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DETECTION OF MOLECULAR INTERACTIONS BY REPORTER SUBUNIT COMPLEMENTATION

机译:通过报告人亚基互补检测分子相互作用

摘要

Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, are provided. The invention allows detection of such interactions in living cells or in vitro. Detection of molecular interactions in living cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. In one embodiment, the method utilizes novel compositions comprising fusion proteins between the molecules of interest and two or more inactive, weakly-complementing β-galactosidase mutants. Association between the molecules of interest brings the complementing β-galactosidase mutants into proximity so that complementation occurs and active β-galactosidase is produced. The active β-galactosidase may be detected by methods well-known in the art. Among the uses of the invention are the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.
机译:提供了用于检测分子相互作用,特别是蛋白质-蛋白质相互作用的方法和组合物。本发明允许在活细胞中或体外检测这种相互作用。活细胞中分子相互作用的检测不仅限于核区室,还可以在细胞质,细胞表面,细胞器或这些实体之间完成。在一个实施方案中,该方法利用了新颖的组合物,其包括在目标分子与两个或多个无活性的,弱互补的β-半乳糖苷酶突变体之间的融合蛋白。目的分子之间的缔合使互补的β-半乳糖苷酶突变体接近,从而发生互补并产生活性的β-半乳糖苷酶。可以通过本领域公知的方法检测活性β-半乳糖苷酶。本发明的用途包括蛋白质-蛋白质相互作用,功能基因组学,激动剂和拮抗剂筛选以及药物发现的研究。

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