Identification of a Mycobacterium tuberculosis putative classical nitroreductase gene whose expression is coregulated with that of the acr aene within macrophages, in standing versus shaking cultures, and under low oxygen conditions.
AuthorsPurkayastha A, McCue LA, McDonough KA,
JournalInfect Immun
PubMed ID11854240
'Tuberculosis remains a leading killer worldwide, and new approaches for its treatment and prevention are urgently needed. This effort will benefit greatly from a better understanding of gene regulation in Mycobacterium tuberculosis, particularly with respect to this pathogen''s response to its host environment. We examined the behavior of two promoters ... More
Expression of the Mycobacterium tuberculosis acr-coregulated genes from the DevR (DosR) regulon is controlled by multiple levels of regulation.
AuthorsVasudeva-Rao HM, McDonough KA,
JournalInfect Immun
PubMed ID18391009
'Little is known about how Mycobacterium tuberculosis regulates gene expression in response to its host environment, despite its importance as a pathogen. We previously characterized 10 acr-coregulated genes (ACGs), all of which belong to the DevR (DosR) ' ... More
Controlling gene expression in mycobacteria with anhydrotetracycline and Tet repressor.
AuthorsEhrt S, Guo XV, Hickey CM, Ryou M, Monteleone M, Riley LW, Schnappinger D,
JournalNucleic Acids Res
PubMed ID15687379
'Gene expression systems that allow the regulation of bacterial genes during an infection are valuable molecular tools but are lacking for mycobacterial pathogens. We report the development of mycobacterial gene regulation systems that allow controlling gene expression in fast and slow-growing mycobacteria, including Mycobacterium tuberculosis, using anhydrotetracycline (ATc) as inducer. ... More
Improved tetracycline repressors for gene silencing in mycobacteria.
AuthorsKlotzsche M, Ehrt S, Schnappinger D,
JournalNucleic Acids Res
PubMed ID19174563
'Tetracycline repressor (TetR)-controlled expression systems have recently been developed for mycobacteria and proven useful for the construction of conditional knockdown mutants and their analysis in vitro and during infections. However, even though these systems allowed tight regulation of some mycobacterial genes, they only showed limited or no phenotypic regulation for ... More
Fluorescence-based detection of lacZ reporter gene expression in intact and viable bacteria including Mycobacterium species.
AuthorsRowland B, Purkayastha A, Monserrat C, Casart Y, Takiff H, McDonough KA
JournalFEMS Microbiol Lett
PubMed ID10518732
A variety of fluorescein di-beta-D-galactopyranoside (FDG)-based substrates were evaluated for measuring beta-galactosidase expression in bacteria. One substrate, 5-acetylamino-FDG (C2FDG), performed well in all bacteria tested, including the slow growing mycobacterium, Mycobacterium bovis BCG. The sensitivity of C2FDG in intact, viable BCG was similar to that of o-nitrophenyl-beta-D-galactopyranoside in cell lysates ... More