title: Nitrile-inducible gene expression in mycobacteria.
abstract: 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.
authors.affiliation: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, MA 01655, USA.
authors.forename: Amit K
authors.initials: AK
authors.lastname: Pandey
authors.forename: Sahadevan
authors.initials: S
authors.lastname: Raman
authors.forename: Rose
authors.initials: R
authors.lastname: Proff
authors.forename: Swati
authors.initials: S
authors.lastname: Joshi
authors.forename: Choong-Min
authors.initials: CM
authors.lastname: Kang
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Rubin
authors.forename: Robert N
authors.initials: RN
authors.lastname: Husson
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Sassetti
mesh_terms: 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
chemical_list: D001426:Bacterial Proteins; D009570:Nitriles; D012097:Repressor Proteins
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