| EB病毒潜伏膜蛋白2A1-119重组质粒的表达及鉴定 |
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| 引用本文： | 张琴,薛向阳,夏克栋,李文姝,陈韶,张丽芳.EB病毒潜伏膜蛋白2A1-119重组质粒的表达及鉴定[J].温州医学院学报,2010,40(4):319-321. |
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| 作者姓名： | 张琴 薛向阳 夏克栋 李文姝 陈韶 张丽芳 |
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| 作者单位： | 温州医学院,微生物学和免疫学教研室、分子病毒和免疫研究所,浙江,温州,325035 |
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| 摘 要： | 目的：构建含EB病毒（Epstein-Barr virus,EBV）潜伏膜蛋白（LMP）2A N端1-119基因的重组质粒,探讨其在原核表达系统中的表达情况。方法：采用RT-PCR方法从B95-8细胞中获得LMP2A1-119外显子基因片段,并克隆到pGEX-4T-1原核表达载体,转化大肠杆菌BL21（DE3）后,经IPTG诱导表达融合蛋白,通过Ni-NTA离子交换柱层析纯化,进一步采用SDS-PAGE和Western blot鉴定。结果：pGEX/LMP2A1-119重组质粒构建成功,并在大肠杆菌中得到表达。SDS-PAGE结果表明表达产物相对分子质量约为50 kD,West-ern blot结果证实其为目的蛋白。结论：EBV LMP2A1-119蛋白可在pGEX-4T-1原核表达系统中表达,为进一步研究其免疫学特性打下了基础。
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| 关 键 词： | 疱疹病毒4型 人 潜伏膜蛋白2A pGEX-4T-1 原核表达 重组 遗传 |
Expression and identification of the gene of LMP2A1-119of Epstein Barr virus |
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| ZHANG Qin,XUE Xiangyang,XIA Kedong,LI Wenshu,CHEN Shao,ZHANG Lifang.Expression and identification of the gene of LMP2A1-119of Epstein Barr virus[J].Journal of Wenzhou Medical College,2010,40(4):319-321. |
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| Authors: | ZHANG Qin XUE Xiangyang XIA Kedong LI Wenshu CHEN Shao ZHANG Lifang |
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| Institution: | ZHANG Qin,XUE Xiangyang,XIA Kedong,LI Wenshu,CHEN Shao,ZHANG Lifang. Institute of Molecular Virology and Immunology,Department of Microbiology and Immunology,Wenzhou Medical College,Wenzhou,325035 |
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| Abstract: | Objective:To construct the recombinant plasmid pGEX/LMP2A1-119 and to probe express situation of the LMP2A1-119 of Epstein-Barr virus(EBV) in Escherichia coli(E.coli) in the prolaryotic expression.Methods:The exon gene fragment encoding 1-119 amino acid of LMP2A was amplified by RT-PCR and was cloned into the expression veteor of pGEX-4T-1 for forming the PGEX/LMP2A1-119 recominant with which is transformed E.coli BL21(DE3).The PGEX/LMP2A1-119 protein was expressed in E.colia,nd then purified with Ni-NTA ag... |
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| Keywords: | pGEX-4T-1 |
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