Isolation of UV-sensitive clones from a haploid frog cell line.
An isolation procedure has been developed which yielded five clones of haploid frog cells which are sensitive to ultraviolet light. This procedure employed a conventional mutagenesis, followed by time for phenotypic expression and then an enrichment for UV-sensitive mutants. The enrichment relies upon the uptake of bromodeoxyuridine (BrdU) by repairing cells following UV-induced damage, rendering repair-proficient cells differentially sensitive to photolysis by black light. The photolysis is potentiated by use of the bisbenzimidazole dye Hoechst 33258. The enriched population was screened for radiation-sensitive isolates resulting in 5 sensitives from 96 tested. No mutants were obtained from 300 isolates tested from a population which had not undergone enrichment.
D000818:Animals; D001001:Anura; D002460:Cell Line; D002999:Clone Cells; D004260:DNA Repair / Q000528:radiation effects; D006238:Haploidy; D009154:Mutation; D010641:Phenotype; D011836:Radiation Tolerance; D014466:Ultraviolet Rays