Hoeijmakers J H, Eker A P, Wood R D, Robins P
Department of Cell Biology and Genetics, Erasmus University, Rotterdam, The Netherlands.
Mutat Res. 1990 Sep-Nov;236(2-3):223-38. doi: 10.1016/0921-8777(90)90007-r.
Elucidation of the molecular mechanism of mammalian nucleotide excision repair requires the availability of purified proteins, DNA substrates with defined lesions and suitable repair assays. Repair assays introduced in recent years vary from testing individual steps and successions of steps in vitro to systems that closely reflect the entire process in vivo. In the first part of this review, an in vivo microinjection system is discussed. The second part of the article reviews an in vitro system for study of repair synthesis promoted by cell extracts. Both systems can be utilized as assays during the purification of protein factors that complement repair-defective xeroderma pigmentosum cells. The effect of purified repair proteins from other organisms on mammalian repair is also considered.
要阐明哺乳动物核苷酸切除修复的分子机制，需要有纯化的蛋白质、带有特定损伤的DNA底物以及合适的修复检测方法。近年来引入的修复检测方法各不相同，从体外测试单个步骤及步骤的连续性到能紧密反映体内整个过程的系统。在本综述的第一部分，将讨论一种体内显微注射系统。文章的第二部分将综述一种用于研究细胞提取物促进的修复合成的体外系统。在纯化能补充修复缺陷型着色性干皮病细胞的蛋白质因子的过程中，这两种系统都可用作检测方法。还会考虑来自其他生物体的纯化修复蛋白对哺乳动物修复的影响。
