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    • 5. 发明申请
    • EXPRESSION VECTOR(S) FOR ENHANCED EXPRESSION OF A PROTEIN OF INTEREST IN EUKARYOTIC OR PROKARYOTIC HOST CELLS
    • 表达载体(S)用于增强在原核细胞或原核细胞中的兴趣蛋白质的表达
    • WO2008000445A1
    • 2008-01-03
    • PCT/EP2007/005650
    • 2007-06-26
    • CHEMOTHERAPEUTISCHES FORSCHUNGSINSTITUT GEORG-SPEYER-HAUSWELS, WinfriedDÄLKEN, Benjamin
    • WELS, WinfriedDÄLKEN, Benjamin
    • C12N15/70C12N15/79
    • C12N15/81C12N15/625
    • The present invention refers to an expression vector (A) for enhanced expression of a protein of interest encoding a fusion protein comprising at least a secretory signal sequence (A1); a solubilizing agent (A2); a proteolytic cleavage site (A3); and a protein of interest (A4), optionally a tag for purification of the protein of interest (A5) and optionally a heterologously expressed protease (A6), capable of cleaving the proteolytic cleavage site (A3) subsequent to or during secretion of the fusion protein. Alternatively, the protease (A6) may be endogenously expressed in the host cell, may be provided to the host cell using a second expression vector (B) and/or may be added extracellularly. The present invention furthermore provides methods for using the inventive expression vectors (A) and, optionally, (B), in vitro and in vivo , and kits, comprising these expression vectors and/or cells comprising same.
    • 本发明涉及用于增强编码融合蛋白的目的蛋白质表达的表达载体(A),其至少包含分泌信号序列(A1); 增溶剂(A2); 蛋白水解切割位点(A3); 和感兴趣的蛋白质(A4),任选地用于纯化目的蛋白质的标签(A5)和任选的异源表达的蛋白酶(A6),其能够在融合分泌之后或期间切割蛋白水解切割位点(A3) 蛋白。 或者,蛋白酶(A6)可以在宿主细胞中内生表达,可以使用第二表达载体(B)提供给宿主细胞和/或可以在细胞外加入。 本发明还提供了在体外和体内使用本发明的表达载体(A)和任选的(B)的方法,以及包含这些表达载体和/或包含它们的细胞的试剂盒。