Rosebrock Adam P
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
Cold Spring Harb Protoc. 2017 Jan 3;2017(1):2017/1/pdb.top080630. doi: 10.1101/pdb.top080630.
Like other eukaryotes, budding yeast temporally separate cell growth and division. DNA synthesis is distinct from chromosome segregation. Storage carbohydrates are accumulated slowly and then rapidly liquidated once per cycle. Cyclin-dependent kinase associates with multiple different transcriptionally and posttranslationally regulated cyclins to drive the cell cycle. These and other crucial events of cellular growth and division are limited to narrow windows of the cell cycle. Many experiments in the yeast laboratory treat a culture of cells as a homogeneous mixture. Measurements of asynchronous cultures are, however, confounded by the presence of cells in various cell cycle stages; measuring a population average in unsynchronized cells provides at best a decreased signal and at worst an artifactual result. A number of experimentally tractable methods have been developed to generate populations of yeast cells that are synchronized with respect to cell cycle phase. Robust methods for determining cell cycle position have also been developed. These methods are introduced here.
与其他真核生物一样，出芽酵母在时间上分离细胞生长和分裂。DNA合成与染色体分离不同。储存碳水化合物缓慢积累，然后每周期迅速消耗一次。细胞周期蛋白依赖性激酶与多种不同的受转录和翻译后调控的细胞周期蛋白结合，以驱动细胞周期。细胞生长和分裂的这些以及其他关键事件仅限于细胞周期的狭窄窗口。酵母实验室中的许多实验将细胞培养物视为均匀混合物。然而，异步培养物的测量会因处于不同细胞周期阶段的细胞的存在而混淆；测量未同步细胞的群体平均值充其量只能提供降低的信号，最坏的情况是得到人为的结果。已经开发出许多易于实验操作的方法来产生在细胞周期阶段同步的酵母细胞群体。也已经开发出用于确定细胞周期位置的可靠方法。这里介绍这些方法。
