title: Characterization of the IncA/C plasmid pSCEC2 from Escherichia coli of swine origin that harbours the multiresistance gene cfr.
abstract: OBJECTIVE
  To determine the complete nucleotide sequence of the multidrug resistance plasmid pSCEC2, isolated from a porcine Escherichia coli strain, and to analyse it with particular reference to the cfr gene region.
  
  
  METHODS
  Plasmid pSCEC2 was purified from its E. coli J53 transconjugant and then sequenced using the 454 GS-FLX System. After draft assembly, predicted gaps were closed by PCR with subsequent sequencing of the amplicons.
  
  
  RESULTS
  Plasmid pSCEC2 is 135 615 bp in size and contains 200 open reading frames for proteins of ≥100 amino acids. Analysis of the sequence of pSCEC2 revealed two resistance gene segments. The 4.4 kb cfr-containing segment is flanked by two IS256 elements in the same orientation, which are believed to be involved in the dissemination of the rRNA methylase gene cfr. The other segment harbours the resistance genes floR, tet(A)-tetR, strA/strB and sul2, which have previously been found on other IncA/C plasmids. Except for these two resistance gene regions, the pSCEC2 backbone displayed >99% nucleotide sequence identity to that of other IncA/C family plasmids isolated in France, Chile and the USA.
  
  
  CONCLUSIONS
  The cfr gene was identified on an IncA/C plasmid, which is well known for its broad host range and transfer and maintenance properties. The location on such a plasmid will further accelerate the dissemination of cfr and co-located resistance genes among different Gram-negative bacteria. The genetic context of cfr on plasmid pSCEC2 underlines the complexity of cfr transfer events and confirms the role that insertion sequences play in the spread of cfr.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agriculture, Harbin 150001, China.
authors.forename: Wan-Jiang
authors.initials: WJ
authors.lastname: Zhang
authors.forename: Xing-Ran
authors.initials: XR
authors.lastname: Xu
authors.forename: Stefan
authors.initials: S
authors.lastname: Schwarz
authors.forename: Xiu-Mei
authors.initials: XM
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Dai
authors.forename: Hua-Jun
authors.initials: HJ
authors.lastname: Zheng
authors.forename: Siguo
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification; D004251:DNA Transposable Elements / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000302:isolation & purification; D008780:Methyltransferases / Q000235:genetics* / Q000302:isolation & purification; D010750:Phosphoproteins / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids / Q000235:genetics*; D013552:Swine
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; C094163:IncA protein, bacteria; D010750:Phosphoproteins; C534716:Cfr protein, E coli; D008780:Methyltransferases
keywords: IS256 element; food-producing animals; oxazolidinone resistance