蓝舌病1型病毒VP7蛋白表(拟)位定位及免疫检测分析

为研究蓝舌病1型病毒VP7蛋白关键性氨基酸表位定位，本试验应用抗蓝舌病1型病毒VP7蛋白单克隆抗体淘选噬菌体展示的7肽随机肽库，对筛选的共有序列噬菌体扩增纯化，通过ELISA、竞争性ELISA分析共有噬菌体拟位与蓝舌1型病毒VP7蛋白单克隆抗体的免疫反应性。结果表明，含有LNWPMVR基序的噬菌体拟位能够与蓝舌病1型病毒VP7蛋白单克隆抗体发生特异性结合，并且此结合能被蓝舌病1型病毒抑制或阻断，表明噬菌体7肽模拟了BTV蛋白上与蓝舌病1型病毒VP7蛋白单克隆抗体结合的抗原决定簇，提示蓝舌病病毒VP7蛋白的第163-189位氨基酸（LNAGARGDVQQIFQGRNDPMMIYLVWR）构成蓝舌病病毒VP7蛋白特异性表位。

Epitope Mapping and Immuno-detection of VP7 Protein of Bluetongue Virus Serotype 1

In this study,in order to map the key amino epitope of VP7 protein of blue tongue virus serotype 1 (BTV1),we used monoclonal antibody against VP7 protein of BTV1 to screen 7 mer phage display random peptide library. The selected phages including VP7 protein consensus sequence were amplified and purified,immunoreactivity between epitope of selected phages and monoclonal antibody against VP7 of BTV1 was analyzed by ELISA and competitive ELISA (c-ELISA).The result showed that the epitope of phages including LNWPMVR sequence could specially combine monoclonal antibody against VP7 of BTV1,and the combination could be inhibited or blocked by BTV1,it demonstrated that 7 mer phage simulated the antigenic determinant of BTV protein which could combine monoclonal antibody against VP7 of BTV1.The results suggested that the amino acides 163 to 189(LNAGARGDVQQIFQGRNDPMMLYLVWR) were the specific epitope of VP7 protein of BTV.