title: Complete sequence of two KPC-harbouring plasmids from Pseudomonas aeruginosa.
abstract: OBJECTIVE
  KPC-producing Pseudomonas aeruginosa are increasingly isolated in the Americas and in the Caribbean islands. Here, we determined the whole-plasmid sequence of two plasmids carrying the blaKPC-2 gene from multidrug-resistant P. aeruginosa clinical isolates from Colombia.
  
  
  METHODS
  The two plasmids, pCOL-1 and pPA-2, were transferred to Escherichia coli recipient strain TOP10 and completely sequenced using high-throughput pyrosequencing for pCOL-1 and classical Sanger sequencing for pPA-2.
  
  
  RESULTS
  Both plasmids could be transferred to E. coli by transformation and displayed no other resistance marker besides KPC. Plasmid pCOL-1 was 31 529 bp in size, contained 31 open reading frames (ORFs) and belonged to the IncP-6 replicon group. It exhibited genes involved in replication, mobilization and partitioning, but none involved in conjugation. Plasmid pPA-2 was 7995 bp in size and contained seven ORFs. It exhibited a replicase gene of IncU, but was lacking genes involved in mobilization, partitioning and conjugation. Only 2072 bp matched Tn4401, including the blaKPC-2 gene, part of ISKpn6 and a 73 bp segment located upstream of the blaKPC-2 gene, containing the P1 promoter. Sequence identity was interrupted by a Tn3 transposon, itself interrupted by an IS26 element inserted within the β-lactamase blaTEM-1 gene.
  
  
  CONCLUSIONS
  Here we present the genetic features of the very first plasmids carrying the blaKPC-2 gene from P. aeruginosa. The emergence of the blaKPC-2 gene on unrelated plasmids, differing in size and in incompatibility group, and harbouring different genetic structures containing the blaKPC-2 genes in P. aeruginosa isolates suggests that this resistance trait may follow a dissemination scheme in P. aeruginosa similar to that seen in Enterobacteriaceae.
authors.affiliation: Service de Bactériologie-Virologie, INSERM U914, Emerging resistance to antibiotics, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine et Université Paris Sud, K-Bicêtre, France. thierry.naas@bct.aphp.fr
authors.forename: Thierry
authors.initials: T
authors.lastname: Naas
authors.forename: Rémy A
authors.initials: RA
authors.lastname: Bonnin
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Cuzon
authors.forename: Maria-Virginia
authors.initials: MV
authors.lastname: Villegas
authors.forename: Patrice
authors.initials: P
authors.lastname: Nordmann
mesh_terms: D003105:Colombia; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; C491478:beta-lactamase KPC-2; D001618:beta-Lactamases
keywords: Gram-negative; KPC; antibiotic resistance; carbapenemases