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裂殖酵母(Schizosaccharomyces pombe)细胞周期同步化方法。

Cell Cycle Synchrony Methods for Fission Yeast, Schizosaccharomyces pombe.

机构信息

Molecular Science Program, Yeates School of Graduate Studies, Toronto Metropolitan University (formerly Ryerson University), Toronto, ON, Canada.

Department of Chemistry and Biology, Toronto Metropolitan University (formerly Ryerson University), Toronto, ON, Canada.

出版信息

Methods Mol Biol. 2022;2579:169-179. doi: 10.1007/978-1-0716-2736-5_13.

Abstract

The fission yeast, Schizosaccharomyces pombe, is a genetically tractable model organism for cell cycle and molecular genetics research. We describe methods to synchronize S. pombe cultures, and the benefits and limitations of each. Drug-induced synchrony is a convenient method to arrest the cell cycle. An example of the drug hydroxyurea is shown, which arrests cells in S-phase. Environmental modulation through media composition or growth conditions may also be used to synchronize cultures, most commonly with nitrogen depletion to arrest in G1-phase. Finally, examples of temperature-sensitive conditional alleles are shown which arrest the cell cycle at key transition points. Each of these methods must be assessed relative to the desired effect and the process being studied, providing the best synchrony with the fewest off-target effects.

摘要

裂殖酵母,Schizosaccharomyces pombe,是细胞周期和分子遗传学研究的一种遗传上易于处理的模式生物。我们描述了同步 S. pombe 培养物的方法,以及每种方法的优缺点。药物诱导的同步是一种方便的方法来阻止细胞周期。以羟基脲为例,它会阻止细胞进入 S 期。通过培养基组成或生长条件的环境调节也可用于同步培养物,最常见的是通过氮饥饿使细胞停滞在 G1 期。最后,还展示了温度敏感条件等位基因的例子,它们可在关键的转换点阻止细胞周期。每种方法都必须根据所需效果和正在研究的过程进行评估,以在最少的脱靶效应下提供最佳的同步性。

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