title: Complete nucleotide sequence of the large conjugative pTC2 multireplicon plasmid encoding the VIM-1 metallo-β-lactamase.
abstract: OBJECTIVE
  To determine the complete nucleotide sequence of the VIM-1-encoding plasmid pTC2, which was isolated from a Greek Providencia stuartii multiresistant strain.
  
  
  METHODS
  The pTC2 plasmid was extracted and sequenced using shotgun and 3 kb paired-end DNA libraries and a 454 sequencing approach. Following assembly into a unique scaffold, gaps were closed by PCR followed by Sanger DNA sequencing. Gene predictions and annotations were performed using the CAAT-Box tool and the Conserved Domain Search service. Sequence comparisons were performed using the Artemis Comparison Tool.
  
  
  RESULTS
  Plasmid pTC2 (180,184 bp) was found to be a multireplicon plasmid (IncA/C and IncR), with a large IncA/C backbone and a mosaic multidrug resistance (MDR) region, in which was inserted a 13 kb IncR fragment. Gene annotation allowed the identification of a complete IncA/C-type transfer system and of several putative maintenance modules, both on the IncA/C backbone and on the IncR fragment. The complex MDR region contained 9 insertion sequences (7 IS26, 1 IS1 and 1 IS6100), 10 resistance genes and a mercury resistance operon integrated into unit transposons, composite transposons or integrons.
  
  
  CONCLUSIONS
  The pTC2 combines a broad host range, transfer and maintenance capacities, plasticity of the MDR region and a wide variety of resistance genes, properties that may contribute to the spreading of resistance determinants.
authors.affiliation: Infection Control Team, Hôpital Charles-Foix, F-94200 Ivry-sur-Seine, France. laurence.drieux-rouzet@psl.aphp.fr
authors.forename: Laurence
authors.initials: L
authors.lastname: Drieux
authors.forename: Dominique
authors.initials: D
authors.lastname: Decré
authors.forename: Lionel
authors.initials: L
authors.lastname: Frangeul
authors.forename: Guillaume
authors.initials: G
authors.lastname: Arlet
authors.forename: Vincent
authors.initials: V
authors.lastname: Jarlier
authors.forename: Wladimir
authors.initials: W
authors.lastname: Sougakoff
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003227:Conjugation, Genetic / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D032023:Integrons / Q000235:genetics; D008628:Mercury; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009876:Operon / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics; D010957:Plasmids / Q000235:genetics* / Q000302:isolation & purification; D011532:Providencia / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D012093:Replicon / Q000235:genetics*; D001618:beta-Lactamases / Q000235:genetics / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C094163:IncA protein, bacteria; D010750:Phosphoproteins; C417264:VIM-1 metallo-beta-lactamase; D001618:beta-Lactamases; D008628:Mercury