Asia1型口蹄疫病毒衣壳蛋白在毕赤酵母中共表达及免疫效果评价  

Co-expression and immunogenicity of capsid proteins of Asia1 type foot and mouth disease virus in Pichia pastoris

作　　者：曹永生[1] 高明春[1] 曹翀[1] 张润祥[1] 李迪[1] 王欢[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030

出　　处：《东北农业大学学报》2013年第9期73-80,共8页Journal of Northeast Agricultural University

基　　金：国家科技支撑计划项目(2012BAD12B03;2012BAD12B05);现代农业(奶牛)产业技术体系项目(CARS-37);黑龙江省"十二五"科技攻关项目(GA09B302)

摘　　要：为研究Asia1型口蹄疫病毒衣壳蛋白在毕赤酵母中共表达及其免疫原性,本研究基于酵母细胞内表达载体pPICZA,利用独立编码三个衣壳蛋白表达盒构建质粒pPICZA-vp031,重组质粒经BglⅡ线性化后进行电转化,获得重组有三个独立表达盒重组酵母菌。SDS-PAGE和Western blot检测结果表明衣壳蛋白均得到正确表达且具有良好抗原性。重组酵母菌经口服免疫小鼠后,机体能够产生血清特异性IgG抗体和黏膜sIgA抗体,淋巴细胞增殖结果显示免疫小鼠淋巴细胞特异性增殖指数显著高于灭活疫苗组,细胞因子检测结果显示经特异性抗原刺激免疫小鼠淋巴细胞分泌IL-4和IL-10与灭活疫苗组无显著差异,显著高于PBS对照组,表明重组菌能够刺激机体产生特异性细胞免疫应答。本研究为口蹄疫基因工程疫苗研制奠定基础。To investigate the immune effects of the co-expressed the capsid proteins of Asia1 type foot and mouth disease virus （FMDV） in Pichia pastoris, based on intra-cellular expression vector pPICZA, three cassettes which encoded the capsid proteins individually were used to construct the plasmid pPICZA-vp031. After being linearized with Bgl II, the plasmid was transformed by electroporation to obtain the Pichia pastoris which contained the three individual cassettes. The results of SDS-PAGE and Western blot showed that the proteins were co-expressed and had good immunoreactivity, The recombinant yeast was able to induce specific serum IgG and mucosal slgA in mice. The specific stimulation index of the vp031group was significantly higher than the inactivated vaccine group. The IL-4 and IL-10 levels of vp031 group had no significant difference with the inactivated vaccine group, higher than negative control group, which indicated the recombinant yeast was able to induce specific cellular immunity. This study lays the foundation for further FMD vaccine research.

关 键 词：Asia1型FMDV衣壳蛋白 毕赤酵母 共表达 抗原性 免疫原性 

分 类 号：S852.4[农业科学—基础兽医学]