Department of Biochemistry, University of Alberta, Edmonton, AB, Canada.
Methods Mol Biol. 2022;2579:145-168. doi: 10.1007/978-1-0716-2736-5_12.
The cell division cycle is a fundamental process required for proliferation of all living organisms. The eukaryotic cell cycle follows a basic template with an ordered series of events beginning with G1 (Gap1) phase, followed successively by S (Synthesis) phase, G2 (Gap 2) phase, and M-phase (Mitosis). The process is tightly regulated in response to signals from both the internal and external milieu. The budding yeast S. cerevisiae is an outstanding model for the study of the cell cycle and its regulatory process. The basic events and regulatory processes of the S. cerevisiae cell cycle are highly conserved with other eukaryotes. The organism grows rapidly in simple medium, has a sequenced annotated genome, well-established genetics, and is amenable to analysis by proteomics and microscopy. Additionally, a range of tools and techniques are available to generate cultures of S. cerevisiae that are homogenously arrested or captured at specific phases of the cell cycle and upon release from that arrest these can be used to monitor cell cycle events as the cells synchronously proceed through a division cycle. In this chapter, we describe a series of commonly used techniques that are used to generate synchronized populations of S. cerevisiae and provide an overview of methods that can be used to monitor the progression of the cells through the cell division cycle.
细胞分裂周期是所有生物增殖所必需的基本过程。真核细胞周期遵循一个基本模板,其中一系列有序的事件始于 G1(Gap1)期,随后依次是 S(合成)期、G2(Gap 2)期和 M 期(有丝分裂)。该过程受到来自内部和外部环境的信号的严格调控。 budding 酵母 S. cerevisiae 是研究细胞周期及其调控过程的出色模型。S. cerevisiae 细胞周期的基本事件和调控过程与其他真核生物高度保守。该生物在简单培养基中生长迅速,具有测序注释的基因组、完善的遗传学,并且易于通过蛋白质组学和显微镜进行分析。此外,还提供了一系列工具和技术来生成 S. cerevisiae 的培养物,这些培养物可以均匀地停滞或在细胞周期的特定阶段被捕获,并且在从该停滞中释放后,可以用于监测细胞周期事件,因为细胞同步地通过分裂周期进行。在本章中,我们描述了一系列常用的技术,用于生成同步的 S. cerevisiae 群体,并概述了可用于监测细胞通过细胞分裂周期进展的方法。