Dove Simon L, Hochschild Ann
Division of Infectious Diseases, Children's Hospital, Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2004;261:231-46. doi: 10.1385/1-59259-762-9:231.
We describe the use of a bacterial two-hybrid system for the study of protein-protein interactions in Escherichia coli. This system is based on transcription activation and involves the synthesis of two fusion proteins within the bacterial cell whose interaction stimulates transcription of a reporter gene. Specifically, one of the fusion proteins can function as a transcription activator when its interaction partner is fused to a subunit of the bacterial RNA polymerase. This bacterial two-hybrid system has been used to study a number of interacting proteins from both prokaryotes and eukaryotes, and can be used to find interacting proteins from complex protein libraries.
我们描述了一种用于研究大肠杆菌中蛋白质-蛋白质相互作用的细菌双杂交系统。该系统基于转录激活,涉及在细菌细胞内合成两种融合蛋白,其相互作用会刺激报告基因的转录。具体而言,当其中一种融合蛋白的相互作用伙伴与细菌RNA聚合酶的一个亚基融合时,该融合蛋白可作为转录激活剂发挥作用。这种细菌双杂交系统已被用于研究来自原核生物和真核生物的多种相互作用蛋白,并且可用于从复杂的蛋白质文库中寻找相互作用蛋白。