Transfer of the human X chromosome to human--Chinese hamster cell hybrids via isolated HeLa metaphase chromosomes.
Evidence is presented for the uptake of the human X chromosome by human-Chinese hamster cell hybrids which lack H P R T activity, following incubation with isolated human HeLa S3 chromosomes. Sixteen independent clonal cell lines were isolated in H A T medium, all of which contained a human X chromosome as determined by trypsin-Giemsa staining. The frequency of H A T-resistant clones was 32 x 10(-6) when 10(7) cells were incubated with 10(8) HeLa chromosomes. Potential reversion of the hybrid cells in H A T medium was less than 5 x 10(-7). The 16 isolated cell lines all contained activity of the human X-linked marker enzymes H P R T, P G K,alpha-Gal A, and G6PD, as determined by electrophoresis. The phenotype of G6PD was G6PD A, corresponding to G6PD A in HeLa cells. The human parental cells used in the fusion to form the hybrids had the G6PD B phenotype. The recipient cells gave no evidence of containing human X chromosomes. These results indicate that incorporation and expression of HeLa X chromosomes is accomplished in human-Chinese hamster hybrids which lack a human X chromosome.
D000818:Animals; D002874:Chromosome Mapping; D002875:Chromosomes; D006224:Cricetinae; D005696:Galactosidases / Q000032:analysis; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis; D006367:HeLa Cells / Q000648:ultrastructure*; D006801:Humans; D006822:Hybrid Cells / Q000648:ultrastructure*; D007041:Hypoxanthine Phosphoribosyltransferase / Q000032:analysis; D066298:In Vitro Techniques; D008938:Mitosis; D010641:Phenotype; D010735:Phosphoglycerate Kinase / Q000032:analysis; D012730:Sex Chromosomes; D013045:Species Specificity
D005954:Glucosephosphate Dehydrogenase; D007041:Hypoxanthine Phosphoribosyltransferase; D010735:Phosphoglycerate Kinase; D005696:Galactosidases