Delaney James C, Essigmann John M
Department of Chemistry and Biological Engineering Division, Massachusetts Institute of Technology, USA.
Methods Enzymol. 2006;408:1-15. doi: 10.1016/S0076-6879(06)08001-3.
DNA damage, if left unrepaired, may hinder translesion synthesis, leading to cytotoxicity, and instruct a DNA polymerase to incorporate an incorrect incipient base opposite the damage, leading to mutagenicity. This chapter describes technology used to measure quantitatively the degree to which a specific type of DNA damage impedes DNA replication. The technology also quantifies the mutation frequency and specificity of such damage after replication within cells. If cells with defined defects in DNA repair are used as hosts for replication, one can pinpoint the specific enzymes or pathways of repair that are operative on specific types of DNA damage.
DNA损伤若不修复，可能会阻碍跨损伤合成，导致细胞毒性，并指令DNA聚合酶在损伤对面掺入错误的起始碱基，从而导致致突变性。本章介绍了用于定量测量特定类型DNA损伤阻碍DNA复制程度的技术。该技术还可量化细胞内复制后此类损伤的突变频率和特异性。如果将具有明确DNA修复缺陷的细胞用作复制宿主，就可以确定对特定类型DNA损伤起作用的具体修复酶或修复途径。
