Site-specific recombination in mammalian cells expressing the Int recombinase of bacteriophage HK022.
The int gene of bacteriophage HK022, coding for the integrase protein, was cloned in a mammalian expression vector downstream of the human cytomegalovirus (CMV) promoter. Green monkey kidney cells (COS-1) and mouse embryo fibroblast cells (NIH3T3) transiently transfected with the recombinant plasmid express the integrase protein. Co-transfection of this plasmid with reporter plasmids for site-specific recombination and PCR analyses show that the integrase promotes site-specific integration as well as excision. These reactions occurred without the need to supply integration host factor and excisionase, the accessory proteins that are required for integrase-promoted site-specific recombination in vitro as well as in the natural host Escherichia coli.
Kolot
M
M
Department of Biochemistry, Tel-Aviv University, Israel.
Silberstein
N
N
Yagil
E
E
D016475:3T3 Cells; D000818:Animals; D001435:Bacteriophages / Q000201:enzymology / Q000235:genetics*; D019556:COS Cells; D002384:Catalysis; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D019426:Integrases / Q000235:genetics*; D051379:Mice; D045522:Recombinases; D011995:Recombination, Genetic / Q000235:genetics*; D014162:Transfection
D045522:Recombinases; D004254:DNA Nucleotidyltransferases; D019426:Integrases; C115006:integron integrase IntI1