重组人白细胞介素-21的原核表达及其复性与纯化  

Expression of recombinant human interleukin-21 in E.coli and the study on its renaturation and purification

作　　者：郑瑞[1] 许晓群[1] 张建华[1] 张彩[1] 刘金生[1] 苏庆红[1] 王郡甫[1] 

机构地区：[1]山东省医学科学院基础医学研究所,山东省肿瘤免疫与基因工程重点实验室,山东省现代医用药物与技术重点实验室,山东济南250062

出　　处：《中国生化药物杂志》2008年第4期233-236,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金项目(No.30371302);山东省优秀中青年科学家科研奖励基金项目(No.2005BS03015)

摘　　要：目的构建工程菌BL21/pET30a(+)-IL-21表达体系,表达并纯化出具有生物学活性的目的蛋白质。方法用基因工程方法构建表达质粒pET30a(+)-IL-21;在BL21大肠杆菌中诱导表达基因重组人白细胞介素-21(rhIL-21);提取包涵体并建立复性条件;用Ni-NTA凝胶纯化出rhIL-21;以NK92为效应细胞,观察rhIL-21对NK92的生物学作用,以此检测其活性。结果构建了重组质粒pET30a(+)-IL-21,工程菌BL21/pET30a(+)-IL-21能大量表达目的蛋白质;经复性和纯化得到具有生物学活性的rhIL-21。结论成功建立了人IL-21的原核表达体系,得到了有生物学活性的rhIL-21蛋白质。Purpose To construct expression vector pET30a（ ＋ ）-IL-21 and express recombinant human interleukin-21 （rhIL-21） in E. coli BL21. Methods Recombinant plasmid pET30a（ ＋ ）-IL-21 was constructed by gene engineering technique and transformed into E. coli BL21. After being expressed with induction of IPTG and purified with Ni-sepharose, the activity of rhIL-21 was tested on NK92 cells. Results rhIL-21 was expressed highly in E. coli BL21 in inclusion bodies. The purified rhIL-21 had synergetic effect on NK92 proliferation after 48 hours incubation with IL-2. Conclusion rhIL-21 was expressed successfully in E. coli BL21 and had a good biological activity after renaturation.

关 键 词：重组人白细胞介素-21 表达 复性 蛋白纯化 

分 类 号：Q78[生物学—分子生物学]