Analysis of DNA Metabolism in Fission Yeast
Analysis of DNA Metabolism in Fission Yeast
复制标题
裂殖酵母 DNA 代谢分析
DOI：
10.1101/pdb.top079863
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发表时间：
2018
影响因子：
--
通讯作者：
Antequera F
中科院分区：
文献类型：
--
作者：
Antequera F
裂殖酵母是研究DNA代谢的理想模式生物，其DNA复制和修复机制在进化上是保守的。在这个介绍中，我们描述了一系列常用的方法来研究裂变酵母中的DNA代谢方面，重点是用于分析基因组稳定性，DNA复制和DNA修复的方法。我们描述了使用一个微型染色体，CH 16，用于监测基因组稳定性的不同方面。我们介绍了二维凝胶电泳和免疫荧光可视化的梳状DNA分子的DNA复制的分析。此外，我们介绍了一种脉冲场凝胶电泳（PFGE）检测，以物理监测染色体的完整性，它可以与DNA双链断裂（DSB）修复检测，以遗传定量不同的DSB修复和误修复的结果，包括总染色体重排，在裂变酵母。
The fission yeast Schizosaccharomyces pombe is an excellent model organism to study DNA metabolism, in which the DNA replication and repair mechanisms are evolutionarily conserved. In this introduction we describe a range of methods commonly used to study aspects of DNA metabolism in fission yeast, focusing on approaches used for the analysis of genome stability, DNA replication, and DNA repair. We describe the use of a minichromosome, Ch 16, for monitoring different aspects of genome stability. We introduce two-dimensional gel electrophoresis and immunofluorescent visualization of combed DNA molecules for the analysis of DNA replication. Further, we introduce a pulsed field gel electrophoresis (PFGE) assay to physically monitor chromosome integrity, which can be used in conjunction with a DNA double-strand break (DSB) repair assay to genetically quantitate different DSB repair and misrepair outcomes, including gross chromosomal rearrangements, in fission yeast.