIG. 2 shows the bacterial load in the lung in the acute phase (A) and in the reactivation phase (B), after prophylactic and therapeutic vaccination respectively. ESAT6 (as previously described by Brandt, 2000) offers protection against acute phase infection at the level of BCG (FIG. 2A). However, neither of the two shows any protective effect against reactivation of the infection when given during latent infection (FIG. 2B). In contrast, Rv2031c, the low oxygen induced antigen, offers no protection against the acute phase of the infection when given as a prophylactic vaccine, but gives some protection against reactivation when given as a therapeutic vaccine. That is, some antigens, here exemplified by ESAT6, though potent as prophylactic vaccines have no effect as therapeutic vaccines. In contrast, other antigens, here exemplified by Rv2031c, can be efficient therapeutic vaccines although they have no effect or only negligible effect as prophylactic vaccines.

Example 3

Low Oxygen Induced Antigens, Rv2031c, as Therapeutic Vaccines

[0152] There is a high variability in bacterial load intrinsic to the reactivation model in the latent and reactivation phase. The analysis of Rv2031c as a therapeutic vaccine was therefore repeated in groups of eight mice. As in the previous experiments the mice were given 3 s.c. immunizations of 10 μg rRv2031c in DDA/MPL. The induced immune responses were analyzed one week post immunization. The mice were partially bled and the PBMC from the blood purified and analyzed for Rv2031c- and ESAT6 specific recall responses. Using ELIspot technique, the frequency of Rv2031c-specific and ESAT6-specific IFN-γ-producing cells were determined in both the rRv2031c immunized and the unimmunized group (FIG. 3). The rRv2031c immunization has increased the frequency of Rv2031c-specific IFN-γ producing cells by a factor of 43 as compared to the unimmunized group. In contrast, the frequency of ESAT6-specific IFN-γ producing cells is significantly higher in the unimmunized group. ESAT6 is an antigen produced in high amounts by the actively-growing M. tuberculosis bacteria. The level of the ESAT6 specific immune response in infected mice could therefore be indicative the degree of actively-growing infection in the animals. Recent reports have in fact demonstrated such a correlation between the level of ESAT6 response and degree of disease in both M. tuberculosis-infected humans and M. bovis-infected cattle (Doherty, 2002, Vordermeier, 2002). Therefore, the higher ESAT6 response in the unimmunized group of latently-infected mice could be indicative of a transition into the reactivation phase, where the bacteria are again beginning to multiply.

[0153] To analyze the epitope recognition pattern of Rv2031c, fourteen overlapping peptides (each 20 amino acids long) covering the whole Rv2031c protein were synthesized. Initially the peptides were analyzed in 4 pools of 3-4 peptides. PBMCs from rRv2031c immunized latently-infected mice were incubated with the peptide pools (5 μg/ml per peptide) for 72 h. The specific peptide-induced IFN-γ production was quantitated in the supernatant in a standard sandwich ELISA using paired anti-murine IFN-γ antibodies (PharMingen) and recombinant IFN-γ (PharMingen) as standard. Both peptide pool 1-4 and 8-10 stimulated a significant IFN-γ response (FIG. 4A). The individual peptides of these two pools were re-analyzed (FIG. 4B). This clearly shows that the response to Rv2031c contains a dominant epitope, peptide 2 (PRSLFPEFSELFAAFPSFAG, aa 11-30 of SEQ ID NO:24), and a subdominant epitope, peptide 9 (RTEQKDFDGRSEFAYGSFVR, aa 81-100 of SEQ ID NO:24).

[0154] The therapeutic effect of the rRv2031c immunizations was analyzed 7 weeks after the last immunization. FIG. 5 shows the bacterial load in the lung (A) and the spleen (B) of both rRv2031c-immunized and unimmunized mice. There is a clear reduction in the level of bacteria in both organs in the rRv2031c-immunized group. That is, the induction of Rv2031c T cell responses can participate in keeping the latent infection in check.

Example 4

Low Oxygen Induced Antigens, Rv0569, as Therapeutic Vaccines

[0155] Rv0569 is also a low oxygen induced antigen described in WO0179274 and illustrates very well the potential as a therapeutic vaccine.

[0156] We have established a murine reactivation model of latent TB [van Pinxteren et al, 2000, 30:3689-98], which is a variant of the so-called Cornell model. An aerosol infection is allowed to be established and at the peak of infection 6 weeks after, the mice receive an 8-week course of anti-mycobacterial drug treatment of isoniazid and rifabutin given in the drinking water. This reduces the bacterial load in spleen and lung to a low level. This latent phase of low chronic infection is stable for 9-10 weeks after which a slow spontaneous reactivation can be detected. This model is used to analyze the protective effect of a post exposure vaccine on reactivation.

[0157] Rv0569, which is highly up regulated under low oxygen growth conditions [Rosenkrands et al, 2002, 184(13): 3485-91], was analyzed for its ability to protect against reactivation given as a therapeutic vaccine in the latent phase of TB infection. Latent infected C57Bl mice were vaccinated with 3 s.c. injections of 3 μg recombinant Rv0569 and for comparison with 3 s.c. injections of 3 μg recombinant ESAT6 or one s.c. injection of BCG. The effect of the vaccine is evaluated 7 weeks after the last immunization. The induced immune responses were analyzed for Rv0569 or ESAT6 specific responses in an in vitro recall assay. Isolated splenocytes were incubated with 1 μg/ml of Rv0569 or 1 μg/ml of ESAT6 for 72 h. The produced IFNγ in the culture supernatant was quantitated in a standard sandwich ELISA. FIG. 6 shows a nice Rv0569 specific IFNγ response induced in the Rv0569 vaccinated group with practically no response in the un-vaccinated group. The ESAT6 vaccination enhanced the ESAT6 specific response though a significant ESAT6 response persisted in the un-vaccinated latent infected group.

[0158] The Rv0569 induced protection against reactivation was determined by enumeration of bacteria in spleen and lung 7 weeks after the last immunization. FIG. 7 shows the bacterial load in the lung and the spleen of both Rv0569-vaccinated, ESAT6-vaccinated, BCG vaccinated and un-vaccinated latently infected mice. There is a clear reduction in the level of viable bacteria in both spleen and lungs of the Rv0569 vaccinated mice, whereas neither ESAT6 nor BCG are able to inhibit the growth of the M. tuberculosis bacteria when given as a vaccine during latent infection. That is, the induction of Rv0569 T cell responses can participate in keeping the latent infection in check.

REFERENCES

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[0162] Cote-Sierra J, et al 1998, Gene October 9; 221(1):25-34

[0163] Doherty T M et al., 2002, J Clin Microbiol. February; 40(2):704-6.

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[0179] Danish Patent application PA 2000 00666 "Nucleic acid fragments and polypeptide fragments derived from M. tuberculosis"

[0180] Danish Patent application PA 1999 01020 (WO 01/23388) "Tuberculosis vaccine and diagnostic based on the Mycobacterium tuberculosis esat-6 gene family".

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[0182] Pearson, W. R. et al. 1988. Proc Natl Acad Sci USA, 85, 2444-2448.

[0183] Ravn, P. et al 1999. J. Infect. Dis. 179:637-645

[0184] Rolph, M. S, and I. A. Ramshaw. 1997. Curr. Opin. Immunol. 9:517-24

[0185] Rosenkrands, I., et al 1998, Infect. Immun 66:6; 2728-2735

[0186] Rosenkrands, I., et al. 2002. Journal of Bacteriology, 184: 3485-3491.

[0187] Sambrook et al Molecular Cloning; A laboratory manual, Cold Spring Harbor Laboratories, NY, 1989

[0188] Sherman, D. R., et al. 2001. PNAS, 98, 7534-7539.

[0189] Sinigaglia F et al. Nature 1988 Dec. 22-29; 336(6201):778-80

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[0191] Stryhn, A., et a/1996 Eur. J. Immunol. 26:1911-1918

[0192] Thompson J., et al Nucleic Acids Res 1994 22:4673-4680

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[0195] Vordermeier, H. M. et al. 2002. Infect. Immun., 70, 3026-3032.

[0196] All documents listed above, and the sequence listing, are incorporated by reference herein. A variety of modifications and variations on the processes, conditions, reagents and compositions described herein will be readily apparent to one of skill in the art given the teachings of the present invention. Such modifications and variations are within the scope of the invention as set forth in the following claims.

Sequence CWU 1

1

1871273PRTMycobacterium tuberculosis 1Val Glu Pro Lys Arg Ser Arg Leu Val Val Cys Ala Pro Glu Pro Ser1 5 10 15His Ala Arg Glu Phe Pro Asp Val Ala Val Phe Ser Gly Gly Arg Ala 20 25 30Asn Ala Ser Gln Ala Glu Arg Leu Ala Arg Ala Val Gly Arg Val Leu 35 40 45Ala Asp Arg Gly Val Thr Gly Gly Ala Arg Val Arg Leu Thr Met Ala 50 55 60Asn Cys Ala Asp Gly Pro Thr Leu Val Gln Ile Asn Leu Gln Val Gly65 70 75 80Asp Thr Pro Leu Arg Ala Gln Ala Ala Thr Ala Gly Ile Asp Asp Leu 85 90 95Arg Pro Ala Leu Ile Arg Leu Asp Arg Gln Ile Val Arg Ala Ser Ala 100 105 110Gln Trp Cys Pro Arg Pro Trp Pro Asp Arg Pro Arg Arg Arg Leu Thr 115 120 125Thr Pro Ala Glu Ala Leu Val Thr Arg Arg Lys Pro Val Val Leu Arg 130 135 140Arg Ala Thr Pro Leu Gln Ala Ile Ala Ala Met Asp Ala Met Asp Tyr145 150 155 160Asp Val His Leu Phe Thr Asp Ala Glu Thr Gly Glu Asp Ala Val Val 165 170 175Tyr Arg Ala Gly Pro Ser Gly Leu Arg Leu Ala Arg Gln His His Val 180 185 190Phe Pro Pro Gly Trp Ser Arg Cys Arg Ala Pro Ala Gly Pro Pro Val 195 200 205Pro Leu Ile Val Asn Ser Arg Pro Thr Pro Val Leu Thr Glu Ala Ala 210 215 220Ala Val Asp Arg Ala Arg Glu His Gly Leu Pro Phe Leu Phe Phe Thr225 230 235 240Asp Gln Ala Thr Gly Arg Gly Gln Leu Leu Tyr Ser Arg Tyr Asp Gly 245 250 255Asn Leu Gly Leu Ile Thr Pro Thr Gly Asp Gly Val Ala Asp Gly Leu 260 265 270Ala2152PRTMycobacterium tuberculosis 2Met Ser Pro Gly Ser Arg Arg Ala Ser Pro Gln Ser Ala Arg Glu Val1 5 10 15Val Glu Leu Asp Arg Asp Glu Ala Met Arg Leu Leu Ala Ser Val Asp 20 25 30His Gly Arg Val Val Phe Thr Arg Ala Ala Leu Pro Ala Ile Arg Pro 35 40 45Val Asn His Leu Val Val Asp Gly Arg Val Ile Gly Arg Thr Arg Leu 50 55 60Thr Ala Lys Val Ser Val Ala Val Arg Ser Ser Ala Asp Ala Gly Val65 70 75 80Val Val Ala Tyr Glu Ala Asp Asp Leu Asp Pro Arg Arg Arg Thr Gly 85 90 95Trp Ser Val Val Val Thr Gly Leu Ala Thr Glu Val Ser Asp Pro Glu 100 105 110Gln Val Ala Arg Tyr Gln Arg Leu Leu His Pro Trp Val Asn Met Ala 115 120 125Met Asp Thr Val Val Ala Ile Glu Pro Glu Ile Val Thr Gly Ile Arg 130 135 140Ile Val Ala Asp Ser Arg Thr Pro145 1503114PRTMycobacterium tuberculosis 3Val Glu Ser Glu Pro Leu Tyr Lys Leu Lys Ala Glu Phe Phe Lys Thr1 5 10 15Leu Ala His Pro Ala Arg Ile Arg Ile Leu Glu Leu Leu Val Glu Arg 20 25 30Asp Arg Ser Val Gly Glu Leu Leu Ser Ser Asp Val Gly Leu Glu Ser 35 40 45Ser Asn Leu Ser Gln Gln Leu Gly Val Leu Arg Arg Ala Gly Val Val 50 55 60Ala Ala Arg Arg Asp Gly Asn Ala Met Ile Tyr Ser Ile Ala Ala Pro65 70 75 80Asp Ile Ala Glu Leu Leu Ala Val Ala Arg Lys Val Leu Ala Arg Val 85 90 95Leu Ser Asp Arg Val Ala Val Leu Glu Asp Leu Arg Ala Gly Gly Ser 100 105 110Ala Thr4344PRTMycobacterium tuberculosis 4Met Pro Ile Ala Thr Pro Glu Val Tyr Ala Glu Met Leu Gly Gln Ala1 5 10 15Lys Gln Asn Ser Tyr Ala Phe Pro Ala Ile Asn Cys Thr Ser Ser Glu 20 25 30Thr Val Asn Ala Ala Ile Lys Gly Phe Ala Asp Ala Gly Ser Asp Gly 35 40 45Ile Ile Gln Phe Ser Thr Gly Gly Ala Glu Phe Gly Ser Gly Leu Gly 50 55 60Val Lys Asp Met Val Thr Gly Ala Val Ala Leu Ala Glu Phe Thr His65 70 75 80Val Ile Ala Ala Lys Tyr Pro Val Asn Val Ala Leu His Thr Asp His 85 90 95Cys Pro Lys Asp Lys Leu Asp Ser Tyr Val Arg Pro Leu Leu Ala Ile 100 105 110Ser Ala Gln Arg Val Ser Lys Gly Gly Asn Pro Leu Phe Gln Ser His 115 120 125Met Trp Asp Gly Ser Ala Val Pro Ile Asp Glu Asn Leu Ala Ile Ala 130 135 140Gln Glu Leu Leu Lys Ala Ala Ala Ala Ala Lys Ile Ile Leu Glu Ile145 150 155 160Glu Ile Gly Val Val Gly Gly Glu Glu Asp Gly Val Ala Asn Glu Ile 165 170 175Asn Glu Lys Leu Tyr Thr Ser Pro Glu Asp Phe Glu Lys Thr Ile Glu 180 185 190Ala Leu Gly Ala Gly Glu His Gly Lys Tyr Leu Leu Ala Ala Thr Phe 195 200 205Gly Asn Val His Gly Val Tyr Lys Pro Gly Asn Val Lys Leu Arg Pro 210 215 220Asp Ile Leu Ala Gln Gly Gln Gln Val Ala Ala Ala Lys Leu Gly Leu225 230 235 240Pro Ala Asp Ala Lys Pro Phe Asp Phe Val Phe His Gly Gly Ser Gly 245 250 255Ser Leu Lys Ser Glu Ile Glu Glu Ala Leu Arg Tyr Gly Val Val Lys 260 265 270Met Asn Val Asp Thr Asp Thr Gln Tyr Ala Phe Thr Arg Pro Ile Ala 275 280 285Gly His Met Phe Thr Asn Tyr Asp Gly Val Leu Lys Val Asp Gly Glu 290 295 300Val Gly Val Lys Lys Val Tyr Asp Pro Arg Ser Tyr Leu Lys Lys Ala305 310 315 320Glu Ala Ser Met Ser Gln Arg Val Val Gln Ala Cys Asn Asp Leu His 325 330 335Cys Ala Gly Lys Ser Leu Thr His 3405113PRTMycobacterium tuberculosis 5Met Gly Glu His Ala Ile Lys Arg His Met Arg Gln Arg Lys Pro Thr1 5 10 15Lys His Pro Leu Ala Gln Lys Arg Gly Ala Arg Ile Leu Val Phe Thr 20 25 30Asp Asp Pro Arg Arg Ser Val Leu Ile Val Pro Gly Cys His Leu Asp 35 40 45Ser Met Arg Arg Glu Lys Asn Ala Tyr Tyr Phe Gln Asp Gly Asn Ala 50 55 60Leu Val Gly Met Val Val Ser Gly Gly Thr Val Glu Tyr Asp Ala Asp65 70 75 80Asp Arg Thr Tyr Val Val Gln Leu Thr Asp Gly Arg His Thr Thr Glu 85 90 95Ser Ser Phe Glu His Ser Ser Pro Ser Arg Ser Pro Gln Ser Asp Asp 100 105 110Leu6380PRTMycobacterium tuberculosis 6Val Ala Gly Asn Pro Asp Val Val Thr Val Leu Leu Gly Gly Asp Val1 5 10 15Met Leu Gly Arg Gly Val Asp Gln Ile Leu Pro His Pro Gly Lys Pro 20 25 30Gln Leu Arg Glu Arg Tyr Met Arg Asp Ala Thr Gly Tyr Val Arg Leu 35 40 45Ala Glu Arg Val Asn Gly Arg Ile Pro Leu Pro Val Asp Trp Arg Trp 50 55 60Pro Trp Gly Glu Ala Leu Ala Val Leu Glu Asn Thr Ala Thr Asp Val65 70 75 80Cys Leu Ile Asn Leu Glu Thr Thr Ile Thr Ala Asp Gly Glu Phe Ala 85 90 95Asp Arg Lys Pro Val Cys Tyr Arg Met His Pro Asp Asn Val Pro Ala 100 105 110Leu Thr Ala Leu Arg Pro His Val Cys Ala Leu Ala Asn Asn His Ile 115 120 125Leu Asp Phe Gly Tyr Gln Gly Leu Thr Asp Thr Val Ala Ala Leu Ala 130 135 140Gly Ala Gly Ile Gln Ser Val Gly Ala Gly Ala Asp Leu Leu Ala Ala145 150 155 160Arg Arg Ser Ala Leu Val Thr Val Gly His Glu Arg Arg Val Ile Val 165 170 175Gly Ser Val Ala Ala Glu Ser Ser Gly Val Pro Glu Ser Trp Ala Ala 180 185 190Arg Arg Asp Arg Pro Gly Val Trp Leu Ile Arg Asp Pro Ala Gln Arg 195 200 205Asp Val Ala Asp Asp Val Ala Ala Gln Val Leu Ala Asp Lys Arg Pro 210 215 220Gly Asp Ile Ala Ile Val Ser Met His Trp Gly Ser Asn Trp Gly Tyr225 230 235 240Ala Thr Ala Pro Gly Asp Val Ala Phe Ala His Arg Leu Ile Asp Ala 245 250 255Gly Ile Asp Met Val His Gly His Ser Ser His His Pro Arg Pro Ile 260 265 270Glu Ile Tyr Arg Gly Lys Pro Ile Leu Tyr Gly Cys Gly Asp Val Val 275 280 285Asp Asp Tyr Glu Gly Ile Gly Gly His Glu Ser Phe Arg Ser Glu Leu 290 295 300Arg Leu Leu Tyr Leu Thr Val Thr Asp Pro Ala Ser Gly Asn Leu Ile305 310 315 320Ser Leu Gln Met Leu Pro Leu Arg Val Ser Arg Met Arg Leu Gln Arg 325 330 335Ala Ser Gln Thr Asp Thr Glu Trp Leu Arg Asn Thr Ile Glu Arg Ile 340 345 350Ser Arg Arg Phe Gly Ile Arg Val Val Thr Arg Pro Asp Asn Leu Leu 355 360 365Glu Val Val Pro Ala Ala Asn Leu Thr Ser Lys Glu 370 375 3807397PRTMycobacterium tuberculosis 7Val Thr Asp His Val Arg Glu Ala Asp Asp Ala Asn Ile Asp Asp Leu1 5 10 15Leu Gly Asp Leu Gly Gly Thr Ala Arg Ala Glu Arg Ala Lys Leu Val 20 25 30Glu Trp Leu Leu Glu Gln Gly Ile Thr Pro Asp Glu Ile Arg Ala Thr 35 40 45Asn Pro Pro Leu Leu Leu Ala Thr Arg His Leu Val Gly Asp Asp Gly 50 55 60Thr Tyr Val Ser Ala Arg Glu Ile Ser Glu Asn Tyr Gly Val Asp Leu65 70 75 80Glu Leu Leu Gln Arg Val Gln Arg Ala Val Gly Leu Ala Arg Val Asp 85 90 95Asp Pro Asp Ala Val Val His Met Arg Ala Asp Gly Glu Ala Ala Ala 100 105 110Arg Ala Gln Arg Phe Val Glu Leu Gly Leu Asn Pro Asp Gln Val Val 115 120 125Leu Val Val Arg Val Leu Ala Glu Gly Leu Ser His Ala Ala Glu Ala 130 135 140Met Arg Tyr Thr Ala Leu Glu Ala Ile Met Arg Pro Gly Ala Thr Glu145 150 155 160Leu Asp Ile Ala Lys Gly Ser Gln Ala Leu Val Ser Gln Ile Val Pro 165 170 175Leu Leu Gly Pro Met Ile Gln Asp Met Leu Phe Met Gln Leu Arg His 180 185 190Met Met Glu Thr Glu Ala Val Asn Ala Gly Glu Arg Ala Ala Gly Lys 195 200 205Pro Leu Pro Gly Ala Arg Gln Val Thr Val Ala Phe Ala Asp Leu Val 210 215 220Gly Phe Thr Gln Leu Gly Glu Val Val Ser Ala Glu Glu Leu Gly His225 230 235 240Leu Ala Gly Arg Leu Ala Gly Leu Ala Arg Asp Leu Thr Ala Pro Pro 245 250 255Val Trp Phe Ile Lys Thr Ile Gly Asp Ala Val Met Leu Val Cys Pro 260 265 270Asp Pro Ala Pro Leu Leu Asp Thr Val Leu Lys Leu Val Glu Val Val 275 280 285Asp Thr Asp Asn Asn Phe Pro Arg Leu Arg Ala Gly Val Ala Ser Gly 290 295 300Met Ala Val Ser Arg Ala Gly Asp Trp Phe Gly Ser Pro Val Asn Val305 310 315 320Ala Ser Arg Val Thr Gly Val Ala Arg Pro Gly Ala Val Leu Val Ala 325 330 335Asp Ser Val Arg Glu Ala Leu Gly Asp Ala Pro Glu Ala Asp Gly Phe 340 345 350Gln Trp Ser Phe Ala Gly Pro Arg Arg Leu Arg Gly Ile Arg Gly Asp 355 360 365Val Arg Leu Phe Arg Val Arg Arg Gly Ala Thr Arg Thr Gly Ser Gly 370 375 380Gly Ala Ala Gln Asp Asp Asp Leu Ala Gly Ser Ser Pro385 390 3958446PRTMycobacterium tuberculosis 8Met Val Glu Pro Gly Asn Leu Ala Gly Ala Thr Gly Ala Glu Trp Ile1 5 10 15Gly Arg Pro Pro His Glu Glu Leu Gln Arg Lys Val Arg Pro Leu Leu 20 25 30Pro Ser Asp Asp Pro Phe Tyr Phe Pro Pro Ala Gly Tyr Gln His Ala 35 40 45Val Pro Gly Thr Val Leu Arg Ser Arg Asp Val Glu Leu Ala Phe Met 50 55 60Gly Leu Ile Pro Gln Pro Val Thr Ala Thr Gln Leu Leu Tyr Arg Thr65 70 75 80Thr Asn Met Tyr Gly Asn Pro Glu Ala Thr Val Thr Thr Val Ile Val 85 90 95Pro Ala Glu Leu Ala Pro Gly Gln Thr Cys Pro Leu Leu Ser Tyr Gln 100 105 110Cys Ala Ile Asp Ala Met Ser Ser Arg Cys Phe Pro Ser Tyr Ala Leu 115 120 125Arg Arg Arg Ala Lys Ala Leu Gly Ser Leu Thr Gln Met Glu Leu Leu 130 135 140Met Ile Ser Ala Ala Leu Ala Glu Gly Trp Ala Val Ser Val Pro Asp145 150 155 160His Glu Gly Pro Lys Gly Leu Trp Gly Ser Pro Tyr Glu Pro Gly Tyr 165 170 175Arg Val Leu Asp Gly Ile Arg Ala Ala Leu Asn Ser Glu Arg Val Gly 180 185 190Leu Ser Pro Ala Thr Pro Ile Gly Leu Trp Gly Tyr Ser Gly Gly Gly 195 200 205Leu Ala Ser Ala Trp Ala Ala Glu Ala Cys Gly Glu Tyr Ala Pro Asp 210 215 220Leu Asp Ile Val Gly Ala Val Leu Gly Ser Pro Val Gly Asp Leu Gly225 230 235 240His Thr Phe Arg Arg Leu Asn Gly Thr Leu Leu Ala Gly Leu Pro Ala 245 250 255Leu Val Val Ala Ala Leu Gln His Ser Tyr Pro Gly Leu Ala Arg Val 260 265 270Ile Lys Glu His Ala Asn Asp Glu Gly Arg Gln Leu Leu Glu Gln Leu 275 280 285Thr Glu Met Thr Thr Val Asp Ala Val Ile Arg Met Ala Gly Arg Asp 290 295 300Met Gly Asp Phe Leu Asp Glu Pro Leu Glu Asp Ile Leu Ser Thr Pro305 310 315 320Glu Ile Ser His Val Phe Gly Asp Thr Lys Leu Gly Ser Ala Val Pro 325 330 335Thr Pro Pro Val Leu Ile Val Gln Ala Val His Asp Tyr Leu Ile Asp 340 345 350Val Ser Asp Ile Asp Ala Leu Ala Asp Ser Tyr Thr Ala Gly Gly Ala 355 360 365Asn Val Thr Tyr His Arg Asp Leu Phe Ser Glu His Val Ser Leu His 370 375 380Pro Leu Ser Ala Pro Met Thr Leu Arg Trp Leu Thr Asp Arg Phe Ala385 390 395 400Gly Lys Pro Leu Thr Asp His Arg Val Arg Thr Thr Trp Pro Thr Ile 405 410 415Phe Asn Pro Met Thr Tyr Ala Gly Met Ala Arg Leu Ala Val Ile Ala 420 425 430Ala Lys Val Ile Thr Gly Arg Lys Leu Ser Arg Arg Pro Leu 435 440 4459210PRTMycobacterium tuberculosis 9Met Ile Ala Thr Thr Arg Asp Arg Glu Gly Ala Thr Met Ile Thr Phe1 5 10 15Arg Leu Arg Leu Pro Cys Arg Thr Ile Leu Arg Val Phe Ser Arg Asn 20 25 30Pro Leu Val Arg Gly Thr Asp Arg Leu Glu Ala Val Val Met Leu Leu 35 40 45Ala Val Thr Val Ser Leu Leu Thr Ile Pro Phe Ala Ala Ala Ala Gly 50 55 60Thr Ala Val Gln Asp Ser Arg Ser His Val Tyr Ala His Gln Ala Gln65 70 75 80Thr Arg His Pro Ala Thr Ala Thr Val Ile Asp His Glu Gly Val Ile 85 90 95Asp Ser Asn Thr Thr Ala Thr Ser Ala Pro Pro Arg Thr Lys Ile Thr 100 105 110Val Pro Ala Arg Trp Val Val Asn Gly Ile Glu Arg Ser Gly Glu Val 115 120 125Asn Ala Lys Pro Gly Thr Lys Ser Gly Asp Arg Val Gly Ile Trp Val 130 135 140Asp Ser Ala Gly Gln Leu Val Asp Glu Pro Ala Pro Pro Ala Arg Ala145 150 155 160Ile Ala Asp Ala Ala Leu Ala Ala Leu Gly Leu Trp Leu Ser Val Ala 165 170 175Ala Val Ala Gly Ala Leu Leu Ala Leu Thr Arg Ala Ile Leu Ile Arg 180 185 190Val Arg Asn Ala Ser Trp Gln His Asp Ile Asp Ser Leu Phe Cys Thr 195 200 205Gln Arg 2101080PRTMycobacterium tuberculosis 10Met Thr Asn Val Gly Asp Gln

Gly Val Asp Ala Val Phe Gly Val Ile1 5 10 15Tyr Pro Pro Gln Val Ala Leu Val Ser Phe Gly Lys Pro Ala Gln Arg 20 25 30Val Cys Ala Val Asp Gly Ala Ile His Val Met Thr Thr Val Leu Ala 35 40 45Thr Leu Pro Ala Asp His Gly Cys Ser Asp Asp His Arg Gly Ala Leu 50 55 60Phe Phe Leu Ser Ile Asn Glu Leu Thr Arg Cys Ala Ala Val Thr Gly65 70 75 8011652PRTMycobacterium tuberculosis 11Val Thr Val Thr Pro Arg Thr Gly Ser Arg Ile Glu Glu Leu Leu Ala1 5 10 15Arg Ser Gly Arg Phe Phe Ile Pro Gly Glu Ile Ser Ala Asp Leu Arg 20 25 30Thr Val Thr Arg Arg Gly Gly Arg Asp Gly Asp Val Phe Tyr Arg Asp 35 40 45Arg Trp Ser His Asp Lys Val Val Arg Ser Thr His Gly Val Asn Cys 50 55 60Thr Gly Ser Cys Ser Trp Lys Ile Tyr Val Lys Asp Asp Ile Ile Thr65 70 75 80Trp Glu Thr Gln Glu Thr Asp Tyr Pro Ser Val Gly Pro Asp Arg Pro 85 90 95Glu Tyr Glu Pro Arg Gly Cys Pro Arg Gly Ala Ala Phe Ser Trp Tyr 100 105 110Thr Tyr Ser Pro Thr Arg Val Arg His Pro Tyr Ala Arg Gly Val Leu 115 120 125Val Glu Met Tyr Arg Glu Ala Lys Ala Arg Leu Gly Asp Pro Val Ala 130 135 140Ala Trp Ala Asp Ile Gln Ala Asp Pro Arg Arg Arg Arg Arg Tyr Gln145 150 155 160Arg Ala Arg Gly Lys Gly Gly Leu Val Arg Val Ser Trp Ala Glu Ala 165 170 175Thr Glu Met Ile Ala Ala Ala His Val His Thr Ile Ser Thr Tyr Gly 180 185 190Pro Asp Arg Val Ala Gly Phe Ser Pro Ile Pro Ala Met Ser Met Val 195 200 205Ser His Ala Ala Gly Ser Arg Phe Val Glu Leu Ile Gly Gly Val Met 210 215 220Thr Ser Phe Tyr Asp Trp Tyr Ala Asp Leu Pro Val Ala Ser Pro Gln225 230 235 240Val Phe Gly Asp Gln Thr Asp Val Pro Glu Ser Gly Asp Trp Trp Asp 245 250 255Val Val Trp Gln Cys Ala Ser Val Leu Leu Thr Tyr Pro Asn Ser Arg 260 265 270Gln Leu Gly Thr Ala Glu Glu Leu Leu Ala His Ile Asp Gly Pro Ala 275 280 285Ala Asp Leu Leu Gly Arg Thr Val Ser Glu Leu Arg Arg Ala Asp Pro 290 295 300Leu Thr Ala Ala Thr Arg Tyr Val Asp Thr Phe Asp Leu Arg Gly Arg305 310 315 320Ala Thr Leu Tyr Leu Thr Tyr Trp Thr Ala Gly Asp Thr Arg Asn Arg 325 330 335Gly Arg Glu Met Leu Ala Phe Ala Gln Thr Tyr Arg Ser Thr Asp Val 340 345 350Ala Pro Pro Arg Gly Glu Thr Pro Asp Phe Leu Pro Val Val Leu Glu 355 360 365Phe Ala Ala Thr Val Asp Pro Glu Ala Gly Arg Arg Leu Leu Ser Gly 370 375 380Tyr Arg Val Pro Ile Ala Ala Leu Cys Asn Ala Leu Thr Glu Ala Ala385 390 395 400Leu Pro Tyr Ala His Thr Val Ala Ala Val Cys Arg Thr Gly Asp Met 405 410 415Met Gly Glu Leu Phe Trp Thr Val Val Pro Tyr Val Thr Met Thr Ile 420 425 430Val Ala Val Gly Ser Trp Trp Arg Tyr Arg Tyr Asp Lys Phe Gly Trp 435 440 445Thr Thr Arg Ser Ser Gln Leu Tyr Glu Ser Arg Leu Leu Arg Ile Ala 450 455 460Ser Pro Met Phe His Phe Gly Ile Leu Val Val Ile Val Gly His Gly465 470 475 480Ile Gly Leu Val Ile Pro Gln Ser Trp Thr Gln Ala Ala Gly Leu Ser 485 490 495Glu Gly Ala Tyr His Val Gln Ala Val Val Leu Gly Ser Ile Ala Gly 500 505 510Ile Thr Thr Leu Ala Gly Val Thr Leu Leu Ile Tyr Arg Arg Arg Thr 515 520 525Arg Gly Pro Val Phe Met Ala Thr Thr Val Asn Asp Lys Val Met Tyr 530 535 540Leu Val Leu Val Ala Ala Ile Val Ala Gly Leu Gly Ala Thr Ala Leu545 550 555 560Gly Ser Gly Val Val Gly Glu Ala Tyr Asn Tyr Arg Glu Thr Val Ser 565 570 575Val Trp Phe Arg Ser Val Trp Val Leu Gln Pro Arg Gly Asp Leu Met 580 585 590Ala Glu Ala Pro Leu Tyr Tyr Gln Ile His Val Leu Ile Gly Leu Ala 595 600 605Leu Phe Ala Leu Trp Pro Phe Thr Arg Leu Val His Ala Phe Ser Ala 610 615 620Pro Ile Gly Tyr Leu Phe Arg Pro Tyr Ile Ile Tyr Arg Ser Arg Glu625 630 635 640Glu Leu Val Leu Thr Arg Pro Arg Arg Arg Gly Trp 645 65012395PRTMycobacterium tuberculosis 12Met Arg Gly Gln Ala Ala Asn Leu Val Leu Ala Thr Trp Ile Ser Val1 5 10 15Val Asn Phe Trp Ala Trp Asn Leu Ile Gly Pro Leu Ser Thr Ser Tyr 20 25 30Ala Arg Asp Met Ser Leu Ser Ser Ala Glu Ala Ser Leu Leu Val Ala 35 40 45Thr Pro Ile Leu Val Gly Ala Leu Gly Arg Ile Val Thr Gly Pro Leu 50 55 60Thr Asp Arg Phe Gly Gly Arg Ala Met Leu Ile Ala Val Thr Leu Ala65 70 75 80Ser Ile Leu Pro Val Leu Ala Val Gly Val Ala Ala Thr Met Gly Ser 85 90 95Tyr Ala Leu Leu Val Phe Phe Gly Leu Phe Leu Gly Val Ala Gly Thr 100 105 110Ile Phe Ala Val Gly Ile Pro Phe Ala Asn Asn Trp Tyr Gln Pro Ala 115 120 125Arg Arg Gly Phe Ser Thr Gly Val Phe Gly Met Gly Met Val Gly Thr 130 135 140Ala Leu Ser Ala Phe Phe Thr Pro Arg Phe Val Arg Trp Phe Gly Leu145 150 155 160Phe Thr Thr His Ala Ile Val Ala Ala Ala Leu Ala Ser Thr Ala Val 165 170 175Val Ala Met Val Val Leu Arg Asp Ala Pro Tyr Phe Arg Pro Asn Ala 180 185 190Asp Pro Val Leu Pro Arg Leu Lys Ala Ala Ala Arg Leu Pro Val Thr 195 200 205Trp Glu Met Ser Phe Leu Tyr Ala Ile Val Phe Gly Gly Phe Val Ala 210 215 220Phe Ser Asn Tyr Leu Pro Thr Tyr Ile Thr Thr Ile Tyr Gly Phe Ser225 230 235 240Thr Val Asp Ala Gly Ala Arg Thr Ala Gly Phe Ala Leu Ala Ala Val 245 250 255Leu Ala Arg Pro Val Gly Gly Trp Leu Ser Asp Arg Ile Ala Pro Arg 260 265 270His Val Val Leu Ala Ser Leu Ala Gly Thr Ala Leu Leu Ala Phe Ala 275 280 285Ala Ala Leu Gln Pro Pro Pro Glu Val Trp Ser Ala Ala Thr Phe Ile 290 295 300Thr Leu Ala Val Cys Leu Gly Val Gly Thr Gly Gly Val Phe Ala Trp305 310 315 320Val Ala Arg Arg Ala Pro Ala Ala Ser Val Gly Ser Val Thr Gly Ile 325 330 335Val Ala Ala Ala Gly Gly Leu Gly Gly Tyr Phe Pro Pro Leu Val Met 340 345 350Gly Ala Thr Tyr Asp Pro Val Asp Asn Asp Tyr Thr Val Gly Leu Leu 355 360 365Leu Leu Val Ala Thr Ala Leu Val Ala Cys Thr Tyr Thr Ala Leu His 370 375 380Ala Arg Glu Pro Val Ser Glu Glu Ala Ser Arg385 390 3951394PRTMycobacterium tuberculosis 13Met Cys Gly Asp Gln Ser Asp His Val Leu Gln His Trp Thr Val Asp1 5 10 15Ile Ser Ile Asp Glu His Glu Gly Leu Thr Arg Ala Lys Ala Arg Leu 20 25 30Arg Trp Arg Glu Lys Glu Leu Val Gly Val Gly Leu Ala Arg Leu Asn 35 40 45Pro Ala Asp Arg Asn Val Pro Glu Ile Gly Asp Glu Leu Ser Val Ala 50 55 60Arg Ala Leu Ser Asp Leu Gly Lys Arg Met Leu Lys Val Ser Thr His65 70 75 80Asp Ile Glu Ala Val Thr His Gln Pro Ala Arg Leu Leu Tyr 85 9014560PRTMycobacterium tuberculosis 14Met Ile Pro Thr Met Thr Ser Ala Gly Trp Ala Pro Gly Val Val Gln1 5 10 15Phe Arg Glu Tyr Gln Arg Arg Trp Leu Arg Gly Asp Val Leu Ala Gly 20 25 30Leu Thr Val Ala Ala Tyr Leu Ile Pro Gln Ala Met Ala Tyr Ala Thr 35 40 45Val Ala Gly Leu Pro Pro Ala Ala Gly Leu Trp Ala Ser Ile Ala Pro 50 55 60Leu Ala Ile Tyr Ala Leu Leu Gly Ser Ser Arg Gln Leu Ser Ile Gly65 70 75 80Pro Glu Ser Ala Thr Ala Leu Met Thr Ala Ala Val Leu Ala Pro Met 85 90 95Ala Ala Gly Asp Leu Arg Arg Tyr Ala Val Leu Ala Ala Thr Leu Gly 100 105 110Leu Leu Val Gly Leu Ile Cys Leu Leu Ala Gly Thr Ala Arg Leu Gly 115 120 125Phe Leu Ala Ser Leu Arg Ser Arg Pro Val Leu Val Gly Tyr Met Ala 130 135 140Gly Ile Ala Leu Val Met Ile Ser Ser Gln Leu Gly Thr Ile Thr Gly145 150 155 160Thr Ser Val Glu Gly Asn Glu Phe Phe Ser Glu Val His Ser Phe Ala 165 170 175Thr Ser Val Thr Arg Val His Trp Pro Thr Phe Val Leu Ala Met Ser 180 185 190Val Leu Ala Leu Leu Thr Met Leu Thr Arg Trp Ala Pro Arg Ala Pro 195 200 205Gly Pro Ile Ile Ala Val Leu Ala Ala Thr Met Leu Val Ala Val Met 210 215 220Ser Leu Asp Ala Lys Gly Ile Ala Ile Val Gly Arg Ile Pro Ser Gly225 230 235 240Leu Pro Thr Pro Gly Val Pro Pro Val Ser Val Glu Asp Leu Arg Ala 245 250 255Leu Ile Ile Pro Ala Ala Gly Ile Ala Ile Val Thr Phe Thr Asp Gly 260 265 270Val Leu Thr Ala Arg Ala Phe Ala Ala Arg Arg Gly Gln Glu Val Asn 275 280 285Ala Asn Ala Glu Leu Arg Ala Val Gly Ala Cys Asn Ile Ala Ala Gly 290 295 300Leu Thr His Gly Phe Pro Val Ser Ser Ser Ser Ser Arg Thr Ala Leu305 310 315 320Ala Asp Val Val Gly Gly Arg Thr Gln Leu Tyr Ser Leu Ile Ala Leu 325 330 335Gly Leu Val Val Ile Val Met Val Phe Ala Ser Gly Leu Leu Ala Met 340 345 350Phe Pro Ile Ala Ala Leu Gly Ala Leu Val Val Tyr Ala Ala Leu Arg 355 360 365Leu Ile Asp Leu Ser Glu Phe Arg Arg Leu Ala Arg Phe Arg Arg Ser 370 375 380Glu Leu Met Leu Ala Leu Ala Thr Thr Ala Ala Val Leu Gly Leu Gly385 390 395 400Val Phe Tyr Gly Val Leu Ala Ala Val Ala Leu Ser Ile Leu Glu Leu 405 410 415Leu Arg Arg Val Ala His Pro His Asp Ser Val Leu Gly Phe Val Pro 420 425 430Gly Ile Ala Gly Met His Asp Ile Asp Asp Tyr Pro Gln Ala Lys Arg 435 440 445Val Pro Gly Leu Val Val Tyr Arg Tyr Asp Ala Pro Leu Cys Phe Ala 450 455 460Asn Ala Glu Asp Phe Arg Arg Arg Ala Leu Thr Val Val Asp Gln Asp465 470 475 480Pro Gly Gln Val Glu Trp Phe Val Leu Asn Ala Glu Ser Asn Val Glu 485 490 495Val Asp Leu Thr Ala Leu Asp Ala Leu Asp Gln Leu Arg Thr Glu Leu 500 505 510Leu Arg Arg Gly Ile Val Phe Ala Met Ala Arg Val Lys Gln Asp Leu 515 520 525Arg Glu Ser Leu Arg Ala Ala Ser Leu Leu Asp Lys Ile Gly Glu Asp 530 535 540His Ile Phe Met Thr Leu Pro Thr Ala Val Gln Ala Phe Arg Arg Arg545 550 555 56015143PRTMycobacterium tuberculosis 15Met Ile Thr Asn Leu Arg Arg Arg Thr Ala Met Ala Ala Ala Gly Leu1 5 10 15Gly Ala Ala Leu Gly Leu Gly Ile Leu Leu Val Pro Thr Val Asp Ala 20 25 30His Leu Ala Asn Gly Ser Met Ser Glu Val Met Met Ser Glu Ile Ala 35 40 45Gly Leu Pro Ile Pro Pro Ile Ile His Tyr Gly Ala Ile Ala Tyr Ala 50 55 60Pro Ser Gly Ala Ser Gly Lys Ala Trp His Gln Arg Thr Pro Ala Arg65 70 75 80Ala Glu Gln Val Ala Leu Glu Lys Cys Gly Asp Lys Thr Cys Lys Val 85 90 95Val Ser Arg Phe Thr Arg Cys Gly Ala Val Ala Tyr Asn Gly Ser Lys 100 105 110Tyr Gln Gly Gly Thr Gly Leu Thr Arg Arg Ala Ala Glu Asp Asp Ala 115 120 125Val Asn Arg Leu Glu Gly Gly Arg Ile Val Asn Trp Ala Cys Asn 130 135 14016905PRTMycobacterium tuberculosis 16Leu Ser Ala Ser Val Ser Ala Thr Thr Ala His His Gly Leu Pro Ala1 5 10 15His Glu Val Val Leu Leu Leu Glu Ser Asp Pro Tyr His Gly Leu Ser 20 25 30Asp Gly Glu Ala Ala Gln Arg Leu Glu Arg Phe Gly Pro Asn Thr Leu 35 40 45Ala Val Val Thr Arg Ala Ser Leu Leu Ala Arg Ile Leu Arg Gln Phe 50 55 60His His Pro Leu Ile Tyr Val Leu Leu Val Ala Gly Thr Ile Thr Ala65 70 75 80Gly Leu Lys Glu Phe Val Asp Ala Ala Val Ile Phe Gly Val Val Val 85 90 95Ile Asn Ala Ile Val Gly Phe Ile Gln Glu Ser Lys Ala Glu Ala Ala 100 105 110Leu Gln Gly Leu Arg Ser Met Val His Thr His Ala Lys Val Val Arg 115 120 125Glu Gly His Glu His Thr Met Pro Ser Glu Glu Leu Val Pro Gly Asp 130 135 140Leu Val Leu Leu Ala Ala Gly Asp Lys Val Pro Ala Asp Leu Arg Leu145 150 155 160Val Arg Gln Thr Gly Leu Ser Val Asn Glu Ser Ala Leu Thr Gly Glu 165 170 175Ser Thr Pro Val His Lys Asp Glu Val Ala Leu Pro Glu Gly Thr Pro 180 185 190Val Ala Asp Arg Arg Asn Ile Ala Tyr Ser Gly Thr Leu Val Thr Ala 195 200 205Gly His Gly Ala Gly Ile Val Val Ala Thr Gly Ala Glu Thr Glu Leu 210 215 220Gly Glu Ile His Arg Leu Val Gly Ala Ala Glu Val Val Ala Thr Pro225 230 235 240Leu Thr Ala Lys Leu Ala Trp Phe Ser Lys Phe Leu Thr Ile Ala Ile 245 250 255Leu Gly Leu Ala Ala Leu Thr Phe Gly Val Gly Leu Leu Arg Arg Gln 260 265 270Asp Ala Val Glu Thr Phe Thr Ala Ala Ile Ala Leu Ala Val Gly Ala 275 280 285Ile Pro Glu Gly Leu Pro Thr Ala Val Thr Ile Thr Leu Ala Ile Gly 290 295 300Met Ala Arg Met Ala Lys Arg Arg Ala Val Ile Arg Arg Leu Pro Ala305 310 315 320Val Glu Thr Leu Gly Ser Thr Thr Val Ile Cys Ala Asp Lys Thr Gly 325 330 335Thr Leu Thr Glu Asn Gln Met Thr Val Gln Ser Ile Trp Thr Pro His 340 345 350Gly Glu Ile Arg Ala Thr Gly Thr Gly Tyr Ala Pro Asp Val Leu Leu 355 360 365Cys Asp Thr Asp Asp Ala Pro Val Pro Val Asn Ala Asn Ala Ala Leu 370 375 380Arg Trp Ser Leu Leu Ala Gly Ala Cys Ser Asn Asp Ala Ala Leu Val385 390 395 400Arg Asp Gly Thr Arg Trp Gln Ile Val Gly Asp Pro Thr Glu Gly Ala 405 410 415Met Leu Val Val Ala Ala Lys Ala Gly Phe Asn Pro Glu Arg Leu Ala 420 425 430Thr Thr Leu Pro Gln Val Ala Ala Ile Pro Phe Ser Ser Glu Arg Gln 435 440 445Tyr Met Ala Thr Leu His Arg Asp Gly Thr Asp His Val Val Leu Ala 450 455 460Lys Gly Ala Val Glu Arg Met Leu Asp Leu Cys Gly Thr Glu Met Gly465 470 475 480Ala Asp Gly Ala Leu Arg Pro Leu Asp Arg Ala Thr Val Leu Arg Ala 485 490 495Thr Glu Met Leu Thr Ser Arg Gly Leu Arg Val Leu Ala Thr Gly Met 500 505 510Gly Ala Gly Ala Gly Thr Pro Asp Asp Phe Asp Glu Asn Val Ile Pro 515 520 525Gly

Ser Leu Ala Leu Thr Gly Leu Gln Ala Met Ser Asp Pro Pro Arg 530 535 540Ala Ala Ala Ala Ser Ala Val Ala Ala Cys His Ser Ala Gly Ile Ala545 550 555 560Val Lys Met Ile Thr Gly Asp His Ala Gly Thr Ala Thr Ala Ile Ala 565 570 575Thr Glu Val Gly Leu Leu Asp Asn Thr Glu Pro Ala Ala Gly Ser Val 580 585 590Leu Thr Gly Ala Glu Leu Ala Ala Leu Ser Ala Asp Gln Tyr Pro Glu 595 600 605Ala Val Asp Thr Ala Ser Val Phe Ala Arg Val Ser Pro Glu Gln Lys 610 615 620Leu Arg Leu Val Gln Ala Leu Gln Ala Arg Gly His Val Val Ala Met625 630 635 640Thr Gly Asp Gly Val Asn Asp Ala Pro Ala Leu Arg Gln Ala Asn Ile 645 650 655Gly Val Ala Met Gly Arg Gly Gly Thr Glu Val Ala Lys Asp Ala Ala 660 665 670Asp Met Val Leu Thr Asp Asp Asp Phe Ala Thr Ile Glu Ala Ala Val 675 680 685Glu Glu Gly Arg Gly Val Phe Asp Asn Leu Thr Lys Phe Ile Thr Trp 690 695 700Thr Leu Pro Thr Asn Leu Gly Glu Gly Leu Val Ile Leu Ala Ala Ile705 710 715 720Ala Val Gly Val Ala Leu Pro Ile Leu Pro Thr Gln Ile Leu Trp Ile 725 730 735Asn Met Thr Thr Ala Ile Ala Leu Gly Leu Met Leu Ala Phe Glu Pro 740 745 750Lys Glu Ala Gly Ile Met Thr Arg Pro Pro Arg Asp Pro Asp Gln Pro 755 760 765Leu Leu Thr Gly Trp Leu Val Arg Arg Thr Leu Leu Val Ser Thr Leu 770 775 780Leu Val Ala Ser Ala Trp Trp Leu Phe Ala Trp Glu Leu Asp Asn Gly785 790 795 800Ala Gly Leu His Glu Ala Arg Thr Ala Ala Leu Asn Leu Phe Val Val 805 810 815Val Glu Ala Phe Tyr Leu Phe Ser Cys Arg Ser Leu Thr Arg Ser Ala 820 825 830Trp Arg Leu Gly Met Phe Ala Asn Arg Trp Ile Ile Leu Gly Val Ser 835 840 845Ala Gln Ala Ile Ala Gln Phe Ala Ile Thr Tyr Leu Pro Ala Met Asn 850 855 860Met Val Phe Asp Thr Ala Pro Ile Asp Ile Gly Val Trp Val Arg Ile865 870 875 880Phe Ala Val Ala Thr Ala Ile Thr Ile Val Val Ala Thr Asp Thr Leu 885 890 895Leu Pro Arg Ile Arg Ala Gln Pro Pro 900 90517258PRTMycobacterium tuberculosis 17Met Ser Phe His Asp Leu His His Gln Gly Val Pro Phe Val Leu Pro1 5 10 15Asn Ala Trp Asp Val Pro Ser Ala Leu Ala Tyr Leu Ala Glu Gly Phe 20 25 30Thr Ala Ile Gly Thr Thr Ser Phe Gly Val Ser Ser Ser Gly Gly His 35 40 45Pro Asp Gly His Arg Ala Thr Arg Gly Ala Asn Ile Ala Leu Ala Ala 50 55 60Ala Leu Ala Pro Leu Gln Cys Tyr Val Ser Val Asp Ile Glu Asp Gly65 70 75 80Tyr Ser Asp Glu Pro Asp Ala Ile Ala Asp Tyr Val Ala Gln Leu Ser 85 90 95Thr Ala Gly Ile Asn Ile Glu Asp Ser Ser Ala Glu Lys Leu Ile Asp 100 105 110Pro Ala Leu Ala Ala Ala Lys Ile Val Ala Ile Lys Gln Arg Asn Pro 115 120 125Glu Val Phe Val Asn Ala Arg Val Asp Thr Tyr Trp Leu Arg Gln His 130 135 140Ala Asp Thr Thr Ser Thr Ile Gln Arg Ala Leu Arg Tyr Val Asp Ala145 150 155 160Gly Ala Asp Gly Val Phe Val Pro Leu Ala Asn Asp Pro Asp Glu Leu 165 170 175Ala Glu Leu Thr Arg Asn Ile Pro Cys Pro Val Asn Thr Leu Pro Val 180 185 190Pro Gly Leu Thr Ile Ala Asp Leu Gly Glu Leu Gly Val Ala Arg Val 195 200 205Ser Thr Gly Ser Val Pro Tyr Ser Ala Gly Leu Tyr Ala Ala Ala His 210 215 220Ala Ala Arg Ala Val Ser Asp Gly Glu Gln Leu Pro Arg Ser Val Pro225 230 235 240Tyr Ala Glu Leu Gln Ala Arg Leu Val Asp Tyr Glu Asn Arg Thr Ser 245 250 255Thr Thr18285PRTMycobacterium tuberculosis 18Val Val Lys Arg Ser Arg Ala Thr Arg Leu Ser Pro Ser Ile Trp Ser1 5 10 15Gly Trp Glu Ser Pro Gln Cys Arg Ser Ile Arg Ala Arg Leu Leu Leu 20 25 30Pro Arg Gly Arg Ser Arg Pro Pro Asn Ala Asp Cys Cys Trp Asn Gln 35 40 45Leu Ala Val Thr Pro Asp Thr Arg Met Pro Ala Ser Ser Ala Ala Gly 50 55 60Arg Asp Ala Ala Ala Tyr Asp Ala Trp Tyr Asp Ser Pro Thr Gly Arg65 70 75 80Pro Ile Leu Ala Thr Glu Val Ala Ala Leu Arg Pro Leu Ile Glu Val 85 90 95Phe Ala Gln Pro Arg Leu Glu Ile Gly Val Gly Thr Gly Arg Phe Ala 100 105 110Asp Leu Leu Gly Val Arg Phe Gly Leu Asp Pro Ser Arg Asp Ala Leu 115 120 125Met Phe Ala Arg Arg Arg Gly Val Leu Val Ala Asn Ala Val Gly Glu 130 135 140Ala Val Pro Phe Val Ser Arg His Phe Gly Ala Val Leu Met Ala Phe145 150 155 160Thr Leu Cys Phe Val Thr Asp Pro Ala Ala Ile Phe Arg Glu Thr Arg 165 170 175Arg Leu Leu Ala Asp Gly Gly Gly Leu Val Ile Gly Phe Leu Pro Arg 180 185 190Gly Thr Pro Trp Ala Asp Leu Tyr Ala Leu Arg Ala Ala Arg Gly Gln 195 200 205Pro Gly Tyr Arg Asp Ala Arg Phe Tyr Thr Ala Ala Glu Leu Glu Gln 210 215 220Leu Leu Ala Asp Ser Gly Phe Arg Val Ile Ala Arg Arg Cys Thr Leu225 230 235 240His Gln Pro Pro Gly Leu Ala Arg Tyr Asp Ile Glu Ala Ala His Asp 245 250 255Gly Ile Gln Ala Gly Ala Gly Phe Val Ala Ile Ser Ala Val Asp Gln 260 265 270Ala His Glu Pro Lys Asp Asp His Pro Leu Glu Ser Glu 275 280 28519285PRTMycobacterium tuberculosis 19Val Val Lys Arg Ser Arg Ala Thr Arg Leu Ser Pro Ser Ile Trp Ser1 5 10 15Gly Trp Glu Ser Pro Gln Cys Arg Ser Ile Arg Ala Arg Leu Leu Leu 20 25 30Pro Arg Gly Arg Ser Arg Pro Pro Asn Ala Asp Cys Cys Trp Asn Gln 35 40 45Leu Ala Val Thr Pro Asp Thr Arg Met Pro Ala Ser Ser Ala Ala Gly 50 55 60Arg Asp Ala Ala Ala Tyr Asp Ala Trp Tyr Asp Ser Pro Thr Gly Arg65 70 75 80Pro Ile Leu Ala Thr Glu Val Ala Ala Leu Arg Pro Leu Ile Glu Val 85 90 95Phe Ala Gln Pro Arg Leu Glu Ile Gly Val Gly Thr Gly Arg Phe Ala 100 105 110Asp Leu Leu Gly Val Arg Phe Gly Leu Asp Pro Ser Arg Asp Ala Leu 115 120 125Met Phe Ala Arg Arg Arg Gly Val Leu Val Ala Asn Ala Val Gly Glu 130 135 140Ala Val Pro Phe Val Ser Arg His Phe Gly Ala Val Leu Met Ala Phe145 150 155 160Thr Leu Cys Phe Val Thr Asp Pro Ala Ala Ile Phe Arg Glu Thr Arg 165 170 175Arg Leu Leu Ala Asp Gly Gly Gly Leu Val Ile Gly Phe Leu Pro Arg 180 185 190Gly Thr Pro Trp Ala Asp Leu Tyr Ala Leu Arg Ala Ala Arg Gly Gln 195 200 205Pro Gly Tyr Arg Asp Ala Arg Phe Tyr Thr Ala Ala Glu Leu Glu Gln 210 215 220Leu Leu Ala Asp Ser Gly Phe Arg Val Ile Ala Arg Arg Cys Thr Leu225 230 235 240His Gln Pro Pro Gly Leu Ala Arg Tyr Asp Ile Glu Ala Ala His Asp 245 250 255Gly Ile Gln Ala Gly Ala Gly Phe Val Ala Ile Ser Ala Val Asp Gln 260 265 270Ala His Glu Pro Lys Asp Asp His Pro Leu Glu Ser Glu 275 280 28520114PRTMycobacterium tuberculosis 20Val Thr Tyr Val Ile Gly Ser Glu Cys Val Asp Val Met Asp Lys Ser1 5 10 15Cys Val Gln Glu Cys Pro Val Asp Cys Ile Tyr Glu Gly Ala Arg Met 20 25 30Leu Tyr Ile Asn Pro Asp Glu Cys Val Asp Cys Gly Ala Cys Lys Pro 35 40 45Ala Cys Arg Val Glu Ala Ile Tyr Trp Glu Gly Asp Leu Pro Asp Asp 50 55 60Gln His Gln His Leu Gly Asp Asn Ala Ala Phe Phe His Gln Val Leu65 70 75 80Pro Gly Arg Val Ala Pro Leu Gly Ser Pro Gly Gly Ala Ala Ala Val 85 90 95Gly Pro Ile Gly Val Asp Thr Pro Leu Val Ala Ala Ile Pro Val Glu 100 105 110Cys Pro21279PRTMycobacterium tuberculosis 21Met Asn Gln Ser His Lys Pro Pro Ser Ile Val Val Gly Ile Asp Gly1 5 10 15Ser Lys Pro Ala Val Gln Ala Ala Leu Trp Ala Val Asp Glu Ala Ala 20 25 30Ser Arg Asp Ile Pro Leu Arg Leu Leu Tyr Ala Ile Glu Pro Asp Asp 35 40 45Pro Gly Tyr Ala Ala His Gly Ala Ala Ala Arg Lys Leu Ala Ala Ala 50 55 60Glu Asn Ala Val Arg Tyr Ala Phe Thr Ala Val Glu Ala Ala Asp Arg65 70 75 80Pro Val Lys Val Glu Val Glu Ile Thr Gln Glu Arg Pro Val Thr Ser 85 90 95Leu Ile Arg Ala Ser Ala Ala Ala Ala Leu Val Cys Val Gly Ala Ile 100 105 110Gly Val His His Phe Arg Pro Glu Arg Val Gly Ser Thr Ala Ala Ala 115 120 125Leu Ala Leu Ser Ala Gln Cys Pro Val Ala Ile Val Arg Pro His Arg 130 135 140Val Pro Ile Gly Arg Asp Ala Ala Trp Ile Val Val Glu Ala Asp Gly145 150 155 160Ser Ser Asp Ile Gly Val Leu Leu Gly Ala Val Met Ala Glu Ala Arg 165 170 175Leu Arg Asp Ser Pro Val Arg Val Val Thr Cys Arg Gln Ser Gly Val 180 185 190Gly Asp Thr Gly Asp Asp Val Arg Ala Ser Leu Asp Arg Trp Leu Ala 195 200 205Arg Trp Gln Pro Arg Tyr Pro Asp Val Arg Val Gln Ser Ala Ala Val 210 215 220His Gly Glu Leu Leu Asp Tyr Leu Ala Gly Leu Gly Arg Ser Val His225 230 235 240Met Val Val Leu Ser Ala Ser Asp Gln Glu His Val Glu Gln Leu Val 245 250 255Gly Ala Pro Gly Asn Ala Val Leu Gln Glu Ala Gly Cys Thr Leu Leu 260 265 270Val Val Gly Gln Gln Tyr Leu 27522339PRTMycobacterium tuberculosis 22Met Thr Glu Pro Ala Ala Trp Asp Glu Gly Lys Pro Arg Ile Ile Thr1 5 10 15Leu Thr Met Asn Pro Ala Leu Asp Ile Thr Thr Ser Val Asp Val Val 20 25 30Arg Pro Thr Glu Lys Met Arg Cys Gly Ala Pro Arg Tyr Asp Pro Gly 35 40 45Gly Gly Gly Ile Asn Val Ala Arg Ile Val His Val Leu Gly Gly Cys 50 55 60Ser Thr Ala Leu Phe Pro Ala Gly Gly Ser Thr Gly Ser Leu Leu Met65 70 75 80Ala Leu Leu Gly Asp Ala Gly Val Pro Phe Arg Val Ile Pro Ile Ala 85 90 95Ala Ser Thr Arg Glu Ser Phe Thr Val Asn Glu Ser Arg Thr Ala Lys 100 105 110Gln Tyr Arg Phe Val Leu Pro Gly Pro Ser Leu Thr Val Ala Glu Gln 115 120 125Glu Gln Cys Leu Asp Glu Leu Arg Gly Ala Ala Ala Ser Ala Ala Phe 130 135 140Val Val Ala Ser Gly Ser Leu Pro Pro Gly Val Ala Ala Asp Tyr Tyr145 150 155 160Gln Arg Val Ala Asp Ile Cys Arg Arg Ser Ser Thr Pro Leu Ile Leu 165 170 175Asp Thr Ser Gly Gly Gly Leu Gln His Ile Ser Ser Gly Val Phe Leu 180 185 190Leu Lys Ala Ser Val Arg Glu Leu Arg Glu Cys Val Gly Ser Glu Leu 195 200 205Leu Thr Glu Pro Glu Gln Leu Ala Ala Ala His Glu Leu Ile Asp Arg 210 215 220Gly Arg Ala Glu Val Val Val Val Ser Leu Gly Ser Gln Gly Ala Leu225 230 235 240Leu Ala Thr Arg His Ala Ser His Arg Phe Ser Ser Ile Pro Met Thr 245 250 255Ala Val Ser Gly Val Gly Ala Gly Asp Ala Met Val Ala Ala Ile Thr 260 265 270Val Gly Leu Ser Arg Gly Trp Ser Leu Ile Lys Ser Val Arg Leu Gly 275 280 285Asn Ala Ala Gly Ala Ala Met Leu Leu Thr Pro Gly Thr Ala Ala Cys 290 295 300Asn Arg Asp Asp Val Glu Arg Phe Phe Glu Leu Ala Ala Glu Pro Thr305 310 315 320Glu Val Gly Gln Asp Gln Tyr Val Trp His Pro Ile Val Asn Pro Glu 325 330 335Ala Ser Pro23681PRTMycobacterium tuberculosis 23Val Leu Met Thr Ala Ala Ala Asp Val Thr Arg Arg Ser Pro Arg Arg1 5 10 15Val Phe Arg Asp Arg Arg Glu Ala Gly Arg Val Leu Ala Glu Leu Leu 20 25 30Ala Ala Tyr Arg Asp Gln Pro Asp Val Ile Val Leu Gly Leu Ala Arg 35 40 45Gly Gly Leu Pro Val Ala Trp Glu Val Ala Ala Ala Leu His Ala Pro 50 55 60Leu Asp Ala Phe Val Val Arg Lys Leu Gly Ala Pro Gly His Asp Glu65 70 75 80Phe Ala Val Gly Ala Leu Ala Ser Gly Gly Arg Val Val Val Asn Asp 85 90 95Asp Val Val Arg Gly Leu Arg Ile Thr Pro Gln Gln Leu Arg Asp Ile 100 105 110Ala Glu Arg Glu Gly Arg Glu Leu Leu Arg Arg Glu Ser Ala Tyr Arg 115 120 125Gly Glu Arg Pro Pro Thr Asp Ile Thr Gly Lys Thr Val Ile Val Val 130 135 140Asp Asp Gly Leu Ala Thr Gly Ala Ser Met Phe Ala Ala Val Gln Ala145 150 155 160Leu Arg Asp Ala Gln Pro Ala Gln Ile Val Ile Ala Val Pro Ala Ala 165 170 175Pro Glu Ser Thr Cys Arg Glu Phe Ala Gly Leu Val Asp Asp Val Val 180 185 190Cys Ala Thr Met Pro Thr Pro Phe Leu Ala Val Gly Glu Ser Phe Trp 195 200 205Asp Phe Arg Gln Val Thr Asp Glu Glu Val Arg Arg Leu Leu Ala Thr 210 215 220Pro Thr Ala Gly Pro Ser Leu Arg Arg Pro Ala Ala Ser Thr Ala Ala225 230 235 240Asp Val Leu Arg Arg Val Ala Ile Asp Ala Pro Gly Gly Val Pro Thr 245 250 255His Glu Val Leu Ala Glu Leu Val Gly Asp Ala Arg Ile Val Leu Ile 260 265 270Gly Glu Ser Ser His Gly Thr His Glu Phe Tyr Gln Ala Arg Ala Ala 275 280 285Met Thr Gln Trp Leu Ile Glu Glu Lys Gly Phe Gly Ala Val Ala Ala 290 295 300Glu Ala Asp Trp Pro Asp Ala Tyr Arg Val Asn Arg Tyr Val Arg Gly305 310 315 320Leu Gly Glu Asp Thr Asn Ala Asp Glu Ala Leu Ser Gly Phe Glu Arg 325 330 335Phe Pro Ala Trp Met Trp Arg Asn Thr Val Val Arg Asp Phe Val Glu 340 345 350Trp Leu Arg Thr Arg Asn Gln Arg Tyr Glu Ser Gly Ala Leu Arg Gln 355 360 365Ala Gly Phe Tyr Gly Leu Asp Leu Tyr Ser Leu His Arg Ser Ile Gln 370 375 380Glu Val Ile Ser Tyr Leu Asp Lys Val Asp Pro Arg Ala Ala Ala Arg385 390 395 400Ala Arg Ala Arg Tyr Ala Cys Phe Asp His Ala Cys Ala Asp Asp Gly 405 410 415Gln Ala Tyr Gly Phe Ala Ala Ala Phe Gly Ala Gly Pro Ser Cys Glu 420 425 430Arg Glu Ala Val Glu Gln Leu Val Asp Val Gln Arg Asn Ala Leu Ala 435 440 445Tyr Ala Arg Gln Asp Gly Leu Leu Ala Glu Asp Glu Leu Phe Tyr Ala 450 455 460Gln Gln Asn Ala Gln Thr Val Arg Asp Ala Glu Val Tyr Tyr Arg Ala465 470 475 480Met Phe Ser Gly Arg Val Thr Ser Trp Asn Leu Arg Asp Gln His Met 485 490 495Ala Gln Thr Leu Gly Ser Leu Leu Thr His Leu Asp

Arg His Leu Asp 500 505 510Ala Pro Pro Ala Arg Ile Val Val Trp Ala His Asn Ser His Val Gly 515 520 525Asp Ala Arg Ala Thr Glu Val Trp Ala Asp Gly Gln Leu Thr Leu Gly 530 535 540Gln Ile Val Arg Glu Arg Tyr Gly Asp Glu Ser Arg Ser Ile Gly Phe545 550 555 560Ser Thr Tyr Thr Gly Thr Val Thr Ala Ala Ser Glu Trp Gly Gly Ile 565 570 575Ala Gln Arg Lys Ala Val Arg Pro Ala Leu His Gly Ser Val Glu Glu 580 585 590Leu Phe His Gln Thr Ala Asp Ser Phe Leu Val Ser Ala Arg Leu Ser 595 600 605Arg Asp Ala Glu Ala Pro Leu Asp Val Val Arg Leu Gly Arg Ala Ile 610 615 620Gly Val Val Tyr Leu Pro Ala Thr Glu Arg Gln Ser His Tyr Leu His625 630 635 640Val Arg Pro Ala Asp Gln Phe Asp Ala Met Ile His Ile Asp Gln Thr 645 650 655Arg Ala Leu Glu Pro Leu Glu Val Thr Ser Arg Trp Ile Ala Gly Glu 660 665 670Asn Pro Glu Thr Tyr Pro Thr Gly Leu 675 68024144PRTMycobacterium tuberculosis 24Met Ala Thr Thr Leu Pro Val Gln Arg His Pro Arg Ser Leu Phe Pro1 5 10 15Glu Phe Ser Glu Leu Phe Ala Ala Phe Pro Ser Phe Ala Gly Leu Arg 20 25 30Pro Thr Phe Asp Thr Arg Leu Met Arg Leu Glu Asp Glu Met Lys Glu 35 40 45Gly Arg Tyr Glu Val Arg Ala Glu Leu Pro Gly Val Asp Pro Asp Lys 50 55 60Asp Val Asp Ile Met Val Arg Asp Gly Gln Leu Thr Ile Lys Ala Glu65 70 75 80Arg Thr Glu Gln Lys Asp Phe Asp Gly Arg Ser Glu Phe Ala Tyr Gly 85 90 95Ser Phe Val Arg Thr Val Ser Leu Pro Val Gly Ala Asp Glu Asp Asp 100 105 110Ile Lys Ala Thr Tyr Asp Lys Gly Ile Leu Thr Val Ser Val Ala Val 115 120 125Ser Glu Gly Lys Pro Thr Glu Lys His Ile Gln Ile Arg Ser Thr Asn 130 135 14025331PRTMycobacterium tuberculosis 25Met Pro Asp Thr Met Val Thr Thr Asp Val Ile Lys Ser Ala Val Gln1 5 10 15Leu Ala Cys Arg Ala Pro Ser Leu His Asn Ser Gln Pro Trp Arg Trp 20 25 30Ile Ala Glu Asp His Thr Val Ala Leu Phe Leu Asp Lys Asp Arg Val 35 40 45Leu Tyr Ala Thr Asp His Ser Gly Arg Glu Ala Leu Leu Gly Cys Gly 50 55 60Ala Val Leu Asp His Phe Arg Val Ala Met Ala Ala Ala Gly Thr Thr65 70 75 80Ala Asn Val Glu Arg Phe Pro Asn Pro Asn Asp Pro Leu His Leu Ala 85 90 95Ser Ile Asp Phe Ser Pro Ala Asp Phe Val Thr Glu Gly His Arg Leu 100 105 110Arg Ala Asp Ala Ile Leu Leu Arg Arg Thr Asp Arg Leu Pro Phe Ala 115 120 125Glu Pro Pro Asp Trp Asp Leu Val Glu Ser Gln Leu Arg Thr Thr Val 130 135 140Thr Ala Asp Thr Val Arg Ile Asp Val Ile Ala Asp Asp Met Arg Pro145 150 155 160Glu Leu Ala Ala Ala Ser Lys Leu Thr Glu Ser Leu Arg Leu Tyr Asp 165 170 175Ser Ser Tyr His Ala Glu Leu Phe Trp Trp Thr Gly Ala Phe Glu Thr 180 185 190Ser Glu Gly Ile Pro His Ser Ser Leu Val Ser Ala Ala Glu Ser Asp 195 200 205Arg Val Thr Phe Gly Arg Asp Phe Pro Val Val Ala Asn Thr Asp Arg 210 215 220Arg Pro Glu Phe Gly His Asp Arg Ser Lys Val Leu Val Leu Ser Thr225 230 235 240Tyr Asp Asn Glu Arg Ala Ser Leu Leu Arg Cys Gly Glu Met Leu Ser 245 250 255Ala Val Leu Leu Asp Ala Thr Met Ala Gly Leu Ala Thr Cys Thr Leu 260 265 270Thr His Ile Thr Glu Leu His Ala Ser Arg Asp Leu Val Ala Ala Leu 275 280 285Ile Gly Gln Pro Ala Thr Pro Gln Ala Leu Val Arg Val Gly Leu Ala 290 295 300Pro Glu Met Glu Glu Pro Pro Pro Ala Thr Pro Arg Arg Pro Ile Asp305 310 315 320Glu Val Phe His Val Arg Ala Lys Asp His Arg 325 33026195PRTMycobacterium tuberculosis 26Met Pro Leu Leu Thr Ile Gly Asp Gln Phe Pro Ala Tyr Gln Leu Thr1 5 10 15Ala Leu Ile Gly Gly Asp Leu Ser Lys Val Asp Ala Lys Gln Pro Gly 20 25 30Asp Tyr Phe Thr Thr Ile Thr Ser Asp Glu His Pro Gly Lys Trp Arg 35 40 45Val Val Phe Phe Trp Pro Lys Asp Phe Thr Phe Val Cys Pro Thr Glu 50 55 60Ile Ala Ala Phe Ser Lys Leu Asn Asp Glu Phe Glu Asp Arg Asp Ala65 70 75 80Gln Ile Leu Gly Val Ser Ile Asp Ser Glu Phe Ala His Phe Gln Trp 85 90 95Arg Ala Gln His Asn Asp Leu Lys Thr Leu Pro Phe Pro Met Leu Ser 100 105 110Asp Ile Lys Arg Glu Leu Ser Gln Ala Ala Gly Val Leu Asn Ala Asp 115 120 125Gly Val Ala Asp Arg Val Thr Phe Ile Val Asp Pro Asn Asn Glu Ile 130 135 140Gln Phe Val Ser Ala Thr Ala Gly Ser Val Gly Arg Asn Val Asp Glu145 150 155 160Val Leu Arg Val Leu Asp Ala Leu Gln Ser Asp Glu Leu Cys Ala Cys 165 170 175Asn Trp Arg Lys Gly Asp Pro Thr Leu Asp Ala Gly Glu Leu Leu Lys 180 185 190Ala Ser Ala 19527272PRTMycobacterium tuberculosis 27Met Ser Gly Arg Gly Glu Pro Thr Met Lys Thr Ile Ile Val Gly Ile1 5 10 15Asp Gly Ser His Ala Ala Ile Thr Ala Ala Leu Trp Gly Val Asp Glu 20 25 30Ala Ile Ser Arg Ala Val Pro Leu Arg Leu Val Ser Val Ile Lys Pro 35 40 45Thr His Pro Ser Pro Asp Asp Tyr Asp Arg Asp Leu Ala His Ala Glu 50 55 60Arg Ser Leu Arg Glu Ala Gln Ser Ala Val Glu Ala Ala Gly Lys Leu65 70 75 80Val Lys Ile Glu Thr Asp Ile Pro Arg Gly Pro Ala Gly Pro Val Leu 85 90 95Val Glu Ala Ser Arg Asp Ala Glu Met Ile Cys Val Gly Ser Val Gly 100 105 110Ile Gly Arg Tyr Ala Ser Ser Ile Leu Gly Ser Thr Ala Thr Glu Leu 115 120 125Ala Glu Lys Ala His Cys Pro Val Ala Val Met Arg Ser Lys Val Asp 130 135 140Gln Pro Ala Ser Asp Ile Asn Trp Ile Val Val Arg Met Thr Asp Ala145 150 155 160Pro Asp Asn Glu Ala Val Leu Glu Tyr Ala Ala Arg Glu Ala Lys Leu 165 170 175Arg Gln Ala Pro Ile Leu Ala Leu Gly Gly Arg Pro Glu Glu Leu Arg 180 185 190Glu Ile Pro Asp Gly Glu Phe Glu Arg Arg Val Gln Asp Trp His His 195 200 205Arg His Pro Asp Val Arg Val Tyr Pro Ile Thr Thr His Thr Gly Ile 210 215 220Ala Arg Phe Leu Ala Asp His Asp Glu Arg Val Gln Leu Ala Val Ile225 230 235 240Gly Gly Gly Glu Ala Gly Gln Leu Ala Arg Leu Val Gly Pro Ser Gly 245 250 255His Pro Val Phe Arg His Ala Glu Cys Ser Val Leu Val Val Arg Arg 260 265 27028393PRTMycobacterium tuberculosis 28Met Arg Asp Ala Ile Pro Leu Gly Arg Ile Ala Gly Phe Val Val Asn1 5 10 15Val His Trp Ser Val Leu Val Ile Leu Trp Leu Phe Thr Trp Ser Leu 20 25 30Ala Thr Met Leu Pro Gly Thr Val Gly Gly Tyr Pro Ala Val Val Tyr 35 40 45Trp Leu Leu Gly Ala Gly Gly Ala Val Met Leu Leu Ala Ser Leu Leu 50 55 60Ala His Glu Leu Ala His Ala Val Val Ala Arg Arg Ala Gly Val Ser65 70 75 80Val Glu Ser Val Thr Leu Trp Leu Phe Gly Gly Val Thr Ala Leu Gly 85 90 95Gly Glu Ala Lys Thr Pro Lys Ala Ala Phe Arg Ile Ala Phe Ala Gly 100 105 110Pro Ala Thr Ser Leu Ala Leu Ser Ala Thr Phe Gly Ala Leu Ala Ile 115 120 125Thr Leu Ala Gly Val Arg Thr Pro Ala Ile Val Ile Ser Val Ala Trp 130 135 140Trp Leu Ala Thr Val Asn Leu Leu Leu Gly Leu Phe Asn Leu Leu Pro145 150 155 160Gly Ala Pro Leu Asp Gly Gly Arg Leu Val Arg Ala Tyr Leu Trp Arg 165 170 175Arg His Gly Asp Ser Val Arg Ala Gly Ile Gly Ala Ala Arg Ala Gly 180 185 190Arg Val Val Ala Leu Val Leu Ile Ala Leu Gly Leu Ala Glu Phe Val 195 200 205Ala Gly Gly Leu Val Gly Gly Val Trp Leu Ala Phe Ile Gly Trp Phe 210 215 220Ile Phe Ala Ala Ala Arg Glu Glu Glu Thr Arg Ile Ser Thr Gln Gln225 230 235 240Leu Phe Ala Gly Val Arg Val Ala Asp Ala Met Thr Ala Gln Pro His 245 250 255Thr Ala Pro Gly Trp Ile Asn Val Glu Asp Phe Ile Gln Arg Tyr Val 260 265 270Leu Gly Glu Arg His Ser Ala Tyr Pro Val Ala Asp Arg Asp Gly Ser 275 280 285Ile Thr Gly Leu Val Ala Leu Arg Gln Leu Arg Asp Val Ala Pro Ser 290 295 300Arg Arg Ser Thr Thr Ser Val Gly Asp Ile Ala Leu Pro Leu His Ser305 310 315 320Val Pro Thr Ala Arg Pro Gln Glu Pro Leu Thr Ala Leu Leu Glu Arg 325 330 335Met Ala Pro Leu Gly Pro Arg Ser Arg Ala Leu Val Thr Glu Gly Ser 340 345 350Ala Val Val Gly Ile Val Thr Pro Ser Asp Val Ala Arg Leu Ile Asp 355 360 365Val Tyr Arg Leu Ala Gln Pro Glu Pro Thr Phe Thr Thr Ser Pro Gln 370 375 380Asp Ala Asp Arg Phe Ser Asp Ala Gly385 39029413PRTMycobacterium tuberculosis 29Met Ala Ser Ser Ala Ser Asp Gly Thr His Glu Arg Ser Ala Phe Arg1 5 10 15Leu Ser Pro Pro Val Leu Ser Gly Ala Met Gly Pro Phe Met His Thr 20 25 30Gly Leu Tyr Val Ala Gln Ser Trp Arg Asp Tyr Leu Gly Gln Gln Pro 35 40 45Asp Lys Leu Pro Ile Ala Arg Pro Thr Ile Ala Leu Ala Ala Gln Ala 50 55 60Phe Arg Asp Glu Ile Val Leu Leu Gly Leu Lys Ala Arg Arg Pro Val65 70 75 80Ser Asn His Arg Val Phe Glu Arg Ile Ser Gln Glu Val Ala Ala Gly 85 90 95Leu Glu Phe Tyr Gly Asn Arg Arg Trp Leu Glu Lys Pro Ser Gly Phe 100 105 110Phe Ala Gln Pro Pro Pro Leu Thr Glu Val Ala Val Arg Lys Val Lys 115 120 125Asp Arg Arg Arg Ser Phe Tyr Arg Ile Phe Phe Asp Ser Gly Phe Thr 130 135 140Pro His Pro Gly Glu Pro Gly Ser Gln Arg Trp Leu Ser Tyr Thr Ala145 150 155 160Asn Asn Arg Glu Tyr Ala Leu Leu Leu Arg His Pro Glu Pro Arg Pro 165 170 175Trp Leu Val Cys Val His Gly Thr Glu Met Gly Arg Ala Pro Leu Asp 180 185 190Leu Ala Val Phe Arg Ala Trp Lys Leu His Asp Glu Leu Gly Leu Asn 195 200 205Ile Val Met Pro Val Leu Pro Met His Gly Pro Arg Gly Gln Gly Leu 210 215 220Pro Lys Gly Ala Val Phe Pro Gly Glu Asp Val Leu Asp Asp Val His225 230 235 240Gly Thr Ala Gln Ala Val Trp Asp Ile Arg Arg Leu Leu Ser Trp Ile 245 250 255Arg Ser Gln Glu Glu Glu Ser Leu Ile Gly Leu Asn Gly Leu Ser Leu 260 265 270Gly Gly Tyr Ile Ala Ser Leu Val Ala Ser Leu Glu Glu Gly Leu Ala 275 280 285Cys Ala Ile Leu Gly Val Pro Val Ala Asp Leu Ile Glu Leu Leu Gly 290 295 300Arg His Cys Gly Leu Arg His Lys Asp Pro Arg Arg His Thr Val Lys305 310 315 320Met Ala Glu Pro Ile Gly Arg Met Ile Ser Pro Leu Ser Leu Thr Pro 325 330 335Leu Val Pro Met Pro Gly Arg Phe Ile Tyr Ala Gly Ile Ala Asp Arg 340 345 350Leu Val His Pro Arg Glu Gln Val Thr Arg Leu Trp Glu His Trp Gly 355 360 365Lys Pro Glu Ile Val Trp Tyr Pro Gly Gly His Thr Gly Phe Phe Gln 370 375 380Ser Arg Pro Val Arg Arg Phe Val Gln Ala Ala Leu Glu Gln Ser Gly385 390 395 400Leu Leu Asp Ala Pro Arg Thr Gln Arg Asp Arg Ser Ala 405 41030120PRTMycobacterium tuberculosis 30Met Ser Thr Gln Arg Pro Arg His Ser Gly Ile Arg Ala Val Gly Pro1 5 10 15Tyr Ala Trp Ala Gly Arg Cys Gly Arg Ile Gly Arg Trp Gly Val His 20 25 30Gln Glu Ala Met Met Asn Leu Ala Ile Trp His Pro Arg Lys Val Gln 35 40 45Ser Ala Thr Ile Tyr Gln Val Thr Asp Arg Ser His Asp Gly Arg Thr 50 55 60Ala Arg Val Pro Gly Asp Glu Ile Thr Ser Thr Val Ser Gly Trp Leu65 70 75 80Ser Glu Leu Gly Thr Gln Ser Pro Leu Ala Asp Glu Leu Ala Arg Ala 85 90 95Val Arg Ile Gly Asp Trp Pro Ala Ala Tyr Ala Ile Gly Glu His Leu 100 105 110Ser Val Glu Ile Ala Val Ala Val 115 12031374PRTMycobacterium tuberculosis 31Met Arg Ser Glu Arg Leu Arg Trp Leu Val Ala Ala Glu Gly Pro Phe1 5 10 15Ala Ser Val Tyr Phe Asp Asp Ser His Asp Thr Leu Asp Ala Val Glu 20 25 30Arg Arg Glu Ala Thr Trp Arg Asp Val Arg Lys His Leu Glu Ser Arg 35 40 45Asp Ala Lys Gln Glu Leu Ile Asp Ser Leu Glu Glu Ala Val Arg Asp 50 55 60Ser Arg Pro Ala Val Gly Gln Arg Gly Arg Ala Leu Ile Ala Thr Gly65 70 75 80Glu Gln Val Leu Val Asn Glu His Leu Ile Gly Pro Pro Pro Ala Thr 85 90 95Val Ile Arg Leu Ser Asp Tyr Pro Tyr Val Val Pro Leu Ile Asp Leu 100 105 110Glu Met Arg Arg Pro Thr Tyr Val Phe Ala Ala Val Asp His Thr Gly 115 120 125Ala Asp Val Lys Leu Tyr Gln Gly Ala Thr Ile Ser Ser Thr Lys Ile 130 135 140Asp Gly Val Gly Tyr Pro Val His Lys Pro Val Thr Ala Gly Trp Asn145 150 155 160Gly Tyr Gly Asp Phe Gln His Thr Thr Glu Glu Ala Ile Arg Met Asn 165 170 175Cys Arg Ala Val Ala Asp His Leu Thr Arg Leu Val Asp Ala Ala Asp 180 185 190Pro Glu Val Val Phe Val Ser Gly Glu Val Arg Ser Arg Thr Asp Leu 195 200 205Leu Ser Thr Leu Pro Gln Arg Val Ala Val Arg Val Ser Gln Leu His 210 215 220Ala Gly Pro Arg Lys Ser Ala Leu Asp Glu Glu Glu Ile Trp Asp Leu225 230 235 240Thr Ser Ala Glu Phe Thr Arg Arg Arg Tyr Ala Glu Ile Thr Asn Val 245 250 255Ala Gln Gln Phe Glu Ala Glu Ile Gly Arg Gly Ser Gly Leu Ala Ala 260 265 270Gln Gly Leu Ala Glu Val Cys Ala Ala Leu Arg Asp Gly Asp Val Asp 275 280 285Thr Leu Ile Val Gly Glu Leu Gly Glu Ala Thr Val Val Thr Gly Lys 290 295 300Ala Arg Thr Thr Val Ala Arg Asp Ala Asp Met Leu Ser Glu Leu Gly305 310 315 320Glu Pro Val Asp Arg Val Ala Arg Ala Asp Glu Ala Leu Pro Phe Ala 325 330 335Ala Ile Ala Val Gly Ala Ala Leu Val Arg Asp Asp Asn Arg Ile Ala 340 345 350Pro Leu Asp Gly Val Gly Ala Leu Leu Arg Tyr Ala Ala Thr Asn Arg 355 360 365Leu Gly Ser His Arg Ser 37032179PRTMycobacterium tuberculosis 32Met Leu His Arg Asp Asp His Ile Asn Pro Pro Arg Pro Arg Gly Leu1 5 10 15Asp Val Pro Cys Ala Arg Leu Arg Ala Thr Asn

Pro Leu Arg Ala Leu 20 25 30Ala Arg Cys Val Gln Ala Gly Lys Pro Gly Thr Ser Ser Gly His Arg 35 40 45Ser Val Pro His Thr Ala Asp Leu Arg Ile Glu Ala Trp Ala Pro Thr 50 55 60Arg Asp Gly Cys Ile Arg Gln Ala Val Leu Gly Thr Val Glu Ser Phe65 70 75 80Leu Asp Leu Glu Ser Ala His Ala Val His Thr Arg Leu Arg Arg Leu 85 90 95Thr Ala Asp Arg Asp Asp Asp Leu Leu Val Ala Val Leu Glu Glu Val 100 105 110Ile Tyr Leu Leu Asp Thr Val Gly Glu Thr Pro Val Asp Leu Arg Leu 115 120 125Arg Asp Val Asp Gly Gly Val Asp Val Thr Phe Ala Thr Thr Asp Ala 130 135 140Ser Thr Leu Val Gln Val Gly Ala Val Pro Lys Ala Val Ser Leu Asn145 150 155 160Glu Leu Arg Phe Ser Gln Gly Arg His Gly Trp Arg Cys Ala Val Thr 165 170 175Leu Asp Val33375PRTMycobacterium tuberculosis 33Val Thr Gln Thr Gly Lys Arg Gln Arg Arg Lys Phe Gly Arg Ile Arg1 5 10 15Gln Phe Asn Ser Gly Arg Trp Gln Ala Ser Tyr Thr Gly Pro Asp Gly 20 25 30Arg Val Tyr Ile Ala Pro Lys Thr Phe Asn Ala Lys Ile Asp Ala Glu 35 40 45Ala Trp Leu Thr Asp Arg Arg Arg Glu Ile Asp Arg Gln Leu Trp Ser 50 55 60Pro Ala Ser Gly Gln Glu Asp Arg Pro Gly Ala Pro Phe Gly Glu Tyr65 70 75 80Ala Glu Gly Trp Leu Lys Gln Arg Gly Ile Lys Asp Arg Thr Arg Ala 85 90 95His Tyr Arg Lys Leu Leu Asp Asn His Ile Leu Ala Thr Phe Ala Asp 100 105 110Thr Asp Leu Arg Asp Ile Thr Pro Ala Ala Val Arg Arg Trp Tyr Ala 115 120 125Thr Thr Ala Val Gly Thr Pro Thr Met Arg Ala His Ser Tyr Ser Leu 130 135 140Leu Arg Ala Ile Met Gln Thr Ala Leu Ala Asp Asp Leu Ile Asp Ser145 150 155 160Asn Pro Cys Arg Ile Ser Gly Ala Ser Thr Ala Arg Arg Val His Lys 165 170 175Ile Arg Pro Ala Thr Leu Asp Glu Leu Glu Thr Ile Thr Lys Ala Met 180 185 190Pro Asp Pro Tyr Gln Ala Phe Val Leu Met Ala Ala Trp Leu Ala Met 195 200 205Arg Tyr Gly Glu Leu Thr Glu Leu Arg Arg Lys Asp Ile Asp Leu His 210 215 220Gly Glu Val Ala Arg Val Arg Arg Ala Val Val Arg Val Gly Glu Gly225 230 235 240Phe Lys Val Thr Thr Pro Lys Ser Asp Ala Gly Val Arg Asp Ile Ser 245 250 255Ile Pro Pro His Leu Ile Pro Ala Ile Glu Asp His Leu His Lys His 260 265 270Val Asn Pro Gly Arg Glu Ser Leu Leu Phe Pro Ser Val Asn Asp Pro 275 280 285Asn Arg His Leu Ala Pro Ser Ala Leu Tyr Arg Met Phe Tyr Lys Ala 290 295 300Arg Lys Ala Ala Gly Arg Pro Asp Leu Arg Val His Asp Leu Arg His305 310 315 320Ser Gly Ala Val Leu Ala Ala Ser Thr Gly Ala Thr Leu Ala Glu Leu 325 330 335Met Gln Arg Leu Gly His Ser Thr Ala Gly Ala Ala Leu Arg Tyr Gln 340 345 350His Ala Ala Lys Gly Arg Asp Arg Glu Ile Ala Ala Leu Leu Ser Lys 355 360 365Leu Ala Glu Asn Gln Glu Met 370 37534371PRTMycobacterium tuberculosis 34Met Arg Val Gly Ile Pro Thr Glu Thr Lys Asn Asn Glu Phe Arg Val1 5 10 15Ala Ile Thr Pro Ala Gly Val Ala Glu Leu Thr Arg Arg Gly His Glu 20 25 30Val Leu Ile Gln Ala Gly Ala Gly Glu Gly Ser Ala Ile Thr Asp Ala 35 40 45Asp Phe Lys Ala Ala Gly Ala Gln Leu Val Gly Thr Ala Asp Gln Val 50 55 60Trp Ala Asp Ala Asp Leu Leu Leu Lys Val Lys Glu Pro Ile Ala Ala65 70 75 80Glu Tyr Gly Arg Leu Arg His Gly Gln Ile Leu Phe Thr Phe Leu His 85 90 95Leu Ala Ala Ser Arg Ala Cys Thr Asp Ala Leu Leu Asp Ser Gly Thr 100 105 110Thr Ser Ile Ala Tyr Glu Thr Val Gln Thr Ala Asp Gly Ala Leu Pro 115 120 125Leu Leu Ala Pro Met Ser Glu Val Ala Gly Arg Leu Ala Ala Gln Val 130 135 140Gly Ala Tyr His Leu Met Arg Thr Gln Gly Gly Arg Gly Val Leu Met145 150 155 160Gly Gly Val Pro Gly Val Glu Pro Ala Asp Val Val Val Ile Gly Ala 165 170 175Gly Thr Ala Gly Tyr Asn Ala Ala Arg Ile Ala Asn Gly Met Gly Ala 180 185 190Thr Val Thr Val Leu Asp Ile Asn Ile Asp Lys Leu Arg Gln Leu Asp 195 200 205Ala Glu Phe Cys Gly Arg Ile His Thr Arg Tyr Ser Ser Ala Tyr Glu 210 215 220Leu Glu Gly Ala Val Lys Arg Ala Asp Leu Val Ile Gly Ala Val Leu225 230 235 240Val Pro Gly Ala Lys Ala Pro Lys Leu Val Ser Asn Ser Leu Val Ala 245 250 255His Met Lys Pro Gly Ala Val Leu Val Asp Ile Ala Ile Asp Gln Gly 260 265 270Gly Cys Phe Glu Gly Ser Arg Pro Thr Thr Tyr Asp His Pro Thr Phe 275 280 285Ala Val His Asp Thr Leu Phe Tyr Cys Val Ala Asn Met Pro Ala Ser 290 295 300Val Pro Lys Thr Ser Thr Tyr Ala Leu Thr Asn Ala Thr Met Pro Tyr305 310 315 320Val Leu Glu Leu Ala Asp His Gly Trp Arg Ala Ala Cys Arg Ser Asn 325 330 335Pro Ala Leu Ala Lys Gly Leu Ser Thr His Glu Gly Ala Leu Leu Ser 340 345 350Glu Arg Val Ala Thr Asp Leu Gly Val Pro Phe Thr Glu Pro Ala Ser 355 360 365Val Leu Ala 37035104PRTMycobacterium tuberculosis 35Met Val Ile Arg Phe Asp Gln Ile Gly Ser Leu Val Leu Ser Met Lys1 5 10 15Ser Leu Ala Ser Leu Ser Phe Gln Arg Cys Leu Arg Glu Asn Ser Ser 20 25 30Leu Val Ala Ala Leu Asp Arg Leu Asp Ala Ala Val Asp Glu Leu Ser 35 40 45Ala Leu Ser Phe Asp Ala Leu Thr Thr Pro Glu Arg Asp Arg Ala Arg 50 55 60Arg Asp Arg Asp His His Pro Trp Ser Arg Ser Arg Ser Gln Leu Ser65 70 75 80Pro Arg Met Ala His Gly Ala Val His Gln Cys Gln Trp Pro Lys Ala 85 90 95Val Trp Ala Val Ile Asp Asn Pro 10036344PRTMycobacterium tuberculosis 36Val Leu Lys Asn Ala Val Leu Leu Ala Cys Arg Ala Pro Ser Val His1 5 10 15Asn Ser Gln Pro Trp Arg Trp Val Ala Glu Ser Gly Ser Glu His Thr 20 25 30Thr Val His Leu Phe Val Asn Arg His Arg Thr Val Pro Ala Thr Asp 35 40 45His Ser Gly Arg Gln Ala Ile Ile Ser Cys Gly Ala Val Leu Asp His 50 55 60Leu Arg Ile Ala Met Thr Ala Ala His Trp Gln Ala Asn Ile Thr Arg65 70 75 80Phe Pro Gln Pro Asn Gln Pro Asp Gln Leu Ala Thr Val Glu Phe Ser 85 90 95Pro Ile Asp His Val Thr Ala Gly Gln Arg Asn Arg Ala Gln Ala Ile 100 105 110Leu Gln Arg Arg Thr Asp Arg Leu Pro Phe Asp Ser Pro Met Tyr Trp 115 120 125His Leu Phe Glu Pro Ala Leu Arg Asp Ala Val Asp Lys Asp Val Ala 130 135 140Met Leu Asp Val Val Ser Asp Asp Gln Arg Thr Arg Leu Val Val Ala145 150 155 160Ser Gln Leu Ser Glu Val Leu Arg Arg Asp Asp Pro Tyr Tyr His Ala 165 170 175Glu Leu Glu Trp Trp Thr Ser Pro Phe Val Leu Ala His Gly Val Pro 180 185 190Pro Asp Thr Leu Ala Ser Asp Ala Glu Arg Leu Arg Val Asp Leu Gly 195 200 205Arg Asp Phe Pro Val Arg Ser Tyr Gln Asn Arg Arg Ala Glu Leu Ala 210 215 220Asp Asp Arg Ser Lys Val Leu Val Leu Ser Thr Pro Ser Asp Thr Arg225 230 235 240Ala Asp Ala Leu Arg Cys Gly Glu Val Leu Ser Thr Ile Leu Leu Glu 245 250 255Cys Thr Met Ala Gly Met Ala Thr Cys Thr Leu Thr His Leu Ile Glu 260 265 270Ser Ser Asp Ser Arg Asp Ile Val Arg Gly Leu Thr Arg Gln Arg Gly 275 280 285Glu Pro Gln Ala Leu Ile Arg Val Gly Ile Ala Pro Pro Leu Ala Ala 290 295 300Val Pro Ala Pro Thr Pro Arg Arg Pro Leu Asp Ser Val Leu Gln Ile305 310 315 320Arg Gln Thr Pro Glu Lys Gly Arg Asn Ala Ser Asp Arg Asn Ala Arg 325 330 335Glu Thr Gly Trp Phe Ser Pro Pro 34037336PRTMycobacterium tuberculosis 37Val Trp Ser Ala Ser Gly Gly Gln Cys Gly Lys Tyr Leu Ala Ala Ser1 5 10 15Met Val Leu Gln Leu Asp Gly Leu Glu Arg His Gly Val Leu Glu Phe 20 25 30Gly Arg Asp Arg Tyr Gly Pro Glu Val Arg Glu Glu Leu Leu Ala Met 35 40 45Ser Ala Ala Ser Ile Asp Arg Tyr Leu Lys Thr Ala Lys Ala Lys Asp 50 55 60Gln Ile Ser Gly Val Ser Thr Thr Lys Pro Ser Pro Leu Leu Arg Asn65 70 75 80Ser Ile Lys Val Arg Arg Ala Gly Asp Glu Val Glu Ala Glu Pro Gly 85 90 95Phe Phe Glu Gly Asp Thr Val Ala His Cys Gly Pro Thr Leu Lys Gly 100 105 110Glu Phe Ala His Thr Leu Asn Leu Thr Asp Val His Ile Gly Trp Val 115 120 125Phe Thr Arg Thr Val Arg Asn Asn Ala Arg Thr His Ile Leu Ala Gly 130 135 140Leu Lys Ala Ser Val Thr Glu Ile Pro His Gly Ile Thr Gly Leu Asp145 150 155 160Phe Asp Asn Gly Thr Val Phe Leu Asn Lys Pro Val Ile Ser Trp Ala 165 170 175Gly Asp Asn Gly Ile Tyr Phe Thr Arg Phe Arg Pro Tyr Lys Lys Asn 180 185 190His Ala Thr Ile Glu Ser Lys Asn Asn His Leu Val Arg Lys Tyr Ala 195 200 205Phe Tyr Tyr Arg Tyr Asp Thr Ala Glu Glu Arg Ala Val Leu Asn Arg 210 215 220Met Trp Lys Leu Val Asn Asp Arg Leu Asn Tyr Leu Thr Pro Thr Ile225 230 235 240Lys Pro Ile Gly Tyr Ala Ser Ser Ala Asp Gly Arg Arg Arg Arg Leu 245 250 255Tyr Asp Ala Pro Gln Thr Pro Leu Asp Arg Pro Leu Ala Ala Arg Val 260 265 270Leu Ser Ala Ala Gln Gln Ala Asp Leu Ile Thr Tyr Arg Asp Ser Leu 275 280 285Asn Pro Ala Gln Ile Gly Arg Lys Ile Ala Asp Leu Gln Asn Arg Leu 290 295 300Leu Ile Leu Ala Lys Glu Lys Thr Glu Gln Leu Tyr Leu Ala Asn Ile305 310 315 320Pro Thr Ala Leu Pro Asp Ile His Lys Gly Ile Leu Ile Lys Ala Gly 325 330 33538110PRTMycobacterium tuberculosis 38Val Val Gln Gly Arg Thr Val Leu Phe Arg Thr Ala Glu Gly Ala Lys1 5 10 15Leu Phe Ser Ala Val Ala Lys Cys Ala Val Ala Phe Glu Ala Asp Asp 20 25 30His Asn Val Ala Glu Gly Trp Ser Val Ile Val Lys Val Arg Ala Gln 35 40 45Val Leu Thr Thr Asp Ala Gly Val Arg Glu Ala Glu Arg Ala Gln Leu 50 55 60Leu Pro Trp Thr Ala Thr Leu Lys Arg His Cys Val Arg Val Ile Pro65 70 75 80Trp Glu Ile Thr Gly Arg His Phe Arg Phe Gly Pro Glu Pro Asp Arg 85 90 95Ser Gln Thr Phe Ala Cys Glu Ala Ser Ser His Asn Gln Arg 100 105 11039463PRTMycobacterium tuberculosis 39Met Asn His Leu Thr Thr Leu Asp Ala Gly Phe Leu Lys Ala Glu Asp1 5 10 15Val Asp Arg His Val Ser Leu Ala Ile Gly Ala Leu Ala Val Ile Glu 20 25 30Gly Pro Ala Pro Asp Gln Glu Ala Phe Leu Ser Ser Leu Ala Gln Arg 35 40 45Leu Arg Pro Cys Thr Arg Phe Gly Gln Arg Leu Arg Leu Arg Pro Phe 50 55 60Asp Leu Gly Ala Pro Lys Trp Val Asp Asp Pro Asp Phe Asp Leu Gly65 70 75 80Arg His Val Trp Arg Ile Ala Leu Pro Arg Pro Gly Asn Glu Asp Gln 85 90 95Leu Phe Glu Leu Ile Ala Asp Leu Met Ala Arg Arg Leu Asp Arg Gly 100 105 110Arg Pro Leu Trp Glu Val Trp Val Ile Glu Gly Leu Ala Asp Ser Lys 115 120 125Trp Ala Ile Leu Thr Lys Leu His His Cys Met Ala Asp Gly Ile Ala 130 135 140Ala Thr His Leu Leu Ala Gly Leu Ser Asp Glu Ser Met Ser Asp Ser145 150 155 160Phe Ala Ser Asn Ile His Thr Thr Met Gln Ser Gln Ser Ala Ser Val 165 170 175Arg Arg Gly Gly Phe Arg Val Asn Pro Ser Glu Ala Leu Thr Ala Ser 180 185 190Thr Ala Val Met Ala Gly Ile Val Arg Ala Ala Lys Gly Ala Ser Glu 195 200 205Ile Ala Ala Gly Val Leu Ser Pro Ala Ala Ser Ser Leu Asn Gly Pro 210 215 220Ile Ser Asp Leu Arg Arg Tyr Ser Ala Ala Lys Val Pro Leu Ala Asp225 230 235 240Val Glu Gln Val Cys Arg Lys Phe Asp Val Thr Ile Asn Asp Val Ala 245 250 255Leu Ala Ala Ile Thr Glu Ser Tyr Arg Asn Val Leu Ile Gln Arg Gly 260 265 270Glu Arg Pro Arg Phe Asp Ser Leu Arg Thr Leu Val Pro Val Ser Thr 275 280 285Arg Ser Asn Ser Ala Leu Ser Lys Thr Asp Asn Arg Val Ser Leu Met 290 295 300Leu Pro Asn Leu Pro Val Asp Gln Glu Asn Pro Leu Gln Arg Leu Arg305 310 315 320Ile Val His Ser Arg Leu Thr Arg Ala Lys Ala Gly Gly Gln Arg Gln 325 330 335Phe Gly Asn Thr Leu Met Ala Ile Ala Asn Arg Leu Pro Phe Pro Met 340 345 350Thr Ala Trp Ala Val Gly Leu Leu Met Arg Leu Pro Gln Arg Gly Val 355 360 365Val Thr Val Ala Thr Asn Val Pro Gly Pro Arg Arg Pro Leu Gln Ile 370 375 380Met Gly Arg Arg Val Leu Asp Leu Tyr Pro Val Ser Pro Ile Ala Met385 390 395 400Gln Leu Arg Thr Ser Val Ala Met Leu Ser Tyr Ala Asp Asp Leu Tyr 405 410 415Phe Gly Ile Leu Ala Asp Tyr Asp Val Val Ala Asp Ala Gly Gln Leu 420 425 430Ala Arg Gly Ile Glu Asp Ala Val Ala Arg Leu Val Ala Ile Ser Lys 435 440 445Arg Arg Lys Val Thr Arg Arg Arg Gly Ala Leu Ser Leu Val Val 450 455 46040332PRTMycobacterium tuberculosis 40Met Asn Thr His Phe Pro Asp Ala Glu Thr Val Arg Thr Val Leu Thr1 5 10 15Leu Ala Val Arg Ala Pro Ser Ile His Asn Thr Gln Pro Trp Arg Trp 20 25 30Arg Val Cys Pro Thr Ser Leu Glu Leu Phe Ser Arg Pro Asp Met Gln 35 40 45Leu Arg Ser Thr Asp Pro Asp Gly Arg Glu Leu Ile Leu Ser Cys Gly 50 55 60Val Ala Leu His His Cys Val Val Ala Leu Ala Ser Leu Gly Trp Gln65 70 75 80Ala Lys Val Asn Arg Phe Pro Asp Pro Lys Asp Arg Cys His Leu Ala 85 90 95Thr Ile Gly Val Gln Pro Leu Val Pro Asp Gln Ala Asp Val Ala Leu 100 105 110Ala Ala Ala Ile Pro Arg Arg Arg Thr Asp Arg Arg Ala Tyr Ser Cys 115 120 125Trp Pro Val Pro Gly Gly Asp Ile Ala Leu Met Ala Ala Arg Ala Ala 130 135 140Arg Gly Gly Val Met Leu Arg Gln Val Ser Ala Leu Asp Arg Met Lys145 150 155 160Ala Ile Val Ala Gln Ala Val Leu Asp His Val Thr Asp Glu Glu Tyr 165 170 175Leu Arg Glu Leu Thr Ile Trp

Ser Gly Arg Tyr Gly Ser Val Ala Gly 180 185 190Val Pro Ala Arg Asn Glu Pro Pro Ser Asp Pro Ser Ala Pro Ile Pro 195 200 205Gly Arg Leu Phe Ala Gly Pro Gly Leu Ser Gln Pro Ser Asp Val Leu 210 215 220Pro Ala Asp Asp Gly Ala Ala Ile Leu Ala Leu Gly Thr Glu Thr Asp225 230 235 240Asp Arg Leu Ala Arg Leu Arg Ala Gly Glu Ala Ala Ser Ile Val Leu 245 250 255Leu Thr Ala Thr Ala Met Gly Leu Ala Cys Cys Pro Ile Thr Glu Pro 260 265 270Leu Glu Ile Ala Lys Thr Arg Asp Ala Val Arg Ala Glu Val Phe Gly 275 280 285Ala Gly Gly Tyr Pro Gln Met Leu Leu Arg Val Gly Trp Ala Pro Ile 290 295 300Asn Ala Asp Pro Leu Pro Pro Thr Pro Arg Arg Glu Leu Ser Gln Val305 310 315 320Val Glu Trp Pro Glu Glu Leu Leu Arg Gln Arg Cys 325 33041578PRTMycobacterium tuberculosis 41Met Thr Thr Gly Gly Leu Val Asp Glu Asn Asp Gly Ala Ala Met Arg1 5 10 15Pro Leu Arg His Thr Leu Ser Gln Leu Arg Leu His Glu Leu Leu Val 20 25 30Glu Val Gln Asp Arg Val Glu Gln Ile Val Glu Gly Arg Asp Arg Leu 35 40 45Asp Gly Leu Val Glu Ala Met Leu Val Val Thr Ala Gly Leu Asp Leu 50 55 60Glu Ala Thr Leu Arg Ala Ile Val His Ser Ala Thr Ser Leu Val Asp65 70 75 80Ala Arg Tyr Gly Ala Met Glu Val His Asp Arg Gln His Arg Val Leu 85 90 95His Phe Val Tyr Glu Gly Ile Asp Glu Glu Thr Val Arg Arg Ile Gly 100 105 110His Leu Pro Lys Gly Leu Gly Val Ile Gly Leu Leu Ile Glu Asp Pro 115 120 125Lys Pro Leu Arg Leu Asp Asp Val Ser Ala His Pro Ala Ser Ile Gly 130 135 140Phe Pro Pro Tyr His Pro Pro Met Arg Thr Phe Leu Gly Val Pro Val145 150 155 160Arg Val Arg Asp Glu Ser Phe Gly Thr Leu Tyr Leu Thr Asp Lys Thr 165 170 175Asn Gly Gln Pro Phe Ser Asp Asp Asp Glu Val Leu Val Gln Ala Leu 180 185 190Ala Ala Ala Ala Gly Ile Ala Val Ala Asn Ala Arg Leu Tyr Gln Gln 195 200 205Ala Lys Ala Arg Gln Ser Trp Ile Glu Ala Thr Arg Asp Ile Ala Thr 210 215 220Glu Leu Leu Ser Gly Thr Glu Pro Ala Thr Val Phe Arg Leu Val Ala225 230 235 240Ala Glu Ala Leu Lys Leu Thr Ala Ala Asp Ala Ala Leu Val Ala Val 245 250 255Pro Val Asp Glu Asp Met Pro Ala Ala Asp Val Gly Glu Leu Leu Val 260 265 270Ile Glu Thr Val Gly Ser Ala Val Ala Ser Ile Val Gly Arg Thr Ile 275 280 285Pro Val Ala Gly Ala Val Leu Arg Glu Val Phe Val Asn Gly Ile Pro 290 295 300Arg Arg Val Asp Arg Val Asp Leu Glu Gly Leu Asp Glu Leu Ala Asp305 310 315 320Ala Gly Pro Ala Leu Leu Leu Pro Leu Arg Ala Arg Gly Thr Val Ala 325 330 335Gly Val Val Val Val Leu Ser Gln Gly Gly Pro Gly Ala Phe Thr Asp 340 345 350Glu Gln Leu Glu Met Met Ala Ala Phe Ala Asp Gln Ala Ala Leu Ala 355 360 365Trp Gln Leu Ala Thr Ser Gln Arg Arg Met Arg Glu Leu Asp Val Leu 370 375 380Thr Asp Arg Asp Arg Ile Ala Arg Asp Leu His Asp His Val Ile Gln385 390 395 400Arg Leu Phe Ala Ile Gly Leu Ala Leu Gln Gly Ala Val Pro His Glu 405 410 415Arg Asn Pro Glu Val Gln Gln Arg Leu Ser Asp Val Val Asp Asp Leu 420 425 430Gln Asp Val Ile Gln Glu Ile Arg Thr Thr Ile Tyr Asp Leu His Gly 435 440 445Ala Ser Gln Gly Ile Thr Arg Leu Arg Gln Arg Ile Asp Ala Ala Val 450 455 460Ala Gln Phe Ala Asp Ser Gly Leu Arg Thr Ser Val Gln Phe Val Gly465 470 475 480Pro Leu Ser Val Val Asp Ser Ala Leu Ala Asp Gln Ala Glu Ala Val 485 490 495Val Arg Glu Ala Val Ser Asn Ala Val Arg His Ala Lys Ala Ser Thr 500 505 510Leu Thr Val Arg Val Lys Val Asp Asp Asp Leu Cys Ile Glu Val Thr 515 520 525Asp Asn Gly Arg Gly Leu Pro Asp Glu Phe Thr Gly Ser Gly Leu Thr 530 535 540Asn Leu Arg Gln Arg Ala Glu Gln Ala Gly Gly Glu Phe Thr Leu Ala545 550 555 560Ser Val Pro Gly Ala Ser Gly Thr Val Leu Arg Trp Ser Ala Pro Leu 565 570 575Ser Gln42268PRTMycobacterium tuberculosis 42Met Ser Asp Pro Arg Pro Ala Arg Ala Val Val Val Gly Ile Asp Gly1 5 10 15Ser Arg Ala Ala Thr His Ala Ala Leu Trp Ala Val Asp Glu Ala Val 20 25 30Asn Arg Asp Ile Pro Leu Arg Leu Val Tyr Val Ile Asp Pro Ser Gln 35 40 45Leu Ser Ala Ala Gly Glu Gly Gly Gly Gln Ser Ala Ala Arg Ala Ala 50 55 60Leu His Asp Ala Ser Arg Lys Val Glu Ala Thr Gly Gln Pro Val Lys65 70 75 80Ile Glu Thr Glu Val Leu Cys Gly Arg Pro Leu Thr Lys Leu Met Gln 85 90 95Glu Ser Arg Ser Ala Ala Met Leu Cys Val Gly Ser Val Gly Leu Asp 100 105 110His Val Arg Gly Arg Arg Gly Ser Val Ala Ala Thr Leu Ala Gly Ser 115 120 125Ala Leu Cys Pro Val Ala Val Ile His Pro Ser Pro Ala Glu Pro Ala 130 135 140Thr Thr Ser Gln Val Ser Ala Val Val Ala Glu Val Asp Asn Gly Val145 150 155 160Val Leu Arg His Ala Phe Glu Glu Ala Arg Leu Arg Gly Val Pro Leu 165 170 175Arg Ala Val Ala Val His Ala Ala Glu Thr Pro Asp Asp Val Glu Gln 180 185 190Gly Ser Arg Leu Ala His Val His Leu Ser Arg Arg Leu Ala His Trp 195 200 205Thr Arg Leu Tyr Pro Glu Val Arg Val Asp Arg Ala Ile Ala Gly Gly 210 215 220Ser Ala Cys Arg His Leu Ala Ala Asn Ala Lys Pro Gly Gln Leu Phe225 230 235 240Val Ala Asp Ser His Ser Ala His Glu Leu Cys Gly Ala Tyr Gln Pro 245 250 255Gly Cys Ala Val Leu Thr Val Arg Ser Ala Asn Leu 260 26543181PRTMycobacterium tuberculosis 43Met Thr Glu Tyr Glu Gly Pro Lys Thr Lys Phe His Ala Leu Met Gln1 5 10 15Glu Gln Ile His Asn Glu Phe Thr Ala Ala Gln Gln Tyr Val Ala Ile 20 25 30Ala Val Tyr Phe Asp Ser Glu Asp Leu Pro Gln Leu Ala Lys His Phe 35 40 45Tyr Ser Gln Ala Val Glu Glu Arg Asn His Ala Met Met Leu Val Gln 50 55 60His Leu Leu Asp Arg Asp Leu Arg Val Glu Ile Pro Gly Val Asp Thr65 70 75 80Val Arg Asn Gln Phe Asp Arg Pro Arg Glu Ala Leu Ala Leu Ala Leu 85 90 95Asp Gln Glu Arg Thr Val Thr Asp Gln Val Gly Arg Leu Thr Ala Val 100 105 110Ala Arg Asp Glu Gly Asp Phe Leu Gly Glu Gln Phe Met Gln Trp Phe 115 120 125Leu Gln Glu Gln Ile Glu Glu Val Ala Leu Met Ala Thr Leu Val Arg 130 135 140Val Ala Asp Arg Ala Gly Ala Asn Leu Phe Glu Leu Glu Asn Phe Val145 150 155 160Ala Arg Glu Val Asp Val Ala Pro Ala Ala Ser Gly Ala Pro His Ala 165 170 175Ala Gly Gly Arg Leu 18044274PRTMycobacterium tuberculosis 44Met Thr Trp Ala Asp Glu Val Leu Ala Gly His Pro Phe Val Val Ala1 5 10 15His Arg Gly Ala Ser Ala Ala Arg Pro Glu His Thr Leu Ala Ala Tyr 20 25 30Asp Leu Ala Leu Lys Glu Gly Ala Asp Gly Val Glu Cys Asp Val Arg 35 40 45Leu Thr Arg Asp Gly His Leu Val Cys Val His Asp Arg Arg Leu Asp 50 55 60Arg Thr Ser Thr Gly Ala Gly Leu Val Ser Thr Met Thr Leu Ala Gln65 70 75 80Leu Arg Glu Leu Glu Tyr Gly Ala Trp His Asp Ser Trp Arg Pro Asp 85 90 95Gly Ser His Gly Asp Thr Ser Leu Leu Thr Leu Asp Ala Leu Val Ser 100 105 110Leu Val Leu Asp Trp His Arg Pro Val Lys Ile Phe Val Glu Thr Lys 115 120 125His Pro Val Arg Tyr Gly Ser Leu Val Glu Asn Lys Leu Leu Ala Leu 130 135 140Leu His Arg Phe Gly Ile Ala Ala Pro Ala Ser Ala Asp Arg Ser Arg145 150 155 160Ala Val Val Met Ser Phe Ser Ala Ala Ala Val Trp Arg Ile Arg Arg 165 170 175Ala Ala Pro Leu Leu Pro Thr Val Leu Leu Gly Lys Thr Pro Arg Tyr 180 185 190Leu Thr Ser Ser Ala Ala Thr Ala Val Gly Ala Thr Ala Val Gly Pro 195 200 205Ser Leu Pro Ala Leu Lys Glu Tyr Pro Gln Leu Val Asp Arg Ser Ala 210 215 220Ala Gln Gly Arg Ala Val Tyr Cys Trp Asn Val Asp Glu Tyr Glu Asp225 230 235 240Ile Asp Phe Cys Arg Glu Val Gly Val Ala Trp Ile Gly Thr His His 245 250 255Pro Gly Arg Thr Lys Ala Trp Leu Glu Asp Gly Arg Ala Asn Gly Thr 260 265 270Thr Arg45248PRTMycobacterium tuberculosis 45Val Ser Asp Gly Glu Gln Ala Lys Ser Arg Arg Arg Arg Gly Arg Arg1 5 10 15Arg Gly Arg Arg Ala Ala Ala Thr Ala Glu Asn His Met Asp Ala Gln 20 25 30Pro Ala Gly Asp Ala Thr Pro Thr Pro Ala Thr Ala Lys Arg Ser Arg 35 40 45Ser Arg Ser Pro Arg Arg Gly Ser Thr Arg Met Arg Thr Val His Glu 50 55 60Thr Ser Ala Gly Gly Leu Val Ile Asp Gly Ile Asp Gly Pro Arg Asp65 70 75 80Ala Gln Val Ala Ala Leu Ile Gly Arg Val Asp Arg Arg Gly Arg Leu 85 90 95Leu Trp Ser Leu Pro Lys Gly His Ile Glu Leu Gly Glu Thr Ala Glu 100 105 110Gln Thr Ala Ile Arg Glu Val Ala Glu Glu Thr Gly Ile Arg Gly Ser 115 120 125Val Leu Ala Ala Leu Gly Arg Ile Asp Tyr Trp Phe Val Thr Asp Gly 130 135 140Arg Arg Val His Lys Thr Val His His Tyr Leu Met Arg Phe Leu Gly145 150 155 160Gly Glu Leu Ser Asp Glu Asp Leu Glu Val Ala Glu Val Ala Trp Val 165 170 175Pro Ile Arg Glu Leu Pro Ser Arg Leu Ala Tyr Ala Asp Glu Arg Arg 180 185 190Leu Ala Glu Val Ala Asp Glu Leu Ile Asp Lys Leu Gln Ser Asp Gly 195 200 205Pro Ala Ala Leu Pro Pro Leu Pro Pro Ser Ser Pro Arg Arg Arg Pro 210 215 220Gln Thr His Ser Arg Ala Arg His Ala Asp Asp Ser Ala Pro Gly Gln225 230 235 240His Asn Gly Pro Gly Pro Gly Pro 24546819DNAMycobacterium tuberculosis 46gtggaaccga aacgcagtcg cctcgtcgta tgtgcacccg agccatcgca cgcgcgggaa 60ttcccggatg tcgccgtatt ctccggcggc cgggctaacg catcccaggc cgaacggttg 120gctcgtgccg tgggtcgcgt gttggccgat cggggcgtca ccgggggtgc tcgggtgcgg 180ctgaccatgg cgaactgcgc cgatgggccg acgctggtgc agataaacct gcaggtaggt 240gacaccccat taagggcgca ggccgccacc gcgggcatcg atgatctgcg acccgcactg 300atcagactgg atcgacagat cgtgcgggcg tcggcacagt ggtgcccccg gccttggccg 360gatcggcccc gccggcgatt gaccacgccg gccgaggcgc tagtcacccg ccgcaaaccg 420gtcgtgctaa ggcgcgcaac cccgttgcag gcgattgccg ctatggacgc catggactac 480gacgtgcatt tgttcaccga cgccgagacg ggggaggacg ctgtggtcta tcgggctgga 540ccgtcggggc tgcggctggc ccgccagcac cacgtatttc ccccaggatg gtcacgttgt 600cgcgccccag ccgggccgcc ggtgccgctg attgtgaatt cgcgtccgac accggttctc 660acggaggccg ccgcggtgga ccgggcgcgc gaacatggac tgccattcct gtttttcacc 720gaccaggcca ccggccgcgg ccagctgctc tactcccgct acgacggcaa cctcgggttg 780atcaccccga ccggtgacgg cgttgccgac ggtctggca 81947819DNAMycobacterium tuberculosis 47gtggaaccga aacgcagtcg cctcgtcgta tgtgcacccg agccatcgca cgcgcgggaa 60ttcccggatg tcgccgtatt ctccggcggc cgggctaacg catcccaggc cgaacggttg 120gctcgtgccg tgggtcgcgt gttggccgat cggggcgtca ccgggggtgc tcgggtgcgg 180ctgaccatgg cgaactgcgc cgatgggccg acgctggtgc agataaacct gcaggtaggt 240gacaccccat taagggcgca ggccgccacc gcgggcatcg atgatctgcg acccgcactg 300atcagactgg atcgacagat cgtgcgggcg tcggcacagt ggtgcccccg gccttggccg 360gatcggcccc gccggcgatt gaccacgccg gccgaggcgc tagtcacccg ccgcaaaccg 420gtcgtgctaa ggcgcgcaac cccgttgcag gcgattgccg ctatggacgc catggactac 480gacgtgcatt tgttcaccga cgccgagacg ggggaggacg ctgtggtcta tcgggctgga 540ccgtcggggc tgcggctggc ccgccagcac cacgtatttc ccccaggatg gtcacgttgt 600cgcgccccag ccgggccgcc ggtgccgctg attgtgaatt cgcgtccgac accggttctc 660acggaggccg ccgcggtgga ccgggcgcgc gaacatggac tgccattcct gtttttcacc 720gaccaggcca ccggccgcgg ccagctgctc tactcccgct acgacggcaa cctcgggttg 780atcaccccga ccggtgacgg cgttgccgac ggtctggca 81948342DNAMycobacterium tuberculosis 48gtggagtccg aaccgctgta caagctcaag gcggagttct tcaaaaccct tgcgcatccg 60gcgcggatca ggattttgga gctgctggtc gagcgggacc gttcggtcgg tgagttgctg 120tcctcggacg tcggcctgga gtcgtcgaac ctgtcccagc agctgggtgt gctacgccgg 180gcgggtgttg tcgcggcacg tcgtgacggc aacgcgatga tctattcgat tgccgcaccc 240gatatcgccg agctgctggc ggtggcacgc aaggtgctgg ccagggtgct cagcgaccgg 300gtggcggtgc tagaggacct ccgcgccggc ggctcggcca cg 342491032DNAMycobacterium tuberculosis 49atgcctatcg caacgcccga ggtctacgcg gagatgctcg gtcaggccaa acaaaactcg 60tacgctttcc cggctatcaa ctgcacctcc tcggaaaccg tcaacgccgc gatcaaaggt 120ttcgccgacg ccggcagtga cggaatcatc cagttctcga ccggtggcgc agaattcggc 180tccggcctcg gggtcaaaga catggtgacc ggtgcggtcg ccttggcgga gttcacccac 240gttatcgcgg ccaagtaccc ggtcaacgtg gcgctgcaca ccgaccactg ccccaaggac 300aagttggaca gctatgtccg gcccttgctg gcgatctcgg cgcaacgcgt gagcaaaggt 360ggcaatcctt tgttccagtc gcacatgtgg gacggctcgg cagtgccaat cgatgagaac 420ctggccatcg cccaggagct gctcaaggcg gcggcggccg ccaagatcat tctggagatc 480gagatcggcg tcgtcggcgg cgaagaggac ggcgtggcga acgagatcaa cgagaagctg 540tacaccagcc cggaggactt cgagaaaacc atcgaggcgc tgggcgccgg tgagcacggc 600aaatacctgc tggccgcgac gttcggcaac gtgcatggcg tctacaagcc cggcaacgtc 660aagcttcgcc ccgacatcct tgcgcaaggg caacaggtgg cggcggccaa gctcggactg 720ccggccgacg ccaagccgtt cgacttcgtg ttccacggcg gctcgggttc gcttaagtcg 780gagatcgagg aggcgctgcg ctacggcgtg gtgaagatga acgtcgacac cgacacccag 840tacgcgttca cccgcccgat cgccggtcac atgttcacca actacgacgg agtgctcaag 900gtcgatggcg aggtgggtgt caagaaggtc tacgacccgc gcagctacct caagaaggcc 960gaagcttcga tgagccagcg ggtcgttcag gcgtgcaatg acctgcactg cgccggaaag 1020tccctaaccc ac 103250339DNAMycobacterium tuberculosis 50atgggtgagc acgccatcaa gcggcacatg cggcaacgga agcctacgaa gcatccccta 60gcccagaaac ggggcgcgcg gattctggtc ttcaccgacg atccccgcag gagcgtcctc 120atagtgcccg gttgccacct ggattccatg cgccgagaaa agaacgcgta ctacttccag 180gacggcaatg cgttggttgg gatggttgtc tcgggcggca cggttgagta cgacgccgac 240gaccgcacat atgtcgtgca gctcaccgac ggaaggcaca ccactgagtc atctttcgaa 300cactcatcgc cgagtcgatc acctcaatcc gatgaccta 339511140DNAMycobacterium tuberculosis 51gtggctggca atcctgatgt ggtgacggtg ctgctgggcg gtgacgtcat gctcggccgt 60ggcgtcgatc agatcctgcc tcatcccggc aaaccgcaat tgcgcgaacg gtatatgcgg 120gatgcgaccg gctatgttcg cctggccgag cgggtgaacg ggcgcattcc gctccccgtg 180gattggcgct ggccctgggg cgaggcgttg gcggtccttg agaacaccgc gaccgacgtc 240tgtttgatca atctggagac gacgatcacc gccgacggtg aattcgccga ccgcaaaccg 300gtctgctacc ggatgcaccc ggataacgtg ccggcgctga cggcattgcg gccgcacgtg 360tgcgcgctgg ccaacaacca cattctcgat ttcggctacc aggggctgac cgatacggtc 420gcggctctcg ccggtgcggg gatccagagt gtcggggcgg gagccgattt gctcgccgct 480cgccgctcgg cgctagtcac ggttggccat gaacgccggg tgatcgtcgg ctcggtagcg 540gcggaatcca gcggcgtccc cgaatcctgg gccgcccgcc gcgaccggcc cggagtgtgg 600ttgatccggg atccggcgca acgcgacgtc gccgacgatg tggcggcaca ggtgctggcg 660gacaaacgcc ccggcgatat cgccatagtc tcgatgcatt ggggatccaa ttggggctat 720gcgaccgcac ccggcgacgt cgcgttcgcg caccgactga tcgacgccgg catcgacatg 780gtccacggac attcctcgca ccatccgcgg ccaatcgaga tatatcgcgg taaaccgatc 840ctgtacggat gcggtgacgt cgttgacgac tacgaaggca tcggcgggca

cgagtcgttc 900cgcagtgaac tgcgactgct gtatctgacc gtcaccgatc ccgccagcgg gaacctgatc 960tcgctgcaga tgcttccact gcgagtgtcg cggatgcgcc tacagcgtgc ctcccagacc 1020gacaccgaat ggctccgcaa caccattgag cgcatcagcc gccggttcgg gattcgagtc 1080gtgactcgac ccgacaacct gctggaggtc gttcccgctg ccaacctaac gagcaaggag 1140521191DNAMycobacterium tuberculosis 52gtgacagacc acgtgcgcga ggcggacgac gcgaacatcg acgatctgtt gggcgacctg 60ggcggtaccg cgcgcgccga gcgtgcgaag cttgtcgagt ggttgctcga gcagggcatc 120acccccgacg agattcgggc gaccaacccg ccgttgctgc tggccacccg ccacctcgtc 180ggcgacgacg gcacctacgt atccgcaagg gagattagcg agaactatgg cgttgacctc 240gagctgctgc agcgggtgca gcgcgctgtc ggtctggcca gagtggatga tcctgacgcg 300gtggtgcaca tgcgtgccga cggtgaggcg gccgcacgcg cacagcggtt cgttgagctg 360gggctgaatc ccgaccaagt cgtgctggtc gtgcgtgtgc tcgccgaggg cttgtcacac 420gccgccgagg ccatgcgcta caccgcgctg gaggccatta tgcggccggg ggctaccgag 480ttggacatcg cgaaggggtc gcaggcgctg gtgagccaga tcgtgccgct gctggggccg 540atgatccagg acatgctgtt catgcagctg cggcacatga tggagacgga ggccgtcaac 600gccggagagc gtgcggccgg caagccgcta ccgggagcgc gacaggtcac cgttgccttc 660gccgacctgg tcggtttcac ccagctaggc gaagtggtgt cggccgaaga gctagggcac 720ctcgccgggc ggctggccgg cctcgcgcgt gacctgaccg ctccgccggt gtggttcatt 780aagacgatcg gcgacgcggt catgttggtc tgtcctgatc cggcgccatt gctggacacc 840gtgctgaagc tggtcgaggt cgtcgacacc gacaacaact ttccccggct gcgagccggc 900gtcgcctccg ggatggcggt tagccgggcc ggcgactggt tcggcagccc ggtcaacgtg 960gcaagccggg tgaccggggt ggcgcgcccg ggtgccgtgc tggtcgcgga ttcggtgcgg 1020gaggcccttg gtgatgcccc cgaagccgac ggatttcagt ggtccttcgc cggcccccgt 1080cgcctcaggg gaatccgggg tgacgtcagg ctttttcgag tccggcgagg ggccactcgc 1140accggctccg gcggcgcggc ccaagacgac gatttggccg gctcgtcacc g 1191531338DNAMycobacterium tuberculosis 53atggtagagc ccggcaattt ggcaggcgcg accggcgccg aatggatcgg ccggccaccg 60cacgaggaat tgcagcgcaa agtgcgcccg ctgctgccat ccgacgatcc gttctacttc 120ccacctgccg gctaccagca tgccgtgccc ggaacggtgt tgcgctcgcg cgatgtcgaa 180ctggcgttta tgggcttgat tccgcagccc gtcaccgcta cccagctgct gtaccggacc 240acgaacatgt acggcaaccc cgaggcgacg gtgaccacgg tgatcgtccc agcggagctt 300gccccgggtc agacctgccc cttgctgtcg taccagtgtg cgatcgatgc catgtcgtcg 360cgctgttttc cgtcatatgc cctgcgacga cgggccaagg ccctggggtc actgacccaa 420atggagctgt tgatgatcag cgccgcactt gccgaaggat gggcggtatc agtacccgac 480catgaagggc cgaaagggct gtgggggtcg ccgtatgaac ccggttaccg agtcctcgac 540ggaatccggg ctgccttgaa ttccgagcgt gtcgggttgt ccccggcaac gccgatcggg 600ctgtggggct actccggcgg cgggctggcc agcgcgtggg ccgccgaagc atgcggcgag 660tacgcaccgg acctagacat cgtcggcgcc gtgctgggat cacccgtcgg tgaccttggt 720cacacgttcc gccggctcaa tggcactctt cttgccggtc tgcccgcgtt ggtggtggcc 780gcgctgcaac acagctaccc cggcctggcc cgggtgatca aggagcacgc caacgacgaa 840ggacgtcagc tgctggagca actgacggag atgacaacgg tagacgcagt gatccggatg 900gccggcaggg acatgggtga cttcctcgac gaaccccttg aggacattct gtcgacgccg 960gaaatttccc atgtcttcgg cgacaccaag ctgggtagcg cggtgcccac cccgccggta 1020ttgatcgtgc aggccgtgca tgactacctc atcgacgtct ctgacatcga cgcgctcgct 1080gacagctata cagccggcgg cgccaacgtc acctaccacc gcgacctgtt cagcgaacat 1140gtgtccctgc acccgctgtc ggccccaatg acgcttcgct ggctcaccga ccggttcgcc 1200ggcaagccac tgaccgacca ccgcgtccgg accacgtggc cgaccatctt caacccgatg 1260acctacgccg gcatggcgag actggccgtg atcgcggcca aggtgatcac cggcaggaag 1320ttgagccgcc gtccgctc 133854630DNAMycobacterium tuberculosis 54atgatcgcca 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 63055240DNAMycobacterium tuberculosis 55atgaccaacg tcggtgacca gggggttgac gcggtcttcg gggtgatcta cccacctcag 60gtcgcgctgg tcagtttcgg caagccggca caacgagttt gcgccgtcga cggcgcgatc 120cacgtcatga cgaccgtgct ggctacgctg cccgctgacc acggctgcag cgatgaccat 180cgcggcgcgc tgttcttcct gtcgatcaac gagctgacgc ggtgcgccgc agtaacagga 240561956DNAMycobacterium tuberculosis 56gtgacggtga caccacggac cggcagccgc atcgaggagc tgcttgcacg cagcggccgg 60ttcttcatcc cgggtgagat ctcggcggat ctgcgtaccg tgacccgccg cggcggccgc 120gacggcgacg tgttctatcg agaccggtgg agccacgaca aggtggtccg ctccacacac 180ggggtgaatt gcaccgggtc gtgttcttgg aagatctacg tcaaagacga catcatcacc 240tgggagacgc aggagaccga ctatccgtcg gtgggcccgg accggcccga gtatgagccc 300cgcggctgcc cgcgcggcgc ggcgttttcc tggtacacgt attcgccgac gcgggtgcgc 360catccgtacg cccgcggcgt gcttgtcgag atgtatcggg aggcgaaggc acgtttgggt 420gatccggtgg cggcctgggc cgacatccag gccgacccgc ggcggcgccg ccgctaccag 480cgcgcccgcg gcaagggcgg gctggtccgg gtcagctggg ccgaggccac cgagatgatc 540gccgccgccc acgtgcacac catctccaca tacggcccgg accgggttgc cggcttctcc 600cccatcccgg cgatgtccat ggtgagccac gccgcggggt cgcggttcgt ggagctaatc 660ggcggggtga tgacgtcgtt ctacgactgg tacgccgacc tgccggtggc ctccccgcag 720gtgttcggcg accagaccga cgtgccggag tccggagatt ggtgggacgt ggtgtggcaa 780tgcgcctcgg tgctgctgac ctacccgaac tcacggcaac tcggcaccgc agaggaattg 840ctggcccaca tcgacggtcc ggccgcggat ctgttggggc gcacggtctc tgagctgcgc 900cgtgccgatc cgctgaccgc ggcgacccgc tacgtcgaca ccttcgacct gcgaggccgc 960gccaccctgt acctgaccta ctggaccgcc ggcgacaccc gcaaccgcgg ccgggagatg 1020ctggccttcg cccagaccta ccgcagcacc gacgtcgcac caccgcgcgg cgagaccccg 1080gacttcctgc cggtggtgct cgaattcgcc gcgaccgtcg accccgaggc ggggcgacgg 1140ttgctgagcg ggtaccgggt gcccatcgcc gcgctgtgca atgccctgac cgaggccgca 1200ttgccatacg cacacacggt ggccgcggta tgccggacgg gtgacatgat gggcgaactc 1260ttctggaccg tcgtgccgta tgtgacgatg acgatcgtcg cggtcggctc ctggtggcgc 1320taccgctatg acaaattcgg ctggaccacc cgctcgtccc agctgtacga gtcgcggctg 1380ctgcggatcg ccagcccgat gtttcatttc ggcatcctgg tggtcatcgt cggccacggt 1440atcgggctcg tgatcccgca gtcgtggact caggccgccg gtttgagcga gggcgcatat 1500cacgtgcagg ccgtcgtgct ggggtcgatc gccggcatca ccaccttggc cggcgttacc 1560ctgctgatct accggcggcg cacccgcggg ccggtgttca tggctaccac cgtcaacgac 1620aaggtgatgt acctcgtgct ggtggcggcg atcgtcgcgg gactgggtgc gacggcgttg 1680ggctccggcg ttgtcggcga ggcgtacaac taccgcgaga cggtgtcggt gtggttccgc 1740tcggtgtggg tactgcaacc gcgcggggac ctgatggccg aggctccgct gtattaccag 1800atccatgtgc tgatcgggtt ggcgttgttc gcgttgtggc cgttcacccg gctggtacac 1860gcgttcagcg ccccgatcgg ctatctgttc cgcccgtaca tcatctaccg cagccgcgag 1920gagctggtgc taacgcggcc gcggcggcgc gggtgg 1956571185DNAMycobacterium tuberculosis 57atgagagggc aagcggccaa tctcgtgctg gccacctgga tctcggtggt caacttctgg 60gcgtggaacc tgatcggccc gctgtcgacc agctacgcgc gtgacatgtc actgtccagc 120gccgaggcgt cgctgctcgt cgccaccccg atcctggtgg gtgcccttgg ccgcatcgtc 180accgggccgc tcaccgaccg cttcggcggg cgcgccatgc tcatcgcggt gacgctggcg 240tcgatcctcc cggtgctcgc ggtcggggtc gcggcaacca tgggctccta cgcgttgctg 300gtgtttttcg ggctcttcct gggcgttgcc ggcacgatct tcgccgtcgg catcccgttc 360gccaacaact ggtaccagcc ggcgcggcgc ggtttctcca ccggcgtgtt cggtatgggc 420atggtcggca ccgcgctctc ggcgttcttc accccgcggt ttgtacggtg gttcggcctg 480ttcaccaccc acgccatcgt cgcggccgcg ctcgcgtcga ccgccgtggt ggccatggtc 540gtgcttcgtg atgcacccta ctttcggccc aacgccgacc cggtgctgcc caggctcaag 600gccgcggcac ggttgccggt gacctgggag atgtcgtttc tgtacgcgat cgtgttcggc 660gggttcgtgg cgttcagcaa ctacctgccc acctacatca ccacgatcta cgggttctcc 720acggtcgacg cgggcgctcg caccgccggg ttcgccctgg cggcggtgct ggcccggccg 780gtgggcgggt ggctctccga ccggatcgca ccgaggcacg tggtgctggc ctcgctcgcc 840gggaccgcgc tgctggcgtt cgccgcggcg ttgcagccgc cgccggaggt gtggtcggcg 900gccaccttca tcaccctggc ggtctgcctc ggcgtgggca ccggcggcgt gttcgcgtgg 960gtggcccgcc gcgccccggc cgcatcggtc ggctcggtca ccggaatcgt cgccgcggca 1020ggcggattgg gcggttactt cccgccgctg gtgatgggcg cgacctacga cccggtcgac 1080aacgactaca cggtcgggtt gctgctgctg gtggcgaccg cgctggtcgc gtgtacctac 1140accgcgctgc acgcgcggga gccggtgagt gaggaggcgt ccagg 118558282DNAMycobacterium tuberculosis 58atgtgcggcg accagtcgga tcacgtgctg cagcactgga ccgtcgacat atcgatcgac 60gaacacgaag gattgactcg ggcgaaggca cggctgcgtt ggcgggaaaa ggaattggtg 120ggtgttggcc tggcaaggct caatccggcc gaccgcaacg tccccgagat cggcgatgaa 180ctctcggtcg cccgagcctt gtccgacttg gggaagcgaa tgttgaaggt gtcgacccac 240gacatcgaag ctgttaccca tcagccggcg cgattgttgt at 282591680DNAMycobacterium tuberculosis 59atgattccca cgatgacatc ggccggctgg gcaccagggg tggtgcagtt ccgcgaatac 60caacggcgtt ggctgcgcgg cgatgtcctc gccggcctga ccgtggccgc ctatctgatc 120ccgcaagcga tggcgtatgc gaccgtggcg ggcctaccgc cggcagccgg gctgtgggcg 180tcgatcgcgc cgcttgccat ttacgcactg ctcggatcgt cccggcagct ttcaatcggc 240ccggaatccg ccaccgcctt gatgacggcg gccgtgctcg ctccgatggc cgccggggat 300cttcgacgct atgccgttct ggcggcaacc ctcggattgc tagtcggcct tatctgccta 360ctcgctggca cggcgcgact aggtttcctc gccagcctgc gatcgcggcc ggtgctcgtc 420ggatacatgg ccggcatcgc gcttgtcatg atctccagcc aactcggcac tatcaccggc 480acctcggtcg aaggcaacga attcttcagc gaagtacact ctttcgcgac tagcgtcacg 540cgagttcact ggccgacttt tgtgttagcc atgtctgtcc tagcgctgct aactatgctc 600acgcggtggg cgccgcgcgc ccccggaccg atcatcgcgg ttcttgcggc cacgatgcta 660gtggccgtta tgtccttgga tgccaaaggt attgcgattg tgggtcggat accttccggt 720ctgccgacgc cgggtgtgcc gcccgtttcg gtggaagact tgcgggcact gatcattccg 780gctgccggga tcgcgattgt taccttcacc gacggtgtgt tgaccgcacg cgccttcgcc 840gctcgtcgag gtcaggaagt caatgccaac gccgagctgc gcgcggtcgg ggcctgcaac 900atcgccgccg ggctgacaca cggttttccg gtgagttcca gcagcagccg taccgccctc 960gccgacgtcg tcggtggccg cacccagctg tactcgctga tcgcgttggg gcttgttgtc 1020atcgtgatgg ttttcgcgag tgggctgctg gccatgtttc cgatcgccgc tctgggcgct 1080ttggtggtat atgccgcgct acgcttgatc gacttgtcag aattccggcg actggcgcgg 1140tttcggcgca gcgaactcat gctggcacta gccaccacag cagccgtgtt aggcctagga 1200gtgttctatg gagtcctcgc cgcggttgcc ctgtccatcc tcgaactgct tcgtcgggtc 1260gcacatccgc atgacagcgt tctcgggttc gtgccgggca ttgccggcat gcacgacatc 1320gatgactatc cgcaggccaa gcgcgtgccc gggctggtgg tgtatcgcta tgacgcgccg 1380ttgtgcttcg ccaatgccga agacttccgc aggcgagcac tgaccgtggt cgatcaggat 1440ccggggcaag tcgagtggtt cgtactcaac gccgaatcca atgtggaggt cgacctgact 1500gcgctggatg cgctcgacca actccgcacc gagctgctgc gtcggggaat agtgttcgcc 1560atggcccggg tcaaacaaga cttgcgtgaa tcactcaggg ccgccagtct tctcgataag 1620attggcgaag accatatctt tatgacattg cctaccgcag tgcaggcgtt ccgtcggcgc 168060429DNAMycobacterium tuberculosis 60atgatcacaa acctccgacg ccgaaccgcg atggcagccg ccggcctagg ggctgctctc 60gggctgggca tcctgctggt tccgacggtg gacgcccatc tcgccaacgg ttcgatgtcg 120gaagtcatga tgtcggaaat tgccgggttg cctatccctc cgattatcca ttacggggcg 180attgcctatg cccccagcgg cgcgtcgggc aaagcgtggc accagcgcac accggcgcga 240gcagagcaag tcgcactaga aaagtgcggt gacaagactt gcaaagtggt tagtcgcttc 300accaggtgcg gcgcggtcgc ctacaacggc tcgaaatacc aaggcggaac cggactcacg 360cgccgcgcgg cagaagacga cgccgtgaac cgactcgaag gcgggcggat cgtcaactgg 420gcgtgcaac 429612715DNAMycobacterium tuberculosis 61ttgtcggcgt cagtgtctgc cacgacggct catcatggct tgccagcaca tgaagtggtg 60ctgctgctgg agagcgatcc atatcacggg ctgtccgacg gcgaggccgc ccaacgacta 120gaacgcttcg ggcccaacac cttggcggtg gtaacgcgcg ctagcttgct ggcccgcatc 180ctgcggcagt ttcatcaccc gctgatctac gttctgctcg ttgccgggac gatcaccgcc 240ggtcttaagg aattcgttga cgccgcagtg atcttcggtg tggtggtgat caatgcgatc 300gtgggtttca ttcaagaatc caaggcagag gccgcactgc agggcctgcg ctccatggtg 360cacacccacg ccaaggtggt gcgcgagggt cacgagcaca caatgccatc cgaagagctg 420gttcccggtg accttgtgct gttagcggcc ggtgacaagg ttcccgccga tttgcggctg 480gtgcgacaga ccggattgag cgtgaacgag tcagcactta ccggcgagtc gacgccggtt 540cacaaggacg aggtggcgtt gccggagggc acaccggtcg ctgatcgtcg caatatcgcg 600tattccggca cattggtaac cgcgggccat ggcgccggga tcgtcgtcgc gaccggcgcc 660gaaaccgaac tcggtgagat tcatcggctc gttggggccg ccgaggttgt cgccacaccg 720ctgaccgcga agctggcgtg gttcagcaag tttctgacca tcgccatcct gggtctggca 780gcgctcacgt tcggcgtggg tttgctgcgc cggcaagatg ccgtcgaaac gttcaccgct 840gcgatcgcgc tggcggtcgg ggcaattccc gaaggtctgc ccaccgccgt gaccatcacc 900ttggccatcg gcatggcccg gatggccaag cgccgcgcgg tcattcgacg tctacccgcg 960gtggaaacgc tgggcagcac cacggtcatc tgcgccgaca agaccggaac gctgaccgag 1020aatcagatga cggtccagtc gatctggaca ccccacggtg agatccgggc gaccggaacg 1080ggctatgcac ccgacgtcct cctgtgcgac accgacgacg cgccggttcc ggtgaatgcc 1140aatgcggccc ttcgctggtc gctgctggcc ggtgcctgca gcaacgacgc cgcactggtt 1200cgcgacggca cacgctggca gatcgtcggc gatcccaccg agggcgcgat gctcgtcgtg 1260gccgccaagg ccggcttcaa cccggagcgg ctggcgacaa ctctgccgca agtggcagcc 1320ataccgttca gttccgagcg gcaatacatg gccaccctgc atcgcgacgg gacggatcat 1380gtggtgctgg ccaagggtgc tgtggagcgc atgctcgacc tgtgcggcac cgagatgggc 1440gccgacggcg cattgcggcc gctggaccgc gccaccgtgt tgcgtgccac cgaaatgttg 1500acttcccggg ggttgcgggt gctggcaacc gggatgggtg ccggcgccgg cactcccgac 1560gacttcgacg aaaacgtgat accaggttcg ctggcgctga ccggcctgca agcgatgagc 1620gatccaccac gagcggccgc ggcatcggcg gtggcggcct gccacagtgc cggcattgcg 1680gtaaaaatga ttaccggtga ccacgcgggc accgccacgg cgatcgcaac cgaggtgggg 1740ttgctcgaca acactgaacc ggcggcaggc tcggtcctga cgggtgccga gctggccgcg 1800ctgagcgcag accagtaccc ggaggccgtg gatacagcca gcgtgtttgc cagggtctct 1860cccgagcaga agctgcggtt ggtgcaagca ttgcaggcca gggggcacgt cgtcgcgatg 1920accggcgacg gcgtcaacga cgccccggcc ttgcgtcagg ccaacattgg cgtcgcgatg 1980ggccgcggtg gcaccgaggt cgccaaggat gccgccgaca tggtgttgac cgacgacgac 2040ttcgccacca tcgaagccgc ggtcgaggaa ggccgcggcg tattcgacaa tctgaccaag 2100ttcatcacct ggacgctgcc caccaacctc ggtgagggcc tagtgatctt ggccgccatc 2160gctgttggcg tcgccttgcc gattctgccc acccaaattc tgtggatcaa catgaccaca 2220gcgatcgcgc tcggactcat gctcgcgttc gagcccaagg aggccggaat catgacccgg 2280ccaccgcgcg accccgacca accgctgctg accggctggc ttgtcaggcg gactcttctg 2340gtttccacct tgctcgtcgc cagcgcgtgg tggctgtttg catgggagct cgacaatggc 2400gcgggcctgc atgaggcgcg cacggcggcg ctgaacctgt tcgtcgtcgt cgaggcgttc 2460tatctgttca gctgccggtc gctgacccga tcggcctggc ggctcggcat gttcgccaac 2520cgctggatca tcctcggcgt cagtgcgcag gccatcgcgc aattcgcgat cacatatcta 2580cccgcgatga atatggtgtt cgacaccgcg ccaatcgata tcggggtgtg ggtgcgcata 2640ttcgctgtcg cgaccgcaat cacgattgtg gtggccaccg acacgctgct gccgagaata 2700cgggcgcaac cgcca 271562774DNAMycobacterium tuberculosis 62atgagtttcc acgatcttca tcaccaaggt gttccgttcg tgttgcccaa cgcctgggat 60gtgccgtcgg ccctggccta cctcgcggag ggcttcacgg ctatcggcac aaccagtttc 120ggggtctcgt ccagcggcgg gcacccggac gggcaccgcg ccactcgcgg cgccaacatc 180gcactggcgg ccgccctggc accgctgcaa tgctacgtca gcgtcgacat cgaggacgga 240tacagcgacg aacccgacgc cattgctgac tacgtcgcac aactgtcgac agccggaatc 300aatatcgagg acagtagcgc cgaaaagctc atcgaccccg ccctggcagc cgctaaaatc 360gttgcgatca aacaacgtaa ccccgaggtg ttcgtcaacg cccgcgtcga cacctattgg 420ttgcgccagc acgccgatac caccagcacg atccagcgcg cacttcgcta cgtcgatgcc 480ggcgccgacg gcgtctttgt cccactggcc aacgatcccg acgaacttgc tgagctcact 540cgcaacattc cgtgcccggt taacacgttg cccgtgcccg gcttgacgat cgccgacctt 600ggtgagctcg gggtggcccg ggtgtcaacc ggttcagtgc cctacagcgc ggggttgtat 660gcagcggccc acgcggctcg ggccgtgagc gacggagagc agctgccacg gtccgtaccg 720tacgccgaac tgcaggcacg cttggttgac tacgagaacc gcacgagtac aacg 77463855DNAMycobacterium tuberculosis 63gtggtcaagc gctctcgggc aacccgactt tcgccgagca tctggtccgg atgggaatca 60cctcagtgtc ggtccattcg ggcgcgattg ctgctacccc ggggtcggtc gcggccgccg 120aacgccgatt gttgctggaa tcagctcgcg gtgacgcctg acacccggat gccggcatcg 180tcggccgccg ggcgcgacgc ggcggcctac gacgcctggt atgactcacc caccgggcgg 240ccgatcctgg cgaccgaggt cgccgcgttg cggccgctca tcgaggtctt tgcccagcca 300cgcttggaaa tcggtgtcgg tacaggacgt ttcgccgacc tgctcggcgt gcggttcgga 360ctcgatccat cccgtgatgc gctgatgttc gcacgccggc gcggcgtcct ggtcgccaat 420gccgtcggcg aggcggtccc tttcgtcagc cggcacttcg gggcggtcct catggcattc 480acgctctgtt tcgtcaccga cccggccgcc atattccggg aaacgcggcg tctgctcgcc 540gacggcggcg gccttgttat cgggttcttg cctcgcggga caccgtgggc cgacctgtac 600gctctgcgcg cggcccgcgg acagccaggc taccgcgacg cccgcttcta caccgcggcc 660gaactcgaac aactgctcgc agactcggga ttccgggtca tcgcccgccg ctgcacgctg 720caccaaccgc cgggactcgc ccggtacgac atcgaagccg cccatgacgg tatccaagcc 780ggcgccggct tcgttgctat ctcggcggtc gaccaagcgc acgagcctaa ggatgatcac 840ccactcgagt cggaa 85564885DNAMycobacterium tuberculosis 64atgtctaaac cccgcaagca gcacggagtt gtcgtcgggg tagatggttc gctcgaatcg 60gatgccgccg cctgttgggg tgccaccgat gcggcgatga ggaacattcc gctgaccgtg 120gtccacgtgg tgaacgccga tgtagcgacg tggccgccga tgccgtatcc ggagacctgg 180ggggtttggc aggaggacga gggtcgccag atcgtcgcca acgccgtcaa gctcgccaaa 240gaggcggttg gagcggatcg aaagctcagc gtaaagagcg agctcgtatt ttccacgccg 300gtacctacca tggttgaaat ctccaacgag gcagagatgg tggtgttggg cagctcgggc 360cggggagcgc tggcccgagg cttgctcggt tcggtcagct cgagcctggt gcgacgcgcc 420gggtgcccgg tcgcggtcat ccacagcgat gatgcggtga tccctgatcc gcagcacgct 480cccgtgctgg tgggaatcga cggttcgccg gtttcggagc ttgcgacggc ggtggcattt 540gacgaggcgt cgcgccgcgg cgtcgaactg atcgccgtgc acgcgtggag tgacgtcgaa 600gtggtggaac ttccgggttt ggacttctcg gctgtacagc aggaagcgga gcttagtctc 660gccgaacgct tggcaggttg gcaagaacgc tatcccgatg tgccggtgag ccgggttgtc 720gtttgcgatc gcccggcgcg gaagctggtg caaaagtcgg cgtccgccca gcttgtcgtc

780gttggcagtc atggccgagg tggcttgacc ggcatgcttc tggggtcggt cagtaacgcg 840gtcttacacg ccgcgcgggt gccagtgatc gtggcacggc agtcg 88565342DNAMycobacterium tuberculosis 65gtgacctatg tgatcggtag tgagtgcgtg gatgtgatgg acaagtcctg tgtgcaggag 60tgtccggtcg actgtatcta tgagggcgcc cgaatgctct acatcaaccc cgacgagtgc 120gtggattgtg gtgcgtgcaa accggcctgc cgcgtcgagg cgatctactg ggaaggcgat 180ctacccgacg atcaacacca gcatctgggg gacaacgccg cctttttcca ccaagtcctg 240ccgggccgag tggctccgct gggttcgccg ggtggtgccg cagcggtggg cccgatcgga 300gtcgacacgc ctctggtcgc ggctatcccg gtggagtgcc ct 34266837DNAMycobacterium tuberculosis 66atgaaccaat cacacaaacc cccatcgatc gtcgtcggta ttgatggctc gaagccggcc 60gtgcaagccg cactgtgggc ggtcgacgag gcagccagcc gtgacatccc gctgcgtctg 120ctgtacgcga tcgaacccga cgatcccggg tacgccgcac acggcgcggc ggctcgcaaa 180ctcgccgccg ccgagaacgc ggtgcgctac gcgttcacag cggtcgaggc ggcggaccgg 240ccggtcaagg tcgaggtgga gatcacccag gagcggccgg tcacctcgtt gatccgcgct 300tcggcggctg ctgccctggt gtgcgttggc gctatcggcg tgcaccactt ccgaccggag 360cgggtgggat ctaccgcagc ggccctggcg ttatcggcgc agtgcccagt ggcgatcgtg 420cgaccccacc gggtccccat cggacgcgac gccgcatgga tcgtcgtcga ggcggacggg 480tcgtccgata tcggtgtttt gctgggggcg gtgatggccg aagcacggct gcgcgactcg 540ccggttcggg tggtcacctg ccggcaatcc ggagtgggcg ataccgggga cgacgtccgt 600gccagcctgg accgctggct tgcccgttgg caaccacggt atcccgatgt gcgggtgcaa 660tcggcggcag tgcacggcga gctgctggat tatctggctg ggctgggtcg atcggtacac 720atggtggtgc tcagcgcgag cgaccaggag catgtggagc aacttgtggg agcgccgggc 780aacgccgtgt tgcaggaggc cggctgcacc ctgctggtcg tcggtcagca gtatctg 837671017DNAMycobacterium tuberculosis 67atgacggagc 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 ctcgcca 1017682043DNAMycobacterium tuberculosis 68gtgctgatga ccgcagcggc tgatgtcacc cggcgctcgc cgcggcgcgt gttccgtgac 60cgccgcgagg ccggccgggt gctggcggaa ttactcgccg cctatcggga ccagccggac 120gtgattgtgc tcggcttggc ccggggtggc ctcccggtcg catgggaggt tgccgcggca 180ctgcatgccc cgctagacgc cttcgtcgtg cgcaaacttg gtgccccggg gcatgacgag 240ttcgccgttg gtgcactggc cagcggcggc cgcgtcgtgg tcaatgacga cgtcgtgcgg 300ggcctgcgga tcacaccgca gcaactgcgc gacatcgccg aacgtgaggg tcgggaactg 360cttcggcgcg agtccgccta ccgcggcgag cgcccgccca ccgatatcac cggcaagacg 420gtcattgtcg tcgatgacgg tttggccacc ggcgcaagca tgttcgcggc ggtacaggca 480ttgcgcgatg cgcaaccagc gcagatcgtg attgccgtgc cggcggcgcc ggagtccacg 540tgccgggagt tcgccggcct cgtcgacgac gttgtgtgcg cgaccatgcc gaccccgttc 600ctggccgtcg gtgagtcgtt ttgggacttc cggcaggtca ccgacgagga ggtccgccgg 660ctcctggcca ccccgaccgc tgggccgtcg ctgcgccggc ccgcggcgtc aacggcggcc 720gatgttctgc gcagagtcgc gatcgacgcc cccgggggtg ttccgacgca cgaggtgttg 780gcggagctgg tcggcgatgc acgaatcgtg ttgatcggcg aaagctcgca cggcacacac 840gagttctacc aggcccgggc cgccatgaca cagtggctga tcgaggagaa gggctttggt 900gcggtagccg ccgaggcgga ctggcccgac gcctaccggg tcaatcggta cgttcgcggc 960ctcggcgagg acaccaacgc tgacgaggcg cttagcggat tcgagcggtt tcccgcctgg 1020atgtggcgca acaccgtggt ccgagatttt gtggaatggc tgcgcacacg caaccagcgc 1080tacgagtcgg gcgcgctgcg gcaagccggc ttctacggtc tggatcttta cagcctgcat 1140cggtcgatcc aagaggtgat cagctatctc gacaaggtcg acccgcgtgc ggcggcacgg 1200gcgcgggccc ggtatgcgtg cttcgaccat gcctgcgccg atgacggtca ggcgtacgga 1260ttcgcggccg cattcggcgc cggtccgtcg tgcgaacgtg aagccgtcga gcaactggtc 1320gacgttcagc gcaatgccct ggcgtatgcg cgccaagacg ggctgcttgc cgaggacgaa 1380ctgttctacg cccagcaaaa cgcgcagacg gtgcgcgacg cagaggtgta ttaccgggcc 1440atgttcagtg gacgcgttac ctcgtggaac ctgcgcgacc agcacatggc gcagaccctt 1500ggcagtttgc tgacgcattt ggaccgacac ctcgatgcgc cgccggcgcg aatagtggtg 1560tgggctcata actcccacgt gggtgacgca cgcgctaccg aggtgtgggc cgacgggcag 1620ctcaccctcg gccagatagt ccgtgagcga tacggtgacg agtcgcgcag catcggattc 1680agcacgtaca cgggcaccgt caccgcggcc agcgaatggg gtggtatcgc ccaacgcaaa 1740gcggttcggc cggcactgca cggcagtgtc gaggagctct tccaccagac tgcagacagt 1800ttcctggtgt cagcgcggct aagccgcgac gccgaagccc cgctggacgt tgtccggttg 1860ggacgtgcca tcggcgtcgt ttatctaccg gcaacggaac ggcaaagtca ctacttgcac 1920gtgcggcccg ccgaccagtt cgacgccatg atccacatcg atcagacccg tgccctggaa 1980cctctcgagg tgacgagccg gtggatcgcc ggcgagaacc cggaaaccta cccgaccggt 2040ctg 204369432DNAMycobacterium tuberculosis 69atggccacca 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 ac 43270993DNAMycobacterium tuberculosis 70atgccggaca 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 cgg 99371585DNAMycobacterium tuberculosis 71atgccactgc taaccattgg cgatcaattc cccgcctacc agctcaccgc tctcatcggc 60ggtgacctgt ccaaggtcga cgccaagcag cccggcgact acttcaccac tatcaccagt 120gacgaacacc caggcaagtg gcgggtggtg ttcttttggc cgaaagactt cacgttcgtg 180tgccctaccg agatcgcggc gttcagcaag ctcaatgacg agttcgagga ccgcgacgcc 240cagatcctgg gggtttcgat tgacagcgaa ttcgcgcatt tccagtggcg tgcacagcac 300aacgacctca aaacgttacc cttcccgatg ctctccgaca tcaagcgcga actcagccaa 360gccgcaggtg tcctcaacgc cgacggtgtg gccgaccgcg tgacctttat cgtcgacccc 420aacaacgaga tccagttcgt ctcggccacc gccggttcgg tgggacgcaa cgtcgatgag 480gtactgcgag tgctcgacgc cctccagtcc gacgagctgt gcgcatgcaa ctggcgcaag 540ggcgacccga cgctagacgc tggcgaactc ctcaaggctt cggcc 58572816DNAMycobacterium tuberculosis 72atgtctggga gaggagagcc gacgatgaaa acaatcattg ttggtatcga tggttcgcac 60gcggcgatta cggccgcatt gtggggggtt gacgaggcca tcagccgagc ggtgccgctg 120cgactggtct cagtgatcaa gccgacacat ccgtccccgg acgactacga ccgcgacctt 180gcgcatgctg aaagatcgct tcgggaagcg cagtccgctg ttgaggccgc gggcaagctc 240gtcaagatcg aaaccgacat cccccgcggg ccagccggcc cggtgcttgt ggaggcatcg 300cgcgacgccg agatgatctg cgtcggctcc gtgggaatcg ggcgctacgc cagctcgatc 360ttgggttcga cggcaaccga gctggccgaa aaggcgcatt gcccggtcgc cgtcatgcgc 420tcaaaagtgg accagccagc gtctgacatc aactggatcg tggtgcgcat gaccgacgca 480ccggataacg aggccgtgct ggaatacgct gcccgggaag cgaagttgcg gcaagcgccc 540atactggcac tcggcgggcg accggaggag ctccgggaga ttccggacgg cgaattcgaa 600cgtcgcgtgc aggattggca ccaccgtcat cccgatgtgc gcgtctaccc gatcaccact 660cacacgggta ttgcccggtt cctggccgac cacgacgagc gcgtacagct ggcagtgatc 720ggcggtggtg aggccggtca gctagcgcgg ctggtcgggc catccggaca tccggtgttc 780cgtcacgccg agtgttcggt gcttgtcgtt cgccgc 816731179DNAMycobacterium tuberculosis 73atgcgtgatg cgatcccgct tgggcggatc gccgggtttg tggtgaacgt ccactggagc 60gtgttggtga tcctgtggtt gttcacctgg agtctggcga ccatgttgcc gggtaccgtc 120ggaggctacc cggccgtggt ctattggctt ctcggcgcag gtggcgcggt catgttgctg 180gcgtcgctgt tggctcatga gctcgcgcac gccgtcgtcg ctcgtcgcgc cggggtatcc 240gttgagagcg tgacgttgtg gctgttcggc ggggtgaccg cgcttggcgg cgaggcaaag 300acgcccaaag ccgctttccg gatcgcgttc gcgggtccgg ctaccagcct ggcgctgtcg 360gcgacattcg gtgcgttggc catcacgctc gccggcgtgc ggaccccggc catcgtgatc 420agcgttgctt ggtggttggc tactgtcaac ctgctgctgg ggctgttcaa tctgctgcct 480ggcgcgccgt tggacggtgg gcggttggtc cgggcctatc tgtggcgccg ccacggcgat 540agtgtgcgcg ccgggatcgg tgcggcgcgg gccggacggg tggttgcgct ggtcttgatc 600gcgttgggat tggccgagtt tgtggctggt ggcctcgtcg gtggggtctg gttagccttc 660attggctggt ttatcttcgc tgccgctcgc gaggaggaga cccggatttc gacccagcag 720ctgtttgccg gggtgcgtgt ggccgatgcg atgaccgccc aaccgcatac ggctcccgga 780tggatcaatg tcgaggattt catccagcgt tacgtgcttg gtgaacggca ctcggcatat 840ccggttgccg atcgggacgg atcgatcacg ggcctggtgg cattgcggca gctgcgcgat 900gttgcgccta gccggcgcag cactaccagc gtaggtgaca ttgcgctgcc gctgcacagc 960gtgccgaccg cccgaccaca agagccgctg accgcgctcc tagagcggat ggcaccgctc 1020ggcccgcgca gccgtgcgct ggtcaccgaa gggagcgcgg tggtcggcat cgtcactccc 1080agcgatgtcg cgcggctgat tgacgtctac cggttggccc agccggaacc gacctttacc 1140acgagtcccc aagatgcgga caggttttcc gatgcgggg 1179741239DNAMycobacterium tuberculosis 74atggcaagtt 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 cgttccgcc 123975360DNAMycobacterium tuberculosis 75atgtccacgc 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 360761122DNAMycobacterium tuberculosis 76atgcgatcag aacgtctccg gtggctggta gccgcagaag gtccgttcgc ctcggtgtat 60ttcgacgact cgcacgacac tcttgatgcc gtcgagcgcc gggaagcgac gtggcgcgat 120gtccggaagc atctcgaaag ccgcgacgcg aagcaggagc tcatcgacag cctcgaagag 180gcggtgcggg attctcgacc ggccgtcggc cagcgtggcc gcgcgctgat cgcgaccggc 240gagcaagtac tggtcaacga gcatctgatc ggcccaccac cggctacggt gattcggctg 300tcggattatc cgtacgtcgt gccattgata gaccttgaga tgcggcgacc gacgtatgta 360tttgccgcgg ttgatcacac cggcgccgac gtcaagctgt atcagggggc caccatcagt 420tccacgaaaa tcgatggggt cggctacccg gtgcacaagc cggtcaccgc cggctggaac 480ggctacggcg acttccagca caccaccgaa gaagccatcc gaatgaactg ccgcgcggtc 540gccgaccatc tcacccgact ggtagacgct gccgaccccg aggtggtgtt cgtgtccggc 600gaggtgcggt cacgcacaga cctgctttcc acattgccgc agcgggtggc ggtccgggtg 660tcgcagctgc atgccggacc gcgcaaaagc gccttagacg aggaagagat ctgggacctg 720acatccgcgg agttcacccg gcggcggtac gccgaaatca ccaatgtcgc acaacaattt 780gaggcggaga tcggacgcgg atcggggctg gcggcccaag ggttggcgga ggtgtgtgcg 840gctctgcgtg acggcgacgt cgacacgctg atcgtcggag agctaggcga ggccaccgtg 900gtcaccggta aagcgcgtac tacggtcgcg cgggatgccg acatgttgtc cgaactcggc 960gaaccggtag atcgcgtggc aagggccgat gaggcgttgc cattcgccgc gatcgcggta 1020ggtgccgcat tggtccgtga cgacaaccgg atcgcgccac tagatggggt gggcgcattg 1080ctgcgttatg ccgccaccaa ccgactcggc agccatagat cc 112277537DNAMycobacterium tuberculosis 77atgctgcacc gcgacgatca catcaatccg ccgcggcccc gcgggttgga tgttccttgc 60gcccgcctac gagcgacaaa tcccctgcgc gccttggcgc gttgcgttca ggcgggcaag 120ccgggcacca gttcagggca tcggtccgtg ccgcatacgg cggacttgcg aatcgaagcc 180tgggcaccga cccgtgacgg ctgtatccgg caggcggtgc tgggtaccgt cgagagcttc 240ctcgacctgg aatccgcgca cgcggtccat acccggctgc gccggctgac cgcggatcgc 300gacgacgatc tactggtcgc ggtgctcgag gaggtcattt atttgctgga caccgtcggt 360gaaacgcctg tcgatctcag gctgcgcgac gttgacgggg gtgtcgacgt cacattcgca 420acgaccgatg cgagtacgct agttcaggtg ggtgccgtgc cgaaggcggt gtcactcaac 480gaacttcggt tctcgcaggg tcgccacggc tggcgatgtg cggtaacgct cgatgtg 537781125DNAMycobacterium tuberculosis 78gtgacgcaaa 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 agatg 1125791113DNAMycobacterium tuberculosis 79atgcgcgtcg gtattccgac cgagaccaaa aacaacgaat tccgggtggc catcaccccg 60gccggcgtcg cggaactaac ccgtcgtggc catgaggtgc tcatccaggc aggtgccgga 120gagggctcgg ctatcaccga cgcggatttc aaggcggcag gcgcgcaact ggtcggcacc 180gccgaccagg tgtgggccga cgctgattta ttgctcaagg tcaaagaacc gatagcggcg 240gaatacggcc gcctgcgaca cgggcagatc ttgttcacgt tcttgcattt ggccgcgtca 300cgtgcttgca ccgatgcgtt gttggattcc ggcaccacgt caattgccta cgagaccgtc 360cagaccgccg acggcgcact acccctgctt gccccgatga gcgaagtcgc cggtcgactc 420gccgcccagg ttggcgctta ccacctgatg cgaacccaag ggggccgcgg tgtgctgatg 480ggcggggtgc ccggcgtcga accggccgac gtcgtggtga tcggcgccgg caccgccggc 540tacaacgcag cccgcatcgc caacggcatg ggcgcgaccg ttacggttct agacatcaac 600atcgacaaac ttcggcaact cgacgccgag ttctgcggcc ggatccacac tcgctactca 660tcggcctacg agctcgaggg tgccgtcaaa cgtgccgacc tggtgattgg ggccgtcctg 720gtgccaggcg ccaaggcacc caaattagtc tcgaattcac ttgtcgcgca tatgaaacca 780ggtgcggtac tggtggatat agccatcgac cagggcggct gtttcgaagg ctcacgaccg 840accacctacg accacccgac gttcgccgtg cacgacacgc tgttttactg cgtggcgaac 900atgcccgcct cggtgccgaa gacgtcgacc tacgcgctga ccaacgcgac gatgccgtat 960gtgctcgagc ttgccgacca tggctggcgg gcggcgtgcc ggtcgaatcc ggcactagcc 1020aaaggtcttt cgacgcacga aggggcgtta ctgtccgaac gggtggccac cgacctgggg 1080gtgccgttca ccgagcccgc cagcgtgctg gcc 111380312DNAMycobacterium tuberculosis 80atggtcatcc ggtttgatca aatagggtca ttggtcctct caatgaaatc ccttgcgtca 60ctgtcgtttc agcggtgtct gcgcgagaat tctagtttgg tcgcggcgct ggaccggctc 120gatgctgcgg tcgatgagct gagcgctttg tcgtttgatg cgttgaccac tccggagcgg 180gatcgcgccc gtcgcgaccg ggaccatcat ccttggtccc gctcccgctc gcagttgtcg 240ccacgaatgg cgcacggtgc agtgcaccaa tgccagtggc cgaaggcggt ttgggctgtc 300attgacaatc ca

312811032DNAMycobacterium tuberculosis 81gtgctcaaga acgcagtctt gctggcatgc cgggcgccgt cggtgcacaa cagccagccc 60tggcgttggg tggccgaaag cggctccgag cacactactg tgcacctgtt cgtcaaccgc 120caccgaacgg tgccggccac cgaccattcc ggccggcaag cgatcatcag ttgcggtgcc 180gtactcgatc accttcgcat cgccatgacg gccgcgcact ggcaggcgaa tatcactcgc 240tttccccagc cgaaccaacc tgaccagttg gccaccgtcg aattcagtcc catcgatcac 300gtcacggcgg gacagcgaaa ccgcgcccag gcgattctgc agcgccgaac cgatcggctt 360ccgtttgaca gcccgatgta ctggcacctg tttgagcccg cgctgcgcga cgccgtcgac 420aaagacgttg cgatgcttga tgtggtatcc gacgaccagc gaacacgact ggtggtagcg 480tcacaactca gcgaagtcct gcggcgggac gatccgtact atcacgccga actcgaatgg 540tggacttcac cgttcgtgct ggcccatggt gtgccgccgg atacgctggc atcagacgcc 600gaacgcttgc gggttgacct gggccgtgac ttcccggtcc ggagctacca gaatcgccgt 660gccgagctag ctgatgaccg atcgaaagtc cttgtgctgt cgacccctag cgacacgcga 720gccgacgcac tgaggtgtgg cgaagtgctg tcgaccatcc tactcgagtg caccatggcc 780ggcatggcta cctgcacgtt gacccatctg atcgaatcca gtgacagtcg tgacatcgtg 840cggggcctga cgaggcagcg aggcgagccg caagccttga tccgggtagg gatagccccg 900ccgttggcag cagttcccgc ccccacacca cggcggccgc tggacagcgt cttgcagatt 960cgccagacgc ccgagaaagg gcgtaatgcc tcagatagaa atgcccgtga aacgggttgg 1020ttcagcccgc ct 1032821011DNAMycobacterium tuberculosis 82gtgtggtccg cctcgggtgg gcagtgcggg aagtatcttg ccgcctcgat ggtgctgcag 60cttgatgggt tggaacgtca cggtgtgttg gagtttgggc gtgaccgcta tggccccgag 120gtgcgtgagg agctgttggc gatgagtgcg gccagcatcg atcgttatct gaagaccgcg 180aaggccaaag accagatatc gggtgtgtcg acgacgaaac cctcaccact gctgcgtaat 240tcgatcaagg ttcgcagggc cggcgatgag gtcgaggcgg agccggggtt cttcgagggc 300gacaccgtcg cccattgcgg tccgacgctc aaaggcgagt tcgcccacac cctgaacttg 360accgacgtgc acatcggatg ggtgttcacc cgcaccgtcc gcaacaacgc ccgtacccac 420atcctcgccg ggctcaaagc ttctgtcacc gagatcccgc atgggataac gggtttagat 480ttcgacaacg gcaccgtgtt tctcaacaag ccggtcatca gctgggccgg cgacaacggt 540atctacttca cccgctttcg cccgtacaag aaaaaccact aggccaccat cgagtccaag 600aacaaccacc tggtccgcaa gtacgcgttc tactaccgct atgacaccgc cgaggaacgc 660gccgtgctca accggatgtg gaagctggtc aacgaccgcc tcaactacct caccccgacc 720atcaaaccga tcgggtatgc cagcagcgcc gacggccgcc gccgacgcct ctacgatgcc 780ccacagacgc cgctggaccg gccactggcc gcaagggtgc tctccgcggc ccagcaggcc 840gacctgatca cctaccgaga cagcctcaac cccgcccaga tcggccgcaa aatcgccgac 900ctgcagaacc gactcctcat cttggccaag gagaaaaccg agcagctcta cctcgctaac 960atccccaccg ccctacccga catccacaaa ggcatcctga tcaaggcggg c 101183330DNAMycobacterium tuberculosis 83gtggtgcaag gccgcaccgt gctgtttcgt accgcggagg gcgccaaatt attttcagcc 60gtcgcgaagt gcgcggtggc tttcgaggcg gacgaccaca acgttgccga gggctggagc 120gtgatcgtca aggttcgcgc ccaggtgctg acgaccgacg cgggggtccg cgaagccgaa 180cgcgcccagt tactaccgtg gaccgcgacg ctgaaacgtc actgtgtgcg ggtgatcccg 240tgggagatca ccggccgcca cttcaggttc ggtccggaac cggaccgcag ccagaccttt 300gcctgcgagg cctcgtcaca caaccagcga 330841389DNAMycobacterium tuberculosis 84atgaatcacc taacgacact tgacgccggg tttctcaagg cagaagacgt ggatcggcac 60gtgagtctgg caatcggcgc tctggcggtc atcgaggggc cggctcccga tcaggaagcc 120ttcttatcgt cgctcgctca acgcctacgt ccctgtaccc ggttcgggca gcggttacgc 180ctgcgcccgt tcgacctcgg tgcacccaaa tgggtggacg atcccgactt cgatcttggc 240cgtcatgtgt ggcgcatcgc cttgccgcgg cctggcaacg aagaccagtt attcgagctg 300atcgccgatc tgatggcgcg tcgtttggac cggggtcgac cgctgtggga ggtctgggtc 360atcgaaggcc tggcggacag caagtgggcg atcctgacca aactgcacca ctgcatggcc 420gacggaatcg cggcgactca cctgctagct gggctctccg atgaaagtat gagcgacagc 480ttcgcgagca acatccacac gaccatgcag tcgcaatccg catctgtgcg gcggggtgga 540ttccgtgtca atccaagcga ggcgttgacc gcgtcgaccg ccgtgatggc aggcatcgtt 600cgcgcggcca agggtgccag tgagatcgcg gccggcgtgc taagtcccgc cgcgtcgtcg 660ttgaacgggc cgatcagtga tttgcgtcgc tacagcgcag caaaggtccc tctcgccgac 720gtcgaacagg tgtgccggaa attcgacgtc accatcaatg atgttgcgct tgccgcgatt 780acggaaagct accgcaacgt cctcatccag cggggtgagc ggcctaggtt tgattcgctg 840cgtacgctag tgccggtctc gacgcgttcc aacagcgctt tgagcaagac cgataaccgt 900gtttcgttaa tgctgcccaa cctgccggtg gatcaagaga acccgctgca gcggctgcgg 960atcgtgcact cgcggctgac tcgggccaag gcggggggac agagacaatt cggaaatact 1020ttgatggcga ttgccaaccg ccttccgttc cccatgaccg catgggcggt cgggctgttg 1080atgcggctgc cgcagcgtgg tgttgtcacc gtggcgacaa atgtgccggg tccacgacgg 1140ccgctgcaga ttatgggcag acgggtgctt gacctatacc cggtttcgcc gatcgcgatg 1200caactgcgca ccagtgtcgc gatgctcagc tacgccgacg acctgtactt cgggatcctg 1260gccgactacg acgtggtagc agatgccggc cagctggcgc gaggaattga agacgccgtc 1320gcacggctgg tggcgatcag taagcggcgc aaggtgactc gcaggcgcgg agcgctatcg 1380ctggttgtg 138985996DNAMycobacterium tuberculosis 85atgaacaccc atttcccgga cgccgaaacc gtgcgaacgg ttctcaccct ggccgtccgg 60gccccctcca tccacaacac gcagccgtgg cggtggcggg tatgcccgac gagtctggag 120ctgttctcta gacccgatat gcagctgcgt agcaccgatc cggacgggcg tgagttgatc 180ctcagctgtg gtgtggcatt gcaccactgc gtcgtcgctt tggcgtcgct gggctggcag 240gccaaggtaa accgtttccc cgatcccaag gaccgctgcc atctggccac catcggggta 300caaccgcttg ttcccgatca ggccgatgtc gccttggcgg cggccatacc gcggcgacgc 360accgatcggc gcgcctacag ttgctggccg gtgccaggag gtgacatcgc gttgatggcc 420gcaagagcag cccgtggcgg ggtcatgctg cggcaggtca gtgccctaga ccgaatgaaa 480gccattgtgg cgcaggctgt cttggaccac gtgaccgacg aggaatatct gcgcgagctc 540accatttgga gtgggcgcta cggttcagtg gccggggttc ccgcccgcaa cgagccgcca 600tcagacccca gtgccccgat ccccggtcgc ctgttcgccg ggcccggtct gtctcagccg 660tccgacgtct tacccgctga cgacggcgcc gcgatcctgg cactaggcac cgagacagac 720gaccggttgg cccggctgcg cgccggcgag gccgccagca tcgtcttgtt gaccgcgacg 780gcaatggggc tggcgtgctg cccgatcacc gaaccgctgg agatcgccaa gacccgcgac 840gcggtccgtg ccgaggtgtt cggcgccggc ggctaccccc agatgctgct gcgagtgggt 900tgggcaccga tcaatgccga cccgttgcca ccgacgccac ggcgcgaact gtcccaggtc 960gttgagtggc cggaagagct actgcgacaa cggtgc 996861734DNAMycobacterium tuberculosis 86atgacaacag ggggcctcgt cgacgaaaac gacggcgccg caatgcgtcc actgcgtcac 60acgctctccc aactacgcct gcacgagctg ctggtcgagg tgcaggaccg ggtcgagcag 120atcgtcgagg gccgggaccg cctcgatggt ctggtggagg ccatgctcgt ggtcacagcg 180ggcctggacc tggaggcaac cctacgcgct atcgtgcatt cagcgaccag ccttgtcgat 240gcgcgctatg gcgctatgga ggtgcacgac cggcagcatc gggtattgca ctttgtctat 300gaaggcatcg acgaggagac cgttcggcgg atcggccacc taccgaaagg cctaggcgtc 360atcgggctgc tcatcgaaga tcccaaaccg ttacggctgg acgatgtttc tgcgcacccg 420gcctcgattg gttttccgcc gtatcatccg ccgatgcgta ccttcctcgg ggtaccggtt 480cgggtgcgcg atgaatcgtt cggcactctg tacctgactg acaagaccaa cgggcaaccg 540ttcagcgacg acgacgaggt tctggtccag gcgctggcgg ccgccgcggg tatcgcagtc 600gcgaatgccc ggctctacca gcaggctaag gcgcgtcagt cgtggatcga ggccacccgt 660gacatcgcca ccgagttgtt gtccggcacc gaacccgcga cggtgttccg gcttgtcgcc 720gcggaggcgc tcaagctgac ggcggctgac gctgccctgg tagccgttcc cgtcgacgag 780gacatgcctg ccgctgacgt gggggagctg ctggtgattg aaacagtcgg cagcgctgtg 840gcttccattg ttgggcgaac gattccggtg gcgggcgcgg tgctgcggga ggtcttcgtc 900aacggcattc cgcgacgggt cgaccgggtc gatttggaag gcctggacga actggccgac 960gcaggtccgg cgctgctgtt gccgctgcgg gccagaggta ccgtagcggg tgtcgttgtt 1020gtgctgagtc aaggcggtcc aggggctttc accgacgaac aactcgagat gatggccgcg 1080ttcgccgacc aggccgcgct ggcttggcaa ttggccactt cgcaacgtcg gatgcgcgaa 1140ctcgacgtac tgaccgaccg ggatcgtatc gcccgtgacc tccatgacca tgtcatccag 1200cggctcttcg cgattggcct ggctttgcag ggtgctgtcc cgcacgaacg taatcctgaa 1260gtgcagcaac gactctcgga cgtggtagac gatctgcaag acgttataca ggaaatccgg 1320accaccattt atgacctgca cggagcatcg cagggtatca ctcggctccg gcagcgaatc 1380gatgcggccg tagcccaatt tgccgactcg gggttgcgca ccagcgttca attcgtgggt 1440ccattgtcgg tggtcgacag cgcgctcgcc gatcaggccg aggcggtggt tcgggaagcg 1500gtcagcaacg cggttcgcca tgcgaaggcc agcacgttga ccgtccgggt caaagtcgac 1560gacgacttgt gcatcgaggt gaccgacaac ggccgcgggc tgcccgacga gttcaccgga 1620agcggcttaa cgaacctgcg gcagcgggca gagcaggccg gcggcgaatt caccctcgcg 1680agcgtaccgg gcgcgagcgg aacagtgctg cgatggtcag caccgttgtc gcag 173487804DNAMycobacterium tuberculosis 87atgagcgatc ctcggccagc tcgggcagtg gtcgttggta tcgacgggtc aagggcggca 60acgcatgcgg cgttgtgggc ggtcgatgag gcggtgaacc gagacattcc gctgcgactg 120gtgtacgtca tcgatccgtc ccaactgtcc gccgccggcg agggcggtgg gcaatcagcg 180gcccgagcgg cgctgcacga cgcctctcgg aaggtcgagg ccaccgggca accggtcaag 240atcgaaacgg aggttctgtg cggcaggccg ctcaccaagc tgatgcagga gtccaggtcc 300gcggcgatgc tgtgcgtcgg ttcggtgggg cttgatcatg tccgcggtcg ccggggttcg 360gtcgcggcga ccctggctgg gtcggcctta tgccccgtgg cggtgattca cccgtcgccg 420gccgagccag cgacaacctc ccaggtcagc gcggttgtcg cggaggtgga caatggtgtg 480gtgctgcggc acgcattcga ggaggccagg ctgcgcggag ttccgctgcg ggccgtggct 540gtccacgctg ctgaaacacc cgatgacgtc gaacagggca gccggttggc gcatgtacac 600ctgagccgtc ggctcgccca ctggacccgg ctctaccccg aggtgcgggt ggatcgggcc 660atcgccggcg gcagtgcgtg ccgtcatctg gccgccaacg caaagccggg tcagctgttc 720gtcgcggact cacactccgc gcacgaattg tgcggtgcat accagcccgg atgcgccgta 780cttacggtac gcagtgccaa cttg 80488543DNAMycobacterium tuberculosis 88atgacagaat acgaagggcc taagacaaaa ttccacgcgt taatgcagga acagattcat 60aacgaattca cagcggcaca acaatatgtc gcgatcgcgg tttatttcga cagcgaagac 120ctgccgcagt tggcgaagca tttttacagc caagcggtcg aggaacgaaa ccatgcaatg 180atgctcgtgc aacacctgct cgaccgcgac cttcgtgtcg aaattcccgg cgtagacacg 240gtgcgaaacc agttcgacag accccgcgag gcactggcgc tggcgctcga tcaggaacgc 300acagtcaccg accaggtcgg tcggctgaca gcggtggccc gcgacgaggg cgatttcctc 360ggcgagcagt tcatgcagtg gttcttgcag gaacagatcg aagaggtggc cttgatggca 420accctggtgc gggttgccga tcgggccggg gccaacctgt tcgagctaga gaacttcgtc 480gcacgtgaag tggatgtggc gccggccgca tcaggcgccc cgcacgctgc cgggggccgc 540ctc 54389822DNAMycobacterium tuberculosis 89atgacatggg ccgacgaggt gctcgccgga catccctttg tggttgctca ccgtggtgcg 60tcggcggctc ggccggagca tacccttgcc gcctacgacc tggcgctcaa agagggcgcc 120gacggcgtgg aatgtgatgt gcggttgacc cgggacgggc atctggtctg tgtgcatgac 180cgccgcctgg accgaacctc gacgggagcc ggcttggtca gcacgatgac gctggcccag 240ctacgcgagc tggagtacgg cgcgtggcac gacagctggc gccccgacgg ttcgcacggc 300gacaccagtc tgctgaccct ggacgcgctt gtttcgctgg ttttggactg gcaccggccg 360gtgaagatct tcgtcgagac caagcatccc gtccgatacg gctcgctggt ggaaaacaag 420ctgctggcgc tgctacaccg gttcggtatt gccgcacccg cctccgcaga tcgatcccgt 480gcggtggtga tgtcgttttc ggccgccgcg gtctggcgga tccggcgggc tgcaccgctg 540ctgccgacgg tgttgctcgg caagaccccc cgatacctga ccagcagtgc ggccacggcg 600gtcggggcaa ccgccgtggg accctcactg cctgcgttaa aggaatatcc gcaactcgtt 660gaccgctcgg cagctcaggg ccgggcggtg tactgctgga acgtcgatga gtacgaggac 720atcgactttt gccgggaggt cggggtggcc tggattggta ctcaccaccc cggccgcacc 780aaggcctggc tggaagacgg gcgggcgaac gggaccactc gc 82290744DNAMycobacterium tuberculosis 90gtgtccgacg gcgaacaagc caaatcacgt cgacgccggg ggcggcgccg cgggcggcgc 60gctgcggcta cagccgagaa tcacatggac gcccaaccgg ccggcgacgc caccccgacc 120ccggcaacgg cgaagcggtc ccggtcccgc tcacctcgtc gcgggtcgac tcggatgcgc 180accgtgcacg aaacatcggc tggagggttg gtcattgacg gtatcgacgg tccacgagac 240gcgcaggtcg cggctctgat cggccgcgtc gaccggcgcg gccggctgct gtggtcgcta 300cccaaggggc acatcgagtt gggcgagacc gccgagcaga ccgccatccg cgaggtcgcc 360gaggagaccg gcatccgcgg cagtgtgctc gccgcgctgg ggcgcatcga ctactggttc 420gtcaccgacg gccggcgggt gcacaagacc gtccaccatt atttgatgcg gtttttaggc 480ggagagctgt ccgacgaaga cctcgaggta gccgaggtag cctgggtgcc gatccgggaa 540ctgccgtctc gactggccta cgccgacgaa cgtcgactag ccgaggtggc cgacgaactg 600atcgacaagc tgcagagcga cggccccgcc gcgcttccgc cgctaccacc cagctcgcct 660cgtcgacggc cgcaaacgca ttcacgcgct cgtcatgccg atgactcagc accgggtcag 720cacaacggtc ccgggccggg gccg 7449188PRTMycobacterium tuberculosis 91Met Lys Ala Lys Val Gly Asp Trp Leu Val Ile Lys Gly Ala Thr Ile1 5 10 15Asp Gln Pro Asp His Arg Gly Leu Ile Ile Glu Val Arg Ser Ser Asp 20 25 30Gly Ser Pro Pro Tyr Val Val Arg Trp Leu Glu Thr Asp His Val Ala 35 40 45Thr Val Ile Pro Gly Pro Asp Ala Val Val Val Thr Ala Glu Glu Gln 50 55 60Asn Ala Ala Asp Glu Arg Ala Gln His Arg Phe Gly Ala Val Gln Ser65 70 75 80Ala Ile Leu His Ala Arg Gly Thr 859224DNAArtificialprimer sequence 92caccgtggaa ccgaaacgca gtcg 249319DNAArtificialprimer sequence 93ttatgccaga ccgtcggca 199419DNAArtificialprimer sequence 94caccatgagc ccgggctcg 199520DNAArtificialprimer sequence 95ttacggcgta cgcgagtcag 209624DNAArtificialprimer sequence 96caccgtggag tccgaaccgc tgta 249717DNAArtificialprimer sequence 97ttacgtggcc gagccgc 179822DNAArtificialprimer sequence 98caccatgcct atcgcaacgc cc 229923DNAArtificialprimer sequence 99ttagtgggtt agggactttc cgg 2310049DNAArtificialprimer sequence 100ggggacaagt ttgtacaaaa aagcaggctt aaaggcaaag gtcggggac 4910150DNAArtificialprimer sequence 101ggggaccact ttgtacaaga aagctgggtc ctacgttccc ctggcatgga 5010222DNAArtificialprimer sequence 102caccatgggt gagcacgcca tc 2210326DNAArtificialprimer sequence 103ttataggtca tcggattgag gtgatc 2610424DNAArtificialprimer sequence 104caccgtggct ggcaatcctg atgt 2410524DNAArtificialprimer sequence 105ttactccttg ctcgttaggt tggc 2410622DNAArtificialprimer sequence 106caccgtgaca gaccacgtgc gc 2210718DNAArtificialprimer sequence 107ttacggtgac gagccggc 1810825DNAArtificialprimer sequence 108caccatggta gagcccggca atttg 2510918DNAArtificialprimer sequence 109ttagagcgga cggcggct 1811022DNAArtificialprimer sequence 110caccatgatc gccacaaccc gc 2211120DNAArtificialprimer sequence 111ttaccgctgc gtgcagaaca 2011224DNAArtificialprimer sequence 112caccatgacc aacgtcggtg acca 2411321DNAArtificialprimer sequence 113ttatcctgtt actgcggcgc a 2111424DNAArtificialprimer sequence 114caccgtgacg gtgacaccac ggac 2411516DNAArtificialprimer sequence 115ttaccacccg cgccgc 1611621DNAArtificialprimer sequence 116caccatgaga gggcaagcgg c 2111722DNAArtificialprimer sequence 117ttacctggac gcctcctcac tc 2211821DNAArtificialprimer sequence 118caccatgtgc ggcgaccagt c 2111921DNAArtificialprimer sequence 119ttaatacaac aatcgcgccg g 2112025DNAArtificialprimer sequence 120caccatgatt cccacgatga catcg 2512117DNAArtificialprimer sequence 121ttagcgccga cggaacg 1712249DNAArtificialprimer sequence 122ggggacaagt ttgtacaaaa aagcaggctt aatcacaaac ctccgacgc 4912351DNAArtificialprimer sequence 123ggggaccact ttgtacaaga aagctgggtc ctagttgcac gcccagttga c 5112424DNAArtificialprimer sequence 124caccttgtcg gcgtcagtgt ctgc 2412517DNAArtificialprimer sequence 125ttatggcggt tgcgccc 1712629DNAArtificialprimer sequence 126caccatgagt ttccacgatc ttcatcacc 2912724DNAArtificialprimer sequence 127ttacgttgta ctcgtgcggt tctc 2412822DNAArtificialprimer sequence 128caccgtggtc aagcgctctc gg 2212922DNAArtificialprimer sequence 129ttattccgac tcgagtgggt ga 2213024DNAArtificialprimer sequence 130caccatgtct aaaccccgca agca 2413119DNAArtificialprimer sequence 131ttacgactgc cgtgccacg 1913230DNAArtificialprimer sequence 132caccgtgacc tatgtgatcg gtagtgagtg 3013320DNAArtificialprimer sequence 133ttaagggcac tccaccggga 2013426DNAArtificialprimer sequence 134caccatgaac caatcacaca aacccc 2613525DNAArtificialprimer sequence 135ttacagatac tgctgaccga cgacc 2513620DNAArtificialprimer sequence 136caccatgacg gagccagcgg 2013718DNAArtificialprimer sequence 137ttatggcgag gcttccgg 1813849DNAArtificialprimer sequence 138ggggacaagt ttgtacaaaa aagcaggctt actgatgacc gcagcggct 4913953DNAArtificialprimer sequence 139ggggaccact ttgtacaaga aagctgggtc ctacagaccg gtcgggtagg ttt 5314048DNAArtificialprimer sequence 140ggggacaagt ttgtacaaaa aagcaggctt

agccaccacc cttcccgt 4814154DNAArtificialprimer sequence 141ggggaccact ttgtacaaga aagctgggtc ctagttggtg gaccggatct gaat 5414222DNAArtificialprimer sequence 142caccatgccg gacaccatgg tg 2214324DNAArtificialprimer sequence 143ttagtgatcc ttagcccgaa cgtg 2414452DNAArtificialprimer sequence 144ggggacaagt ttgtacaaaa aagcaggctt aatgccactg ctaaccattg gc 5214552DNAArtificialprimer sequence 145ggggaccact ttgtacaaga aagctgggtc ctaggccgaa gccttgagga gt 5214625DNAArtificialprimer sequence 146caccatgtct gggagaggag agccg 2514721DNAArtificialprimer sequence 147ttagcgaacg acaagcaccg a 2114848DNAArtificialprimer sequence 148ggggacaagt ttgtacaaaa aagcaggctt acgtgatgcg atcccgct 4814950DNAArtificialprimer sequence 149ggggaccact ttgtacaaga aagctgggtc ctaccccgca tcggaaaacc 5015051DNAArtificialprimer sequence 150ggggacaagt ttgtacaaaa aagcaggctt aatggcaagt tctgcgagcg a 5115151DNAArtificialprimer sequence 151ggggaccact ttgtacaaga aagctgggtc ctaggaacgg tcgcgctgtg t 5115222DNAArtificialprimer sequence 152caccatgtcc acgcaacgac cg 2215321DNAArtificialprimer sequence 153ttaaccgcaa cggcaatctc a 2115424DNAArtificialprimer sequence 154caccatgcga tcagaacgtc tccg 2415523DNAArtificialprimer sequence 155ttaggatcta tggctgccga gtc 2315621DNAArtificialprimer sequence 156caccatgctg caccgcgacg a 2115723DNAArtificialprimer sequence 157ttacacatcg agcgttaccg cac 2315848DNAArtificialprimer sequence 158ggggacaagt ttgtacaaaa aagcaggctt agtgacgcaa accggcaa 4815952DNAArtificialprimer sequence 159ggggaccact ttgtacaaga aagctgggtc ctacatctcc tggttctcgg cc 5216049DNAArtificialprimer sequence 160ggggacaagt ttgtacaaaa aagcaggctt acgcgtcggt attccgacc 4916148DNAArtificialprimer sequence 161ggggaccact ttgtacaaga aagctgggtc ctacacgctg gcgggctc 4816228DNAArtificialprimer sequence 162caccatggtc atccggtttg atcaaata 2816324DNAArtificialprimer sequence 163ttatggattg tcaatgacag ccca 2416426DNAArtificialprimer sequence 164caccgtgctc aagaacgcag tcttgc 2616521DNAArtificialprimer sequence 165ttaaggcggg ctgaaccaac c 2116620DNAArtificialprimer sequence 166caccgtgtgg tccgcctcgg 2016720DNAArtificialprimer sequence 167ttagcccgcc ttgatcagga 2016820DNAArtificialprimer sequence 168caccgtggtg caaggccgca 2016922DNAArtificialprimer sequence 169ttatcgctgg ttgtgtgacg ag 2217029DNAArtificialprimer sequence 170caccatgaat cacctaacga cacttgacg 2917123DNAArtificialprimer sequence 171ttacacaacc agcgatagcg ctc 2317223DNAArtificialprimer sequence 172caccatgaac acccatttcc cgg 2317323DNAArtificialprimer sequence 173ttagcaccgt tgtcgcagta gct 2317423DNAArtificialprimer sequence 174caccatgaca acagggggcc tcg 2317522DNAArtificialprimer sequence 175ttactgcgac aacggtgctg ac 2217622DNAArtificialprimer sequence 176caccatgagc gatcctcggc ca 2217723DNAArtificialprimer sequence 177ttacaagttg gcactgcgta ccg 2317832DNAArtificialprimer sequence 178ccggctgaga tctatgacag aatacgaagg gc 3217924DNAArtificialprimer sequence 179ccccgccagg gaactagagg cggc 2418022DNAArtificialprimer sequence 180caccatgaca tgggccgacg ag 2218120DNAArtificialprimer sequence 181ttagcgagtg gtcccgttcg 2018222DNAArtificialprimer sequence 182caccgtgtcc gacggcgaac aa 2218315DNAArtificialprimer sequence 183ttacggcccc ggccc 151848DNAArtificialreference sequence 184agtcagtc 81858DNAArtificialreference sequence 185aatcaatc 81866DNAArtificialreference sequence 186agtgtc 61878DNAArtificialreference sequence 187agtcagtc 8

Claims

1. A method for inducing an immune response in an animal to tuberculosis, comprising delivering a composition comprising a mycobacteria polypeptide having amino acid sequence SEQ ID NO: 33 and a pharmaceutically acceptable polymeric carrier bound to the polypeptide or a pharmaceutically acceptable adjuvant.

2. The method according to claim 1, wherein said polypeptide is fused to at least one fusion partner which is an antigen expressed by bacteria within the mycobacteria family, wherein said at least one fusion partner is heterologous to said polypeptide.

3. The method according to claim 1, wherein said polypeptide is lipidated thereby providing a self-adjuvanting effect of the polypeptide.

4. The method according to claim 1, which comprises a polymeric carrier bound by covalent or non-covalent interactions to said polypeptide.

5. The method according to claim 4, wherein the polymeric carrier is a polystyrene.

6. The method according to claim 2, wherein the fusion partner is selected from the polypeptides of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, and 45.

7. The method according to claim 6, wherein the polypeptides are recombinant or synthetic and are delivered in a delivery system comprising an adjuvant.

8. The method according to claim 2, wherein said at least one fusion partner is an antigen expressed by bacteria within the mycobacteria family other than a polypeptide selected from SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, and 45.

9. The method according to claim 8, wherein said at least one fusion partner is selected from the group consisting of ESAT-6, ESAT-6-Ag85B, TB10.4, CFP10, RD1-ORF5, RD1-ORF2, Rv1036, MPB64, MPT64, Ag85A, Ag85B (MPT59), MPB59, Ag85C, 19 kDa lipoprotein, and MPT32.

10. The method according to claim 8, wherein the polypeptides are recombinant or synthetic and are delivered in a delivery system comprising an adjuvant.

11. The method according to claim 1, wherein said animal is a mammal.

12. The method according to claim 11, wherein said mammal is a human.

13. A method for inducing an immune response in an animal to tuberculosis, comprising delivering one or more mycobacteria polypeptides which are upregulated or expressed during the latent stage of the mycobacteria infection which is characterized by low-oxygen tension in the microenvironment of the mycobacteria and (i) a pharmaceutically acceptable polymeric carrier bound to the one or more polypeptides or (ii) a pharmaceutically acceptable adjuvant, wherein said one or more polypeptides has the amino acid sequence of SEQ ID NO: 33, wherein said one or more polypeptides is fused to at least one mycobacteria fusion partner.

14. The method according to claim 13, where the polypeptides are recombinant or synthetic and are delivered in a delivery system comprising an adjuvant.

15. The method according to claim 13, wherein the fusion partner is selected from SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, and 45.

16. The method according to claim 15, wherein the fusion partner is an antigen expressed by bacteria within the mycobacteria family other than a polypeptide selected from SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, and 45.

17. The method according to claim 16, wherein the fusion partner is selected from the group consisting of ESAT-6, ESAT-6-Ag85B, TB10.4, CFP10, RD1-ORF5, RD1-ORF2, Rv1036, MPB64, MPT64, Ag85A, Ag85B (MPT59), MPB59, Ag85C, 19 kDa lipoprotein, and MPT32.

18. A method for inducing an immune response in an animal to latent tuberculosis, comprising delivering a composition comprising one or more mycobacteria polypeptides which are upregulated or expressed during the latent stage of the mycobacteria infection, wherein one polypeptide has the amino acid sequence of SEQ ID NO: 33.

19. The method according to claim 18, wherein the polypeptides are recombinant or synthetic and are delivered in a delivery system comprising an adjuvant.

20. The method according to claim 18, wherein the fusion partner is selected from SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, and 45.

Images