O型口蹄疫病毒样颗粒编码基因的真核表达载体构建及表达  

Construction and Expression of Eukaryotic Expression Vector of O Type Foot-and-mouth Disease Virus-like Particles Encoding Gene

作　　者：李淑萍 孙世琪[2] 莫亚霞 胡永浩[1] 郭慧琛[2] LI Shu-ping;SUN Shi-qi;MO Ya-xia;HU Yong-hao;GUO Hui-chen(Colledge of Veterinary Medicine, Gansu Agriculture University,Lanzhou,Gansu,730070,China;State Key Laboratory of Veterinary Etiological Biology/National Foot-and-Mouth Disease Reference Laboratory/ Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu,730046, China)

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家畜疾病病原生物学国家重点实验室OIE/国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《动物医学进展》2019年第1期1-6,共6页Progress In Veterinary Medicine

基　　金：动物重大疫病新概念防控产品研发项目(2017YFD0501100);牛羊重要疫病免疫防控新技术研究项目(2017YFD0500900)

摘　　要：为了构建O型口蹄疫病毒(FMDV)病毒样颗粒编码基因的真核表达载体,将FMDV结构蛋白VP0、VP1、VP3的基因分别克隆到真核表达载体pVAX1中,再分别通过带有BsmBⅠ、EcoRⅠ和SalⅠ酶切位点的引物,分别对重组质粒pVAX-VP0、pVAX-VP1和pVAX-VP3扩增后进行酶切,用T4连接酶连接得到重组的真核表达载体pVAX-VP0VP1VP3。对重组真核表达载体pVAX-VP0VP1VP3进行PCR鉴定并测序。用脂质体将重组真核表达载体pVAX-VP0VP1VP3转染BHK-21细胞,用间接免疫荧光试验(IFA)及Western blot检测重组真核表达载体pVAX-VP0VP1VP3在细胞中的表达情况。结果表明,成功构建了重组真核表达载体pVAX-VP0VP1VP3,并在BHK-21细胞中得到表达。To construct and express the eukaryotic expression vector of serotype O foot-and-mouth disease virus-like particles encoding gene,VP0,VP1 and VP3 genes were cloned into the eukaryotic expression vector pVAX1 through the known FMDV structural protein gene sequences in this paper.Then,the recombinant plasmids pVAX-VP0,pVAX-VP1,pVAX-VP3 were amplified by using primers with Bsm B Ⅰ, Eco R Ⅰ and Sal Ⅰ restriction sites respectively,the recombinant eukaryotic expression vector pVAX-VP0VP1VP3 was obtained by ligating with T4 ligase.The recombinant eukaryotic expression vector pVAX-VP0VP1VP3 was identified by PCR and sent to Suzhou Jinzhizhi Biotechnology Co.,Ltd. for sequencing.The recombinant eukaryotic expression vector pVAX-VP0VP1VP3 was transfected into BHK cells by lipofectin transfection method and the expression of recombinant eukaryotic expression vector pVAX-VP0VP1VP3 in cells was detected by indirect immunofluorescence assay test (IFAT) and Western blot.The results showed that the recombinant eukaryotic expression vector pVAX-VP0VP1VP3 was constructed and expressed successfully in BHK-21 cells.

关 键 词：口蹄疫病毒 重组真核表达载体 pVAX-VP0VP1VP3构建和表达 病毒样颗粒 

分 类 号：S852.659.6[农业科学—基础兽医学] Q789[农业科学—兽医学]