Nitrile-inducible gene expression in mycobacteria.
The ability to ectopically control gene expression is a fundamental tool for the study of bacterial physiology and pathogenesis. While many efficient inducible expression systems are available for Gram-negative bacteria, few are useful in phylogenetically distant organisms, such as mycobacteria. We have adapted a highly-inducible regulon of Rhodococcus rhodochrous to artificially regulate gene expression in both rapidly-growing environmental mycobacteria and slow-growing pathogens, such as Mycobacterium tuberculosis. We demonstrate that this artificial regulatory circuit behaves as a bistable switch, which can be manipulated regardless of growth phase in vitro, and during intracellular growth in macrophages. High-level overexpression is also possible, facilitating biochemical and structural studies of mycobacterial proteins produced in their native host.
Pandey
Amit K
AK
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, MA 01655, USA.
Raman
Sahadevan
S
Proff
Rose
R
Joshi
Swati
S
Kang
Choong-Min
CM
Rubin
Eric J
EJ
Husson
Robert N
RN
Sassetti
Christopher M
CM
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001431:Bacteriological Techniques; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009570:Nitriles / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics; D012240:Rhodococcus / Q000235:genetics
D001426:Bacterial Proteins; D009570:Nitriles; D012097:Repressor Proteins
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