Identification of Mycobacterium tuberculosis signal sequences that direct the export of a leaderless beta-lactamase gene product in Escherichia coli.

Microbiology
Anthony J ChubbMario R W Ehlers

Abstract

Proteins secreted by Mycobacterium tuberculosis may play a key role in virulence and may also constitute antigens that elicit the host immune response. However, the M. tuberculosis protein export machinery has not been characterized. A library of M. tuberculosis H37Rv genomic DNA fragments ligated into a signal sequence selection vector that contained a leaderless beta-lactamase gene and an upstream Tac promoter was constructed. Transformation of Escherichia coli with the M. tuberculosis DNA library and selection on plates containing 50-100 micrograms ampicillin ml-1 resulted in the identification of 15 Ampr clones out of a total of 14,000 transformants. Twelve of the beta-lactamase gene fusions conferred high levels of Ampr (up to 1 mg ampicillin ml-1); insert sizes ranged from 350 to 3000 bp. Of ten inserts that were completely sequenced, two were identified as fragments of the genes for M. tuberculosis antigens 85A and 85C, which are the major secreted proteins of this pathogen. Seven of the remaining inserts were > or = 97% identical to hypothetical ORFs in the M. tuberculosis genome, one of which encoded a protein with 35% identity to a low-affinity penicillin-binding protein (PBP) from Streptomyces clavuligerus. Four of t...Continue Reading

References

Sep 1, 1992·Molecular & General Genetics : MGG·G Perez-MartinezS Bron
Jan 1, 1992·Molecular Microbiology·D B YoungJ E Thole
Oct 1, 1990·Molecular Microbiology·J K Broome-SmithY Zhang
Jul 1, 1995·The Journal of Clinical Investigation·B Y Lee, M A Horwitz
Sep 1, 1994·Molecular Microbiology·J W Izard, D A Kendall
Feb 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·M A HorwitzG Harth
Mar 1, 1993·Microbiological Reviews·A P Pugsley
Aug 1, 1993·Journal of Bacteriology·S K Das GuptaA K Tyagi
Jan 1, 1993·Trends in Biochemical Sciences·G P Salmond, P J Reeves
Jun 1, 1993·The Journal of Infectious Diseases·I M OrmeW H Boom
Mar 1, 1996·Infection and Immunity·M J Fenton, M W Vermeulen
Nov 22, 1996·The Journal of Biological Chemistry·J LiuH Nikaido
Feb 12, 1998·Science·T J Silhavy
Jan 24, 1998·Science·N Williams

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Citations

Apr 24, 2001·Research in Microbiology·J L JestinG Winter
Sep 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·J E Graham, J E Clark-Curtiss
Sep 24, 2004·The International Journal of Biochemistry & Cell Biology·Edith E MachowskiValerie Mizrahi
Mar 1, 2000·Tubercle and Lung Disease : the Official Journal of the International Union Against Tuberculosis and Lung Disease·F TekaiaS T Cole
Feb 3, 2000·Tubercle and Lung Disease : the Official Journal of the International Union Against Tuberculosis and Lung Disease·M Daffé, G Etienne

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