中国畜牧兽医2017，Vol.44Issue(7)：2119-2125,7.DOI:10.16431/j.cnki.1671-7236.2017.07.030
蓝舌病1型病毒VP2蛋白的原核表达及免疫反应性分析
Prokaryotic Expression and Immunoreactivity Analysis of VP2 Protein of Bluetongue Virus Serotype 1
摘要
Abstract
The study was aimed to test the immunoreactivity of the VP2 protein of bluetongue virus serotype 1 (BTV-1) in vitro.Based on the published BTV-1 L2 gene of Y863 strain, specific cloning PCR primers were designed and synthesized.The L2 gene was amplified through RT-PCR method and then was purified and cloned into the expressing vector pEASY-Blunt E1.The cloned recombinant plasmids were identified.The positive recombinant L2 plasmid was cloned into BL21(DE3) competent cells to express VP2 protein.The acquired purified recombinant BTV-1 VP2 protein was analyzed through the methods of Western blotting, ELISA and blocking ELISA.The results showed that: BTV-1 VP2 protein was expressed as the inclusion bodies in the pEASY-Blunt E1 vector;160 and 200 mmol/L glyoxaline were the best condition to wash down the expressed protein.The molecular weight of this purified recombinant protein with N-terminal His-tag was about 105 ku.Through the results of Western blotting, ELISA and blocking ELISA, it had been proved that this recombinant protein could combine with BTV-1 specific antibody and this combination could be blocked by BTV-1 virus.The study showed that the recombinant BTV-1 VP2 protein, expressed through the prokaryotic expression vector pEASY-Blunt E1, possessed good immunoreactivity and this study had established foundation for locating the serotypic epitopes of the BTV-1 VP2 protein.关键词
蓝舌病/VP2蛋白/表达/免疫反应性Key words
bluetongue/VP2 protein/expression/immunoreactivity分类
农业科技引用本文复制引用
宋建领,李华春..蓝舌病1型病毒VP2蛋白的原核表达及免疫反应性分析[J].中国畜牧兽医,2017,44(7):2119-2125,7.基金项目
国家自然科学基金(31260612) （31260612）
公益性行业(农业)科研专项(201303035) （农业）
