Microcell-mediated transfer of a single human chromosome complements xeroderma pigmentosum group A fibroblasts.
Chromosomes from an immortalized aneuploid human fibroblast cell line were randomly tagged with the selectable marker neo by transfection with the plasmid pSV2neo. Somatic cell fusions between transfected human cells and mouse A9 cells generated pools of G418-resistant human-mouse hybrid clones containing various numbers of human chromosomes. Microcell-mediated chromosome transfer from the hybrid pools to xeroderma pigmentosum complementation group A (XP-A) cells in culture and selection for G418-resistant colonies resulted in the identification of XP cells with enhanced resistance to ultraviolet radiation. Screening of subclones from selected pools of human-mouse hybrids facilitated the identification of hybrids containing a single neo-tagged human chromosome. Transfer of this chromosome to XP-A cells (but not to XP-F or XP-C cells) results in enhanced resistance to ultraviolet light and enhanced excision repair capacity. The identification of a single human chromosome that complements the phenotype of XP-A cells in culture provides the potential for genetic mapping of the complementing gene and for its isolation by molecular cloning.
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D002871:Chromosome Banding; D002877:Chromosomes, Human / Q000528:radiation effects; D005816:Genetic Complementation Test; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D007621:Karyotyping; D008677:Metaphase; D051379:Mice; D010641:Phenotype; D013539:Simian virus 40 / Q000235:genetics; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics* / Q000473:pathology