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Protease from Streptomyces griseus

来自灰色链霉菌(Streptomyces griseus)的蛋白酶

Company: Sigma-Aldrich
Catalog#: P6911
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Synchronization of Saccharomyces cerevisiae Cells in G1 Phase of the Cell Cycle
[Abstract]  The baker’s yeast, Saccharomyces cerevisiae is a widely used model organism in molecular biology because of the high homology it shares with human cells in many basic cellular processes such as DNA replication, repair, recombination, transcription, and because of the ease its genome can be manipulated. Other advantages of working with yeast are its fast production rate which is comparable to bacteria’s, and its cheap maintenance.

To examine certain phenomena, for example whether a mutation affects the passage through a cell cycle phase, it can be necessary to work with a yeast culture, in which all the cells are in the same phase of the cell cycle. Yeasts can be arrested and kept in different phases of the cell cycle. Here we describe the method that allows ...
[摘要]  面包酵母,酿酒酵母(Saccharomyces cerevisiae)是分子生物学中广泛使用的模式生物体,因为它在许多基本细胞过程如DNA复制,修复,重组,转录中与人细胞具有高同源性,并且因为的易于其基因组可以操纵。使用酵母的其他优点是其快速的生产速率与细菌相当,并且其廉价的维护。
为了检查某些现象,例如突变是否影响通过细胞周期阶段,可能有必要与酵母培养物一起工作,其中所有细胞处于细胞周期的相同阶段。酵母可以被捕获并保持在细胞周期的不同阶段。在这里我们描述了允许同步和保持酵母细胞在细胞周期的G1期与交配信息素,α因子的方法。只有MATa细胞可以与由MATα细胞产生的α因子同步。强烈建议使用MATa bar1 缺失菌株。 BAR1 基因编码胞外蛋白酶,通过切割它使α因子失活(MacKay等人,1988)。为了在使用BAR1细胞时抵消蛋白酶活性,相比于bar1 缺失细胞,需要100-1000倍的α因子(α-因子是相当昂贵的!) ,同步仍然是瞬态的。相反,bar1 缺失细胞可以在具有α因子的G1期保持几小时,并且同步程度通常高于使用BAR1 株。此外,由于分泌的Bar1蛋白酶的活性,仅在低细胞密度下,即使在高细胞密度下, bar1 缺失细胞也可以同步,而BAR1 细胞。