O型口蹄疫病毒代表性抗原VP1基因的人工合成与表达及其免疫原性检测  

Synthesis,expression of representative VP1 gene of FMDV type O and immunogenicity of recombinant protein

作　　者：李润成[1] 余兴龙[1] 白霞[1] 侯强红[1] 罗维[1] 刘忠华[1] 向卫军[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2008年第3期333-337,共5页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：湖南省科技重大专项(05NK1001-03);湖南省教育厅重点项目(04A025)

摘　　要：通过对GenBank近10年内收录的O型口蹄疫(FMD)病毒VP1基因序列的同源性比较分析,并根据中国、老挝、缅甸等国家FMDV流行毒株的基因序列,设计并合成了有群内抗原代表性的结构蛋白VP1全基因序列.将合成的序列克隆到原核表达载体pET-28a(+)中,构建了VP1的表达质粒pETVP1.SDS-PAGE和Western-blot分析表明,该质粒的BL21(DE3)LysS转化菌在IPTG诱导下可特异性表达VP1蛋白,该重组蛋白以包涵体的形式存在,相对分子质量约30 000,能与口蹄疫病毒(FMDV)阳性血清发生特异性反应.动物接种试验表明,重组蛋白能诱导小白鼠产生特异性抗FMDV抗体.The homology of VP1 gene sequence accessed in GeneBank was analyzed. The VP1 gene of FMDV type O was designed and synthesized artificially according to gene sequence of FMDV isolated in China and its neighboring countries. Plasmid pETVP 1, expressing the VP1 of FMDV, was obtained by the technology of genetic engineering. The expression pETVP1 was transformed into BL21（DE3）LysS. 30 000 recombinant VP1 could be experessed in BL21（DE3）lyss amounting to 28 % of the total protein of the induced recombinant bacteria at the presence of IPTG as proved by means of SDS-PAGE. Western-blot analysis showed recombinant VP1 was FMDV-specific. Animal inoculation experiments showed that the recombinant protein can induce a specific anti-FMDV antibody.

关 键 词：口蹄疫病毒 VP1基因 合成 表达 免疫原性 

分 类 号：S852.659.6[农业科学—基础兽医学]