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We generated an <jats:italic>Escherichia coli<\/jats:italic> plasmid genomic library that was transformed into <jats:italic>E. coli.<\/jats:italic> These transformants were selected step by step using 0.25\u00d7 to 2\u00d7 minimum\u2009inhibitory concentrations for ceftriaxone, gentamicin, levofloxacin, tetracycline or trimethoprim. Plasmids were isolated at each selection step and subjected to Illumina sequencing. By searching for genomic loci whose sequencing coverage increased with antibiotic pressure we were able to detect 48 different genomic loci that were enriched by at least one antibiotic. Fifteen of these loci were studied functionally, and we showed that 13 can decrease the susceptibility of <jats:italic>E. coli<\/jats:italic> to antibiotics when overexpressed. These genes coded for drug targets, transcription factors, membrane proteins and resistance factors. The technique of Plas-Seq is expediting the discovery of genes associated with the mode of action or resistance to antibiotics and led to the isolation of a novel gene influencing drug susceptibility. It has the potential for being applied to novel molecules and to other microbial species.<\/jats:p>","DOI":"10.1099\/mgen.0.000148","type":"journal-article","created":{"date-parts":[[2018,1,10]],"date-time":"2018-01-10T16:13:11Z","timestamp":1515600791000},"source":"Crossref","is-referenced-by-count":6,"title":["Coupling next-generation sequencing to dominant positive screens for finding antibiotic cellular targets and resistance mechanisms in Escherichia coli"],"prefix":"10.1099","volume":"4","author":[{"given":"H\u00e9l\u00e8ne","family":"Gingras","sequence":"first","affiliation":[{"name":"Centre de Recherche en Infectiologie, Universit\u00e9 Laval, Qu\u00e9bec, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B\u00e9dis","family":"Dridi","sequence":"additional","affiliation":[{"name":"Centre de Recherche en Infectiologie, Universit\u00e9 Laval, Qu\u00e9bec, 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