申请人:——
公开号:US20020048778A1
公开(公告)日:2002-04-25
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 &bgr;-galactosidase mutants. Association between the molecules of interest brings the complementing &bgr;-galactosidase mutants into proximity so that complementation occurs and active &bgr;-galactosidase is produced. The active &bgr;-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.
本发明提供了检测分子相互作用,特别是蛋白质-蛋白质相互作用的方法和组合物。本发明可在活细胞或体外检测此类相互作用。活细胞中分子相互作用的检测并不局限于核区,也可在细胞质、细胞表面、细胞器或这些实体之间进行。在一个实施方案中,该方法利用了新型组合物,其中包括相关分子与两种或两种以上无活性、弱互补的&bgr;-半乳糖苷酶突变体之间的融合蛋白。相关分子之间的结合使互补的&bgr;-半乳糖苷酶突变体接近,从而发生互补,产生活性&bgr;-半乳糖苷酶。活性&bgr;-半乳糖苷酶可通过本领域众所周知的方法检测。本发明的用途包括蛋白质-蛋白质相互作用研究、功能基因组学、激动剂和拮抗剂筛选以及药物发现。