一种胸腺肽α1的原核表达方法

摘要

In order to obtain a economic method of producing recombinant thymosin α1（Tα1）. The chemically synthesized Tctl and ELP were inserted into pET41a（＋） vector. The RimJ was amplified by PCR and inserted into pACYCDuetl vector. Then the recombinant plasmids were cotransformed into BL21. After expression in self-induced medium, the fusion protein was purified by precipitation under a temperature higher than transition temperature and cleavage with hydroxylamine. The results show that the yield of fusion protein was 240mg from 1 liter of self-induced medium. The fusion protein was cleaved with hydroxylamine, and 29.5mg of target protein （purity： 98%） was obtained. A production method of thymosin α1 was established by non-chromatographic purification and affinity purification.%采用融合表达方式表达Tα1基因，通过人工合成Tα1和ELP基因，构建Tα1-G—ELPn／pET41a（＋），同时构建RimJ-pA-CYCDuet1载体，共转BL21，采用自诱导表达培养基进行蛋白表达，表达蛋白通过变温自动聚合沉淀获得高纯度融合蛋白，将融合蛋白羟胺裂解，变温沉淀，收集上清，最终获得目标蛋白．结果显示，1L培养基最终获得纯度为98％的目标蛋白29．5mg．建立了一种不需色谱和亲和纯化的生产胸腺肽α1的方法。