Two-step method for curing Escherichia coli of ColE1-derived plasmids
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Published on: May 29, 2016
1Section for Biochemical Genetics, Department of Molecular Biology, University of Copenhagen, 5 Ole Maaløes Vej, Copenhagen N, Denmark. hove@mermaid.molbio.ku.dk
Abstract:
To cure Escherichia coli for plasmids derived from the ColE1 replicon advantage is taken of the fact that maintenance of this replicon requires a wild-type allele of polA, encoding DNA polymerase I. Curing is achieved by cotransduction of a mutant polA allele with metE::Tn10, fadAB::Tn10 or other transposon insertions near polA. Reciprocal transduction to Met(+) Pol(+) or to Fad(+) Pol(+) ensures reestablishment of the original genotype except for loss of the plasmid. A set of useful bacterial strains is provided.
