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Survival and DNA Repair of Somatic Cell Hybrids after Ultraviolet Irradiation
Authors:S GOLDSTEIN  C C LIN
Institution:1.Departments of Medicine and Pediatrics,McMaster University,Hamilton
Abstract:THE technique of somatic cell hybridization has opened up studies on genetic regulation1 and human genetic analysis2–5. Hybrid cells are isolated in conditions that select against parental cells while allowing hybrids to survive by genomic complementation. In xeroderma pigmentosum (XP), a human disease with an autosomal recessive defect in an early stage of DNA repair6, the skin is extremely sensitive to sunlight in vivo7 and skin fibroblasts show sharply reduced survival following ultraviolet irradiation in vitro8,9. This communication concerns the use of ultraviolet irradiation in combination with a chemical method to produce hybrids between fibroblasts from XP and a hamster line, followed by analysis of these cells for their capacity to survive and repair DNA after exposure to ultraviolet. Methods for initiation and propagation of skin fibroblasts from two subjects, male and female siblings with XP, have been described8. Details on the origin of the TG2 line of golden hamster fibroblasts, which has a non-reverting mutation in the gene for hypoxanthine-guanine phosphoribosyltransferase (HGPRT), the general hybridization procedure10 and methods for cell survival and DNA repair by unscheduled synthesis8 were also described previously. Hybrids were produced by fusion with Sendai virus and selected by ultraviolet irradiation followed by culture on HAT medium (Fig. 1).
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