鸭坦布苏病毒E蛋白抗原表位的鉴定及其表位交叉反应性分析  

Antigenic epitopes identification of duck tembusu virus envelope protein and evaluation of cross reactivity

作　　者：李晨曦[1] 华荣虹[1] 张云[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150069

出　　处：《中国预防兽医学报》2018年第1期60-65,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：十三五重点研发项目(2016YFD0500100);国家自然科学基金(31670153);中国农业科学院协同创新项目(CAAS-XTCX2016011-01)

摘　　要：为鉴定鸭坦布苏病毒(DTMUV)E蛋白的抗原表位,本研究以纯化的抗DTMUV E蛋白单克隆抗体(MAb)3B6为固相筛选分子,对噬菌体随机12肽库进行淘选,通过对阳性噬菌体克隆测序分析,确定DTMUV E蛋白的模拟表位氨基酸序列,将模拟表位序列与GST蛋白融合表达并进行western blot验证。利用DNAStar对表位序列进行同源性分析,并通过dot blotting方法分析抗原表位在黄病毒阳性血清间的交叉反应性。结果鉴定了DTMUV E蛋白的一个线性抗原表位EVEPPFG,并且该表位在DTMUV与其它黄病毒中均具有高度的保守性。Dot blotting结果显示抗原表位EVEPPFG在黄病毒阳性血清间具有交叉反应性。因此,EVEPPFG为黄病毒共享型的抗原表位,这对于黄病毒E蛋白的抗原结构功能的研究以及研制表位疫苗和诊断抗原等具有重要意义。To identify the epitopes of Duck tembusu virus （DTMUV） E protein, a phage-displayed 12-mer random peptide library was used to screen B-cell epitopes on E protein with purified monoclonal antibodies （MAb） against DTMUV E protein. The amino acid sequence of the E protein was determined according to the positive clones sequence and expressed with GST protein. Western blot assay indicated that the amino acid sequence EVEPPFG was a linear antigenic epitope on E protein. Sequence alignment revealed that epitope EVEPPFG was highly conserved. Dot blotting with corresponding flaviviruses positive sera showed that cross-reaction might be existed among DTMUV and other flaviviruses at this epitopes position, suggesting that the epitope EVEPPFG was a cross-reactive epitope in flaviviruses. These findings provids useful information for understanding the structure and organization of epitopes on E protein of flaviviruses and might be valuable in the development of epitope-based serological diagnostic tests for DTMUV and other flaviviruses infection.

关 键 词：鸭坦布苏病毒 噬菌体表面展示技术 E蛋白 抗原表位 黄病毒 

分 类 号：S852.65[农业科学—基础兽医学]