Transformation of Leuconostoc carnosum 4010 and evidence for natural competence of the organism

Publication: Research - peer-reviewJournal article – Annual report year: 2004

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Plasmid transformation in Leuconostoc carnosum 4010 was analyzed. A successful transformation protocol for L. carnosum was established by modifying an existing protocol for Lactococcus lactis. Several parameters, including the number of generations that the cells had grown at the time of harvest, glycine concentration, the time of incubation for phenotypic expression, and the electrical field strength, were investigated and proved to have influence on the transformation frequency. Electrocompetence was found to be transient and to peak in the early exponential growth phase. Optimized conditions resulted in transformation frequencies of up to 6.7 x 10(5) transformants per microgram of plasmid DNA. A total of five plasmids in L. carnosum were successfully introduced and maintained. Interestingly, we discovered that DNA uptake was of a frequency of 3 x 10(-6) to 19 x 10(-6) transformants per CFU in the absence of an applied electrical field. We concluded that L. carnosum is naturally competent.
Original languageEnglish
JournalApplied and Environmental Microbiology
Publication date2004
Volume70
Journal number6
Pages3695-3699
ISSN0099-2240
DOIs
StatePublished
CitationsWeb of Science® Times Cited: 10
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