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Updated: Jun 16, 2026
Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
Published on: March 16, 2012
A chimeric vector for efficient chromosomal modification in Enterococcus faecalis and other lactic acid bacteria
1Instituto 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.
Aim:
To construct a chimeric vector named pBVGh for quickly generating gene modifications in Enterococcus faecalis.
Methods And 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.
Significance And Impact Of The Study:
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.
