The construction of Streptomyces cyaneus genomic libraries in Escherichia coli is dependent upon the use of Mcr-deficient strains.
Streptomyces cyaneus genomic DNA ligated into either lambda phage or plasmid vectors was very inefficiently cloned into standard Escherichia coli host strains. However, the same material could be efficiently cloned using Mcr-deficient E. coli strains. These results suggest that the S. cyaneus genome contains 5-methylcytosine residues, some of which occur within the recognition sequences of the E. coli Mcr restriction system.
Wang
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Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology, UK.
Harvey
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SS
Sims
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Broda
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P
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial; D015257:DNA-Cytosine Methylases / Q000378:metabolism; D015253:Deoxyribonucleases, Type I Site-Specific / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015698:Genomic Library; D010957:Plasmids; D013302:Streptomyces / Q000235:genetics*
D004269:DNA, Bacterial; D015257:DNA-Cytosine Methylases; D004262:DNA Restriction Enzymes; C029602:endodeoxyribonuclease EcoK; D015253:Deoxyribonucleases, Type I Site-Specific