Quantitative evaluation of Escherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants

D. M. Woodcock, P. J. Crowther, J. Doherty, S. Jefferson, E. Decruz, M. Noyer-Weidner, S. S. Smith, M. Z. Michael, M. W. Graham

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Abstract

Many strains of E.coli K12 restrict DNA containing cytosine methylation such as that present in plant and animal genomes. Such restriction can severely inhibit the efficiency of cloning genomic DNAs. We have quantitatively evaluated a total of 39 E.coli strains for their tolerance to cytosine methylation in phage and plasmid cloning systems. Quantitative estimations of relative tolerance to methylation for these strains are presented, together with the evaluation of the most promising strains in practical recombinant cloning situations. Host strains are recommended for different recombinant cloning requirements. These data also provide a rational basis for future construction of 'ideal' hosts combining optimal methylation tolerance with additional advantageous mutations.

Original languageEnglish
Pages (from-to)3469-3478
Number of pages10
JournalNucleic Acids Research
Volume17
Issue number9
DOIs
Publication statusPublished - 11 May 1989
Externally publishedYes

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    Woodcock, D. M., Crowther, P. J., Doherty, J., Jefferson, S., Decruz, E., Noyer-Weidner, M., Smith, S. S., Michael, M. Z., & Graham, M. W. (1989). Quantitative evaluation of Escherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants. Nucleic Acids Research, 17(9), 3469-3478. https://doi.org/10.1093/nar/17.9.3469