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重组人IL-21融合蛋白在大肠杆菌中的可溶性形式表达、纯化及体外对T细胞增殖的影响

Soluble expression and purification of recombinant human interleukin-21 in escherichia coli
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摘要 目的为重组人白细胞介素-21(rhIL-21)的生物学活性和功能研究奠定基础。方法构建重组表达质粒pGEX4T-2/IL-21,通过降低培养温度等优化条件于大肠杆菌中表达出可溶性形式的rhIL-21融合蛋白,纯化后以不同质量浓度作用于T细胞,观察其对T细胞增殖的影响。结果大肠杆菌表达出可溶性形式的rhIL-21融合蛋白,纯化纯度达95%;纯化后的rhIL-21蛋白作用后T细胞增殖增加,且随rhIL-21蛋白质量浓度的增加,T细胞增殖率升高。结论大肠杆菌中获得rhIL-21的可溶性表达形式,纯化后的蛋白对T细胞增殖有促进作用;此为rhIL-21的生物活性及功能研究奠定了基础。 Objective To investigate the soluble expression and purification of recombinant human interleukin-21 （rhIL-21） in escheriehia coli（E, eoli） ,and analyze its immunologic activity in vitro. Methods The E. eoli DI45α containing recombinant plasmid pGEX4T-2 / IL-21 was induced to express protein by IPTG. Supematants of lysate was purified by affinity chromatography, thrombin digestion and cation chromatography sequentially. The purified products were an- alyzed by SDS-PAGE. The effect of rhIL-21 on proliferation of T lymphocyte was studied by using MTT method. Results The target protein was soluble in the supematant. Purity of rhIL-21 was 95%. The proliferation of T lymphocyte increased after treated by rhIL-21 in vitro. Conclusion Soluble rhIL-21 can be successfully expressed and purified;rhIL-21 can promote the proliferation of T lymphocyte.
作者 刘明军 鲁晓晴 王斌 钱冬萌
机构地区 青岛大学医学院附属医院 青岛大学医学院
出处 《山东医药》 CAS 北大核心 2008年第26期3-5,共3页 Shandong Medical Journal
基金 山东省自然科学基金项目(E99C02) 青岛市科技计划资助项目(04-2-J2-95)
关键词 人白细胞介素-21 大肠杆菌 可溶性表达 纯化 human Interleukin-21 escherichia coli soluble expression purification
分类号 Q78 [生物学—分子生物学]
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参考文献9

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二级参考文献9

  • 1Habib T,Senadheera S,Weinberg K,et al.The common gamma chain (gamma C) is a required signaling component of the IL-21 receptor and supports IL-21-induced cell proliferation via JAK3[].Biochemistry.2002
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共引文献5

山东医药
2008年 第26期

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