浙江农业学报 ›› 2016, Vol. 28 ›› Issue (9): 1476-1484.DOI: 10.3969/j.issn.1004-1524.2016.09.04

• 动物科学 • 上一篇    下一篇

奶牛乳房炎重要候选疫苗蛋白的抗原表位分析及三联重组表位疫苗的氨基酸序列设计

刘祥1, 孔智翔1, 殷书平2, 殴莎莎1, 刘成艳1, 同韩虎1, 吴三桥1, 陈琛1, 陈春琳1   

  1. 1.陕西理工大学 中德天然产物研究与开发利用联合研究所,陕西 汉中 723001;
    2.陕西医药控股集团生物制品有限公司,陕西 汉中 723000
  • 收稿日期:2015-12-22 出版日期:2016-09-15 发布日期:2016-11-23
  • 作者简介:刘祥(1982—),男,安徽合肥人,博士,讲师,研究方向为蛋白质组学与免疫学。E-mail: liuxiang888525@163.com
  • 基金资助:
    国家大学生创新创业训练计划项目(201510720556); 陕西省农业科技创新与攻关项目(2016NY-088); 陕西理工大学校级科研项目(SLGKY16-13)

Epitope analysis of cow mastitis important vaccine candidate proteins and design of amino acid sequence of triple recombinant epitope vaccine

LIU Xiang1, KONG Zhi-xiang1, YIN Shu-ping2, OU Sha-sha1, LIU Cheng-yan1, TONG Han-hu1, WU San-qiao1, CHEN Chen1, CHEN Chun-lin1   

  1. 1. Chinese-German Joint Institute for Natural Product Research, Shaanxi University of Technology, Hanzhong 723001, China;
    2. Shaanxi Pharmaceutical Holding Group Co., Ltd, Hanzhong 723000, China
  • Received:2015-12-22 Online:2016-09-15 Published:2016-11-23

摘要: 为设计奶牛乳房炎三联重组表位多肽疫苗,选取奶牛乳房炎3种主要感染菌的候选疫苗蛋白:金黄色葡萄球菌的Ebps与ClfA,大肠埃希菌的OmpA与OmpC,链球菌的SIP与PGK,通过ABCpred和BepiPred方案,预测获得每种蛋白各有2个优势B细胞表位;利用神经网络与量化矩阵法预测蛋白的CTL表位。结果显示,SIP蛋白无CTL表位,其余蛋白均存在1个CTL细胞表位;采用MHC-Ⅱ类分子结合肽在线程序预测蛋白的Th表位,结果发现每种蛋白各有1个Th细胞表位。使用DNASTAR软件分析6个蛋白的二级结构,结果发现,预测获得的B/T细胞表位大多处于蛋白易于产生表位的暴露表面、无规则卷曲与转角等位置。再通过Protean程序重组拼接获得的B/T细胞抗原表位,最终设计获得抗原性较好的三联表位疫苗氨基酸序列。为新型奶牛乳房炎三联表位疫苗的研制奠定基础。

关键词: 奶牛乳房炎, 表位疫苗, 抗原分析

Abstract: In order to design cow mastitis triple recombinant epitope polypeptide vaccine, six candidate vaccine proteins were selected, which were from three main pathogens of cow mastitis, concluding Ebps and ClfA of Staphylococcus aureus, OmpA and OmpC of Escherichia coli, SIP and PGK of Streptococcus. Using ABCpred and BepiPred prediction program, each protein had 2 B cell epitopes. CTL cell epitopes were obtained by the prediction method of quantitative matrix and artificial neural network, the results showed that the SIP protein had no CTL cell epitope, and there was 1 CTL cell epitope to other proteins. Th cell epitopes were gained using an online server prediction for MHC class Ⅱ peptide binding affinity, the results showed that each protein had 1 Th cell epitope. DNASTAR software was used to analyze the secondary structure for the six proteins, and the results showed that most of the B/T cell epitopes were located on the exposed surface, the random coil and the corner. The B/T cell epitopes were recomposed by DNASTAR Protean software, and triple epitope vaccine amino acid sequence holding better antigen was designed finally. These laid the foundation for the novel triple epitope vaccine of cow mastitis.

Key words: cow mastitis, epitope vaccine, antigenic analysis

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