title: ISEcp1-mediated transposition of linked blaCTX-M-3 and blaTEM-1b from the IncI1 plasmid pEK204 found in clinical isolates of Escherichia coli from Belfast, UK.
abstract: OBJECTIVE
  Linked bla(CTX-M-3)-bla(TEM-1b) genes, as found on the IncI1 plasmid pEK204 prevalent in Belfast, also occur on plasmids belonging to other rep types in Escherichia coli isolated from nursing-home residents in Belfast. We investigated the mechanisms for their joint dissemination among diverse plasmids.
  
  
  METHODS
  Plasmid pEK204 was transferred by electroporation into E. coli DH5α harbouring derivative pBAD Myc-His vectors. Transposition experiments were then performed at 37°C. Transposition of bla genes onto the derivative pBAD Myc-His vector was confirmed by sequencing.
  
  
  RESULTS
  ISEcp1 mediated transposition of bla(CTX-M-3) alone from pEK204, as well as both bla(CTX-M-3) and bla(TEM-1b) jointly. The 5' and 3' termini of the transposed fragments were identical to or resembled the ISEcp1 IR(L) and IR(R), respectively, thereby replicating the environments previously found in various Belfast clinical plasmids harbouring bla(CTX-M-3).
  
  
  CONCLUSIONS
  Simultaneous dissemination of bla(CTX-M-3) and bla(TEM-1b) among plasmids in Belfast's nursing homes is facilitated by ISEcp1-mediated transposition of these bla genes from pEK204-like and other plasmids. Such transposition events are of public health concern, as they potentially allow wider dissemination of CTX-M-3 enzyme than would be possible through the spread of extended-spectrum β-lactamase-encoding plasmids and strains alone.
authors.affiliation: Antibiotic Resistance Monitoring and Reference Laboratory, Microbiology Services-Colindale, Health Protection Agency, 61 Colindale Avenue, London NW9 5EQ, UK. hiran.dhanji@hpa.org.uk
authors.forename: Hiran
authors.initials: H
authors.lastname: Dhanji
authors.forename: Michel
authors.initials: M
authors.lastname: Doumith
authors.forename: Russell
authors.initials: R
authors.lastname: Hope
authors.forename: David M
authors.initials: DM
authors.lastname: Livermore
authors.forename: Neil
authors.initials: N
authors.lastname: Woodford
mesh_terms: D001483:Base Sequence; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D018274:Electroporation; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000188:drug therapy / Q000382:microbiology; D018014:Gene Transfer Techniques; D008040:Genetic Linkage; D005822:Genetic Vectors; D006801:Humans; D007494:Ireland; D010957:Plasmids; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: C417271:beta-lactamase CTX-M-3; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1