Two-step method for curing Escherichia coli of ColE1-derived plasmids

TY - JOUR

T1 - Two-step method for curing Escherichia coli of ColE1-derived plasmids

AU - Hove-Jensen, Bjarne

N1 - Keywords: Bacterial Proteins; DNA Polymerase I; DNA Replication; DNA Transposable Elements; Escherichia coli; Mutagenesis, Insertional; Plasmids; Transduction, Genetic

PY - 2008

Y1 - 2008

N2 - 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.

AB - 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.

U2 - 10.1016/j.mimet.2007.11.020

DO - 10.1016/j.mimet.2007.11.020

M3 - Journal article

C2 - 18164774

SN - 0167-7012

VL - 72

SP - 208

EP - 213

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

IS - 2

ER -