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    • 8. 发明申请
    • RECOMBINANT SECRETABLE FUSION PROTEINS
    • 重组可溶性融合蛋白
    • WO1993002198A1
    • 1993-02-04
    • PCT/FI1991000225
    • 1991-07-23
    • TECHNICAL RESEARCH CENTRE OF FINLANDTEERI, Tuula, T.TAKKINEN, KristiinaLAUKKANEN, Marja-LeenaALFTHAN, KaijaSIZMANN, DorotheaKNOWLES, Jonathan, K., C.
    • TECHNICAL RESEARCH CENTRE OF FINLAND
    • C12N15/62
    • C07K16/00
    • A method for producing a single chain fusion protein, comprises the steps of: (a) constructing an rDNA molecule coding for a secretable single chain fusion protein having a plurality of functional proteins or protein domains joined by spacer peptides, wherein each such spacer peptide comprises a flexible hinge region of a naturally secreted multidomain protein or an analogue thereof, each adjacent pair of functional proteins or protein domains is joined by a spacer peptide which is not naturally continuous with either functional protein or protein domain to which it is joined, and at least two of the functional proteins or protein domains each independently exhibit physicochemical or biological activity or interact to form an assembly that exhibits physicochemical or biological activity; (b) inserting the rDNA molecule into an expression vector capable of transforming a host organism; (c) transforming a host organism with the resultant vector, and isolating transformants; and (d) culturing the transformants and recovering thesingle chain fusion protein.
    • 一种制备单链融合蛋白的方法,包括以下步骤:(a)构建编码具有通过间隔肽连接的多个功能蛋白或蛋白结构域的可分离单链融合蛋白的rDNA分子,其中每个这样的间隔肽包含 天然分泌的多结构域蛋白质或其类似物的柔性铰链区域,每个相邻的功能蛋白质或蛋白质区域对通过间隔肽连接,所述间隔子肽与其所连接的功能性蛋白质或蛋白质结构域不是天然连续的,并且在 功能性蛋白质或蛋白质结构域中至少有两个独立地表现出物理化学或生物活性或相互作用以形成具有物理化学或生物活性的组合物; (b)将rDNA分子插入能够转化宿主生物的表达载体中; (c)用所得载体转化宿主生物体,并分离转化体; (d)培养转化体并回收单链融合蛋白。