Cellular reporter screens for inhibitors of Burkholderia pseudomallei targets in Pseudomonas aeruginosa

Donald T. Moir, Ming Di, Richard A. Moore, Herbert P. Schweizer, Donald E. Woods

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Summary: To facilitate the discovery of new therapeutics for Burkholderia pseudomallei infections, we have developed cellular reporter screens for inhibitors of B. pseudomallei targets in the surrogate host Pseudomonas aeruginosa. Pseudomonas aeruginosa strains carrying deletions of essential genes were engineered to be dependent on the isopropyl-β-D-thiogalactopyranoside (IPTG)-regulated expression of their B. pseudomallei orthologues on a broad-host-range plasmid. Pseudomonas aeruginosa genes which are upregulated in response to depletion of each target gene product, were fused to the Photorhabdus luminescens luxCDABE operon via pGSV3-lux-SpR to generate reporter strains with increased bioluminescence upon target inhibition. A total of 11 of 19 B. pseudomallei genes complemented deletions of their orthologues in P. aeruginosa. The dependence of growth on IPTG levels varied from complete dependence (ftsQ, gyrA, glmU, secA) to slower growth in the absence of IPTG (coaD, efp, mesJ), to apparently normal growth in the absence of IPTG (ligA, lpxA, folA, ipk). Reporter screening strains have been constructed for three gene targets (gyrA, glmU, secA), and one (gyrA) has been applied to 68,000 compounds resulting in a primary hit rate of 0.5% and a confirmed hit rate of 0.06%, including several fluoroquinolones. These results provide proof of principle for surrogate cellular reporter screens as a useful approach to identify inhibitors of essential gene products.

Original languageEnglish (US)
Pages (from-to)S152-S162
JournalTransactions of the Royal Society of Tropical Medicine and Hygiene
Issue numberSUPPL. 1
StatePublished - Dec 2008
Externally publishedYes


  • Bioluminescence
  • Burkholderia pseudomallei
  • Pseudomonas aeruginosa
  • Reporter
  • Screen
  • Target

ASJC Scopus subject areas

  • Parasitology
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases


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