A recombinant Mycobacterium tuberculosis in vitro transcription system

被引：33

作者：

Jacques, JF

Rodrigue, S ^{[1
]}

Brzezinski, R

Gaudreau, L

机构：

[1] Univ Sherbrooke, Dept Biol, Ctr Rech Mecanismes Fonct Cellulaire, Sherbrooke, PQ J1K 2R1, Canada

[2] Univ Sherbrooke, Dept Biol, Ctr Etude & Valorisat Divers Microbienne, Sherbrooke, PQ J1K 2R1, Canada

来源：

FEMS MICROBIOLOGY LETTERS | 2006年 / 255卷 / 01期

关键词：

RNA polymerase; sigma factors; Mycobacterium tuberculosis; in vitro transcription; reconstitution;

D O I：

10.1111/j.1574-6968.2005.00071.x

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

In vitro transcription constitutes an important tool in the study of the regulation of gene expression. Here, we present a fast and easy procedure to prepare Mycobacterium tuberculosis RNA polymerase for in vitro transcription assays. RNA polymerase is assembled from recombinant proteins expressed in Escherichia coli, thus eliminating the need for biosafety containment facilities, and is mixed with any of the 13 M. tuberculosis sigma factors. We show that the recombinant RNA polymerase is free from contaminating sigma factors, produces transcriptional start sites matching those characterized in vivo and allows the formal identification of sigma factors involved in the expression of genes of interest.

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收藏

页码：140 / 147

页数：8

相关论文

共 26 条

[1] RNA polymerases from Bacillus subtilis and Escherichia coli differ in recognition of regulatory signals in vitro [J].

Artsimovitch, I ;

Svetlov, V ;

Anthony, L ;

Burgess, RR ;

Landick, R .

JOURNAL OF BACTERIOLOGY, 2000, 182 (21) :6027-6035

[2] Novel Mycobacterium tuberculosis anti-σ factor antagonists control σF activity by distinct mechanisms [J].

Beaucher, J ;

Rodrigue, S ;

Jacques, PÉ ;

Smith, I ;

Brzezinski, R ;

Gaudreau, L .

MOLECULAR MICROBIOLOGY, 2002, 45 (06) :1527-1540

[3] Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence [J].

Cole, ST ;

Brosch, R ;

Parkhill, J ;

Garnier, T ;

Churcher, C ;

Harris, D ;

Gordon, SV ;

Eiglmeier, K ;

Gas, S ;

Barry, CE ;

Tekaia, F ;

Badcock, K ;

Basham, D ;

Brown, D ;

Chillingworth, T ;

Connor, R ;

Davies, R ;

Devlin, K ;

Feltwell, T ;

Gentles, S ;

Hamlin, N ;

Holroyd, S ;

Hornby, T ;

Jagels, K ;

Krogh, A ;

McLean, J ;

Moule, S ;

Murphy, L ;

Oliver, K ;

Osborne, J ;

Quail, MA ;

Rajandream, MA ;

Rogers, J ;

Rutter, S ;

Seeger, K ;

Skelton, J ;

Squares, R ;

Squares, S ;

Sulston, JE ;

Taylor, K ;

Whitehead, S ;

Barrell, BG .

NATURE, 1998, 393 (6685) :537-+

[4] A stationary-phase stress-response sigma factor from Mycobacterium tuberculosis [J].

DeMaio, J ;

Zhang, Y ;

Ko, C ;

Young, DB ;

Bishai, WR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (07) :2790-2794

[5] Global burden of tuberculosis - Estimated incidence, prevalence, and mortality by country [J].

Dye, C ;

Scheele, S ;

Dolin, P ;

Pathania, V ;

Raviglione, RC .

JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION, 1999, 282 (07) :677-686

[6] Escherichia coli RNA polymerase subunit ω and its N-terminal domain bind full-length β′ to facilitate incorporation into the α2β subassembly [J].

Ghosh, P ;

Ishihama, A ;

Chatterji, D .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2001, 268 (17) :4621-4627

[7] Multiple sigma subunits and the partitioning of bacterial transcription space [J].

Gruber, TM ;

Gross, CA .

ANNUAL REVIEW OF MICROBIOLOGY, 2003, 57 :441-466

[8] MtbRegList, a database dedicated to the analysis of transcriptional regulation in Mycobacterium tuberculosis [J].

Jacques, PÉ ;

Gervais, AL ;

Cantin, M ;

Lucier, JF ;

Dallaire, G ;

Drouin, G ;

Gaudreau, L ;

Goulet, J ;

Brzezinski, R .

BIOINFORMATICS, 2005, 21 (10) :2563-2565

[9] Genetic analysis of the Mycobacterium smegmatis rpsL promoter [J].

Kenney, TJ ;

Churchward, G .

JOURNAL OF BACTERIOLOGY, 1996, 178 (12) :3564-3571

[10] Transcriptional activation of Agrobacterium tumefaciens virulence gene promoters in Escherichia coli requires the A-tumefaciens rpoA gene, encoding the alpha subunit of RNA polymerase [J].

Lohrke, SM ;

Nechaev, S ;

Yang, H ;

Severinov, K ;

Jin, SJ .

JOURNAL OF BACTERIOLOGY, 1999, 181 (15) :4533-4539