鹅细小病毒VP3与禽分枝杆菌副结核亚种hsp65融合基因真核表达载体的构建及鉴定  

Construction and Identification of Eukaryotic Expression Vectors of Fusion Gene of Goose Parvovirus VP3 and Mycobacterium avium Subspecies paratuberculosis hsp65

作　　者：张岩[1,2] 姜秀云[3] 马红霞[1,2] 高云航[1,2] 徐凤宇[1,2] 

机构地区：[1]吉林农业大学动物科技学院,长春130118 [2]吉林农业大学动物生产及产品质量安全教育部重点实验室,长春130118 [3]吉林农业大学生命科学学院,长春130118

出　　处：《中国兽药杂志》2013年第5期6-9,共4页Chinese Journal of Veterinary Drug

基　　金：吉林省教育厅"十二五"科学技术研究项目(201149;201139);国家现代农业产业技术体系建设专项(CARS-43);国家星火计划课题(2012GA660003);吉林省科技厅项目(20040206-2-2)

摘　　要：为了构建鹅细小病毒(GPV)的VP3与禽分枝杆菌副结核亚种(MAP)的hsp65融合基因重组真核表达载体,试验克隆了VP3和hsp65基因,并将二者先后插入真核表达载体pVAX1中,构建重组质粒pVAX1-VP3和pVAX1-hsp65-VP3。酶切和PCR鉴定表明表达载体构建正确,用脂质体将二者分别转染入Vero细胞中,间接免疫荧光检测其在细胞中的表达。结果显示,转染细胞表面可见绿色荧光,说明hsp65、VP3基因表达成功。试验为GPV的DNA疫苗研制及hsp65分子佐剂在动物医学中的应用奠定基础。The aim of study was to construct recombinant eukaryotic expression vectors for goose parvovirus （GPV） VP3 and Mycobacterium avium subspecies paratuberculosis （MAP） hsp65 fusion gene. VP3 and hsp65 genes were cloned and the recombinant expression vector pVAX1 -VP3 and pVAX1 -hsp65 -VP3 were constructed. Restriction enzymes digestion analysis and PCR results indicated the recombinant expression vectors pVAX1 -hsp65- VP3 and pVAX1 -VP3 have been constructed successfully. The pVAX1 -hsp65 -VP3 and pVAX1- VP3 were transfected into Vero cells respectively by lipofectamine and the expressed products were detected by indirect immunofluorescenee. Results show that green fluorescence was observed on the surface of transfected cells. It is illustrated that hsp65 and VP3 had been expressed in Vero cells. Laid the foundation for preparation GPV DNA vaccines and application of MAP hsp65 in animal medicine.

关 键 词：鹅细小病毒 VP3 hs P65 基因表达 

分 类 号：S852.65[农业科学—基础兽医学]