Recombinant Mycobacterium aurum expressing Escherichia coli beta-galactosidase in high throughput screening of antituberculosis drugs

Biochem Biophys Res Commun. 1997 Nov 26;240(3):536-9. doi: 10.1006/bbrc.1997.7689.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antitubercular Agents / pharmacology*
  • DNA, Recombinant
  • Drug Evaluation, Preclinical / methods
  • Escherichia coli / genetics*
  • Fluoroquinolones*
  • Lac Operon / genetics
  • Microbial Sensitivity Tests / methods*
  • Mycobacterium / drug effects*
  • Mycobacterium / enzymology
  • Mycobacterium / genetics*
  • Mycobacterium / growth & development
  • Quinolones / pharmacology
  • Recombinant Proteins
  • beta-Galactosidase / biosynthesis*
  • beta-Galactosidase / genetics

Substances

  • Antitubercular Agents
  • DNA, Recombinant
  • Fluoroquinolones
  • Quinolones
  • Recombinant Proteins
  • beta-Galactosidase
  • sparfloxacin