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

104 Citations (Scopus)

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 - Jan 1 1993
Externally publishedYes

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Firefly Luciferases
Anti-Infective Agents
Mycobacterium tuberculosis
Luciferases
Genetic Vectors
Electroporation
Octoxynol
Bacillus
Shock
Plasmids
Cell Count
Hot Temperature
Light
Genes

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

A rapid method for screening antimicrobial agents for activities against a strain of Mycobacterium tuberculosis expressing firefly luciferase. / Cooksey, R. C.; Crawford, J. T.; Jacobs, W. R.; Shinnick, T. M.

In: Antimicrobial Agents and Chemotherapy, Vol. 37, No. 6, 01.01.1993, p. 1348-1352.

Research output: Contribution to journalArticle

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