A chimeric vector for efficient chromosomal modification in Enterococcus faecalis and other lactic acid bacteria.
OBJECTIVE
To construct a chimeric vector named pBVGh for quickly generating gene modifications in Enterococcus faecalis.


RESULTS
The constructed plasmid pBVGh carries the pG(+)host replicon (a thermosensitive (TS) derivative of pWV01), allowing a simple generation of mutants by growing colonies first at the permissive temperature and then switching the culture to the nonpermissive temperature. Additionally, this vector facilitates the screening of mutants by a rapid colorimetric blue-white discrimination of plasmid-free bacteria.


CONCLUSIONS
The pBVGh vector allows a straightforward inactivation or modification of target genes as well as a fast selection of enterococcal mutant strains.


CONCLUSIONS
The broad range of the TS replicon utilized in this plasmid permits the easy establishment and the efficient generation of food-grade mutant strains in Ent. faecalis and several other gram-positive bacteria.
Blancato
V S
VS
Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET) and Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, Rosario, Argentina.
Magni
C
C
D002876:Chromosomes, Bacterial / Q000235:genetics*; D013293:Enterococcus faecalis / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000235:genetics* / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism
D019344:Lactic Acid