| p62Dok PTB结构域融合蛋白的表达和纯化 |
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| 引用本文： | 阎志勇,吴爱群,张勇,由振东,路长林,周明明,何成.p62Dok PTB结构域融合蛋白的表达和纯化[J].郑州大学学报(医学版),2001,36(4):404-407. |
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| 作者姓名： | 阎志勇 吴爱群 张勇 由振东 路长林 周明明 何成 |
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| 作者单位： | 1. 第二军医大学神经生物学教研室
2. 河南医科大学解剖学教研室 |
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| 基金项目： | 国家自然科学基金资助项目 30 0 70 16 7，海外青年学者合作研究基金资助项目 30 0 2 80 0 3，国家 973基金资助项目 G19990 5 40 0 0 |
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| 摘 要： | 目的 :制备p6 2DokPTB结构域的GST融合蛋白。方法 :用PCR方法扩增编码p6 2DokPTB结构域的cD NA ,并将其克隆入表达载体pGEX 4T 3中 ,转化大肠杆菌BL 2 1,构建成表达GST PTB融合蛋白的菌株。经IPTG诱导表达后 ,经Glutathion Sepharose 4B亲和层析柱纯化 ,用SDS PAGE进行分析 ,该融合蛋白的表达量超过菌体总蛋白的 2 5 %。结果 :获得重组GST PTB融合蛋白 ,纯度在 95 %以上 ,产物得率约 75 %。结论 :成功制备高纯度p6 2DokPTB结构域的GST融合蛋白 ,为进一步研究p6 2DokPTB结构域的结构和生物学功能奠定了基础。
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| 关 键 词： | p62Dok PTB结构域 重组蛋白 |
| 修稿时间： | 2001年2月21日 |
Expression and purification of the GST fusion protein of p62Dok PTB domain |
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| YAN Zhiyong ,WU Aiqun ,ZHANG Yong ,YOU Zhendong ,LU Changlin ,ZHOU Mingming ,HE Cheng.Expression and purification of the GST fusion protein of p62Dok PTB domain[J].Journal of Zhengzhou University: Med Sci,2001,36(4):404-407. |
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| Authors: | YAN Zhiyong WU Aiqun ZHANG Yong YOU Zhendong LU Changlin ZHOU Mingming HE Cheng |
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| Institution: | YAN Zhiyong 1,2),WU Aiqun 2),ZHANG Yong 1),YOU Zhendong 1),LU Changlin 1),ZHOU Mingming 1,3),HE Cheng 1) 1)Department of Neurobiology,the Second Military Medical University,Shanghai 200433 2)Department of Human Anato |
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| Abstract: | Aim:To produce the GST PTB domain fusion protein Methods:The cDNA encoding p62Dok PTB domain was amplified by using PCR method, and cloned into pGEX 4T 3 vector The expression plasmid was introduced into E coli strain BL 21 The GST PTB fusion protein accounted for over 25% of the total bacterial protein after IPTG induction, It was purified to homogeneity from the cell lysates via affinity chromatography and analyzed by SDS PAGE gel Results:The recombinant GST PTB fusion proteins were prepared with high purity, more than 95%, and high recovery, about 75% Conclusion: The established method for preparation large quantity recombinant GST PTB fusion proteins would promote the structure function analysis of the PTB domain of p62 Dok protein |
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| Keywords: | p62Dok PTB domain recombinant protein |
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