中国兽医科学2017，Vol.47Issue(12)：1475-1480,6.DOI:10.16656/j.issn.1673-4696.2017.12.001
基因Ⅰ型乙脑病毒EDⅢ结构域蛋白的原核表达及其抗体制备与鉴定
Identification of EDⅢ protein from the genotype Ⅰ Japanese encephalitis virus
摘要
Abstract
The studywas conducted to identify the immunogenicity of EDⅢprotein and the inhibitory effects on the invasion of JEV produced by the polyclonal antibodies against the EDⅢprotein.The EDⅢ gene from the genotype Ⅰ JEV was amplified by RT-PCR and constructed into the expression vector PET-30a.The recombinant plasmids verified by sequencing were transformed into E.coli BL21 and the positive clone was induced for expression and purified.New Zealand white rabbits were immunized with the purified recombinant EDⅢprotein(rEDⅢ).The specific reactivity between the rabbit polyclonal antibodies against rEDⅢ and JEV was determined by Western-blot and the indirect immunofluorescence test,and the neutralizing antibody titer of the rabbit polyclonal antibodies against rEDⅢ was measured by the plaque-reduction neutralization test(PRNT50).The results showed that the rEDⅢwas expressed in a inclusion body form.Rabbit polyclonal antibodies against the rEDⅢ specifically recognize the native antigen of JEV and its neutralizing antibody titer was 133±46.2.These results indicated that EDⅢproteinofgenotype Ⅰ JEVGS strainhad strong immunity andplayed the critical roles in the invasion of JEV.From the above,the EDⅢ protein could be a better candidate antigen for JEV subunit vaccines.关键词
乙脑病毒/EDⅢ结构域/原核表达/多克隆抗体Key words
Japanese encephalitis virus/EDⅢ domain/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
于瑞明,田占成,独军政,高闪电,刘光远,罗建勋,殷宏..基因Ⅰ型乙脑病毒EDⅢ结构域蛋白的原核表达及其抗体制备与鉴定[J].中国兽医科学,2017,47(12):1475-1480,6.基金项目
国家自然科学基金项目(31201899) （31201899）
中国农业科学院农业科技创新工程项目(2014ZL010) （2014ZL010）
