Identification of a DNA segment containing the human DNA polymerase alpha gene.
We isolated a temperature-sensitive mutant from mouse FM3A cells, designated as tsFT20, the DNA polymerase alpha activity of which is heat-labile. A hybrid clone (M6-39 cells) between human cells and tsFT20 cells contained one or two human chromosomes. M6-39 cells (primary hybrid) were exposed to gamma-ray and re-fused with tsFT20, after which we isolated two temperature-resistant secondary hybrids, both of which retained an identical minute portion of the human chromosome, 400-500 kilobase pairs (kbp). Immunological studies demonstrated that this secondary hybrid expressed human DNA polymerase alpha. Thus, the human DNA polymerase alpha gene was located within a DNA region of 400-500 kbp.
D000818:Animals; D000911:Antibodies, Monoclonal; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D004257:DNA Polymerase II / Q000235:genetics*; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D009693:Nucleic Acid Hybridization
D000911:Antibodies, Monoclonal; D004247:DNA; D004257:DNA Polymerase II