beta-Galactosidase reporter system in mycobacteria and its application in rapid antimycobacterial drug screening

Biochem Biophys Res Commun. 1997 Jun 27;235(3):602-5. doi: 10.1006/bbrc.1997.6837.

Abstract

Pathogenic mycobacteria are generally slow growing organisms and it takes several weeks to evaluate inhibitors of growth. Therefore, for rapid screening of the inhibitors of mycobacterial growth, a beta-galactosidase reporter system has been described which utilises a recombinant Mycobacterium smegmatis mc(2)155 expressing E. coli lacZ gene as the test organism. The assay is based on production of beta-galactosidase in presence of drugs during growth. A correlation between beta-galactosidase production and colony forming ability of mycobacteria was obtained. beta-galactosidase production was inhibited within 6 h by front line standard antimycobacterial drugs like streptomycin, rifampicin, isoniazid, ethambutol, pyrazinamide and ofloxacin at their reported MICs. The assay was performed on mycobacterial cells permeabilized with chloroform and sodium dodecyl sulfate.

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Antitubercular Agents / pharmacology*
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Genes, Reporter
  • Isoniazid / pharmacology
  • Kinetics
  • Microbial Sensitivity Tests / methods*
  • Mycobacterium / drug effects*
  • Mycobacterium / enzymology
  • Mycobacterium / growth & development
  • Ofloxacin / pharmacology
  • Plasmids
  • Pyrazinamide / pharmacology
  • Recombinant Proteins / biosynthesis
  • Rifampin / pharmacology
  • Streptomycin / pharmacology
  • beta-Galactosidase / biosynthesis*

Substances

  • Anti-Bacterial Agents
  • Antitubercular Agents
  • Recombinant Proteins
  • Pyrazinamide
  • Ofloxacin
  • beta-Galactosidase
  • Isoniazid
  • Rifampin
  • Streptomycin