表达猪流行性腹泻病毒S1蛋白的重组乳酸乳球菌的构建及其免疫原性的研究  

Construction and immunogenicity of recombinant Lactococcus lactis expressing S1 protein of porcine epidemic diarrhea virus

作　　者：王莉平 韩先杰[1] 王孝彬 盖春云 李军伟 单虎[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109

出　　处：《中国兽医科学》2017年第9期1118-1123,共6页Chinese Veterinary Science

基　　金：山东省现代农业产业技术体系项目(SDAIT-06-022-08);青岛市民生科技计划项目(16-6-2-42-nsh)

摘　　要：为探究表达猪流行性腹泻病毒(PEDV)纤突糖蛋白(S1)的重组乳酸乳球菌口服免疫动物诱导的特异性免疫应答,本研究从PEDV SDLY株中扩增得到S1基因序列,构建pMG36e-S1重组质粒并将其电转至乳酸乳球菌MG1363感受态细胞中,制备表达猪流行性腹泻病毒S1蛋白的重组乳酸乳球菌。应用SDS-PAGE、Western-blot鉴定目的蛋白的表达。将重组菌口服免疫新西兰大白兔,中和试验检测小肠黏膜和血清中的中和抗体效价,间接ELISA方法测定血清中特异性IgG。结果显示,该重组菌能够有效引起动物体产生较高水平血清IgG和一定量的黏膜免疫中和抗体,表明该重组菌表达系统能刺激动物系统产生免疫应答,具有作为口服疫苗的潜在应用价值。To evaluate the specific immune responses induced by recombinant Lactococcus lactis （L.lactis） which expresses porcine epidemic diarrhea virus （PEDV） S1 protein as an oral vaccine,the spike gene fragment of PEDV was amplified from the genome and then was linked to the expression vector p MG36e.The recombinant plasmid was electro-transferred into competent L.lactis MG1363.Expression of objective protein was confirmed by SDS-PAGE and Western-blot.New Zealand White rabbits were orally administered with the recombinant strain,then the titer of mucosal immune antibody in intestinal mucosa and IgG in serum was detected by neutralizing test,and the titer of IgG was evaluated by indirect ELISA.The result showed that the high level of IgG in serum and limited mucosal immune antibody could be detected.The recombinant strain is qualified to be a potential oral vaccine because it could successfully evoke both humoral and mucosal immune responses against PEDV.

关 键 词：猪流行性腹泻病毒 S蛋白 pMG36e表达载体 乳酸乳球菌 免疫反应 

分 类 号：S852.65[农业科学—基础兽医学]