RECONSTITUTION OF MAMMALIAN EXCISION-REPAIR ACTIVITY WITH MUTANT CELL-FREE-EXTRACTS AND XPAC AND ERCC1 PROTEINS EXPRESSED IN ESCHERICHIA-COLI

被引：18

作者：

PARK, CH ^{[1
]}

SANCAR, A ^{[1
]}

机构：

[1] UNIV N CAROLINA, SCH MED, DEPT BIOCHEM & BIOPHYS, CHAPEL HILL, NC 27599 USA

来源：

NUCLEIC ACIDS RESEARCH | 1993年 / 21卷 / 22期

关键词：

D O I：

10.1093/nar/21.22.5110

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Nucleotide excision repair in humans involves the co-ordinated actions of 8 - 10 proteins. To understand the roles of each of these proteins in excision it is necessary to develop an in vitro excision repair system reconstituted entirely from purified proteins. Towards this goal we have expressed in E.coli two of the 8 genes known to be essential for the excision reaction. XPAC and ERCC1 were expressed as fusion proteins with the Escherichia coli maltose binding protein (MBP) and purified to > 80% homogeneity by affinity chromatography. The purified proteins either as fusions or after cleavage from the MBP were able to complement the CFE of cells with mutations in the corresponding genes in an excision assay with thymine dimer containing substrate.

引用

页码：5110 / 5116

页数：7

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