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    Interaction trap systems for detecting protein interactions
    20.
    发明申请
    Interaction trap systems for detecting protein interactions 审中-公开
    用于检测蛋白质相互作用的相互作用捕获系统

    公开(公告)号：US20090130676A1

    公开(公告)日：2009-05-21

    申请号：US12079358

    申请日：2008-03-26

    申请人： Roger Brent ,  John M. McCoy ,  Timm H. Jessen ,  Chanxing Wilson Xu

    发明人： Roger Brent ,  John M. McCoy ,  Timm H. Jessen ,  Chanxing Wilson Xu

    IPC分类号： C12Q1/68

    CPC分类号： C12N9/0036 ,  C07K2319/00 ,  C07K2319/35 ,  C07K2319/70 ,  C07K2319/71 ,  C07K2319/80 ,  C12N15/1055 ,  C12Q1/6897

    摘要： Disclosed herein is a method of determining whether a first protein is capable of physically interacting with a second protein, involving: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a gene activating moiety and being conformationally-constrained; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Also disclosed are methods for assaying protein interactions, and identifying antagonists and agonists of protein interactions. Proteins isolated by these methods are also discussed. Finally, populations of eukaryotic cells are disclosed, each cell having a recombinant DNA molecule encoding a conformationally-constrained intracellular peptide.

    摘要翻译： 本文公开了一种确定第一蛋白质是否能够与第二蛋白质物理相互作用的方法，涉及：（a）提供宿主细胞，其包含（i）可操作地连接到蛋白质结合位点的报告基因; （ii）表达第一融合蛋白的第一融合基因，所述第一融合蛋白包括共价键合到能够特异性结合蛋白结合位点的结合部分的第一蛋白; 和（iii）表达第二融合蛋白的第二融合基因，所述第二融合蛋白包括共价键合到基因激活部分并被构象约束的第二蛋白; 和（b）测量报告基因的表达作为第一和第二蛋白质之间相互作用的量度。 还公开了用于测定蛋白质相互作用的方法，以及鉴定蛋白质相互作用的拮抗剂和激动剂。 还讨论了通过这些方法分离的蛋白质。 最后，公开了真核细胞群，每个细胞具有编码构象约束细胞内肽的重组DNA分子。