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    • 2. 发明申请
    • COMPOSITIONS AND METHODS FOR THE IDENTIFICATION OF PROTEIN INTERACTIONS IN VERTEBRATE CELLS
    • 用于鉴定VERTEBRATE细胞中蛋白质相互作用的组合物和方法
    • WO2002086450A2
    • 2002-10-31
    • PCT/US2002/013008
    • 2002-04-22
    • PRESIDENT AND FELLOWS OF HARVARD COLLEGEMCKEON, FrankYANG, Annie
    • MCKEON, FrankYANG, Annie
    • G01N
    • G01N33/5035G01N33/5008G01N33/5041G01N33/5076G01N33/542G01N33/6803G01N2500/02
    • The present invention relates to a two-hybrid system for studying protein-protein interactions in mammalian host cells. The invention provides methods, reagents and kits for carrying out the two-hybrid screen. Accordingly the invention provides a bait construct that is capable of targeting a bait protein to a specific subcellular locale within the host cell. The invention also provides a prey construct that contains a detection sequence fused to a prey protein. The bait and prey constructs are introduced into a host cell under conditions that promote expression of the bait and prey constructs. A positive bait/prey interaction can be detected by comparing the subcellular localization of the prey construct in relation to the bait construct. The invention also provides methods for screening for compounds capable of disrupting protein-protein interaction and methods for detecting interactions between proteins and small molecule compounds.
    • 本发明涉及用于研究哺乳动物宿主细胞中的蛋白质 - 蛋白质相互作用的双杂交系统。 本发明提供用于进行双杂交筛选的方法,试剂和试剂盒。 因此,本发明提供了能够将诱饵蛋白靶向宿主细胞内的特定亚细胞区域的诱饵构建体。 本发明还提供了含有与猎物蛋白融合的检测序列的猎物构建体。 诱饵和猎物构建体在促进诱饵和猎物构建体的条件下被引入宿主细胞。 通过比较猎物构建体相对于诱饵构建体的亚细胞定位,可以检测到正的诱饵/猎物相互作用。 本发明还提供了筛选能够破坏蛋白质 - 蛋白质相互作用的化合物的方法以及检测蛋白质与小分子化合物之间相互作用的方法。