title: Construction of an inducible system for the analysis of essential genes in Yersinia pestis.
abstract: 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.
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK. Electronic address: dcford@dstl.gov.uk.
authors: D C
authors: DC
authors: Ford
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK.
authors: P M
authors: PM
authors: Ireland
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK.
authors: H L
authors: HL
authors: Bullifent
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK.
authors: R J
authors: RJ
authors: Saint
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK.
authors: E V
authors: EV
authors: McAlister
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK.
authors: M
authors: M
authors: Sarkar-Tyson
authors: Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK.
authors: P C F
authors: PC
authors: Oyston
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D055786:Gene Knockout Techniques; D020043:Genes, Essential; D008807:Mice, Inbred BALB C; D050296:Microbial Viability; D009703:Nucleoside-Phosphate Kinase / Q000235:genetics; D010930:Plague; D010957:Plasmids; D014774:Virulence; D015010:Yersinia pestis / Q000235:genetics* / Q000502:physiology
chemical_list: C023276:uridine monophosphate kinase; D009703:Nucleoside-Phosphate Kinase
keywords: Antibiotic targets; Conditional lethal; DeoD; Essential genes; PyrH; Yersinia pestis