Open AccessTransformation of Lactobacillus delbrückii ssp. lactis by electroporation and cloning of origins of replication by use of a positive selection vector
Author(s) -
Zink Andreas,
Klein Jürgen Robert,
Plapp Roland
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04444.x
Subject(s) - electroporation , transformation (genetics) , plasmid , lactobacillus casei , biology , cloning (programming) , molecular cloning , shuttle vector , microbiology and biotechnology , cloning vector , strain (injury) , lactobacillus , lactococcus lactis , bacteriophage , dna , bacteria , vector (molecular biology) , genetics , recombinant dna , escherichia coli , gene , peptide sequence , lactic acid , anatomy , computer science , programming language
Summary In this communication we report the first successful transformation of Lactobacillus delbrückii ssp. lactis WS97 with plasmid DNA by means of electroporation and describe the optimization of the transformation procedure for this strain. Efficiencies of electroporation varied between 10 2 to 10 4 transformants per μg pGK12, depending on the strain from which the DNA was isolated. The application of electroporation in molecular cloning was achieved by using the newly constructed origin screening vector, pAZ8. The replication origins of two cryptic plasmids were cloned. These plasmids were isolated from a thermophilic Lactobacillus strain Lb. delbrückii ssp. lactis WS97 and a mesophilic Lactobacillus strain Lb. casei NCDO151 which are both used in the dairy industry as starter cultures.