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Complete Nucleotide Sequence of the pCTX-M3 Plasmid and Its Involvement in Spread of the Extended-Spectrum ß-Lactamase Gene bla_CTX-M-3

M. Gobiewski,^1, I. Kern-Zdanowicz,^1* M. Zienkiewicz,^1, M. Adamczyk,¹ J. yliska,¹ A. Baraniak,² M. Gniadkowski,² J. Bardowski,¹ and P. Cegowski¹

Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences,¹ National Medicines Institute, Warsaw, Poland²

Received 3 April 2007/ Returned for modification 12 May 2007/ Accepted 3 August 2007

Here we report the nucleotide sequence of pCTX-M3, a highly conjugative plasmid that is responsible for the extensive spread of the gene coding for the CTX-M-3 extended-spectrum ß-lactamase in clinical populations of the family Enterobacteriaceae in Poland. The plasmid belongs to the IncL/M incompatibility group, is 89,468 bp in size, and carries 103 putative genes. Besides bla_CTX-M-3, it also bears the bla_TEM-1, aacC2, and armA genes, as well as integronic aadA2, dfrA12, and sul1, which altogether confer resistance to the majority of ß-lactams and aminoglycosides and to trimethoprim-sulfamethoxazole. The conjugal transfer genes are organized in two blocks, tra and trb, separated by a spacer sequence where almost all antibiotic resistance genes and multiple mobile genetic elements are located. Only bla_CTX-M-3, accompanied by an ISEcp1 element, is placed separately, in a DNA fragment previously identified as a fragment of the Kluyvera ascorbata chromosome. On the basis of sequence analysis, we speculate that pCTX-M3 might have arisen from plasmid pEL60 from plant pathogen Erwinia amylovora by acquiring mobile elements with resistance genes. This suggests that plasmids of environmental bacterial strains could be the source of those plasmids now observed in bacteria pathogenic for humans.

Published ahead of print on 13 August 2007.

Present address: Department of Biotechnology, Nicolaus Copernicus University, Toru, Poland.

Present address: Department of Plant Physiology, Warsaw University, Warsaw, Poland.

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