Corynebacterium glutamicum DNA is subjected to methylation-restriction in Escherichia coli.
Efficient electroporation of Escherichia coli with plasmid DNA isolated from Corynebacterium glutamicum depends on the use of Mcr-deficient E. coli strains. The transformation frequency increased nearly 800-fold when the Mcr-deficient E. coli DH5 alpha MCR was used instead of E. coli DH5 alpha. We used E. coli strains with different mutations in the methyl-specific restriction systems to show that McrBC-deficiency is sufficient to generate this effect. The results imply that C. glutamicum DNA contains methylcytosine in specific sequences recognized by the E. coli McrBC system.
Tauch
A
A
Department of Genetics, University of Bielefeld, Germany.
Kirchner
O
O
Wehmeier
L
L
Kalinowski
J
J
Pühler
A
A
D003001:Cloning, Molecular; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005822:Genetic Vectors; D008745:Methylation; D009154:Mutation / Q000235:genetics; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; C120301:McrBC endonuclease