V S Blancato1, C Magni. 1. 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.
Abstract
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.
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.
Authors: Guillermo D Repizo; Víctor S Blancato; Pablo Mortera; Juke S Lolkema; Christian Magni Journal: Appl Environ Microbiol Date: 2013-02-22 Impact factor: 4.792
Authors: Ingrid Quintana; Martín Espariz; Silvina R Villar; Florencia B González; Maria F Pacini; Gabriel Cabrera; Iván Bontempi; Estefanía Prochetto; Jörg Stülke; Ana R Perez; Iván Marcipar; Victor Blancato; Christian Magni Journal: Front Microbiol Date: 2018-09-04 Impact factor: 5.640