Nucleic Acids Research, 1989, Vol. 17, No. 9 3469-3478
© 1989
MOLECULAR BIOLOGY |
Quantitative evaluation of Escherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants
Molecular Science Group, Peter MacCallum Cancer Institute 481 Little Lonsdale Street, Melbourne, Victoria 3000 1Calgene Pacific Pty Ltd 16 Gipps St., Collingwood, Victoria 3066, Australia 2Max-Planck-Institut für Molekulare Genetik Ihnestraße 73, D-1000 Berlin-33, FRG 3Molecular Surgery, City of Hope National Medical Center 1500 E. Duarte Road, Duarte, CA 91010-0269, USA
*To whom correspondence should be addressed
Received December 15, 1988. Revised April 3, 1989. Accepted April 3, 1989.
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.
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