Efficient system for directed integration into the Lactobacillus acidophilus and Lactobacillus gasseri chromosomes via homologous recombination.
An efficient method is described for the generation of site-specific chromosomal integrations in Lactobacillus acidophilus and Lactobacillus gasseri. The strategy is an adaptation of the lactococcal pORI system (K. Leenhouts, G. Venema, and J. Kok, Methods Cell Sci. 20:35-50, 1998) and relies on the simultaneous use of two plasmids. The functionality of the integration strategy was demonstated by the insertional inactivation of the Lactobacillus acidophilus NCFM lacL gene encoding beta-galactosidase and of the Lactobacillus gasseri ADH gusA gene encoding beta-glucuronidase.
Russell
W M
WM
Department of Microbiology, Southeast Dairy Foods Research Center, North Carolina State University, Raleigh, North Carolina 27695, USA.
Klaenhammer
T R
TR
D001709:Biotechnology / Q000379:methods; D002876:Chromosomes, Bacterial / Q000235:genetics*; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics; D007778:Lactobacillus / Q000235:genetics*; D007779:Lactobacillus acidophilus / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D001616:beta-Galactosidase; D005966:Glucuronidase