Method for improving solubility and folding efficiency of target proteins using RNA as molecular chaperone
    1.
    发明授权
    Method for improving solubility and folding efficiency of target proteins using RNA as molecular chaperone 有权
    使用RNA作为分子伴侣来提高靶蛋白的溶解度和折叠效率的方法

    公开(公告)号:US07759303B2

    公开(公告)日:2010-07-20

    申请号:US11482401

    申请日:2006-07-07

    IPC分类号: C07K14/00 C07H21/02 C12N15/00

    摘要: Disclosed is a method for improving folding efficiency and solubility of a target protein linked to a RNA-binding protein by using RNA molecule as a molecular chaperone, wherein the RNA molecule interacts with the RNA-binding protein. More particularly, the present invention discloses method for improving folding efficiency and solubility of a target protein by transformation of a host cell with a expression vector comprising a polynucleotide encoding the target protein linked to an RNA-binding protein; culturing the transformed host cell in an appropriate culture medium under the condition that an RNA molecule either resident inside the host cell or provided by cotransformation of the host cell with polynucleotide encoding the RNA molecule interacts with the RNA-binding protein; and purifying the soluble protein from host cell lysate. The method of the present invention is very useful for production of soluble proteins for therapeutic, prophylactic and diagnostic applications.

    摘要翻译: 公开了通过使用RNA分子作为分子伴侣来提高与RNA结合蛋白质连接的靶蛋白的折叠效率和溶解度的方法,其中RNA分子与RNA结合蛋白相互作用。 更具体地,本发明公开了通过用包含编码与RNA结合蛋白连接的靶蛋白的多核苷酸的表达载体转化宿主细胞来提高靶蛋白的折叠效率和溶解度的方法。 在宿主在宿主细胞内的RNA分子或通过宿主细胞与编码RNA分子的多核苷酸共转化的RNA分子与RNA结合蛋白相互作用的条件下,在适当的培养基中培养转化的宿主细胞; 并从宿主细胞裂解液纯化可溶性蛋白质。 本发明的方法对于用于治疗,预防和诊断应用的可溶性蛋白质的生产非常有用。