Positive selection of recombinant plasmids based on the EcoK restriction activity of Escherichia coli K-12.
We have constructed a pTZ19R-derived vector which allows efficient positive selection of recombinant plasmids. The system uses the EcoK restriction activity of Escherichia coli K-12 to select against non-recombinant plasmids. The vector contains an EcoK site which, if deleted or disrupted by ligating a DNA fragment, yields recombinant plasmids that are no longer susceptible to EcoK restriction when transformed into a restriction-proficient E. coli host.
Ludwig Institute for Cancer Research, Brussels Branch, Belgium.
O
O
De Backer
P
P
Chomez
E
E
De Plaen
D001483:Base Sequence; D004274:DNA, Recombinant / Q000032:analysis; D015253:Deoxyribonucleases, Type I Site-Specific / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids / Q000032:analysis*
D004274:DNA, Recombinant; C029602:endodeoxyribonuclease EcoK; D015253:Deoxyribonucleases, Type I Site-Specific