{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T05:05:01Z","timestamp":1791003901711,"version":"4.1.0"},"reference-count":26,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2012,9,1]],"date-time":"2012-09-01T00:00:00Z","timestamp":1346457600000},"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":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2012,9]]},"abstract":"ABSTRACT<\/jats:title>\n \n The carbapenemase-encoding\n bla<\/jats:italic>\n KPC<\/jats:sub>\n gene, which is rapidly spreading in Gram-negative rods, is located on a Tn\n 3<\/jats:italic>\n -based transposon, Tn\n 4401<\/jats:italic>\n , which carries a polymorphic region giving rise to five isoforms (\n a<\/jats:italic>\n ,\n b<\/jats:italic>\n ,\n c<\/jats:italic>\n ,\n d<\/jats:italic>\n , and\n e<\/jats:italic>\n ) that is located immediately upstream of the\n bla<\/jats:italic>\n KPC<\/jats:sub>\n gene and thus likely involved in its expression. Using 5\u2032 rapid amplification of cDNA ends (5\u2032RACE), we identified three potential promoter sequences (P1, P2, and P3) upstream of the\n bla<\/jats:italic>\n KPC<\/jats:sub>\n gene, of which only P1 (absent from isoforms\n c<\/jats:italic>\n and\n d<\/jats:italic>\n ) and P2 (present in all isoforms, with a \u221235 box located inside the right inverted repeat of IS\n Kpn7<\/jats:italic>\n ) were shown to be true promoters involved in expression. One representative of each different promoter combination of Tn\n 4401<\/jats:italic>\n , i.e., P2 alone (isoform\n c<\/jats:italic>\n ), P1-P2 (isoform\n a<\/jats:italic>\n ), and P1-P2-P3 (isoform\n b<\/jats:italic>\n ), was cloned into an\n Escherichia coli<\/jats:named-content>\n plasmid vector. Using reverse transcription-PCR (RT-PCR), the highest level of expression was obtained with isoform\n a<\/jats:italic>\n (P1 and P2), which is also the most commonly encountered form in enterobacterial clinical isolates, followed by isoforms\n b<\/jats:italic>\n (P1, P2, and P3) and\n c<\/jats:italic>\n (P2 only). These differences in expression led to slight differences in MIC values of carbapenems.\n In silico<\/jats:italic>\n analysis of the DNA sequence of isoform\n b<\/jats:italic>\n revealed a stem-loop structure that is likely responsible for strong stops observed in 5\u2032RACE experiments and for decreased expression compared to that with isoform\n a<\/jats:italic>\n (P1 and P2). In addition, such structures could also be at the origin for the deletions observed in isoforms\n a<\/jats:italic>\n and\n c<\/jats:italic>\n . Taken together, these results indicate that the P1 and P2 promoters both contribute to the expression of the\n bla<\/jats:italic>\n KPC<\/jats:sub>\n gene and that the construct with the highest level of expression is that possessing isoform\n a<\/jats:italic>\n , which is also the most commonly encountered form in clinical isolates.\n <\/jats:p>","DOI":"10.1128\/aac.00334-12","type":"journal-article","created":{"date-parts":[[2012,6,27]],"date-time":"2012-06-27T03:48:33Z","timestamp":1340768913000},"page":"4753-4759","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":116,"title":["Role of IS\n Kpn7<\/i>\n and Deletions in\n bla<\/i>\n KPC<\/sub>\n Gene Expression"],"prefix":"10.1128","volume":"56","author":[{"given":"Thierry","family":"Naas","sequence":"first","affiliation":[{"name":"Service de Bact\u00e9riologie-Virologie, INSERM U914, Emerging Resistance to Antibiotics, H\u00f4pital de Bic\u00eatre, Assistance Publique-H\u00f4pitaux de Paris, Facult\u00e9 de M\u00e9decine, Universit\u00e9 Paris-Sud, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaelle","family":"Cuzon","sequence":"additional","affiliation":[{"name":"Service de Bact\u00e9riologie-Virologie, INSERM U914, Emerging Resistance to Antibiotics, H\u00f4pital de Bic\u00eatre, Assistance Publique-H\u00f4pitaux de Paris, Facult\u00e9 de M\u00e9decine, Universit\u00e9 Paris-Sud, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ha-Vy","family":"Truong","sequence":"additional","affiliation":[{"name":"Service de Bact\u00e9riologie-Virologie, INSERM U914, Emerging Resistance to Antibiotics, H\u00f4pital de Bic\u00eatre, Assistance Publique-H\u00f4pitaux de Paris, Facult\u00e9 de M\u00e9decine, Universit\u00e9 Paris-Sud, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Nordmann","sequence":"additional","affiliation":[{"name":"Service de Bact\u00e9riologie-Virologie, INSERM U914, Emerging Resistance to Antibiotics, H\u00f4pital de Bic\u00eatre, Assistance Publique-H\u00f4pitaux de Paris, Facult\u00e9 de M\u00e9decine, Universit\u00e9 Paris-Sud, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.7.1636"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1086\/421495"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro787"},{"key":"e_1_3_2_5_2","volume-title":"CLSI M100-S21","author":"Clinical and Laboratory Standards Institute","year":"2012","unstructured":"Clinical and Laboratory Standards Institute. 2012. Performance standards for antimicrobial susceptibility testing; 21st informational supplement. CLSI M100-S21. Clinical and Laboratory Standards Institute, Wayne, PA."},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1609.091389"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.05202-11"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00297-11"},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1111\/j.1574-6968.2009.01796.x","article-title":"Characterization of regulatory element Rp3 of regulation of beta-lactamases from Ralstonia pickettii","volume":"301","author":"Girlich D","year":"2009","unstructured":"GirlichD KolbA NaasT NordmannP. 2009. Characterization of regulatory element Rp3 of regulation of beta-lactamases from Ralstonia pickettii. FEMS Microbiol. Lett. 301:50\u201356.","journal-title":"FEMS Microbiol. 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DNA determinants of promoter selectivity in Escherichia coli. Cold Spring Harbor Symp. Quant. Biol. 47:477\u2013481.","journal-title":"Cold Spring Harbor Symp. Quant. 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