四川农业大学学报 ›› 2016, Vol. 34 ›› Issue (02): 234-238.doi: 10.16036/j.issn.1000-2650.2016.02.019

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PRRSV-GP5基因的克隆及其B细胞抗原表位预测与分析

尹国友1,2, 王林嵩1   

  1. 1. 河南师范大学生命科学学院, 河南 新乡 453007;
    2. 河南城建学院生命科学与工程学院, 河南 平顶山 467036
  • 收稿日期:2015-12-30 出版日期:2016-06-30 发布日期:2017-02-15
  • 通讯作者: 王林嵩,教授,主要研究方向为生物化学与分子生物学,E-mail:041008@htu.cn。 E-mail:041008@htu.cn
  • 作者简介:尹国友,博士研究生。
  • 基金资助:
    河南省教育厅项目(14A180009)

PRRSV-GP5 Gene Cloning and Analysis of B Cell Antigen Epitope

YIN Guo-you1,2, WANG Lin-song1   

  1. 1. Department of Bioscience, Henan Normal University, Xinxiang 453002, Henan, China;
    2. School of Life Science and Engineer, Henan University of Urban Construction, Pingdingshan 467036, Henan, China
  • Received:2015-12-30 Online:2016-06-30 Published:2017-02-15

摘要: [目的] 研究PRRSV-GP5基因B细胞抗原表位性质。[方法] 根据Gen Bank上发表猪繁殖与呼吸综合征病毒(PRRSV)ATCC VR-2332的全基因序列,设计并合成引物,采用RT-PCR技术,对阳性病料扩增回收,并将其克隆入pMD18-T载体中,送上海生工测序。应用生物信息学同源模型化的方法建立其PRRSV-GP5蛋白3D结构,并根据生物信息学软件DNAStar预测PRRSV-GP5蛋白的二级结构、表面可及性、抗原指数、亲水性及柔韧性,综合分析预测其B细胞抗原表位。[结果] 结果表明,其肽段30-36、50-55、140-142、146-151和196-198可能是其优势B细胞抗原表位区域。[结论] 该基因高度保守,PRRSV-GP5蛋白呈现较规则的空间结构,同时该结果将为PRRSV-GP5蛋白体外表达产物的应用和基因工程疫苗的研究提供理论依据。

关键词: PRRSV-GP5, 克隆, 抗原表位

Abstract: [Objective] The objective of this experiment was to study the B cell epitopes of PRRSV-GP5 gene. [Method] Primers for RT-PCR amplification of PRRSV isolates from Heilongjiang were designed according to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published in the GenBank, and sequences were cloned into pMD18-T vector respectively and sequenced in Shanghai. The 3D model of PRRSV-GP5 protein was obtained by homology modeling of bioinfor-matics, and DNAStar was used to predict the secondary structure, surface probability, antigenic index, hydrophilicity and flexibility region. [Result] The peptides of PRRSV-GP5 including 30-36, 50-55, 140-142, 146-151 and 196-198 were larvaceous dominant peptide. [Conclusion] The results indicat-ed that the target gene of PRRSV-GP5 was highly conservative and space structure of PRRSV-GP5 was regular. The results will provide theoretical evidence for application of expression product and engineering vaccine of PRRSV-GP5 protein.

Key words: PRRSV-GP5, clone, antigen epitope

中图分类号: 

  • Q518.2