Induced recombination between duplicated neo genes stably integrated in the genome of CHO cells.
Homologous recombination between 2 truncated neo genes stably integrated in the genome of Chinese hamster ovary (CHO) cells was studied. A vector containing a functional gpt gene and 2 tandemly arranged G418 resistance (neo) gene fragments with about 400 bp of sequence homology was transfected into CHO cells. Clonal cell lines were established from transfected cultures and the spontaneous frequency of G418-resistant revertants was found to range between 1 x 10(-4) and 5 x 10(-4). The ability of the alkylating agents MMS and HN2 to induce recombination of the transfected neo genes was studied in 2 of the cell lines. After treatment with MMS at doses that reduced survival to 10% of the control these cell lines showed a dose-dependent increase in the frequency of G418-resistant revertants. No effect was observed after treatment with HN2. All G418-resistant subclones contained a new restriction fragment indicating that a whole neo gene had been formed by rearrangement in pairs of truncated neo genes. Hence, this system can be used to study molecular mechanisms and chemical inducibility of homologous recombination in mammalian cells.
Hellgren
D
D
Department of Clinical Genetics, Karolinska Hospital, Stockholm, Sweden.
Luthman
H
H
Lambert
B
B
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004351:Drug Resistance; D005798:Genes, Bacterial; D005839:Gentamicins / Q000494:pharmacology; D019868:Kanamycin Kinase; D008466:Mechlorethamine / Q000494:pharmacology; D008741:Methyl Methanesulfonate / Q000633:toxicity; D009154:Mutation / Q000187:drug effects*; D010770:Phosphotransferases / Q000235:genetics; D010957:Plasmids; D011995:Recombination, Genetic
D005839:Gentamicins; D008466:Mechlorethamine; C010680:antibiotic G 418; D008741:Methyl Methanesulfonate; D010770:Phosphotransferases; D019868:Kanamycin Kinase