Complete sequence of two KPC-harbouring plasmids from Pseudomonas aeruginosa

J Antimicrob Chemother. 2013 Aug;68(8):1757-62. doi: 10.1093/jac/dkt094. Epub 2013 Apr 7.


Objectives: 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.

Keywords: Gram-negative; KPC; antibiotic resistance; carbapenemases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Colombia
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Humans
  • Molecular Sequence Data
  • Plasmids*
  • Pseudomonas Infections / microbiology
  • Pseudomonas aeruginosa / enzymology*
  • Pseudomonas aeruginosa / genetics*
  • Pseudomonas aeruginosa / isolation & purification
  • Sequence Analysis, DNA
  • beta-Lactamases / genetics*
  • beta-Lactamases / metabolism


  • DNA, Bacterial
  • beta-lactamase KPC-2
  • beta-Lactamases

Associated data

  • GENBANK/KC609322
  • GENBANK/KC609323
  • RefSeq/NC_022345
  • RefSeq/NC_022346