›› 2016, Vol. 43 ›› Issue (6): 1630-1634.doi: 10.16431/j.cnki.1671-7236.2016.06.035

• 疾病防治 • 上一篇    下一篇

水貂细小病毒NS1与VP2蛋白的原核表达及免疫原性分析

王洋1, 牛登云2, 刘昊1, 胡博1, 马凡舒1, 鲁荣光1, 吕爽1, 闫喜军1   

  1. 1. 中国农业科学院特产研究所, 长春 130112;
    2. 青岛农业大学动物科技学院, 青岛 266109
  • 收稿日期:2015-12-02 出版日期:2016-06-20 发布日期:2016-07-11
  • 通讯作者: 闫喜军 E-mail:tcsyxj@126.com
  • 作者简介:王洋(1990-),男,吉林公主岭人,硕士生,研究方向:预防兽医学,E-mail:wang691665@126.com
  • 基金资助:
    吉林省科技发展计划项目(20130206026NY)

Prokaryotic Expression and Immunogenicity Analysis of Mink Parvovirus NS1 and VP2 Protein

WANG Yang1, NIU Deng-yun2, LIU Hao1, HU Bo1, MA Fan-shu1, LU Rong-guang1, LV Shuang1, YAN Xi-jun1   

  1. 1. Institute of Special Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, Jilin 130112, China;
    2. College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
  • Received:2015-12-02 Online:2016-06-20 Published:2016-07-11

摘要: 为了比较VP2和NS1两种蛋白的免疫原性,选择免疫原性较好的蛋白进行亚单位疫苗制备。本试验分别扩增了水貂细小病毒(mink enteritis virus,MEV)NS1与VP2基因,连接pET-32a表达载体并进行表达,对表达产物进行SDS-PAGE及Western blotting分析。以His-Bind亲和层析柱纯化目的蛋白,将纯化后的蛋白免疫小鼠,分析目的蛋白的免疫原性。经SDS-PAGE与Western blotting鉴定,表明NS1与VP2蛋白大小分别为83 和67 ku,且均具有生物学活性;免疫小鼠后,目的蛋白NS1和VP2均可诱导小鼠产生抗MEV特异性抗体,且VP2蛋白诱导小鼠产生的抗体滴度要高于NS1蛋白。与NS1蛋白比较,VP2蛋白更适合亚单位疫苗的制备。

关键词: 水貂细小病毒; 原核表达; 重组蛋白

Abstract: In order to develop subunit vaccine of mink enteritis virus,the immunogenicity of mink parvovirus protein NS1 and VP2 had been evaluated.Two pairs of primers were designed,and the full-length NS1 and VP2 genes had been amplificated,and then prokaryotic expression vector pET-32a-NS1,PET-32a-VP2 were constructed.After the analysis of SDS-PAGE and Western blotting,target proteins had been purified by His-Bind affinity chromatography.The immunogenicity of purified protein NS1 and VP2 were evaluated by serum ELISA testing,after inoculated BALB/c mouse.The results showed that the molecular mass of NS1 and VP2 protein were 83 and 67 ku by SDS-PAGF and Western blotting;Although both target protein NS1 and VP2 had the ability to induce BALB/c mouse to produce anti-MEV specific antibodies,the level of antibodies induced by the protein VP2 was higher than protein NS1.Mink parvovirus protein VP2 was more suitable for the development of subunit vaccine.

Key words: mink parvovirus; prokaryotic expression; recombinant protein

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