对活酵母中的实时基因表达进行成像。
Imaging real-time gene expression in living yeast.
作者信息
Zenklusen Daniel, Wells Amber L, Condeelis John S, Singer Robert H
机构信息
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
出版信息
CSH Protoc. 2007 Nov 1;2007:pdb.prot4870. doi: 10.1101/pdb.prot4870.
INTRODUCTIONThis protocol describes the application of the MS2 system to the yeast Saccharomyces cerevisiae. ASH1 mRNA tagged with six MS2 repeats (6MBSs) is used to follow the localization of the ASH1 mRNA particles to the bud tip of a haploid yeast cell. W303 yeast cells transformed with pG14-MS2-GFP and pGAL-lacZ-MS2-ASH1 are grown on select medium lacking tryptophan and leucine. RNA expression is induced by the addition of galactose, and a time-lapse movie is then acquired.
引言
本方案描述了MS2系统在酿酒酵母中的应用。用六个MS2重复序列（6MBSs）标记的ASH1 mRNA用于追踪ASH1 mRNA颗粒在单倍体酵母细胞芽尖的定位。用pG14-MS2-GFP和pGAL-lacZ-MS2-ASH1转化的W303酵母细胞在缺乏色氨酸和亮氨酸的选择培养基上生长。通过添加半乳糖诱导RNA表达，然后获取延时电影。
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