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粟酒裂殖酵母细胞周期分析

Analysis of the Schizosaccharomyces pombe Cell Cycle.

作者信息

Hagan Iain M, Grallert Agnes, Simanis Viesturs

机构信息

CRUK Cell Division Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester M20 4BX, United Kingdom;

EPFL SV ISREC UPSIM, SV2.1830, Station 19, CH-1015 Lausanne, Switzerland.

出版信息

Cold Spring Harb Protoc. 2016 Sep 1;2016(9):2016/9/pdb.top082800. doi: 10.1101/pdb.top082800.

Abstract

Schizosaccharomyces pombe cells are rod shaped, and they grow by tip elongation. Growth ceases during mitosis and cell division; therefore, the length of a septated cell is a direct measure of the timing of mitotic commitment, and the length of a wild-type cell is an indicator of its position in the cell cycle. A large number of documented stage-specific changes can be used as landmarks to characterize cell cycle progression under specific experimental conditions. Conditional mutations can permanently or transiently block the cell cycle at almost any stage. Large, synchronously dividing cell populations, essential for the biochemical analysis of cell cycle events, can be generated by induction synchrony (arrest-release of a cell cycle mutant) or selection synchrony (centrifugal elutriation or lactose-gradient centrifugation). Schizosaccharomyces pombe cell cycle studies routinely combine particular markers, mutants, and synchronization procedures to manipulate the cycle. We describe these techniques and list key landmarks in the fission yeast mitotic cell division cycle.

摘要

粟酒裂殖酵母细胞呈杆状,通过顶端延伸生长。有丝分裂和细胞分裂期间生长停止;因此,分隔细胞的长度是有丝分裂承诺时间的直接度量,野生型细胞的长度是其在细胞周期中位置的指标。大量已记录的阶段特异性变化可作为标志,用于在特定实验条件下表征细胞周期进程。条件突变几乎可在任何阶段永久或暂时阻断细胞周期。通过诱导同步化(细胞周期突变体的阻滞释放)或选择同步化(离心淘洗或乳糖梯度离心)可产生大量同步分裂的细胞群体,这对于细胞周期事件的生化分析至关重要。粟酒裂殖酵母细胞周期研究通常结合特定标记、突变体和同步化程序来操纵细胞周期。我们描述这些技术,并列出裂殖酵母有丝分裂细胞分裂周期中的关键标志。

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