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Molecular Cloning of the Human Nucleotide-Excision-Repair Gene ERCC4

by Weber, Christine A. , Michael J . Siciliano , Sancar, Aziz , Brookman, Kerry W. , Salazar, Edmund P. , Reardon, Joyce T. , Deng, Zuoming , Thompson, Larry H.

in Animals / Cell Line / chromosome 6 / Chromosomes / Chromosomes, Human, Pair 16 / Cloning / Cloning, Molecular / Complementary DNA / Cosmids / Cricetinae / DNA / DNA repair / DNA Repair - genetics / DNA-Binding Proteins - genetics / Drug Resistance - genetics / ERCC4 gene / Genes / Genetic engineering / Genetic Linkage / Genomics / Human genetics / Humans / Hybrid Cells / Libraries / man / mapping / Molecules / Neomycin - pharmacology / Somatic cells / Transfection / Transformation, Genetic / Ultraviolet radiation

1994

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Molecular Cloning of the Human Nucleotide-Excision-Repair Gene ERCC4

by Weber, Christine A. , Michael J . Siciliano , Sancar, Aziz , Brookman, Kerry W. , Salazar, Edmund P. , Reardon, Joyce T. , Deng, Zuoming , Thompson, Larry H.

1994

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Molecular Cloning of the Human Nucleotide-Excision-Repair Gene ERCC4

by Weber, Christine A. , Michael J . Siciliano , Sancar, Aziz , Brookman, Kerry W. , Salazar, Edmund P. , Reardon, Joyce T. , Deng, Zuoming , Thompson, Larry H.

1994

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Journal Article

Molecular Cloning of the Human Nucleotide-Excision-Repair Gene ERCC4

Weber, Christine A.,

Michael J . Siciliano,

Sancar, Aziz,

Brookman, Kerry W.,

Salazar, Edmund P.,

Reardon, Joyce T.,

Deng, Zuoming,

Thompson, Larry H.

1994

Overview

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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Publisher

National Academy of Sciences of the United States of America,National Acad Sciences,National Academy of Sciences

Subject

/ Chromosomes, Human, Pair 16

/ Cloning

/ Cloning, Molecular