Complete sequence of broad-host-range plasmid pNOR-2000 harbouring the metallo-β-lactamase gene blaVIM-2 from Pseudomonas aeruginosa.
OBJECTIVE
Metallo-β-lactamases (MBLs) are increasingly reported not only in Enterobacteriaceae but also in Pseudomonas spp. These enzymes hydrolyse all β-lactams, including carbapenems, and are not inhibited by β-lactamase inhibitors. The aim of this study was to fully characterize a plasmid bearing the blaVIM-2 MBL gene identified in a Pseudomonas aeruginosa isolate.


METHODS
This plasmid was fully sequenced by high-density pyrosequencing and annotated using the GenDB version 2.0 annotation tool. The evaluation of the broad-host-range replication of the pNOR-2000 replication initiation gene was assessed using electro-transformation and conjugation assays and the distribution of this replicase gene was evaluated using an international collection of VIM-producing Pseudomonas spp.


RESULTS
Analysis of the 21 880 bp sequence of pNOR-2000 revealed a truncated and non-functional transfer operon, in addition to novel genes encoding a serine protease and toxin/antitoxin addiction systems. This broad-host-range plasmid shares high gene synteny with part of the mobile genomic island pKLC102 identified in P. aeruginosa strain C.


CONCLUSIONS
We report here the complete nucleotide sequence of plasmid pNOR-2000 from a P. aeruginosa clinical isolate harbouring the integron-located MBL gene blaVIM-2.
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.
Rémy A
RA
Bonnin
Laurent
L
Poirel
Patrice
P
Nordmann
Felix G
FG
Eikmeyer
Daniel
D
Wibberg
Alfred
A
Pühler
Andreas
A
Schlüter
D016470:Bacteremia / Q000382:microbiology; D019295:Computational Biology; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018014:Gene Transfer Techniques; D022761:Gene Transfer, Horizontal; D005798:Genes, Bacterial; D058507:Host Specificity; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000302:isolation & purification*; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D017422:Sequence Analysis, DNA; D014169:Transformation, Bacterial; D001618:beta-Lactamases / Q000235:genetics*
D004269:DNA, Bacterial; C425108:beta-lactamase bla(vim-2); D001618:beta-Lactamases