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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

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【作者】 王新华胡敬东孔娜李宏梅赵宏坤

【Author】 WANG Xin-hua,HU Jing-dong,KONG Na,LI Hong-mei,ZHAO Hong-kun(College of Animal Science and Veterinary Medicine,Shandong Agricultural University,Tai’an,Shandong 271018,China)

【机构】 山东农业大学动物科技学院动物传染病实验室

【摘要】 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。

【Abstract】 The recombinant plasmid of mChIL-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 different 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.

【基金】 国家“十一五”科技支撑计划重大项目(2006BAK02A21);山东省科技攻关重大项目(SDSP2005041005);山东省科技攻关资助项目(2007GG20009001);山东省优秀中青年科学家科研奖励基金项目资助(2006BS06015)
  • 【文献出处】 中国兽医学报 ,Chinese Journal of Veterinary Science , 编辑部邮箱 ,2009年07期
  • 【分类号】S831
  • 【被引频次】1
  • 【下载频次】114
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