| 重组人磷脂爬行酶1的原核表达及纯化 |
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| 引用本文： | 朱向前,杨静,丁晓然,王升启.重组人磷脂爬行酶1的原核表达及纯化[J].生物技术通讯,2011,22(6):809-813. |
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| 作者姓名： | 朱向前 杨静 丁晓然 王升启 |
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| 作者单位： | 1. 河南中医学院,河南郑州450003;军事医学科学院放射与辐射研究所,北京100850
2. 军事医学科学院放射与辐射研究所,北京,100850 |
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| 基金项目： | 国家传染病防治重大专项，国家自然科学基金 |
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| 摘 要： | 目的:建立重组人磷脂爬行酶1(hPLSCR1)原核表达及纯化工艺。方法:PCR扩增hPLSCR1编码基因并连接到原核表达载体pET-28a,转化大肠杆菌BL21(DE3)并进行诱导表达,Western印迹鉴定所表达蛋白;优化表达条件后,Ni2+柱亲和层析纯化重组蛋白。结果:构建了pET-28a-PLSCR1重组质粒,诱导表达后,经SDS-PAGE分析目的蛋白的表达量达32%,Ni2+金属螯合法纯化目的蛋白后纯度达到95%以上,Western印迹验证了融合蛋白的特异性。结论:建立了高效稳定的His-PLSCR1表达体系,获得大规模生产His-PLSCR1的分离纯化工艺,为进一步研究其蛋白功能奠定了基础。
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| 关 键 词： | 重组人磷脂爬行酶1 原核表达 纯化 |
Prokaryotic Expression and Purification of Recombinant Human Phospholipid Scramblase 1 |
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| ZHU Xiang-Qian,YANG Jing,Ding Xiao-Ran,WANG Sheng-Qi.Prokaryotic Expression and Purification of Recombinant Human Phospholipid Scramblase 1[J].Letters in Biotechnology,2011,22(6):809-813. |
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| Authors: | ZHU Xiang-Qian YANG Jing Ding Xiao-Ran WANG Sheng-Qi |
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| Institution: | ZHU Xiang-Qian1,2,YANG Jing2,Ding Xiao-Ran2,WANG Sheng-Qi2 1.Triditional Chinese Medcine of Henan University,Zhengzhou 450003,2.Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China |
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| Abstract: | Objective: To express and purify the recombinant human phospholipid scramblase 1(hPLSCR1).Methods: The coding sequence of hPLSCR1 was amplified by PCR,and it was cloned into the vector of pET-28a to construct the recombinant plasmid pET-28a-PLSCR1.The recombinant E.coli BL21(DE3)/pET-28a-PLSCR1 was induced by IPTG,the recombinant protein was identified by Western blot.Optimized the terms of expressiom,the fusion protein was purified by nickel-chelating chromatography.Results: Recombinant plasmid pET-28a-PLS... |
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| Keywords: | recombinant human phospholipid scramblase 1 prokaryotic expression purification |
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