Construction of DNA Vaccine for FMDV P1 Gene and Immunization Experiment  

FMDV P1基因DNA疫苗构建及免疫实验(英文)

作　　者：史秋梅[1] 高桂生[1] 张艳英[1] 高光平[1] 张东林[1] 

机构地区：[1]河北科技师范学院动物科技学院,河北省预防兽医学重点实验室,河北昌黎066600

出　　处：《Agricultural Science & Technology》2013年第8期1069-1071,共3页农业科学与技术（英文版）

基　　金：科技部农业成果转化项目(2012GB2A200045);农业部转基因专项(2011ZX08006-001);2012年河北科技师范学院动物疫病防控科研创新团队项目~~

摘　　要：[Objective] This study aimed to construct DNA vaccine of foot-and-mouth disease （FMD）.[Method] Plasmid carriers plESZP1 and pUTK3CP1 with PRV were constructed for FMDV P1 gene expression.Mice were immunized,and their antibody level was detected.The two eukaryotic expression plasmids constructed were transfected into Vero cells.PCR,IFA and Westem-blot were carried out to detect the transcription and expression of the objective gene.Balb/C mice were intramuscularly inoculated with the DNA plasmid which expressed the target gene correctly,and the antibody level in mice was detected by the means of ELISA and serum neutralization （SN）.[Result] DNA plasmid carrying P1 gene which encodes FMDV capsid protein caused specific body fluid immunoreaction in mice,and the antibody level of anti-FMDV had no difference in the mice induced by the two recombinant plasmids.[Conclusion] This study lays a foundation for evaluating the genetically modified vaccine by immunizing animals with recombinant PRV containing the FMDV P1 gene and recombinant virus.[目的]构建口蹄疫DNA疫苗。[方法]构建以PRV为载体的表达FMDV P1基因的质粒pIESZP1和pUTK3CP1,免疫小鼠并检测了免疫后小鼠的抗体水平。将构建的2个真核表达质粒分别转染Vero细胞,通过PCR、IFA、Western-blot等方法检测目的基因的转录和表达。将正确表达目的基因的DNA质粒肌肉接种3周龄的Balb/C小鼠,采用ELISA和SN方法检测了免疫小鼠体内的抗体水平。[结果]携带有FMDV衣壳蛋白P1基因的DNA质粒能引起小鼠产生特异性的体液免疫反应,两重组质粒诱导小鼠产生抗FMDV的抗体水平无差异。[结论]该研究为含有FMDV P1基因重组PRV以及重组病毒的动物免疫试验以评价基因工程疫苗的免疫效果奠定了基础。

关 键 词：FMDV P1 gene DNA plasmid Immunization experiment 

分 类 号：S852.5[农业科学—基础兽医学]