Two-step method for curing Escherichia coli of ColE1-derived plasmids.
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.
Hove-Jensen
Bjarne
B
Section for Biochemical Genetics, Department of Molecular Biology, University of Copenhagen, 5 Ole Maaløes Vej, Copenhagen N, Denmark. hove@mermaid.molbio.ku.dk
D001426:Bacterial Proteins / Q000235:genetics*; D004256:DNA Polymerase I / Q000235:genetics; D004261:DNA Replication; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics*; D016254:Mutagenesis, Insertional; D010957:Plasmids; D014161:Transduction, Genetic
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C042833:ROM protein, Bacteria; D004256:DNA Polymerase I