Yadav Vipin Kumar, Awasthi Poorwa, Kumar Amit
Genome & Cell Integrity Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR-Indian Institute of Toxicology Research, Lucknow, India.
Methods Mol Biol. 2019;2031:313-322. doi: 10.1007/978-1-4939-9646-9_17.
Ultraviolet rays induce interstrand and intrastrand DNA cross-links, usually thymine-thymine cyclobutane dimer (T-T) and thymine-thymine pyrimidine-pyrimidone (6-4) photoproduct (T (6-4) T). These DNA cross-links, if left unrepaired, increase the risk of these mutation being incorporated in the genetic material (i.e., DNA). Numerous studies have reported the mutagenic potential of above mentioned DNA adducts in prokaryotes, yeast and mammalian cells. Different techniques have been developed to identify such DNA adducts such as immuno-Southern blotting. This is a routinely used quantitative method to determine especially the amount of thymine dimers formed, following irradiation. In this chapter, the detailed methodology to identify thymine dimers formation is provided, using specific antibody against these adducts.
紫外线会诱导链间和链内DNA交联，通常形成胸腺嘧啶-胸腺嘧啶环丁烷二聚体（T-T）和胸腺嘧啶-胸腺嘧啶嘧啶-嘧啶酮（6-4）光产物（T（6-4）T）。这些DNA交联如果不进行修复，会增加这些突变整合到遗传物质（即DNA）中的风险。许多研究报道了上述DNA加合物在原核生物、酵母和哺乳动物细胞中的诱变潜力。已经开发出不同的技术来鉴定此类DNA加合物，如免疫Southern印迹法。这是一种常规使用的定量方法，尤其用于测定照射后形成的胸腺嘧啶二聚体的数量。在本章中，将提供使用针对这些加合物的特异性抗体来鉴定胸腺嘧啶二聚体形成的详细方法。
