马疱疹病毒1型UL56蛋白的原核表达及多克隆抗体的制备

Prokaryotic Expression and Polyclonal Antibody Preparation of UL56 Protein of Equid Alphaherpesvirus 1

为制备马疱疹病毒1型(EHV-1)UL56蛋白(pUL56)的多克隆抗体。该研究根据GenBank中EHV-1 YM2019株全基因组序列,采用Ni-NTA亲和层析纯化蛋白,与弗氏佐剂乳化后免疫BALB/c小鼠并制备pUL56多克隆抗体,经Western blot与间接ELISA试验测定多克隆抗体反应性。结果显示,纯化后的pUL56可以与EHV-1阳性血清发生反应;pUL56多克隆抗体的效价为1∶128 000,并能与pUL56特异性结合。本研究成功制备具有良好反应性的pUL56多克隆抗体,为pUL56功能特性及EHV-1致病机制的研究奠定理论基础。

The purpose of this study is to prepare polyclonal antibody of pUL56 ofEquidalphaherpesvirus 1.Based on the whole genome sequence of EHV-1 YM2019 strain in GenBank.The pUL56 was purified by Ni-NTA affinity chromatography,and BALB/c mice were immunized with Freund's adjuvant and pUL56 polyclonal antibody was prepared.The polyclonal antibody of the protein was measured by enzyme linked immunosorbent assay and Western blot.The results showed that the purified pUL56 could react with EHV-1 positive serum.The anti-pUL56 polyclonal antibody had a potency of 1∶128000 and bound specifically to pUL56.In this study,the mouse anti-pUL56 polyclonal antibody had good reactogenicity,which laid the theoretical foundation for functional properties of pUL56 and the pathogenic mechanisms of virus.