【摘要】 以无活性人胰岛素原突变体为分子支架,在C肽位置嵌合特异拮抗血小板GPⅡbⅢa受体的去整合素活性结构位点序列,构建嵌合人胰岛素原突变体.该突变体相对分子质量大小为7.0×103,与钙调素的C末端融合后形成新型重组融合蛋白质,相对分子质量为24.7×103.构建该重组融合蛋白质的大肠杆菌BL21(DE3)表达系统,尝试在大肠杆菌表达体系中,对该重组融合蛋白质进行高效表达.结果表明,该重组融合蛋白质的表达率约占菌体全蛋白的30%,优化后的表达条件为25℃,50μmol.L-1诱导表达16 h.更多还原

【Abstract】 Chimeric antithrombotic proinsulin mutant(Mr=7.0×103) was constructed based on inactive proinsulin scaffold with GPⅡbⅢa receptor antagonist disintegrin sequence replacing connecting peptide of proinsulin and was further fused with camodulin at C-terminal region.The fusion protein(Mr=24.7×103) was expressed at high-level in an E.coli BL21(DE3) host-cell system.Variations in temperature,culture medium,and induction method were considered for optimal expression.更多还原