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Expression vector(s) for enhanced expression of a protein of interest in eukaryotic or prokaryotic host cells

机译:用于在真核或原核宿主细胞中增强目的蛋白表达的表达载体

摘要

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.
机译:本发明涉及一种表达载体(A),用于增强目的蛋白的表达,所述目的蛋白编码至少包含分泌信号序列(A1)的融合蛋白。增溶剂(A2);蛋白水解切割位点(A3);以及目的蛋白(A4),可选地是用于纯化目的蛋白的标签(A5),以及可选地是异源表达的蛋白酶(A6),能够在融合蛋白分泌后或分泌过程中裂解蛋白水解切割位点(A3)蛋白。备选地,蛋白酶(A6)可以在宿主细胞中内源表达,可以使用第二表达载体(B)提供给宿主细胞和/或可以在细胞外添加。本发明还提供了使用本发明表达载体(A)和任选地( B ),体外和体内 的方法。和试剂盒,其包含这些表达载体和/或包含它们的细胞。

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