Yersinia pestis, a Gram negative bacterium, causes bubonic and pneumonic plague. Emerging antibiotic resistance in clinical isolates is driving a need to develop novel antibiotics to treat infection by this transmissible and highly virulent pathogen. Proteins required for viability, so called essential genes, are attractive potential therapeutic targets, however, confirmation of essentiality is problematic. For the first time, we report the development of a system that allows the rapid determination of Y. pestis gene essentiality through mutagenesis and inducible expression of a plasmid borne copy of the target gene. Using this approach, we have confirmed the uridine monophosphate kinase PyrH as an essential protein in Y. pestis. This methodology and the tools we have developed will allow the confirmation of other putative essential genes in this dangerous pathogen, and facilitate the identification of novel targets for antimicrobial development. © 2014.
Ford, D. C., Ireland, P. M., Bullifent, H. L., Saint, R. J., Mcalister, E. V., Sarkar-Tyson, M., & Oyston, P. C. F. (2014). Construction of an inducible system for the analysis of essential genes in Yersinia pestis. Journal of Microbiological Methods, 100(1), 1-7. https://doi.org/10.1016/j.mimet.2014.01.017