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当前位置: 首页 > 文章 > 日本脑炎病毒囊膜蛋白的基因原核表达及抗原性分析 动物医学进展 2010,31 (6) 19-25
Position: Home > Articles > Prokaryotic Expression of Japanese Encephalitis Virus Envelope Protein and Antigenic Analysis Progress in Veterinary Medicine 2010,31 (6) 19-25

日本脑炎病毒囊膜蛋白的基因原核表达及抗原性分析

作  者：
张宁;金洪涛;夏志平;张瑞岩;刘雯;李勇;薛慧亮;李丹;丁壮
单  位：
吉林大学动物科技学院;解放军军事医学科学院军事兽医研究所
关键词：
日本脑炎病毒;E蛋白;原核表达;抗原性
摘  要：
参照GenBank中的日本脑炎病毒序列设计一对特异性引物,采用RT-PCR法扩增E基因得到cDNA,克隆至pMD-18T载体,经测序证实后亚克隆至原核表达载体pET-28a中,转化入BL21(DE3),IPTG诱导表达,对表达产物进行SDS-PAGE电泳分析。结果表明,成功构建了重组质粒pET-28a/JEV E,目的蛋白主要以包涵体形式表达。Western blot检测表明,表达产物与兔抗JEV多克隆抗体呈阳性反应,在ELISA试验中,用表达出的E蛋白包被,能够很明显地区分出猪日本脑炎阴性、阳性血清。
译  名：
Prokaryotic Expression of Japanese Encephalitis Virus Envelope Protein and Antigenic Analysis
作  者：
ZHANG Ning1,JIN Hong-tang2,XIA Zhi-ping2,ZHANG Rui-yan2,LIU Wen1,LI Yong1,XUE Hui-liang2,LI Dan2,DING Zhuang1(1.College of Animal Science and Veterinary Medicine,Jilin University,Changchun,Jilin,130062,China;2.Military Veterinary Institute,Academy of Military Sciences of PLA,Changchun,Jilin,130062,China)
关键词：
Japanese encephalitis virus;E protren;prokaryotic expression;antigenicity
摘  要：
In order to construct and express E protein of Japanese encephalitis virus(JEV),and analyze antigenicity of its products.A special pairs of primers of Japanese encephalitis virus were designed according to the GenBank,the gene encoding E protein was amplified using RT-PCR and then was cloned into pET-28a(+) to construct the recombinant plasmid pET-28a/JEV E,which was then transformed into BL21(DE3).The E protein expressed in BL21(ED3) was determined by SDS-PAGE.The result showed that the recombinant plasmid pET-28a/JEV E was successfully constructed and the expressed product was mainly in the form of inclusion body.Western blot analysis showed that the recombinant protein had a positive reaction with polyclonal antibody of rabbit against JEV,in the ELISA tests the positive serum of swine should be identified from negative ones obviously.This research made a foundation for further research on the serological diagnosis and the construction and function analysis of E protein.

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