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Efficient transformation of Lactococcus lactis IL1403 and generation of knock-out mutants by homologous recombination

✍ Scribed by Simon D. Gerber; Marc Solioz


Book ID
102390779
Publisher
John Wiley and Sons
Year
2007
Tongue
English
Weight
177 KB
Volume
47
Category
Article
ISSN
0233-111X

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✦ Synopsis


Abstract

Lactococcus lactis IL1403 is a Gram‐positive bacterium of great biotechnological interest for food grade applications. Its use is however hampered by the difficulty to efficiently transform this strain. We here describe a detailed, optimized electrotransformation protocol which yields a transformation efficiency of 10^6^ cfu/μg of DNA with the two E. coli Gram‐positive shuttle vectors pC3 and pVA838. The utility of the protocol was demonstrated by the generation of single‐ and double‐knock‐out mutants by homologous recombination. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)