Recombinant Mycobacterium aurum expressing Escherichia coli beta-galactosidase in high throughput screening of antituberculosis drugs.
Mycobacterium aurum is considered a surrogate of M. tuberculosis and recently has been proposed as test organism in high throughput screening of antituberculosis drugs (3). In this investigation, we suggest use of a recombinant M. aurum expressing E. coli lacZ gene, in which beta-galactosidase production is the reporter system as recently reported by us (6). The assay is based on production of beta-galactosidase in presence of drugs during growth. Enzyme production was inhibited within 4 h by frontline antimycobacterial drugs like streptomycin, rifampicin, isoniazid, ethambutol, ofloxacin, and sparfloxacin at their MICs. The assay could be performed conveniently in 96-well microtiter plate format.
Srivastava
R
R
Division of Microbiology, Central Drug Research Institute, Lucknow, India.
Kumar
D
D
Srivastava
B S
BS
D000995:Antitubercular Agents / Q000494:pharmacology*; D004274:DNA, Recombinant; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D024841:Fluoroquinolones; D007763:Lac Operon / Q000235:genetics; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009161:Mycobacterium / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D015363:Quinolones / Q000494:pharmacology; D011994:Recombinant Proteins; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
D000995:Antitubercular Agents; D004274:DNA, Recombinant; D024841:Fluoroquinolones; D015363:Quinolones; D011994:Recombinant Proteins; D001616:beta-Galactosidase; C061363:sparfloxacin