| EB病毒基因Z2A原核表达载体的构建与表达(英文) |
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| 引用本文： | 薛庆节,王晖,李秀真,吕厚东,陈廷,朱伟,李士根.EB病毒基因Z2A原核表达载体的构建与表达(英文)[J].中国自然医学杂志,2011(4). |
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| 作者姓名： | 薛庆节 王晖 李秀真 吕厚东 陈廷 朱伟 李士根 |
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| 作者单位： | 济宁医学院医学微生物学教研室; |
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| 基金项目： | 山东省卫生厅基金(2007HW035); 济宁医学院青年基金项目资助 |
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| 摘 要： | 目的构建EB病毒基因LMP2A、BZLF1重组分泌性表达质粒Ag85B,然后将它成功转化成卡介苗。方法分别用PCR扩增Ag85B信号序列和LMP2A和BZLF1融合基因,将Ag85B信号序列与大肠杆菌-卡介苗穿梭质粒pMV261重组构建获得质粒pMVS。然后将LMP2A和BZLF1融合基因Z2A与质粒pMVS重组获得质粒pMVZ2A,将其电转化卡介苗,用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)对其结果进行分析。结果信号序列与LMP2A和BZLF1融合基因被正确插入分泌表达载体pMV261。重组质粒经限制性内切酶双酶切、PCR扩增、基因测序等鉴定。蛋白纯化后可得到一条带。结论 pMVZ2A在BCG中能够分泌表达,本研究为获得抗结核杆菌和EB病毒的双价疫苗奠定了基础。
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| 关 键 词： | 人类疱疹病毒4型 卡介苗 基因融合 基因载体 Z2A |
Construction and expression of prokaryotic expression vector containing EB virus gene Z2A |
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| Institution: | XUE Qing-jie,WANG Hui,LI Xiu-zhen,Lü Hou-dong,CHEN Ting,ZHU Wei,LI Shi-gen Department of Pathogenic Biology,Jingning Medical College,Jining 272013,China |
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| Abstract: | Objective To construct a recombinant secretory plasmid of Bacillus Calmette-Guerin(BCG) Ag85B-fused EB virus LMP2A and BZLF1 genes,and then transform it into BCG.Methods BCG-Ag85B signal sequence and Z2A gene were amplified from the genome of BCG and Z2A by PCR,respectively.BCG-Ag85B signal sequence was cloned into E.coli-BCG shuttle-vector pMV261 to obtain pMVS.The new recombinant plasmid of pMVZ2A was constructed by inserting the Z2A gene into pMVS and transformed into BCG by electrotransformation.The rec... |
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| Keywords: | Human herpesvirus type 4 BCG Gene fusion Genetic vectors Z2A |
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