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This study aimed to identify genes that are important for growth of\n                    <jats:italic>E. coli<\/jats:italic>\n                    during aminoglycoside exposure, since such genes may be targeted to re-sensitize resistant\n                    <jats:italic>E. coli<\/jats:italic>\n                    to treatment. We constructed three transposon mutant libraries each containing\u2009&gt;\u2009230.000 mutants in\n                    <jats:italic>E. coli<\/jats:italic>\n                    MG1655 strains harboring streptomycin (\n                    <jats:italic>aph(3\u2033)-Ib\/aph(6)-Id<\/jats:italic>\n                    ), gentamicin (\n                    <jats:italic>aac(3)-IV<\/jats:italic>\n                    ), or neomycin (\n                    <jats:italic>aph(3\u2033)-Ia<\/jats:italic>\n                    ) resistance gene(s). Transposon Directed Insertion-site Sequencing (TraDIS), a combination of transposon mutagenesis and high-throughput sequencing, identified 56 genes which were deemed important for growth during streptomycin, 39 during gentamicin and 32 during neomycin exposure. Most of these fitness-genes were membrane-located (n\u2009=\u200955) and involved in either cell division, ATP-synthesis or stress response in the streptomycin and gentamicin exposed libraries, and enterobacterial common antigen biosynthesis or magnesium sensing\/transport in the neomycin exposed library. For validation, eight selected fitness-genes\/gene-clusters were deleted (\n                    <jats:italic>minCDE<\/jats:italic>\n                    <jats:italic>,<\/jats:italic>\n                    <jats:italic>hflCK<\/jats:italic>\n                    <jats:italic>,<\/jats:italic>\n                    <jats:italic>clsA<\/jats:italic>\n                    and\n                    <jats:italic>cpxR<\/jats:italic>\n                    associated with streptomycin and gentamicin resistance, and\n                    <jats:italic>phoPQ<\/jats:italic>\n                    ,\n                    <jats:italic>wecA<\/jats:italic>\n                    <jats:italic>,<\/jats:italic>\n                    <jats:italic>lpp<\/jats:italic>\n                    and\n                    <jats:italic>pal<\/jats:italic>\n                    associated with neomycin resistance), and all mutants were shown to be growth attenuated upon exposure to the corresponding antibiotics. In summary, we identified genes that are advantageous in aminoglycoside-resistant\n                    <jats:italic>E. coli<\/jats:italic>\n                    during antibiotic stress. In addition, we increased the understanding of how aminoglycoside-resistant\n                    <jats:italic>E. coli<\/jats:italic>\n                    respond to antibiotic exposure.\n                  <\/jats:p>","DOI":"10.1038\/s41598-024-54169-8","type":"journal-article","created":{"date-parts":[[2024,2,20]],"date-time":"2024-02-20T03:03:10Z","timestamp":1708398190000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Genome-wide identification of fitness-genes in aminoglycoside-resistant Escherichia coli during antibiotic stress"],"prefix":"10.1038","volume":"14","author":[{"given":"Sandra Marina","family":"Wellner","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mosaed Saleh A.","family":"Alobaidallah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao","family":"Fei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Herrero-Fresno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John Elmerdahl","family":"Olsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,2,20]]},"reference":[{"key":"54169_CR1","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1016\/S0140-6736(21)02724-0","volume":"399","author":"CJ Murray","year":"2022","unstructured":"Murray, C. 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