Efficient transformation of Lactococcus lactis IL1403 and generation of knock-out mutants by homologous recombination.
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/microg 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.
Gerber
Simon D
SD
Department of Clinical Pharmacology, University of Berne, Berne, Switzerland.
Solioz
Marc
M
D018274:Electroporation / Q000379:methods*; D005822:Genetic Vectors; D013294:Lactococcus lactis / Q000235:genetics*; D009154:Mutation; D014169:Transformation, Bacterial