Kotova Elena, Serebriiskii Ilya, Coleman Thomas
Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA.
Curr Protoc Mol Biol. 2002 Nov;Chapter 20:Unit 20.7. doi: 10.1002/0471142727.mb2007s60.
The yeast two-hybrid system, or interaction trap, is one of the most versatile methods available with which to identify and establish protein-protein interactions. The dual bait system is one adaptation of the classic approach. This system facilitates the simultaneous comparison of two distinct baits with one prey. One protein of interest is expressed as a fusion to the DNA binding protein LexA (bait 1), while a second protein of interest is expressed as a fusion to the DNA binding protein cI (bait 2). Strains of yeast engineered for screening of these dual baits possess four separate reporter genes. A plasmid expressing an activation domain-fused protein (prey), which can be either a defined protein interactor or a cDNA library, is also expressed to allow dual hybrid-mediated transcriptional activation.
酵母双杂交系统，或称相互作用陷阱，是可用于识别和建立蛋白质-蛋白质相互作用的最通用方法之一。双诱饵系统是经典方法的一种改进。该系统便于同时将两个不同的诱饵与一个猎物进行比较。一种感兴趣的蛋白质与DNA结合蛋白LexA融合表达（诱饵1），而另一种感兴趣的蛋白质与DNA结合蛋白cI融合表达（诱饵2）。为筛选这些双诱饵而构建的酵母菌株拥有四个独立的报告基因。还表达一个表达激活域融合蛋白（猎物）的质粒，该猎物可以是一个确定的蛋白质相互作用分子或一个cDNA文库，以实现双杂交介导的转录激活。
