表达猪传染性胃肠炎病毒SLN基因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 piglets expressing SLN gene of transmissible gastroenteritis virus

作　　者：宋子杰[1] 商纪娟 盖春云 解倩倩 张洪亮[1] 宫照欣 孙鑫鹏[1] 单虎[1] 黄娟[1] SONG Zi-jie;SHANG Ji-juan;GAI Chun-yun;XIE Qian-qian;ZHANG Hong-liang;GONG Zhao-xin;SUN Xin-peng;SHAN Hu;HUANG Juan(College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China;Shandong Vocational Animal Science and Veterinary College, Weifang 261000, China;Shandong Dezheng Dairy Co. ,Ltd., Weihai 264200, China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]山东畜牧兽医职业学院,山东潍坊261000 [3]山东德正乳业股份有限公司,山东威海264200

出　　处：《中国兽医科学》2018年第7期838-843,共6页Chinese Veterinary Science

基　　金：山东省重点研发计划项目:猪传染性胃肠炎基因工程疫苗的研制与开发(2015GNC111013);山东省现代农业产业技术体系项目(SDAIT-08-01);威海市产学研战略合作“特聘专家”资助计划项目(2015TP2J9)

摘　　要：为探究表达猪传染性胃肠炎病毒SLN基因的重组乳酸乳球菌口服免疫动物诱导的特异性免疫应答,本研究构建了p MG36e-SLN重组质粒并将其电转化至乳酸乳球菌感受态细胞MG1363中,制备表达猪传染性胃肠炎病毒SLN基因的重组乳酸乳球菌,经SDS-PAGE、Western-blot分析鉴定目的基因表达,并将重组菌口服免疫新西兰大白兔,利用间接ELISA方法测定家兔小肠黏膜和血清中抗SLN蛋白特异性Ig G和Ig A。结果显示,表达重组SLN蛋白的重组菌经口服能够有效引起动物体产生较高水平抗SLN蛋白Ig G和一定量的抗SLN蛋白Ig A,表明该重组菌表达系统能刺激动物免疫系统产生应答,具有作为口服疫苗的潜在应用价值。In order to explore the specific immune response induced by oral immunization of recombinant Lactococcus lactis expressing SLN gene of swine transmissible gastroenteritis virus（TGEV）,the recombinant plasmid p MG36 e-SLN was constructed and electro-transferred into Lactococcus lactis competent cells MG1363.A recombinant Lactococcus lactis,which can express the transmissible gastroenteritis virus SLN gene,was prepared,and expression of the objective gene was confirmed by SDS-PAGE and Western-blot. New Zealand White rabbits were orally administered with the recombinant strain,and the titers of anti-SLN Ig G and anti-SLN Ig A were evaluated by indirect ELISA. In result,the high level of Ig G in serum and limited Ig A could be detected. The results indicated the recombinant strain was qualified to be a potential oral vaccine because it could successfully evoke both humoral and mucosal immune responses against TGEV.

关 键 词：猪传染性胃肠炎病毒 SLN基因 免疫原性 ELISA 

分 类 号：S852.659.6[农业科学—基础兽医学]