Abstract
DNA from a repair competent hamster cell line, ligated to a dominant selectable marker, was transfected into immortalised xeroderma D cells. Two rounds of transfection resulted in the isolation of a cell line with UV-induced excision repair capability similar to that of a primary transfectant but with few integrated copies of both marker and hamster DNA sequences. Cosmid rescue of the dominant marker from secondary transfectant DNA gave rise to several clones, two of which contain hamster DNA sequences and confer increased survival to UV on transfection into two, independently-immortalised xeroderma D cell lines, but not on transfection into a xeroderma A line. Restriction enzyme analysis revealed a 13 kilobase overlap between these 2 cosmids; no similarities to the previously isolated UV excision repair genes ERCC1 and 2 were evident. A non-repetitive portion of the overlap region, when used to probe total cellular RNA, showed that the gene is expressed to varying degrees in all cell lines tested.
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© 1989 Plenum Press, New York
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Arrand, J.E., Bone, N.M., Johnson, R.T. (1989). The Cloning and Characterization of a Candidate Gene for the Correction of the Xeroderma D Defect. In: Lambert, M.W., Laval, J. (eds) DNA Repair Mechanisms and Their Biological Implications in Mammalian Cells. NATO ASI Series, vol 182. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-1327-4_46
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DOI: https://doi.org/10.1007/978-1-4684-1327-4_46
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