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Mycobacteria: Genomics, BACs, Deletions , Duplications and Resistance
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Bacterial Artificial Chromosome Libraries of Mycobacterium tuberculosis H37Rv and M. bovis BCG Pasteur

Ordered M. tuberculosis H37Rv and M. bovis BCG Pasteur 1173 Bacterial Artificial Chromosome (BAC) maps, originally overlaid on the M. tuberculosis H37Rv DraI and AsnI physical/integrated map, (established by Philipp et al., 1996). 'Rv' clones are M. tuberculosis pBeloBAC11 clones, 'X' clones indicate M. bovis BCG pBeloBAC11 clones, 'XE' clones are M. bovis BCG pBACe3.6 clones. The location of each BAC clone relative to the genomic position on the M. tuberculosis genome (in kilobases) is marked by small numbers.

Protocol for DNA-preparation in agarose plugs

Set of selected BACs, which is distributed by our laboratory after receipt of a Material Transfer Agreement (MTA).

The Bacterial Artificial Chromosome (BAC) cloning system is capable of stably propagating large, complex DNA inserts in Escherichia coli.
As part of the Mycobacterium tuberculosis H37Rv genome sequencing project, we constructed a BAC library in collaboration with A. Billault from the CEPH (Centre d'Etude de Polymorphisme Humain), Paris, France, in the pBeloBAC11 vector (kindly provided by Dr. Shizuya at the California Institute of Technology) and used it for genome mapping, confirmation of sequence assembly, and sequencing (Brosch et al., Infect. Immun. 1998; 66 (5):2221-9).

The library contains about 5,000 BAC clones, with inserts ranging in size from 25 to 104 kb, representing theoretically a 70-fold coverage of the M. tuberculosis genome (4.4 Mb). A seleted set of clones covering almost all of the genome of M. tuberculosis is shown in the BAC-MAP above.

This set of clones was also used for the development of BAC arrays, in an attempt to identify polymorphic regions in the genomes of tubercle bacilli.

These clones are listed with their coordinates on the H37Rv genome below.
They are also shown, superimposed on a physical DraI map in the figure below.


In addition, a second BAC library with inserts from the vaccine strain of Mycobacterium bovis BCG Pasteur was constructed and used for comparative genomics of members of the M. tuberculosis complex.(Mol. Microbiol. 32: 643-656).

BAC-Clone numbers and their position on the M. tuberculosis genome (MTBH37RV)

BAC-Rv13 4367940..4411529 and 1. . 27227
BAC-Rv60 27227..122825
BAC-Rv5 135871..175332
BAC-Rv165 175332..251596
BAC-Rv329 258028..341170
BAC-Rv240 295559..375851
BAC-Rv167 341170..419767
BAC-Rv66 419767..489081
BAC-Rv59 441632..494134
BAC-Rv313 489081..566737
BAC-Rv265 513402..599515
BAC-Rv308 599515..664643
BAC-Rv220 664643..735596
BAC-Rv258 664643..714742
BAC-Rv339 742460..811258
BAC-Rv418 814625..894677
BAC-Rv45 866545..908666
BAC-Rv134 894666..970935
BAC-Rv217 934302..1000547
BAC-Rv17 970935..1028020
BAC-Rv103 1034667..1103616
BAC-Rv270 1103616..1155297
BAC-Rv267 1124621..1169811
BAC-Rv174 1171447..1238409
BAC-Rv44 1238409..1289201
BAC-Rv71 1276186..1322584
BAC-Rv7 1322584..1401546
BAC-Rv230 1336764..1411979
BAC-Rv407 1411979..1464436
BAC-Rv128 1429521..1502945
BAC-Rv39 1502945..1555030
BAC-Rv106 1519109..1591385
BAC-Rv255 1555030..1640681
BAC-Rv268 1623721..1689338
BAC-Rv58 1640681..1727235
BAC-Rv264 1689338..1755770
BAC-Rv417 1755770..1830362
BAC-Rv401 1830262..1934621
BAC-Rv302 1934621..2003728
BAC-Rv81 1998001..2040500
BAC-Rv281 2040500..2116467
BAC-Rv221 2115612..2198604
BAC-Rv420 2201807..2238065
BAC-Rv175 2238065..2310745
BAC-Rv412 2310745..2380808
BAC-Rv73 2380808..2407395
BAC-Rv269 2400863..2467639
BAC-Rv214 2467639..2543372
BAC-Rv42 2543372..2610983
BAC-Rv143 2611923..2679685
BAC-Rv234 2847864..2928420
BAC-Rv351 2928420..2964335
BAC-Rv35 2986777..3057845
BAC-Rv415 3049118..3127011
BAC-Rv404 3127011..3214529
BAC-Rv209 3214529..3281413
BAC-Rv228 3263223..3342558
BAC-Rv30 3281413..3356541
BAC-Rv280 3342541..3417299
BAC-Rv177 3417299..3441794
BAC-Rv48 3441794..3484616
BAC-Rv151 3481838..3554642
BAC-Rv92 3554642..3632390
BAC-Rv3 3632390..3669398
BAC-Rv403 3669398..3754791
BAC-Rv330 3749809..3814590
BAC-Rv285 3814590..3862021
BAC-Rv233 3862021..3939512
BAC-Rv416 3939512..4018320
BAC-Rv67 3975788..4051198
BAC-Rv222 4018320..4085317
BAC-Rv149 4054019..4122411
BAC-Rv254 4119595..4194053
BAC-Rv273 4122411..4194053
BAC-Rv266 4166694..4207176
BAC-Rv25 4207176..4291638
BAC-Rv136 4266694..4342605
BAC-Rv414 4291638..4367942

Further information and services concerning M. tuberculosis genes and predicted proteins can be obtained via the TubercuList server at the Institut Pasteur and the Mycobacterium tuberculosis project www-page at the Sanger Centre.

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