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    Mammalian protein interaction cloning system
    1.
    发明授权
    Mammalian protein interaction cloning system 有权
    哺乳动物蛋白质相互作用克隆系统

    公开(公告)号：US06316223B1

    公开(公告)日：2001-11-13

    申请号：US09359081

    申请日：1999-07-22

    Applicant: Donald Payan ,  Ying Luo ,  Betty Huang

    Inventor： Donald Payan ,  Ying Luo ,  Betty Huang

    IPC: C12P2106

    CPC classification number: C12N15/1055 ,  C12Q2565/201

    Abstract: The present invention is directed to compositions and methods for a genetic system of detecting protein-protein interactions in a mammalian host cell. Two fusion proteins are made in the host cell. The first fusion protein contains a DNA binding domain which is fused to a so-called bait protein. The second fusion protein consists of a transcriptional activation domain fused to a so-called test protein. The DNA binding domain binds to an operator sequence which controls expression of one or more reporter genes. The transcriptional activation domain is recruited to the promoter through the functional interaction between the bait protein and the test protein. Subsequently the transcriptional activation domain interacts with the basal transcription machinery to activate expression of one or more reporter genes which can be identified and characterized. The individual compositions are useful for analyzing protein-protein interactions between known proteins and to isolate, clone and characterize unknown proteins. The individual compositions can be used to express the fusion proteins either transiently or stably. In addition, the present invention is directed to methods for screening for candidate bioactive agents that modulate the protein-protein interaction between a bait protein and a test protein and thus are useful for the identification of novel therapeutic drugs and the like.

    Abstract translation: 本发明涉及用于在哺乳动物宿主细胞中检测蛋白质 - 蛋白质相互作用的遗传系统的组合物和方法。 在宿主细胞中制备两种融合蛋白。 第一个融合蛋白含有与所谓诱饵蛋白融合的DNA结合结构域。 第二种融合蛋白由与所谓的测试蛋白融合的转录激活结构域组成。 DNA结合域与控制一个或多个报道基因表达的操纵子序列结合。 通过诱饵蛋白质和测试蛋白质之间的功能相互作用将转录激活结构域引入启动子。 随后，转录激活结构域与基础转录机制相互作用以激活一种或多种可鉴定和表征的报告基因的表达。 单独的组合物可用于分析已知蛋白质之间的蛋白质 - 蛋白质相互作用并分离，克隆和表征未知蛋白质。 单独的组合物可用于瞬时或稳定地表达融合蛋白。 此外，本发明涉及筛选调节诱饵蛋白质和测试蛋白质之间的蛋白质 - 蛋白质相互作用的候选生物活性剂的方法，因此可用于鉴定新的治疗药物等。

    Mammalian protein interaction cloning system
    2.
    发明授权
    Mammalian protein interaction cloning system 失效
    哺乳动物蛋白质相互作用克隆系统

    公开(公告)号：US6114111A

    公开(公告)日：2000-09-05

    申请号：US50863

    申请日：1998-03-30

    Applicant: Ying Luo ,  Betty Huang ,  Donald Payan

    Inventor： Ying Luo ,  Betty Huang ,  Donald Payan

    IPC: C12N15/10 ,  C12N15/12 ,  C12Q1/70

    CPC classification number: C12N15/1055 ,  C12Q2565/201

    Abstract: The present invention is directed to compositions and methods for a genetic system of detecting protein--protein interactions in a mammalian host cell. Two fusion proteins are made in the host cell. The first fusion protein contains a DNA binding domain which is fused to a so-called bait protein. The second fusion protein consists of a transcriptional activation domain fused to a so-called test protein. The transcriptional activation domain is recruited to the promoter through the functional interaction between the bait protein and the test protein. Subsequently the transcriptional activation domain interacts with the basal transcription machinery to activate expression of one or more reporter genes which can be identified and characterized. The individual compositions are useful for analyzing protein--protein interactions between known proteins and to isolate, clone and characterize unknown proteins. The individual compositions can be used to express the fusion proteins either transiently or stably.

    Abstract translation: 本发明涉及用于在哺乳动物宿主细胞中检测蛋白质 - 蛋白质相互作用的遗传系统的组合物和方法。 在宿主细胞中制备两种融合蛋白。 第一个融合蛋白含有与所谓诱饵蛋白融合的DNA结合结构域。 第二种融合蛋白由与所谓的测试蛋白融合的转录激活结构域组成。 通过诱饵蛋白质和测试蛋白质之间的功能相互作用将转录激活结构域引入启动子。 随后，转录激活结构域与基础转录机制相互作用以激活一种或多种可鉴定和表征的报告基因的表达。 单独的组合物可用于分析已知蛋白质之间的蛋白质 - 蛋白质相互作用并分离，克隆和表征未知蛋白质。 单独的组合物可用于瞬时或稳定地表达融合蛋白。

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