4.3 Article

High-density transposon libraries utilising outward-oriented promoters identify mechanisms of action and resistance to antimicrobials

期刊

FEMS MICROBIOLOGY LETTERS
卷 367, 期 22, 页码 -

出版社

OXFORD UNIV PRESS
DOI: 10.1093/femsle/fnaa185

关键词

bacteria; antibiotic; development; resistance; mechanism; mutagenesis

资金

  1. BBSRC [BBS/E/F/000PR10349] Funding Source: UKRI

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The use of bacterial transposon mutant libraries in phenotypic screens is a well-established technique for determining which genes are essential or advantageous for growth in conditions of interest. Standard, inactivating, transposon libraries cannot give direct information about genes whose over-expression gives a selective advantage. We report the development of a system wherein outward-oriented promoters are included in mini-transposons, generation of transposon mutant libraries in Escherichia coli and Pseudomonas aeruginosa and their use to probe genes important for growth under selection with the antimicrobial fosfomycin, and a recently-developed leucyl-tRNA synthase inhibitor. In addition to the identification of known mechanisms of action and resistance, we identify the carbon-phosphorous lyase complex as a potential resistance liability for fosfomycin in E. coli and P. aeruginosa. The use of this technology can facilitate the development of novel mechanism-of-action antimicrobials that are urgently required to combat the increasing threat worldwide from antimicrobial-resistant pathogenic bacteria.

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