Pabla Ritu, Pawar Vaibhav, Zhang Hong, Siede Wolfram
Department of Cell Biology and Genetics, University of North Texas Health Science Center, Fort Worth, USA.
Methods Enzymol. 2006;409:101-17. doi: 10.1016/S0076-6879(05)09006-3.
In spite of certain special features of its cell cycle, the yeast Saccharomyces cerevisiae has proved to be an excellent and widely used model to study eukaryotic checkpoint responses to DNA damage. This chapter primarily summarizes selected cytological methods that are useful for initial characterization of cell cycle responses. These can be useful in order to study mutants, conditions, or selected DNA damaging agents and experimental examples are given. We have also included protocols for flow-cytometric cell cycle analysis and for determination of Rad53 phosphorylation, a commonly used indicator of checkpoint activation.
尽管酿酒酵母的细胞周期具有某些特殊特征，但它已被证明是研究真核细胞对DNA损伤的检查点反应的一种优秀且广泛使用的模型。本章主要总结了一些有助于初步表征细胞周期反应的细胞学方法。这些方法对于研究突变体、条件或特定的DNA损伤剂可能有用，并给出了实验示例。我们还纳入了流式细胞术细胞周期分析和Rad53磷酸化测定的方案，Rad53磷酸化是检查点激活的常用指标。
