应用噬菌体展示7肽文库技术进行蓝舌病1型病毒VP2蛋白表(拟)位分析

Epitope mapping of VP2 of bluetongue virus serotype 1 using 7-mer random peptide phage display

为了对蓝舌病1型病毒(BTV1)VP2蛋白表位进行定位,本研究利用BTV1型特异性抗体淘选噬菌体展示的7肽随机肽库,进行VP2蛋白表(拟)位分析。对筛选获得共有序列噬菌体,采用ELISA和竞争性ELISA分析不同噬菌体拟位与BTV1型特异性抗体的免疫反应性,结果显示含有TTPNNLS基序的噬菌体拟位能够与BTV1型特异性抗体发生特异性结合,并且该结合能够被BTV1抑制或阻断,表明拟位多肽真实模拟病毒蛋白上与BTV1型特异性抗体结合的抗原决定簇或表位。本研究揭示了BTV1 VP2蛋白的865THPNKCLVA873位氨基酸构成BTV1 VP2蛋白特异性表位,为该病的免疫机理研究奠定了基础。

To locate the epitope of the VP2 protein of bluetongue virus （BTV） serotype 1 （BTV1）, this study screened 7-mer random peptide phage display library with the serotype-specific antibody against BTV1. The consensus sequence were then analyzed through ELISA and competitive ELISA approaches to identify the immunoreactivities of different phage mimotopes to the antibody against BTV1. The study indicated that mimotopes with basic amino acid sequence of TTPNNLS was able to specifically bind with the antibody against BTV1. And this binding was blocked or inhibited by BTV1. The result shows that mimotope peptides could simulate the determinants or linear epitope on the viral protein that bind with BTV1 specific antibody. This result indicates that BTV1 VP2 protein specific epitope locate from the 865th to 873th amino acid （THPNKCLVA）.