A rapid method for screening antimicrobial agents for activities against a strain of Mycobacterium tuberculosis expressing firefly luciferase

R. C. Cooksey, J. T. Crawford, W. R. Jacobs, T. M. Shinnick

Research output: Contribution to journalArticle

106 Scopus citations

Abstract

We developed a rapid method to screen the efficacy of antimicrobial agents against Mycobacterium tuberculosis. A restriction fragment carrying a promoterless firefly luciferase gene was cloned into a 4,488-bp shuttle vector, pMV261, and luciferase was expressed under the control of a mycobacterial heat shock promoter. The resulting plasmid, pLUC10, was introduced by electroporation into the avirulent strain M. tuberculosis H37Ra. Luciferase assays of sonic lysates of Triton X-100-treated cells of M. tuberculosis H37Ra(pLUC10) yielded bioluminescence in excess of 1,000 relative light units/~109 tubercle bacilli, compared with 0.0025 for the same number of parental cells. A 48-h microdilution antimicrobial agent- screening assay using this strain was developed.

Original languageEnglish (US)
Pages (from-to)1348-1352
Number of pages5
JournalAntimicrobial agents and chemotherapy
Volume37
Issue number6
DOIs
StatePublished - 1993

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

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