华北农学报 ›› 2009, Vol. 24 ›› Issue (5): 81-85. doi: 10.7668/hbnxb.2009.05.018

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口蹄疫病毒结构蛋白VP1上B细胞表位的筛选鉴定

张昱, 王永录, 张永光, 方玉珍, 潘丽, 蒋守田, 吕建亮, 刘力宽, 张中旺, 张淑刚, 李正丰, 杜进鑫   

  1. 中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 农业部畜禽病毒学重点开放实验室, 甘肃兰州 730046
  • 收稿日期:2009-05-12 出版日期:2009-10-28
  • 通讯作者: 王永录(1964-),男,甘肃临洮人,研究员,主要从事口蹄疫病毒免疫学与分子生物学研究。
  • 作者简介:张昱(1983-),男,甘肃静宁人,在读硕士,主要从事动物病毒免疫学与分子生物学研究。
  • 基金资助:
    国家支撑计划项目(2006BAD06A06)

Screening and Identification of B Cell Epitopes of VP1 of Foot-and-mouth Disease Virus

ZHANG Yu, WANG Yong-lu, ZHANG Yong-guang, FANG Yu-zhen, PAN Li, JIANG Shou-tian, LU Jian-liang, LIU Li-kuan, ZHANG Zhong-wang, ZHANG Shu-gang, LI Zheng-feng, DU Jin-xin   

  1. Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
  • Received:2009-05-12 Published:2009-10-28

摘要: 以口蹄疫病毒株AF72 RNA为模板,反转录并扩增结构蛋白VP1基因,PCR纯化产物与pGEM-T easy载体连接并转化JM109菌株,对经凝胶电泳、PCR和EcoRⅠ酶切法鉴定为阳性的重组质粒进行测序,通过序列比对获得AF72VP1的核苷酸序列和推导氨基酸序列,综合分析结构蛋白VP1的亲水性、可塑性、抗原指数以及表面可能性等参数,预测其潜在B细胞抗原表位并人工合成表位肽段,利用间接ELISA对潜在表位肽段进行筛选鉴定,结果显示,表位VP1a和VP1d为病毒株AF72结构蛋白VP1的优势B细胞表位,该结果为进一步的FMDV多表位疫苗研究提供有价值的参考依据。

关键词: 口蹄疫病毒, 结构蛋白VP1, 间接ELISA, B细胞表位

Abstract: Foot-and-mouth disease virus strain AF72 RNAs were used as templates for RT-PCR to amplify the VP1 gene.The purified PCR products were cloned into pGEM-T easy vectors and transformed into E.coli JM109.The positive recombinant plasmids identified by electrophoresis,PCR,and EcoR ivcleavage were sequenced.The nucleotide and were obtained by comparing with the ful-llength sequence of the other reference strains.Potential B-cell epitopes of VP1 were predicted and epitope peptide segments were synthesized.They were ident ified by indirect ELISA.The result showed that VP1a and VP1d were predominant B-cell epitope of VP1,which provided valuable information for further study on the vaccine of FMD.

Key words: FMDV, Structural protein VP1, Indirect ELISA, B-cell epitope

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张昱, 王永录, 张永光, 方玉珍, 潘丽, 蒋守田, 吕建亮, 刘力宽, 张中旺, 张淑刚, 李正丰, 杜进鑫. 口蹄疫病毒结构蛋白VP1上B细胞表位的筛选鉴定[J]. 华北农学报, 2009, 24(5): 81-85. doi: 10.7668/hbnxb.2009.05.018.

ZHANG Yu, WANG Yong-lu, ZHANG Yong-guang, FANG Yu-zhen, PAN Li, JIANG Shou-tian, LU Jian-liang, LIU Li-kuan, ZHANG Zhong-wang, ZHANG Shu-gang, LI Zheng-feng, DU Jin-xin. Screening and Identification of B Cell Epitopes of VP1 of Foot-and-mouth Disease Virus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(5): 81-85. doi: 10.7668/hbnxb.2009.05.018.

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