The aim was to study the method of purification of GST-tagged proteins from inclusion bodies. The vector PGEX-4T1-Rpfd expressed Rpfd -GST fusion protein was transformed into E. Coli DH5a and protein expression was induced with IPTG. The effect of these factors is observed on the impact of protein purification by comparing lysis with guanidine hydrochloride and pretreatment with Novagen Protein Refolding Kit, and adjusting the different types of protein refolding solution. The active GST fusion protein can be efficiently purified, through removing soluble protein after ultrasonic treatment, and then using the reaction mixture containing 20 mmol Tris-HCl and 0.1 mmol DTT for rehabilitation. To remove soluble protein expression and change the formula of protein refolding solutions can, effectively improve protein purification of GST-tagged proteins from inclusion bodies.%探讨GST融合蛋白包涵体纯化的方法.将表达Rpfd与GST融合蛋白的质粒PGEX-4T1 -Rpfd转入大肠杆菌DH5a,IPTG诱导表达目的蛋白.通过比较盐酸胍裂解和Novagen蛋白折叠试剂盒预处理,及调整不同类型蛋白复性液,观察上述因素对GST融合蛋白包涵体纯化效果的影响.结果:超声后先去除可溶性表达,再使用20 mmolris-HC1和0.1 mmol DTT进行复性,可获得高效纯化,具有活性的GST融合蛋白.对于GST包涵体蛋白进行纯化时,去除可溶性表达蛋白,改变蛋白复性液的成分,可有效提高蛋白纯化效果.
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