Molecular cloning in Lactobacillus helveticus by plasmid pSA3::pVA797 co-integrate formation and conjugal transfer.
A gene encoding beta-glucanase activity from Bacillus amyloliquefaciens was subcloned in both orientations into plasmid shuttle vector pSA3. In only one orientation could a co-integrate be generated with the conjugative plasmid pVA797. The plasmid co-integrate was conjugated into Lactobacillus helveticus strain CNRZ450, where it was stably maintained without antibiotic selection and exhibited beta-glucanase activity. This method of introducing cloned DNA into thermophilic lactobacilli will facilitate the study of heterologous gene expression in non-transformable species.
Food and Agricultural Microbiology Research Division, Department of Agriculture for Northern Ireland, Belfast, UK.
K
K
Thompson
M
M
Collins
D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D015870:Gene Expression; D007778:Lactobacillus / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids; D001617:beta-Glucosidase / Q000235:genetics
D001617:beta-Glucosidase