| Vif-ΔN28和Cullin5-N138蛋白的表达及纯化 |
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| 引用本文： | 张文艳,楼朝平,朱可彤,张帆,姜春来,吴永革,于湘晖,孔维. Vif-ΔN28和Cullin5-N138蛋白的表达及纯化[J]. 中国生物制品学杂志, 2006, 0(5) |
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| 作者姓名： | 张文艳 楼朝平 朱可彤 张帆 姜春来 吴永革 于湘晖 孔维 |
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| 作者单位： | 吉林大学生命科学学院疫苗研究中心 长春130012 |
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| 基金项目： | 国家自然科学基金资助项目(30570363)，吉林省科技发展计划杰出青年资助项目(20050112)资助 |
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| 摘 要： | 目的获得高纯度的Vif-ΔN28和Cullin5-N138蛋白。方法以Vif/VR1012质粒为模板,PCR扩增出Vif-ΔN28基因并插入pRSETB质粒。将构建的原核表达载体Vif-ΔN28/pRSETB以及已有的Cullin5-N138/pRSETB质粒分别转化大肠杆菌BL21(DE3),经IPTG诱导表达,产物经Ni-NTA离子纯化柱纯化、复性。结果所表达的Vif-ΔN28和Cullin5-N138蛋白约占各自菌体总蛋白的20%,纯化后蛋白纯度均可达90%以上。结论已成功构建了Vif-ΔN28原核表达质粒,并在大肠杆菌中高效表达,获得了可溶性的纯化的Vif-ΔN28和Cullin5-N138蛋白。
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| 关 键 词： | 人免疫缺陷病毒 Vif蛋白 Cullin5蛋白 |
Expression and Purification of Vif-ΔN28 and Cullin5-N138 Proteins |
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| ZHANG Wen-yan,LOU Chao-ping,ZHU Ke-tong,et al. Expression and Purification of Vif-ΔN28 and Cullin5-N138 Proteins[J]. Chinese Journal of Bilogicals, 2006, 0(5) |
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| Authors: | ZHANG Wen-yan LOU Chao-ping ZHU Ke-tong et al |
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| Abstract: | Objective To obtain highly purified Vif-ΔN28 and Cullin5-N138 proteins.Methods Amplify Vif-ΔN28 gene by PCR using plasmid Vif/VR1012 as template and insert into plasmid pRSETB.Transform the constructed recombinant plasmid Vif-ΔN28/pRSETB,as well as recombinant plasmid Cullin5-N138/pRSETB constructed in advance,to E.coli BL21(DE3) for expression under induction of IPTG.The expressed product was purified by Ni-NTA ion exchange chromatography and re-naturalized.Results Either the expressed Vif-ΔN28 or Cullin5-N138 contained 20% of total somatic protein and reached a purity of more than 90% after purification.Conclusion Recombinant plasmid Vif-ΔN28/pRSETB was successfully constructed and highly expressed in E.coli.Both purified Vif-ΔN28 and Cullin5-N138 proteins in soluble forms were obtained. |
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| Keywords: | Human immunodeficiency virus(HIV) Vif protein Cullin5 protein |
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