【摘要】 利用基因工程表达的方法 ,在大肠杆菌中通过与GST蛋白融合的方式高效表达了胸腺素α1前体基因 ,随后经亲和层析和SP强阳离子树脂纯化相结合的方式 ,得到了胸腺素前体肽段 31肽和N 端未经乙酰化修饰的 2 8肽。融合蛋白表达量达到菌体总蛋白的 35 %～ 40 % ,样品肽的产量也达到了约 2 0 0mg L(肽 发酵液 )的产量。经质谱测定 ,分子量分别为 336 6和 30 6 6。BalB C小鼠脾脏淋巴细胞体外测活表明 ,所构建的GST Tα1融合蛋白和纯化后的产物对于淋巴细胞具有比较明显的增殖作用 ,其中N 端未经乙酰化的 2 8肽产物与 31肽产物活性相近 ,均对淋巴细胞具有明显的刺激增殖作用更多还原

【Abstract】 Engineering E.coli strain, BL21 (DE3)/pGEX-4T-human Thymosin α1, was constructed by oligonucleotide annealing and PCR amplifying the target gene, then ligating it with pGEX-4T-3 vector and transferring into BL21 host. The yield of fusion protein of GST-Thymosin α1 expressed from BL21(DE3)/pGEX-4T-thymosin α1 is about 35%～40% of total protein after fermentation. Following the simple cut of thrombin or CNBr, about 0.2g/L thymosin α 1 can be harvested. The product is checked by MS and activity test, which indicates that the recombinant product has full biological activity of native thymosin α1.更多还原