title: Gene replacement in Lactobacillus helveticus.
abstract: An efficient method for gene replacement in Lactobacillus helveticus CNRZ32 was developed by utilizing pSA3 as an integration vector. This plasmid is stably maintained in CNRZ32 at 37 degrees C but is unstable at 45 degrees C. This method consisted of a two-step gene-targeting technique: (i) chromosomal integration of a plasmid carrying an internal deletion in the gene of interest via homologous recombination and (ii) excision of the vector and the wild-type gene via homologous recombination, resulting in gene replacement. By using this procedure, the chromosomal X-prolyl dipeptidyl aminopeptidase gene (pepXP) of CNRZ32 was successfully inactivated.
authors.affiliation: Department of Food Science, University of Wisconsin-Madison 53706.
authors.forename: T
authors.initials: T
authors.lastname: Bhowmik
authors.forename: L
authors.initials: L
authors.lastname: Fernández
authors.forename: J L
authors.initials: JL
authors.lastname: Steele
mesh_terms: D002876:Chromosomes, Bacterial; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000235:genetics; D018274:Electroporation; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D005821:Genetic Techniques; D005822:Genetic Vectors; D007778:Lactobacillus / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids; D011995:Recombination, Genetic; D015183:Restriction Mapping
chemical_list: D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4