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Epitope tagging of yeast genes using a PCR-based strategy: More tags andimproved practical routines

机译:使用基于PCR的策略对酵母基因进行表位标记:更多标记和改进的常规程序

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摘要

Epitope tagging of proteins as a strategy for the analysis of function, interactions and the subcellular distribution of proteins has become widely used. In the yeast Saccharomyces cerevisiae, molecular biological techniques have been developed that use a simple PCR-based strategy to introduce epitope tags to chromosomal loci (Wach et al., 1994). To further employ the power of this strategy, a variety of novel tags was constructed. These tags were combined with different selectable marker genes, resulting in PCR amplificable modules. Only one set of primers is required for the amplification of any module. Furthermore, convenient laboratory techniques are described that facilitate the genetic manipulations of yeast strains, as well as the analysis of the epitope-tagged proteins. Copyright (C) 1999 John Wiley & Sons, Ltd.
机译:蛋白质的表位标记作为蛋白质功能,相互作用和亚细胞分布分析的一种策略已被广泛使用。在酿酒酵母中,已经开发了分子生物学技术,该技术使用基于PCR的简单策略将表位标签引入染色体位点(Wach等,1994)。为了进一步利用这种策略的力量,构建了各种新颖的标签。这些标签与不同的选择标记基因结合在一起,形成了PCR扩增模块。任何模块的扩增仅需要一组引物。此外,描述了便利的实验室技术,其促进酵母菌株的遗传操作以及表位标记的蛋白质的分析。版权所有(C)1999 John Wiley&Sons,Ltd.

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