Spontaneous and induced homologous recombination between lacZ chromosomal direct repeats in CV-1 cells.
A lacZ substrate for intrachromosomal homologous recombination was generated at a specific site within the genome of CV-1 cells by FLP recombinase-mediated gene targeting. A histochemical stain was used to detect cells that contained recombined lacZ genes. The spontaneous rate of homologous recombination was approximately 1 x 10(-5) events per cell generation. Recombination was induced 30-fold in cells following exposure to mitomycin C (MMC) and by serum starvation. These results demonstrate the utility of the FLP recombinase in modifying the genome of mammalian cells in a predetermined manner and show that homologous recombination between direct repeats is increased in cells as a result of the withdrawal of serum growth factors.
Ludwig
D L
DL
Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati Medical Center, Ohio 45267-0524.
Stringer
J R
JR
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D002875:Chromosomes; D016895:Culture Media, Serum-Free; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D016685:Mitomycin / Q000494:pharmacology; D011995:Recombination, Genetic / Q000187:drug effects; D012091:Repetitive Sequences, Nucleic Acid; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D016895:Culture Media, Serum-Free; D016685:Mitomycin; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase; D001616:beta-Galactosidase