福建农林大学学报（自然科学版）2013，Vol.42Issue(3)：307-310,4.
猪日本乙型脑炎病毒NS5原核表达和多克隆抗体制备
Prokaryotic expression and polyclone antibody preparation of nonstructural protein 5 of Japanese encephalitis virus from pig
摘要
Abstract
The cDNA of Japanese encephalitis virus (JEV) was synthensized from viral genome by RT-PCR.The JEV-NS5 gene was cloned from cDNA by PCR.The JEV-NS5 gene was subcloned into pET-28(a) plasmid.The recombinant plasmid pET-28(a)-JEV-NS5 was transformed into E.coli.BL21 (DE3),The recombinant JEV-NS5 protein was expressed by IPTG induction.SDS-PAGE results showed that its molecular weight is 103 ku.The protein was mainly expressed in the form of inclusion bodies.The recombinant JEV-NS5 was more than 35％ of the total cell protein by TLC scanning analysis.A large number of high-purity recombinant proteins were obtained by his-band Ni+ purification.The proportion of recombinant JEV-NS5 reached more than 75％ after purification.The recombinant JEV-NS5 proteins were immunized into mouse,and mouse anti-JEV NS5 antibody was prepared.The antibody titer is 3 × 104.The specific of antibody was detected by western-blotting.This study was a tool and foundation of JEV NS5 function and virus replication mechanism.关键词
日本乙型脑炎病毒/非结构蛋白NS5/原核表达/纯化Key words
Japanese encephalitis virus/ nonstructural protein 5 / prokaryotic expression/ purification分类
农业科技引用本文复制引用
余风艳,韩秀杰,沈红霞,张保新,赵凡凡,王晓杜..猪日本乙型脑炎病毒NS5原核表达和多克隆抗体制备[J].福建农林大学学报（自然科学版）,2013,42(3):307-310,4.基金项目
浙江省自然科学基金资助项目(Y3110124) （Y3110124）
浙江农林大学人才启动基金项目(2010FR080). （2010FR080）
