传染性法氏囊病病毒JS株VP2基因真核表达载体的构建及其应用  

Construction of VP2 eukaryotic expression vector of infectious bursal disease virus and its application

作　　者：刘红梅[1] 秦爱建[1] 许小琴[1] 李迎晓[1] 陈鸿军[1] 叶建强[1] 金文杰[1] 刘岳龙[1] 邵红霞[1] 

机构地区：[1]扬州大学江苏省动物预防医学重点实验室

出　　处：《中国预防兽医学报》2006年第4期461-465,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家863高技术项目(863-2002AA245051);全国博士学位论文作者专项资金(200256)

摘　　要：应用RT-PCR方法从鸡传染性法氏囊病病毒(IBDV)JS株中扩增出VP2基因,并克隆入T-easy载体。序列测定分析结果表明IBDVJS株的VP2基因与国际标准强毒株的核苷酸序列同源性达98%,氨基酸序列同源性达99%以上。随后将VP2基因克隆入真核表达载体pcDNA3.1/zeo(+),构建成功真核表达质粒pcD-VP2,pcD-VP2体外转染COS-1细胞,能在COS-1细胞中表达。利用pcD-VP2质粒进行动物实验,结果表明雏鸡免疫14d后在体内可检测到特异性抗体,pcD-VP2的真核表达质粒二次免疫诱导鸡产生对IBDV强毒攻击的保护率为67%,这一结果提示VP2基因具有重要开发应用价值。The cDNA fragment of protective antigen VP2 gene of infectious bursal disease virus JS strain was amplified by RT-PCR, subcloned into pGEM-T easy vector and then into pcDNA3.1/zeo （＋） vector, designated as pcDVP2. By sequencing, the VP2 gene has high homology to that of very virulent IBDV strains. The transiently expression of VP2 in COS-1 cells was detected with the Mab specific to IBDV by the immunflurescent assay. After injection with pcDVP2 DNA to SPF chicken, the results showed that the specific anti-IBDV antibody could be detected in 14 days, 67 % of the chickens were protected after challenge with IBDV JS strain.

关 键 词：传染性法氏囊病病毒 VP2 表达 

分 类 号：S852.65[农业科学—基础兽医学]