| 重组人白细胞介素-21的原核表达及其复性与纯化 |
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| 引用本文： | ZHENG Rui,许晓群,ZHANG Jian-hua,张彩,LIU Jin-sheng,苏庆红,WANG Jun-fu.重组人白细胞介素-21的原核表达及其复性与纯化[J].中国生化药物杂志,2008,29(4). |
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| 作者姓名： | ZHENG Rui 许晓群 ZHANG Jian-hua 张彩 LIU Jin-sheng 苏庆红 WANG Jun-fu |
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| 作者单位： | 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 |
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| 基金项目： | 国家自然科学基金
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山东省优秀中青年科学家科研奖励基金 |
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| 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。
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| 关 键 词： | 重组人白细胞介素-21 表达 复性 蛋白纯化 |
Expression of recombinant human interleukin-21 in E. coli and the study on its renaturation and purification |
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| ZHENG Rui,XU Xiao-qun,ZHANG Jian-hua,ZHANG Cai,LIU Jin-sheng,SU Qing-hong,WANG Jun-fu.Expression of recombinant human interleukin-21 in E. coli and the study on its renaturation and purification[J].Chinese Journal of Biochemical Pharmaceutics,2008,29(4). |
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| Authors: | ZHENG Rui XU Xiao-qun ZHANG Jian-hua ZHANG Cai LIU Jin-sheng SU Qing-hong WANG Jun-fu |
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| Abstract: | 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. |
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| Keywords: | rhIL-21 expression renaturation protein purification |
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