Asia 1型口蹄疫病毒抗原表位突变株的构建及其生物学特性分析  

Construction and characterization of an epitope-mutated Asia 1 type foot-and-mouth disease virus

作　　者：张岩[1] 胡永浩[1] 杨帆[2] 杨波[2] 王松豪[2] 朱紫祥[2] 郑海学[2] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所口蹄疫国家参考实验室家畜疫病病原学国家重点实验室,甘肃兰州730046

出　　处：《生物工程学报》2015年第1期96-104,共9页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划(863计划)(No.2011AA10A211-1);国家自然科学基金(No.31302118);甘肃省科技重大专项(No.1302NKDA027)资助~~

摘　　要：为了研制口蹄疫抗原表位突变标记疫苗,本研究以含有Asia 1型口蹄疫病毒(FMDV)c DNA全长的感染性克隆p Asia 1-FMDV作为骨架,将3D蛋白中第27位氨基酸的H和31位的氨基酸N分别突变成Y和R,从而突变3D蛋白的一个抗原表位,将构建的带有突变表位的重组质粒转染BHK-21细胞,成功拯救出一株突变FMDV。经比较后发现,重组病毒的生物学特性与亲本毒株相似。病毒中和试验结果显示,抗重组病毒的血清与亲本病毒有良好的反应性。Western blotting结果表明重组病毒诱导的抗体能与突变的表位合成肽反应而不与野生型病毒的表位合成肽发生反应,从而区分重组病毒与亲本病毒。综上所述,这株抗原表位突变FMDV有望作为口蹄疫标记疫苗候株进一步评估。To generate an epitope-mutated foot-and-mouth disease virus （FMDV） as a marker vaccine, the infectious clone pAsia 1-FMDV containing the complete genomic eDNA of Asia 1 type FMDV was used as backbone, the residues at positions 27 and 31 in the 3D gene were mutated （H27Y and N31R）. The resulting plasmid pAsia 1-FMDV-3DM encoding a mutated epitope was transfected into BHK-21 cells and the recombinant virus rAsia 1-3DM was rescued. The recombinant virus showed similar biological characteristics comparable with the parental virus. In serological neutralization test the antisera against recombine virus have a good reactivity with parental virus. The antisera against the mutant virus were shown to be reactive with the mutated epitope but not the wild-type one. The results indicated that the two virus strains could be distinguished by western blotting using synthetic peptides. This epitope-mutated FMDV strain will be evaluated as a potential marker vaccine against FMDV infections.

关 键 词：口蹄疫病毒 感染性cDNA克隆 负标记 表位突变 

分 类 号：S852.65[农业科学—基础兽医学]