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    • 2. 发明授权
    • Periplasmic membrane-bound system for detecting protein-protein
interactions
    • 用于检测蛋白质 - 蛋白质相互作用的周质膜结合系统
    • US5744314A
    • 1998-04-28
    • US642552
    • 1996-05-03
    • Rolf MenzelScott T. Taylor
    • Rolf MenzelScott T. Taylor
    • G01N33/566C07K14/035C07K14/16C07K14/28C07K14/71C07K19/00C12N1/21C12N15/00C12N15/09C12N15/62C12P21/02C12Q1/02C12Q1/68C12R1/19C12R1/63C12N15/12
    • C12Q1/6897C07K14/005C07K14/28C07K14/71C12N15/62C07K2319/00C07K2319/02C07K2319/03C07K2319/61C07K2319/715C07K2319/73C12N2710/16622C12N2740/16222
    • The present invention relates to: (1) a fusion protein having a dimerizing domain with or without a ligand-binding region and toxR DNA-binding and hydrophobic transmembrane regions; (2) host cells comprising the fusion protein and a nucleic acid molecule having a reporter gene operatively linked to the ctx operon, wherein dimerization (ligand-dependent or -independent) is signaled by expression of the reporter gene; (3) a nucleic acid molecule coding for the fusion protein; (4) an expression vector comprising a coding region for the fusion protein;(5) a process for detecting dimer formation (ligand dependent or ligand independent) of the fusion protein, which comprises treating a culture of the host cells with a ligand, ligand mimetic, or dimerization inhibitor, and screening for expression of the reporter gene. The present invention can be used to generate a signal from a variety of ligand-binding domains, allowing ligand binding to be indicated by a simple colorimeteric test or antibiotic resistance. The fusion proteins could include therapeutically relevant domains, so that biologically pertinent interactions can be indicated by a readily measurable signal.
    • 本发明涉及:(1)具有具有或不具有配体结合区和toxR DNA结合和疏水跨膜区的二聚结构域的融合蛋白; (2)包含融合蛋白的宿主细胞和具有可操作地连接到ctx操纵子的报告基因的核酸分子,其中通过报告基因的表达来发出二聚(配体依赖或不依赖)信号通路; (3)编码融合蛋白的核酸分子; (4)包含融合蛋白的编码区的表达载体; (5)检测融合蛋白的二聚体形成(配体依赖性或配体相关性)的方法,其包括用配体,配体模拟物或二聚抑制剂处理宿主细胞的培养物,以及筛选报道基因的表达。 本发明可用于产生来自各种配体结合结构域的信号,允许通过简单的色度计测试或抗生素抗性来指示配体结合。 融合蛋白可以包括治疗相关的结构域,使得生物相关的相互作用可以通过容易测量的信号来指示。
    • 5. 发明授权
    • Periplasmic membrane-bound system for detecting protein-protein
interactions
    • 用于检测蛋白质 - 蛋白质相互作用的周质膜结合系统
    • US5521066A
    • 1996-05-28
    • US121201
    • 1993-09-13
    • Rolf MenzelScott T. Taylor
    • Rolf MenzelScott T. Taylor
    • G01N33/566C07K14/035C07K14/16C07K14/28C07K14/71C07K19/00C12N1/21C12N15/00C12N15/09C12N15/62C12P21/02C12Q1/02C12Q1/68C12R1/19C12R1/63C12N15/12
    • C12Q1/6897C07K14/005C07K14/28C07K14/71C12N15/62C07K2319/00C07K2319/02C07K2319/03C07K2319/61C07K2319/715C07K2319/73C12N2710/16622C12N2740/16222
    • The present invention relates to:(1) a fusion protein having a dimerizing domain with or without a ligand-binding region and toxR DNA-binding and hydrophobic transmembrane regions;(2) host cells comprising the fusion protein and a nucleic acid molecule having a reporter gene operatively linked to the ctx operon, wherein dimerization (ligand-dependent or -independent) is signaled by expression of the reporter gene;(3) a nucleic acid molecule coding for the fusion protein;(4) an expression vector comprising a coding region for the fusion protein;(5) a process for detecting dimer formation (ligand dependent or ligand independent) of the fusion protein, which comprises treating a culture of the host cells with a ligand, ligand mimetic, or dimerization inhibitor, and screening for expression of the reporter gene.The present invention can be used to generate a signal from a variety of ligand-binding domains, allowing ligand binding to be indicated by a simple colorimetric test or antibiotic resistance. The fusion proteins could include therapeutically relevant domains, so that biologically pertinent interactions can be indicated by a readily measurable signal.
    • 本发明涉及:(1)具有具有或不具有配体结合区和toxR DNA结合和疏水跨膜区的二聚结构域的融合蛋白; (2)包含融合蛋白的宿主细胞和具有可操作地连接到ctx操纵子的报告基因的核酸分子,其中通过报告基因的表达来发出二聚(配体依赖或不依赖)信号通路; (3)编码融合蛋白的核酸分子; (4)包含融合蛋白的编码区的表达载体; (5)检测融合蛋白的二聚体形成(配体依赖性或配体相关性)的方法,其包括用配体,配体模拟物或二聚抑制剂处理宿主细胞的培养物,以及筛选报道基因的表达。 本发明可用于产生来自各种配体结合结构域的信号,允许通过简单的比色测试或抗生素抗性来指示配体结合。 融合蛋白可以包括治疗相关的结构域,使得生物相关的相互作用可以通过容易测量的信号来指示。