弹性蛋白样多肽融合胸腺肽α1的原核表达方法

Prokaryotic expression of recombinant thymosinα1 by ELP fusion protein

目的建立一种低成本生产胸腺肽α1(Tα1)的方法。方法采用融合表达方式表达Tα1基因,通过人工合成Tα1和弹性蛋白多肽(ELP)基因,构建Tα1-G-ELPn/pET41a(+),同时构建RimJ-pACYCDuet1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 1L培养基最终获得纯度为98%的目标蛋白29.5mg。结论建立了一种不需色谱和亲和纯化的生产胸腺肽α1的方法。

Objective In order to obtain a economic method of producing recombinant thymosinal （ Tα1）. Methods The chemically synthesized Tal 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. Results The yield of fusion protein was from 1 liter of self- induced medium. The fusion protein was cleaved with hydroxylamine, and 29.5mg of target protein （purity： 98%） was obtained. Conclusion A production method of thymosinal was established by non- chromatographic purification and affinity purification.