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    Multiple Gene Expression including sORF Constructs and Methods with Polyproteins, Pro-Proteins, and Proteolysis
    5.
    发明申请
    Multiple Gene Expression including sORF Constructs and Methods with Polyproteins, Pro-Proteins, and Proteolysis 审中-公开
    多基因表达包括sORF构建体和多聚蛋白，前蛋白和蛋白水解的方法

    公开(公告)号：US20070065912A1

    公开(公告)日：2007-03-22

    申请号：US11459098

    申请日：2006-07-21

    申请人： Gerald CARSON ,  Jochen SALFELD ,  Dean REGIER ,  Jijie GU ,  Wendy GION ,  Yune KUNES

    发明人： Gerald CARSON ,  Jochen SALFELD ,  Dean REGIER ,  Jijie GU ,  Wendy GION ,  Yune KUNES

    IPC分类号： C12P21/08 ,  C07H21/04 ,  C12N5/06 ,  C12N5/08 ,  C12N1/18 ,  C07K16/18

    CPC分类号： C12N15/67 ,  C07K16/00 ,  C07K2319/50 ,  C07K2319/92 ,  C12N15/1055 ,  C12P21/02 ,  C12P21/06

    摘要： Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and/or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

    摘要翻译： 公开了使用在重组宿主细胞内处理的主要翻译产物来表达蛋白质的有用构建体和方法。 描述了包含单个开放阅读框（sORF）的构建体用于蛋白质表达，包括多个多肽的表达。 初级翻译产物（前蛋白质或多蛋白质）含有多肽，例如内含肽或刺猬蛋白家族自身处理结构域或其变体，其插入到感兴趣的多个蛋白质亚基之间。 初级产物还可以含有切割序列，如其他蛋白水解切割或蛋白酶识别位点，或含有信号肽酶识别序列的信号肽，分离多个蛋白质亚基中的至少两个。 可以操纵插入的自动处理多肽或切割位点的序列，以提高分离的多个蛋白质亚基的表达效率。 还公开了进行蛋白质例如免疫球蛋白的有效表达，分泌和/或多聚体装配的独立方面。 当多蛋白含有免疫球蛋白重链和轻链片段或能够进行抗原识别的片段时，在一个实施方案中，可选择的化学计量比为每个重链段的轻链片段的至少两个拷贝，结果是正确折叠和组装 制备功能性抗体。 描述了包含免疫球蛋白轻链的修饰的信号肽。