| EBV-LMP2蛋白的表达和初步纯化 |
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| 引用本文： | 王湛,马晶,周玲,张晓光,杜海军,杨松梅,曾毅. EBV-LMP2蛋白的表达和初步纯化[J]. 中华实验和临床病毒学杂志, 2010, 24(1): 168-170. DOI: 10.3760/cma.j.issn.1003-9279.2010.03.003 |
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| 作者姓名： | 王湛 马晶 周玲 张晓光 杜海军 杨松梅 曾毅 |
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| 作者单位： | 中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室,100052;北京工业大学生命科学与生物工程学院; |
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| 摘 要： | 目的 应用杆状病毒表达载体表达并纯化EB病毒LMP2蛋白.方法 利用杆状病毒Bac-to-Bae杆状病毒表达系统,将EBV-LMP2基因插入到质粒pFastBacTMHT B中,获得携带EBV-LMP2基因的重组杆状病毒Bac-LMP2.重组病毒感染Sf-9细胞,表达N端携带6个组氨酸(6 X His)的LMP2融合蛋白His-LMP2.经镍离子亲和层析纯化,获得纯化蛋白.结果 SDS-PAGE及Western-Blot检测表达的蛋白相对分子质量大小与预计结果一致.HPLC分析纯化后蛋白的纯度可达86%.结论 利用杆状病毒表达系统表达EB病毒LMP2基因并经过初步纯化后,可以获得较好纯度的LMP2蛋白.
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| 关 键 词： | 基因,LMP2 杆状病毒科 基因表达 |
Expression and purification of EBV-LMP2 protein |
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| Abstract: | Objective To obtain a second Epstein-Barr virus membrane protein (LMP2) in insect cells.Methods The fuIl length EBV-LMP2 gene was inserted into baculovirus expression transfer vector pFastBacTM HT B to obtain the recombinant baculoviruses Bac-LMP2.And generation of recombinant baculoviruses was followed by transfection of the recombinant Bac-LMP2 into insect cells,then the recombinant LMP2 protein was recognized by SDS-PAGE and western blot.The expressed LMP2 protein was purified by one step with Ni-NTA metal chelation chromatography.Results The expressed LMP2 protein was confirmed by SDS-PAGE and western blot.The purity of purified LMP2 protein is up to 86% by HPLC analysis.Conclusion The EBV-LMP2 was expressed in insect cells.and the purified LMP2 protein was obtained. |
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| Keywords: | Genes LMP2BaculoviridaeGene expression |
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