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| TRAF6-GST融合蛋白在大肠杆菌中的表达及纯化 | |
| 引用本文： | 孙利,王晓辉,关薇,唐刘君,王建,杨晓明,汪思应. TRAF6-GST融合蛋白在大肠杆菌中的表达及纯化[J]. 生物技术通讯, 2011, 22(5): 679-682. DOI: 10.3969/j.issn.1009-0002.2011.05.018 |
| 作者姓名： | 孙利 王晓辉 关薇 唐刘君 王建 杨晓明 汪思应 |
| 作者单位： | 1. 安徽医科大学基础医学院,安徽合肥230032;军事医学科学院放射与辐射医学研究所,北京100850 2. 军事医学科学院放射与辐射医学研究所,北京,100850 3. 安徽医科大学基础医学院,安徽合肥,230032 |
| 基金项目： | 国家重点基础研究发展计划 |
| 摘 要： | 目的:在大肠杆菌中表达肿瘤坏死因子受体相关因子6(TRAF6)与GST的融合蛋白并进行纯化。方法:采用PCR方法从肝文库中扩增编码TRAF6的DNA片段,将其插入原核表达载体pGEX-4T-2,构建GST-TRAF6原核表达载体,并转入大肠杆菌BL21(DE3)中,用IPTG诱导表达;用谷胱甘肽-琼脂糖珠亲和纯化表达的GST-TRAF6融合蛋白。结果:酶切鉴定和测序分析显示,长为1569 bp的TRAF6 DNA片段在pGEX-4T-2-TRAF6中的碱基序列、插入位点及读框正确,且位于表达载体的GST序列下游;经IPTG最佳浓度0.5 mmol/L诱导表达、亲和纯化后,获得了相对分子质量约85×103的GST-TRAF6融合蛋白。结论:构建了重组GST-TRAF6原核表达载体,获得了GST-TRAF6的大肠杆菌BL21表达菌株及GST-TRAF6融合蛋白,利于深入研究TRAF6的功能。 |
| 关 键 词： | 肿瘤坏死因子受体相关因子6 原核表达 蛋白纯化 |
Expression in Escherichia coli and Purification of GST tag TRAF6 |
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| SUN Li,WANG Xiao-Hui,GUAN Wei,TANG Liu-Jun,WANG Jian,YANG Xiao-Ming,WANG Si-Ying. Expression in Escherichia coli and Purification of GST tag TRAF6[J]. Letters in Biotechnology, 2011, 22(5): 679-682. DOI: 10.3969/j.issn.1009-0002.2011.05.018 | |
| Authors: | SUN Li WANG Xiao-Hui GUAN Wei TANG Liu-Jun WANG Jian YANG Xiao-Ming WANG Si-Ying |
| Affiliation: | SUN Li 1,2,WANG Xiao-Hui 2,GUAN Wei 2,TANG Liu-Jun 2,WANG Jian 2,YANG Xiao-Ming 1,WANG Si-Ying 1 1.Department of Pathophysiology,Anhui Medical University,Hefei 230032,2.Beijing Institute of Radiation Medicine,Beijing 100850,China |
| Abstract: | Objective: To express tumor necrosis factor receptor associated factor-6(TRAF6) fused with GST tag in E.coli BL21(DE3) and to purify it.Methods: The gene of TRAF-6 was amplified from liver gene library by PCR method,and was inserted into prokaryotic expression plasmid pGEX-4T-2 to construct prokaryotic expression vector of GST-TRAF6.Induced by IPTG,the recombinant GST-TRAF6 was expressed.At last,we purified recom-binant protein of GST-TRAF6 with glutathione-agarose beans.Results: Recombinant expression plas... |
| Keywords: | tumor necrosis factor receptor associated factor-6 prokaryotic expression protein purification |
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