Sister Chromatid Exchange as a Genotoxic Stress Marker
Sister Chromatid Exchange as a Genotoxic Stress Marker
复制标题
姐妹染色单体交换作为基因毒性应激标记
DOI：
10.1007/978-1-4939-9432-8_7
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Kato Takamitsu A.
中科院分区：
文献类型：
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作者：
Sunada Shigeaki;Haskins Jeremy S.;Kato Takamitsu A.
姐妹染色单体交换（SCE）是DNA复制过程中部分DNA交换的现象。SCE检测是通过激发DNA的保守复制特性而发展起来的。胸苷类似物如5′-溴脱氧尿苷（BrdU）和乙炔基脱氧尿苷（EdU）掺入新合成的DNA中，持续两个细胞周期。在有丝分裂时，向培养块和传代细胞中加入秋水仙胺，并进行常规细胞遗传学制备。使用Hoechst染料和Giemsa（荧光+Giemsa染色）的差异染色方法，针对BrdU的抗体检测，或针对EdU的高度特异性Click反应，可以识别染色单体内新合成的DNA。SCE代表DNA合成过程中DNA模板交换的一个点，通过差异染色单体染色或丑角染色体可视化。本章将介绍三种基本的SCE显微分析方法，包括非荧光法和荧光法。SCE是复制过程中基因毒性应激的一个非常敏感的标记。
Sister chromatid exchange (SCE) is the phenomenon of partial DNA exchange during DNA replication. SCE detection has been developed through eliciting DNA’s semiconservative replicative nature. Thymidine analogues such as 5′-bromodeoxyuridine (BrdU) and ethynyldeoxyuridine (EdU) are incorporated into the newly synthesized DNA for two cell cycles. The addition of Colcemid to the culture blocks and synchronizes cells at mitosis, and conventional cytogenetic preparations are made. Differential staining methods with Hoechst dye and Giemsa (Fluorescence Plus Giemsa staining), antibody detection against BrdU, or highly specific Click reaction to EdU, allow the newly synthesized DNA within a chromatid to be recognized. SCEs represent a point of DNA template exchange during DNA synthesis, visualized by differential chromatid staining or harlequin chromosomes. We will introduce three basic protocols in this chapter including non-fluorescence and fluorescence methods for SCE microscopic analysis. SCE is a very sensitive marker of genotoxic stress during replication.