犬细小病毒VP2基因编码主要抗原表位区的核酸免疫及其免疫原性研究  

Immunogenicity with gene encoding the main antigen domain of canine parvovirus VP2

作　　者：吴植[1,2] 曹斌[1] 贺生中[1] 戴建华[1] 吴迪[1] 

机构地区：[1]江苏农牧科技职业学院,江苏泰州225300 [2]南京农业大学动物医学学院,江苏南京210095

出　　处：《中国预防兽医学报》2015年第6期473-476,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：江苏农牧科技职业学院院级课题(NSFYB1302)

摘　　要：为评价犬细小病毒(CPV)VP2主要抗原表位的免疫原性,本研究对CPV YZ株VP2的氨基酸序列进行分析,确定了主要抗原表位区域(VP2-228),通过PCR扩增相应的表位编码区域基因片段(700 bp),将其克隆于真核表达载体p VAX1中构建重组真核表达质粒p VAX1-VP2-228。Western blot结果显示VP2-228能够在COS-7细胞中正确表达。将该重组质粒免疫小鼠,利用血凝抑制试验检测不同时期的抗体水平,以MTT法检测免疫35 d时淋巴细胞的增殖活性。结果表明p VAX1-VP2-228能够诱导小鼠产生较高的抗体水平;淋巴细胞增殖试验表明免疫小鼠的淋巴细胞刺激指数显著高于对照组(p<0.05)。本研究为开展CPV DNA疫苗的研究提供了实验依据。To evaluate the immunogenicity of the main antigen domain of canine parvovirus （CPV） VP2, the encoding sequence of aa293-aa520 in VP2 of CPV YZ strain was prodicted to be the main antigen domain analyzed by relevant software and named VP2-228. Then, the encoding sequence of VP2-228 was amplified by PCR, and cloned into pVAX1 to construct eukaryotic expression recombinant plasmid of pVAX1-VP2-228. Western blot showed that VP2-228 was expressed in pVAX1- VP2-228 transfected COS-7 ceils in vitro. Moreover, the higher titer of HI antibody was induced from pVAX1-VP2-228 immunized mice and the lymphocyte stimulation index was also significantly （p〈0.05） higher than that of the control, which demonstrated that the pVAX1-VP2-228 was efficiently stimulate both humoral and cellular immunity.

关 键 词：犬细小病毒 VP2蛋白 主要抗原表位 免疫原性 

分 类 号：S852.65[农业科学—基础兽医学]