| 乙型肝炎病毒全-X基因的克隆及原核表达 |
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| 引用本文： | 薛红安,杨倩,刘拉羊,成军,董菁.乙型肝炎病毒全-X基因的克隆及原核表达[J].第四军医大学学报,2004,25(23):2133-2135. |
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| 作者姓名： | 薛红安 杨倩 刘拉羊 成军 董菁 |
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| 作者单位： | 西安交通大学第二医院感染科,陕西,西安,710004;解放军302医院传染病研究所基因治疗研究中心,北京,100039 |
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| 摘 要： | 目的:构建乙型肝炎病毒全-X基因原核表达载体并在细菌中表达、纯化. 方法:利用基因重组技术,将全-X基因定向克隆于原核表达载体pET-32a( )多克隆位点中,限制性酶切和测序鉴定. 细菌表达产物及纯化后的产物经SDS-PAGE和Western免疫印迹分析. 结果:经限制性酶切分析及测序鉴定,证实成功构建全-X原核表达质粒. SDS-PAGE和Western免疫印迹分析表明重组质粒在大肠杆菌中表达. 结论:本研究将全-X基因定向克隆于原核表达载体,并在大肠杆菌中成功表达、纯化. 为进一步研究全-X的功能及制备相应抗体奠定了基础.
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| 关 键 词： | 全-X基因 乙型肝炎病毒 克隆 原核表达 |
| 文章编号： | 1000-2790(2004)23-2133-03 |
| 修稿时间： | 2004年9月7日 |
Cloning and prokaryotic expression of HBV Whole-X gene |
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| Abstract: | AIM: To construct the prokaryotic expression plasmid pET-32a(+)-Whole-X. METHODS: The Whole-X gene was cloned into EcoR I/Sal I site of pET-32a(+) and was sequenced. The fusion protein was expressed in E. coli with IPTG induction. After the induction and purification, the protein was analyzed in SDS-PAGE and identified by Western blotting. RESULTS: Endonucleases digesting and sequencing confirmed that the Whole-X gene was correctly inserted into the vector. The Whole-X gene was successfully expressed in E.coli. The fusion protein was confirmed by SDS-PAGE and Western blotting. CONCLUSION: The prokaryotic expression plasmid pET-32a(+)-Whole-X has been successfully reconstructed, which provides the experimental base for study on its biological functions and antibody preparation. |
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| Keywords: | whole-X gene hepatitis B clone prokaryotic expression |
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