{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T01:39:14Z","timestamp":1783993154288,"version":"3.55.0"},"reference-count":38,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2006,7,1]],"date-time":"2006-07-01T00:00:00Z","timestamp":1151712000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2006,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            We report the construction and application of a novel insertion element for transposase-mediated mutagenesis in gram-negative bacteria. Besides Km\n            <jats:sup>r<\/jats:sup>\n            as a selectable marker, the insertion element InsTet\n            <jats:sup>G\u2212<\/jats:sup>\n            1 carries the anhydrotetracycline (atc)-regulated outward-directed P\n            <jats:sub>A<\/jats:sub>\n            promoter so that atc-dependent conditional gene knockouts or knockdowns are generated. The complex formed between the purified hyperactive transposase and InsTet\n            <jats:sup>G\u2212<\/jats:sup>\n            1 was electroporated into\n            <jats:italic>Escherichia coli<\/jats:italic>\n            or\n            <jats:italic>Salmonella enterica<\/jats:italic>\n            serovar Typhimurium, and mutant pools were collected. We used\n            <jats:italic>E. coli<\/jats:italic>\n            strains with either TetR or the reverse variant revTetR\n            <jats:sup>r2<\/jats:sup>\n            , while only TetR was employed in\n            <jats:italic>Salmonella<\/jats:italic>\n            . Screening of the InsTet\n            <jats:sup>G\u2212<\/jats:sup>\n            1 insertion mutant pools revealed 15 atc-regulatable auxotrophic mutants for\n            <jats:italic>E. coli<\/jats:italic>\n            and 4 atc-regulatable auxotrophic mutants for\n            <jats:italic>Salmonella<\/jats:italic>\n            . We have also screened one\n            <jats:italic>Salmonella<\/jats:italic>\n            mutant pool in murine macrophage-like J774-A.1 cells using ampicillin enrichment. Two mutants with the InsTet\n            <jats:sup>G\u2212<\/jats:sup>\n            1 insertion in the gene\n            <jats:italic>pyrE<\/jats:italic>\n            or\n            <jats:italic>argA<\/jats:italic>\n            survived this procedure, indicating a reduced intracellular growth rate in J774-A.1 cells. The nature of the mutants and the modes of their regulation are discussed.\n          <\/jats:p>","DOI":"10.1128\/aem.00492-06","type":"journal-article","created":{"date-parts":[[2006,7,7]],"date-time":"2006-07-07T01:51:13Z","timestamp":1152237073000},"page":"4717-4725","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Generating Tetracycline-Inducible Auxotrophy in\n            <i>Escherichia coli<\/i>\n            and\n            <i>Salmonella enterica<\/i>\n            Serovar Typhimurium by Using an Insertion Element and a Hyperactive Transposase"],"prefix":"10.1128","volume":"72","author":[{"given":"Martin","family":"Ko\u0308stner","sequence":"first","affiliation":[{"name":"Lehrstuhl fu\u0308r Mikrobiologie, Friedrich Alexander Universita\u0308t Erlangen-Nu\u0308rnberg, D-91058 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Britta","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Lehrstuhl fu\u0308r Mikrobiologie, Friedrich Alexander Universita\u0308t Erlangen-Nu\u0308rnberg, D-91058 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ralph","family":"Bertram","sequence":"additional","affiliation":[{"name":"Lehrstuhl fu\u0308r Mikrobiologie, Friedrich Alexander Universita\u0308t Erlangen-Nu\u0308rnberg, D-91058 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang","family":"Hillen","sequence":"additional","affiliation":[{"name":"Lehrstuhl fu\u0308r Mikrobiologie, Friedrich Alexander Universita\u0308t Erlangen-Nu\u0308rnberg, D-91058 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.17.7996"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gni154"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1159\/000084565"},{"key":"e_1_3_2_5_2","first-page":"9","volume":"8","year":"1986","unstructured":"Bethesda Research Laboratories. 1986. 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