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. 1994 Jul 19;91(15):6855–6859. doi: 10.1073/pnas.91.15.6855

Molecular cloning of the human nucleotide-excision-repair gene ERCC4.

L H Thompson 1, K W Brookman 1, C A Weber 1, E P Salazar 1, J T Reardon 1, A Sancar 1, Z Deng 1, M J Siciliano 1
PMCID: PMC44296  PMID: 8041709

Abstract

ERCC4 was previously identified in somatic cell hybrids as a human gene that corrects the nucleotide-excision-repair deficiency in mutant hamster cells. The cloning strategy for ERCC4 involved transfection of the repair-deficient hamster cell line UV41 with a human sCos-1 cosmid library derived from chromosome 16. Enhanced UV resistance was seen with one cosmid-library transformant and two secondary transformants of UV41. Cosmid clones carrying a functional ERCC4 gene were isolated from a library of a secondary transformant by selecting in Escherichia coli for expression of a linked neomycin-resistance gene that was present in the sCos-1 vector. The cosmids mapped to 16p13.13-p13.2, the location assigned to ERCC4 by using somatic cell hybrids. Upon transfection into UV41, six cosmid clones gave partial correction ranging from 30% to 64%, although all appeared to contain the complete gene. The capacity for in vitro excision of thymine dimers from a plasmid by transformant cell extracts correlated qualitatively with enhanced UV resistance.

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Selected References

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