Genetic Dissection of a Prevalent Plasmid-Encoded Conjugation System in Lactococcus lactis.

ABSTRACT: Plasmid pNP40, which was first identified nearly 40 years ago in Lactococcus lactis subsp. lactis biovar diacetylactis DRC3, encodes functions such as heavy metal-, bacteriophage-, and nisin-resistance, as well as plasmid transfer ability by conjugation. Here, we report an optimized conjugation protocol for this plasmid, yielding a transfer frequency that is approximately 4,000-fold higher than those previously reported in literature, while we also observed high-frequency plasmid co-mobilization. Individual mutations in 18 genes that encompass the presumed conjugation cluster of pNP40 were generated using ssDNA recombineering to evaluate the role of each gene in the conjugation process. A possible transcriptional repressor of this conjugation cluster, the product of the tr

SUBMITTER: Ortiz Charneco G 

PROVIDER: S-EPMC8194271 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature