Development of an E. coli strain for highthroughput screening of YidC inhibitors

Development of a strain that is useful for screening compounds that inhibit YidC in E. coli and identification of compounds with antibacterial activity.

The Need

In bacteria, the YidC membrane protein functions in the insertation of proteins into the cytoplasmic membrane of E. coli. This protein is essential for life and is evolutionarily conserved.

The rise of drug-resistant bacterial pathogens has created an urgent demand for novel antibiotic treatments that can act through new ways or against new targets. The bacterial YidC provides one such target. However, there is no effective way to screen compounds that inhibit YidC in E.coli or identify compounds with antibacterial activity.

The Technology

Dr. Ross E. Dalbey developed a modified JS7131 strain that has a luciferase operon under a pspA promoter. Because the PspA protein is switched on when YidC is inactivated, the strain will fluoresce upon inhibition of YidC by a compound. In addition, whole-cell measurements and live-cell readings can be conducted without a need to lyse the cells prior to obtaining fluorescence results.

Commercial Applications

  • Highthroughput screening of YidC inhibitors

Benefits/Advantages

  • The strain should be very useful for screening compounds that inhibit YidC in E. coli and identification of compounds with antibacterial activity by utilizing its fluorescence activity upon activation
  • Able to perform whole-cell measurements plus live-cell reading without the need to destroy the cell prior to reading the fluorescence

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