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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, particularlyprotein-protein interactions, are provided. The invention allows detection ofsuch interactions in living cells or in vitro. Detection of molecularinteractions in living cells is not limited to the nuclear compartment, butcan be accomplished in the cytoplasm, cell surface, organelles, or betweenthese entities. In one embodiment, the method utilizes novel compositionscomprising fusion proteins between the molecules of interest and two or moreinactive, weakly-complementing .beta.-galactosidase mutants. Associationbetween the molecules of interest brings the complementing .beta.-galactosidase mutants into proximity so that complementation occurs and active.beta.-galactosidase is produced. The active .beta.-galactosidase may bedetected by methods well-known in the art. Among the uses of the invention arethe study of protein-protein interactions, functional genomics, agonist andantagonist screening and drug discovery.
机译:用于检测分子相互作用特别是分子相互作用的方法和组合物提供了蛋白质-蛋白质相互作用。本发明允许检测在活细胞或体外的这种相互作用。分子检测活细胞中的相互作用不仅限于核区室可以在细胞质,细胞表面,细胞器或细胞之间完成这些实体。在一个实施方案中,该方法利用了新颖的组合物在目标分子和两个或多个分子之间包含融合蛋白非活性的,弱互补的β-半乳糖苷酶突变体。协会感兴趣的分子之间产生互补的β-半乳糖苷酶突变体接近,从而发生互补并具有活性产生β-半乳糖苷酶。活性β-半乳糖苷酶可以是通过本领域众所周知的方法检测。本发明的用途是蛋白质间相互作用,功能基因组学,激动剂和拮抗剂筛选和药物发现。

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