山羊痘病毒P32基因与绵羊CD58基因共表达载体的构建及其免疫活性分析  

Construction of co-expression vector containing goatpox virus P32 gene and ovine CD58 gene and immune activity analysis of the expressed product

作　　者：芦晓立[1] 吴国华[2] 颜新敏[2] 张强[2] 

机构地区：[1]商洛学院生物医药与食品工程学院,陕西商洛726000 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室,甘肃兰州730046

出　　处：《中国兽医科学》2015年第3期298-303,共6页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2012AA101304);商洛学院院级项目(10sky1005)

摘　　要：为了增强山羊痘病毒P32蛋白的免疫效果,发挥CD58的免疫佐剂作用,将山羊痘病毒P32基因和绵羊CD58基因构建在同一载体上,转染BHK21细胞,通过直接免疫荧光试验和间接免疫荧光试验分别间接检测P32和直接检测CD58在BHK21中的表达。结果显示,P32和CD58在BHK21细胞中都有表达。以构建的P32基因与绵羊CD58基因共表达载体为DNA疫苗,通过肌肉注射免疫BALB/c小鼠,定期采血,用ELISA检测免疫抗体水平,用MTT法检测细胞免疫情况;结果表明,共表达载体可诱导机体产生体液免疫和细胞免疫。In order to enhance the immune effect of the expression of goatpox virus antigen P32 and to play the role of CD58 in immunological adjuvant,ovine CD58 gene and P32 gene of goatpox virus were cloned into the eukaryotic plasmid pBudCE4.1,and the positive plasmid was named pBudCE4.1-CD58-P32.After the plasmid was purified,the acquired recombinant plasmid was transfected into BHK21 cells with liposome mediated method.The expressions of P32 and CD58proteins were tested by direct immunofluorescence assay(IFA)or indirect immunofluorescence assay,respectively.The results showed that P32 and CD58were successfully expressed in BHK21 cells.BALB/c mice were injected intramuscularly with the coexpression vector.ELISA and MTT assays results showed that humoral and cellular immune responses were elicited by pBudCE4.1-CD58-P32.

关 键 词：山羊痘病毒 P32基因 CD58基因 共表达载体 体液免疫 细胞免疫 

分 类 号：S852.654[农业科学—基础兽医学]