| PAP-a/CD81-LEL融合蛋白表达载体的构建 |
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| 引用本文： | 潘延凤 贺永文 刘薇 郭劲松 王华. PAP-a/CD81-LEL融合蛋白表达载体的构建[J]. 中国现代医学杂志, 2005, 15(19): 2909-2911 |
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| 作者姓名： | 潘延凤 贺永文 刘薇 郭劲松 王华 |
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| 作者单位： | 华中科技大学同济医学院附属协和医院传染病科,湖北武汉430022 |
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| 基金项目： | 湖北省科技攻关计划2001ABB148 |
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| 摘 要： | 目的 探讨将商陆抗病毒蛋白-a（PAP-a）与CD81的胞外大环（CD81-LEL）融合,并在大肠杆菌内表达.方法 采用重叠延伸PCR方法,将PAP-a和CD81-LEL的基因拼接成PAP-a/CD81-LEL融合基因,并在两基因的连接处引入柔性肽段linker（GSGGSG）的核苷酸片断,然后将此融合基因克隆到原核表达载体PET-28a上,并在大肠杆菌BL21中进行表达.结果 所获得的PAP-a/CD81-LEL融合基因经测序正确,并可以在大肠杆菌BL21中表达,经SDS-PAGE分析,重组蛋白的相对分子量为50 KD.结论 PAP-a/CD81-LEL融合基因表达载体的成功构建与表达为进一步研究生物活性打下基础.
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| 关 键 词： | 商陆抗病毒蛋白 CD81 融合蛋白 |
| 文章编号： | 1005-8982（2005）19-2909-03 |
| 收稿时间： | 2004-10-27 |
| 修稿时间： | 2004-10-27 |
Construction of expression vector of PAP-a/CD81-LEL fusion protein |
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| PAN Yan-feng, HE Yong-wen, LIU Wei, GUO Jin-song, WANG Hua. Construction of expression vector of PAP-a/CD81-LEL fusion protein[J]. China Journal of Modern Medicine, 2005, 15(19): 2909-2911 |
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| Authors: | PAN Yan-feng HE Yong-wen LIU Wei GUO Jin-song WANG Hua |
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| Affiliation: | Department of Infectious Diseases, Union Hospital, Tongji Medical College, Central China Science and Technology University, Wuhan, Hubei 430022, P.R.China |
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| Abstract: | [Objective] To link pokeweed antiviral protein(PAP-a) gene to CD81-LEL gene by a mediate sequence GSGGSG and express in E. coli. [Methods] Overlap extend PCR was used to construct PAP-a -linker-CD81-LEL(PAP-A/CD81-LEL) fusion gene, the fusion gene was cloned into PET-28a and the vector was expressed in E. coli BL21. [Results] The sequence of the fusion protein obtained was correct. Recombinant PAP-a/CD81-LEL protein was expressed in E.coli BL21(DE3) by IPTG introduction. [Conclusion] The successful construction and expression of the fusion gene laid a foundation of study on biological activity of it. |
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| Keywords: | cloning and expression prokaryotic expression pokeweed antiviral protein CD81 |
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