Prokaryotic two-hybrid system
    3.
    发明授权
    Prokaryotic two-hybrid system 失效
    原核双杂交系统

    公开(公告)号:US6051381A

    公开(公告)日:2000-04-18

    申请号:US987965

    申请日:1997-12-10

    摘要: A two-hybrid system that can detect homo- and heterodimeric protein interactions in E. coli and other cells. This system is useful for the same applications as a yeast two-hybrid system; i.e. interaction cloning, mapping protein interaction domains, analyzing protein interactions, detecting protein interactions and detecting modulators thereof. The invention concerns a prokaryotic host cell comprising:(a) a fusion protein having (i) a first DNA-binding domain and (ii) a first interacting domain;(b) a fusion protein having (i) a second DNA-binding domain and (ii) a second interacting domain capable of binding to the first interacting domain; and(c) a nucleic acid molecule having a reporter gene operatively linked to (i) a promoter, (ii) a first operator site capable of binding to the first DNA-binding domain, located upstream of the promoter, and (iii) a second operator site capable of binding the second DNA-binding domain, located downstream of the promoter of the reporter gene;wherein binding of the first interacting domain to the second interacting domain is signaled by altered expression of the reporter gene.

    摘要翻译: 可以检测大肠杆菌和其他细胞中的同源和异二聚体蛋白质相互作用的双杂交系统。 该系统对于与酵母双杂交系统相同的应用是有用的; 即相互作用克隆,测定蛋白质相互作用区域,分析蛋白质相互作用,检测蛋白质相互作用并检测其调节剂。 本发明涉及原核宿主细胞,其包含:(a)融合蛋白,其具有(i)第一DNA结合结构域和(ii)第一相互作用结构域; (b)融合蛋白,其具有(i)第二DNA结合结构域和(ii)能够结合第一相互作用结构域的第二相互作用结构域; 和(c)具有与(i)启动子有效连接的报告基因的核酸分子,(ii)能够结合位于启动子上游的第一DNA结合结构域的第一操纵位点,和(iii) 能够结合位于报告基因启动子下游的第二DNA结合结构域的第二操作部位; 其中所述第一相互作用结构域与所述第二相互作用结构域的结合通过所述报道基因的表达改变来表示。