人工分子伴侣系统辅助鸡IL-18重组蛋白复性过程中研究的影响因素

Influencing factors in the refolding process of artificial molecular chaperone assisting chicken IL-18 recombination protein

将重组原核表达质粒pGEX-mChIL-18转化宿主细胞大肠杆菌BL21(DE3),并用IPTG于37℃诱导培养获得表达。表达的包涵体经超声波破碎、洗涤后以6 mol/L的盐酸胍溶解,使蛋白彻底变性。然后按照试验设计,考察不同浓度组成的人工分子伴侣体系对不同浓度鸡IL-18重组蛋白复性率的影响。试验结果表明,利用人工分子伴侣系统辅助鸡IL-18重组蛋白的复性存在最佳的条件,优化该条件能够提高该融合蛋白的复性率,且产物都具有良好的生物学活性,为鸡IL-18重组蛋白的进一步应用研究奠定了基础。

The recombinant plasmid of mChlL-18 prokaryotic expression was transformed into E. coli BL21 （DE3） strain and then induced by IPTG at 37℃. After crushed and washed, the expressing inclusion bodies were thoroughly denatured with 6 mol/L guanidine hydrochloride, Then according to experiment design, the effects of rChIL-18 protein refolding yield at different densities were investigated by the systems of artificial chaperone at dif- ferent densities. Experiment results indicate that there is a optimal condition on assiting rChIL-18 protein by using the artificial chaperone technique. The optimal condition can improve the refolding yield of rChIL-18 protein, and then the expressed product of fusion chicken IL-18 gene in E. coli has a relativity high bioactivity.