FN Archimer Export Format
PT J
TI Construction of a Vibrio splendidus Vsm metalloprotease mutant using a novel counter-selectable suicide vector
BT 
AF LEROUX, Frederique
   BINESSE, Johan
   SAULNIER, Denis
   MAZEL, Didier
AS 1:1,2;2:1;3:2;4:1;
FF 1:PDG-DOP-DCN-AGSAE-LGP;2:;3:PDG-DOP-DCN-AGSAE-LGP;4:;
C1 Inst Pasteur, Unite Postulante Plasticite Genome Bacterien, CNRS URA 2171, F-75015 Paris, France.
   Lab Genet & Pathol, F-17390 La Tremblade, France.
C2 INST PASTEUR, FRANCE
   IFREMER, FRANCE
SI LA TREMBLADE RONCE
   LA TREMBLADE
SE PDG-DOP-DCN-AGSAE-LGP
IN WOS Ifremer jusqu'en 2018
   copubli-france
IF 4.004
TC 176
UR https://archimer.ifremer.fr/doc/2007/publication-3292.pdf
LA English
DT Article
AB splendidus is a dominant culturable Vibrio in seawater, but strains related to this group are also associated with mortalities in a variety of marine animals. The determinants encoding the pathogenic properties of these strains are still poorly known, however, the recent sequencing of the genome of V. splendidus LGP32, an oyster pathogen, offers the opportunity to decipher the basis of the virulence properties by the disruption of candidate genes. We have developed a novel suicide vector based on the pir-dependant R6K replicative origin, which can be transferred by RP4 based conjugation to, potentially, any Vibrio strain, and which also carries the plasmid F toxin ccdB gene under the control of the PBAD promoter. We demonstrate that this genetic system allows the efficient counter-selection of integrated plasmids in presence of arabinose, both in V. splendidus and V. cholerae, and as such permits the efficient marker-less allelic replacement in these species. We used this technique to construct several mutants of V. splendidus LGP32, including a derivative deleted of a secreted metalloprotease gene, vsm. We show that this gene is essential for the LGP32 extracellular products toxicity when injected in oysters, but is not necessary for the virulence of bacteria in the model of oyster infection after bacterial injection.
PY 2007
PD FEB
SO Applied and environmental microbiology
SN 0099-2240
PU American society for microbiology
VL 73
IS 3
UT 000244263800014
DI 10.1128/AEM.02147-06
ID 3292
ER

EF