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用于分析粟酒裂殖酵母DNA复制的细胞周期同步化

Cell-cycle synchrony for analysis of S. pombe DNA replication.

作者信息

Luche Douglas D, Forsburg Susan L

机构信息

Department of Molecular and Computational Biology, University of Southern California, Los Angeles, CA, USA.

出版信息

Methods Mol Biol. 2009;521:437-48. doi: 10.1007/978-1-60327-815-7_24.

Abstract

Analysis of S phase events requires the ability to synchronize the cell cycle for subsequent physiological or molecular analysis. The cell cycle can be arrested at different stages, using drugs or temperature-sensitive mutations, and then released (block and release). These methods offer high levels of synchrony. Synchrony is lower in methods that fractionate the cells (lactose gradient synchrony), which avoids any cell-cycle perturbation. The degree of synchronization is assessed by monitoring cell morphology or DNA content. The choice of method is influenced by the needs of the experiment and the requirements of any mutations already in the strain of interest.

摘要

对S期事件的分析需要具备使细胞周期同步化的能力,以便进行后续的生理学或分子分析。利用药物或温度敏感突变,细胞周期可停滞在不同阶段,然后再解除阻滞(阻断与解除)。这些方法可实现高度同步化。通过对细胞进行分级分离的方法(乳糖梯度同步化)同步化程度较低,该方法可避免任何细胞周期扰动。通过监测细胞形态或DNA含量来评估同步化程度。方法的选择受实验需求以及所研究菌株中已有任何突变的要求影响。

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