Tools
ToolsYasir, Muhammad, Keith Turner, A., Bastkowski, Sarah, Baker, David, Page, Andrew J., Telatin, Andrea, Phan, Minh Duy, Monahan, Leigh, Savva, George M., Darling, Aaron, Webber, Mark A. and Charles, Ian G. (2020) TraDIS-Xpress: a high-resolution whole-genome assay identifies novel mechanisms of triclosan action and resistance. Genome Research, 30 (2). pp. 239-249. ISSN 1088-9051
Preview |
PDF (Published_Version)
- Published Version
Available under License Creative Commons Attribution. Download (8MB) | Preview |
Abstract
Understanding the genetic basis for a phenotype is a central goal in biological research. Much has been learnt about bacterial genomes by creating large mutant libraries and looking for conditionally important genes. However, current genome-wide methods are largely unable to assay essential genes which are not amenable to disruption. To overcome this limitation, we developed a new version of “TraDIS” (transposon directed insertion-site sequencing) that we term “TraDIS-Xpress” that combines an inducible promoter into the transposon cassette. This allows controlled overexpression and repression of all genes owing to saturation of inserts adjacent to all open reading frames as well as conventional inactivation. We applied TraDIS-Xpress to identify responses to the biocide triclosan across a range of concentrations. Triclosan is endemic in modern life, but there is uncertainty about its mode of action with a concentration-dependent switch from bacteriostatic to bactericidal action unexplained. Our results show a concentration-dependent response to triclosan with different genes important in survival between static and cidal exposures. These genes include those previously reported to have a role in triclosan resistance as well as a new set of genes, including essential genes. Novel genes identified as being sensitive to triclosan exposure include those involved in barrier function, small molecule uptake, and integrity of transcription and translation. We anticipate the approach we show here, by allowing comparisons across multiple experimental conditions of TraDIS data, and including essential genes, will be a starting point for future work examining how different drug conditions impact bacterial survival mechanisms.
| Item Type: | Article |
|---|---|
| Additional Information: | Funding Information: We gratefully acknowledge the support of the Biotechnology and Biological Sciences Research Council (BBSRC); S.B., A.K.T., M.Y., M.A.W., and I.G.C. were supported by the BBSRC Institute Strategic Programme Microbes in the Food Chain BB/R012504/1 and its constituent project BBS/E/F/000PR10349. A.J.P. and G.M.S. were supported by the Quadram Institute Bioscience BBSRC funded Core Capability Grant (project number BB/ CCG1860/1). Genomic analysis used the Medical Research Council (MRC) “CLIMB” cloud computing environment supported by grant MR/L015080/1. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2020 Yasir et al. This article, published in Genome Research, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/. |
| Uncontrolled Keywords: | bacteria,discovery,escherichia-coli,package,genetics,genetics(clinical) ,/dk/atira/pure/subjectarea/asjc/1300/1311 |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 20 Feb 2020 07:58 |
| Last Modified: | 28 Oct 2024 11:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/74265 |
| DOI: | 10.1101/gr.254391.119 |
Downloads
Downloads per month over past year
Actions (login required)
 |
View Item |