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    • 4. 发明公开
    • 세포 내 절단 시스템을 이용하여 수용성 외래 단백질을 제조하는 방법
    • 使用特定细胞内清除系统生产可溶性外源蛋白的方法
    • KR1020130020208A
    • 2013-02-27
    • KR1020110082686
    • 2011-08-19
    • 연세대학교 산학협력단
    • 성백린최성일임거흔유지은
    • C12N15/62C12N15/70C07K19/00
    • PURPOSE: A method for preparing a foreign protein is provided to effectively remove a fusion tail from a fusion protein and to produce the foreign protein. CONSTITUTION: A method for preparing a foreign protein comprises: a step of preparing a first expression vector containing a gene encoding a soluble expression promoting protein and a gene encoding the foreign protein which are linked by a first linker; a step of preparing a second expression vector containing the gene encoding the soluble expression promoting gene and a gene encoding a proteolytic cleavage enzyme which are linked by a second linker; a step of transducing the first and second expression vectors into a host cell and preparing a transformant; and a step of culturing the transformant and cleaving the soluble expression promoting protein. The second linker is denoted by an amino acid sequence, DDDDK. The soluble expression promoting protein is LysRS or RBD. The foreign protein is BMP-2. The proteolytic cleavage enzyme is TEV.
    • 目的:提供一种制备外源蛋白的方法,以有效地从融合蛋白中除去融合尾部并产生外源蛋白。 构成:制备外源蛋白质的方法包括:制备含有编码可溶性表达促进蛋白质的基因的第一表达载体和编码由第一接头连接的外源蛋白质的基因的步骤; 制备含有编码可溶性表达促进基因的基因的第二表达载体和通过第二接头连接的编码蛋白水解切割酶的基因的步骤; 将第一和第二表达载体转导入宿主细胞并制备转化体的步骤; 以及培养转化体并切割可溶性表达促进蛋白的步骤。 第二个接头由氨基酸序列DDDDK表示。 可溶性表达促进蛋白是LysRS或RBD。 外源蛋白是BMP-2。 蛋白水解切割酶是TEV。