浙江农业学报 ›› 2017, Vol. 29 ›› Issue (12): 2009-2014.DOI: 10.3969/j.issn.1004-1524.2017.12.08

• 动物科学 • 上一篇    下一篇

鸭坦布苏病毒E蛋白线性化B细胞抗原表位的鉴定

张琳1, 2, 余斌1, *, 倪征1, 陈柳1, 云涛1, 叶伟成1, 华炯钢1, 崔言顺2, 张存1   

  1. 1.浙江省农业科学院 畜牧兽医研究所,浙江 杭州 210021;
    2.山东农业大学 动物科技学院,山东 泰安 271018
  • 收稿日期:2016-08-22 出版日期:2017-12-20 发布日期:2018-01-08
  • 通讯作者: 余斌,E-mail:biogiant@126.com
  • 作者简介:张琳(1986—),女,黑龙江大兴安岭人,硕士研究生,主要从事动物检疫与食品检验研究。E-mail:zhanglin_0109@126.com
  • 基金资助:
    国家重点研发计划(2016YFD0500100); 浙江省自然科学基金(LY15C18002); 浙江省公益技术研究农业项目(2016C32070)

Identification of linear B cell epitope for E protein of duck Tembusu virus

ZHANG Lin1, 2, YU Bin1, *, NI Zheng1, CHEN Liu1, YUN Tao1, YE Weicheng1, HUA Jionggang1, CUI Yanshun2, ZHANG Cun1   

  1. 1.Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
    2 College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China
  • Received:2016-08-22 Online:2017-12-20 Published:2018-01-08

摘要: 为鉴定鸭坦布苏病毒(duck Tembusu virus,DTMUV)的B细胞抗原表位,以纯化的DTMUV YY5株为抗原制备了2株单克隆抗体AE4和BD10,研究证实BD10为线性化表位,其结合的抗原表位位于E蛋白的第三结构域(Domain Ⅲ,D)。将DTMUV E蛋白D结构域基因截短为具有末端相互重叠的15段,与MBP融合表达后以单克隆抗体BD10进行肽库扫描,结果筛选出DTMUV一个B细胞线性表位385LVGSGKGQI393(EP385)。该表位原核融合表达产物免疫小鼠制备的抗体,能够和DTMUV E蛋白反应,表明EP385表位具有良好的免疫原性,可为DTMUV多肽疫苗的研制及特异血清学诊断方法的开发奠定基础。

关键词: 鸭坦布苏病毒, 单克隆抗体, 肽库扫描, 抗原表位

Abstract: This experiment was conducted to identify B cell epitope of E protein of duck Tembusu virus (DTMUV). In this study, a linear B cell epitope 385LVGSGKGQI393 (EP385) for DTMUV E protein were identified by truncated expressing the third domain of E protein (Domain III, DIII), which is recognized by monoclonal antibody BD10 prepared by the purified DTMUV YY5 strain as antigen, and then by scanning the peptide library which containing 15 overlapping segment of truncated and fused expressed DIII with BD10. Then, the immunogenicity of EP385 was verified by fusion expressed in E. coli and immunized mice to prepare a polyclonal antibody which was confirmed to be capable of recognizing viral E protein. The identification of B cell epitope for the E protein of DTMUV may lead to the preparation of a DTMUV peptide vaccine and the development of specific serological diagnostic methods.

Key words: duck Tembusu virus, monoclonal antibody, peptide library scanning, epitope

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