Hess M T, Gunz D, Naegeli H
Institute of Pharmacology and Toxicology, University of Zürich-Tierspital, Zürich, Switzerland.
Nucleic Acids Res. 1996 Mar 1;24(5):824-8. doi: 10.1093/nar/24.5.824.
We developed a competition assay to compare, in a quantitative manner, the ability of human nucleotide excision repair (NER) to recognise structurally different forms of DNA damage. This assay uses a NER substrate consisting of M13 double-stranded DNA with a single and uniquely located acetylaminofluorene (AAF) adduct, and measures the efficiency by which multiply damaged plasmid DNA competes for excision repair of the site-directed modification. To validate this assay, we tested competitor DNA containing defined numbers of either AAF adducts or UV radiation products. In both cases, repair of the site-directed NER substrate was inhibited in a damage-specific and dose-dependent manner. We then exploited this competition assay to determine the susceptibility of bulky adozelesin-DNA adducts to human NER.
我们开发了一种竞争检测法，以定量方式比较人类核苷酸切除修复（NER）识别结构不同形式DNA损伤的能力。该检测法使用一种NER底物，其由含有单个且位置独特的乙酰氨基芴（AAF）加合物的M13双链DNA组成，并测量多重损伤的质粒DNA竞争修复定点修饰的效率。为验证该检测法，我们测试了含有特定数量的AAF加合物或紫外线辐射产物的竞争DNA。在这两种情况下，定点NER底物的修复均以损伤特异性和剂量依赖性方式受到抑制。然后，我们利用这种竞争检测法来确定大分子阿多来新-DNA加合物对人类NER的敏感性。
