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    Recombinant expression vectors for expression of heterologous proteins
    4.
    发明授权
    Recombinant expression vectors for expression of heterologous proteins 失效
    用于表达异源蛋白质的重组表达载体

    公开(公告)号：US5846819A

    公开(公告)日：1998-12-08

    申请号：US472045

    申请日：1995-06-06

    申请人： Mark H. Pausch ,  Bradley A. Ozenberger ,  John R. Hadcock ,  Laura A. Price ,  Eileen M. Kajkowski ,  Donald R. Kirsch ,  Deborah T. Chaleff

    发明人： Mark H. Pausch ,  Bradley A. Ozenberger ,  John R. Hadcock ,  Laura A. Price ,  Eileen M. Kajkowski ,  Donald R. Kirsch ,  Deborah T. Chaleff

    IPC分类号： C12N15/09 ,  C07K14/72 ,  C12N1/15 ,  C12N1/19 ,  C12N15/12 ,  C12P21/02 ,  C12Q1/02 ,  C12R1/865 ,  C12N15/63 ,  C07K14/705

    CPC分类号： C07K14/723 ,  C07K2319/00

    摘要： The present invention is directed to a expression vectors and yeast cells transformed therewith containing a first heterologous nucleotide sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second nucleotide sequence which codes for all or a portion of a G protein .alpha..beta..gamma. complex. Said heterologous protein is physically expressed in a host cell membrane in proper orientation for both stereoselective binding of ligands, as well as functional interaction with G proteins on the cytoplasmic side of the cell membrane. In some embodiments, a nucleotide sequence encoding a heterologous or chimeric G.alpha. protein is expressed in conjunction with nucleotide sequences from the yeast G protein .beta..gamma. subunits. A second aspect of the present invention provides expression vectors and transformed yeast cells encoding chimeric yeast/heterologous G protein coupled receptors. A third aspect of the present invention is directed to methods of assaying compounds using these expression constructs and yeast cell expression systems to determine the effects of ligand binding to the heterologous receptors expressed in the systems.

    摘要翻译： 本发明涉及转化的表达载体和酵母细胞，其含有编码G蛋白偶联受体（例如生长抑素受体）的第一异源核苷酸序列，以及编码全部或一部分的第二核苷酸序列 一个G蛋白αβγ复合物。 所述异源蛋白质在适当取向的宿主细胞膜中物理表达，用于配体的立体选择性结合，以及与细胞膜细胞质侧上的G蛋白的功能相互作用。 在一些实施方案中，编码异源或嵌合Gα蛋白的核苷酸序列与来自酵母G蛋白βγ亚基的核苷酸序列一起表达。 本发明的第二方面提供了编码嵌合酵母/异源G蛋白偶联受体的表达载体和转化的酵母细胞。 本发明的第三方面涉及使用这些表达构建体和酵母细胞表达系统测定化合物以确定配体与系统中表达的异源受体结合的作用的方法。