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Antimicrob. Agents Chemother. doi:10.1128/AAC.00772-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Mosaic structure of p1658/97, a 125-kilobase plasmid harbouring an active amplicon with the extended-spectrum -lactamase gene bla_SHV-5

M. Zienkiewicz^*, I. Kern-Zdanowicz, M. Gobiewski, J. yliska, P. Mieczkowski, M. Gniadkowski, J. Bardowski, and P. Cegowski

Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics of Polish Academy of Sciences, National Institute of Public Health, Warsaw, Poland

^* To whom correspondence should be addressed. Email: maximus{at}biol.uw.edu.pl.

	Abstract

Escherichia coli isolates recovered from patients during a clonal outbreak in one of Warsaw hospitals in 1997 produced different levels of an extended-spectrum -lactamase (ESBL) of the SHV type. The -lactamase hyperproduction correlated with the multiplication of ESBL gene copies within a plasmid. Here, we present the complete nucleotide sequence of the plasmid p1658/97 carried by the isolates recovered during the outbreak. The plasmid is 125491 bp in size and shows a mosaic structure where all modules constituting the plasmid core are homologous to those found in plasmids F and R100 and are separated by segments of homology to other known regions (plasmid R64, Providencia rettgeri genomic island R391, Vibrio cholerae STX transposon, Klebsiella pneumoniae or E. coli chromosomes). The p1658/97 plasmid bears two replication systems, IncFII and IncFIB; we demonstrated that both are active in E. coli. The presence of an active partition system (sopABC locus) and two post-segregational killing systems (pemIK and hok/sok) indicates that the plasmid should be stably maintained in E. coli populations. The conjugative transfer is ensured by the operon of tra and trb genes. We also demonstrate that the plasmidic segment undergoing amplification contains the bla_SHV-5 gene and is homologous to a 7.9 kb fragment of the K. pneumoniae chromosome. The amplicon displays the structure of a composite transposon of type I.

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