牛乳头瘤病毒2型L1基因真核表达载体的构建及真核表达  

Construction and Expression of Bovine Papillomavirus Type 2 L1 Gene Eukaryotic Expression Vector

作　　者：贺志昊[1] 乔军[1] 孟庆玲[1] 杨海波[1] 刘昱成[1] 王国超[1] 才学鹏[2] 陈创夫[1] 都曼.努尔坎杰 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]中国农业科学院兰州兽医研究所,兰州730046 [3]沙湾县畜牧兽医站,新疆沙湾832100

出　　处：《西北农业学报》2014年第10期18-21,共4页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：兵团国际科技合作项目(2012BC006);农业公益行业专项子课题(201303037-5)

摘　　要：为在真核细胞中表达牛乳头瘤病毒2型(BPV2)衣壳蛋白L1基因。采用PCR方法从奶牛皮肤肿瘤病料中扩增BPV2沙湾分离株L1基因,克隆入pMD18-T载体中,测序后进行序列分析。以pcDNA 3.1(+)为真核表达载体构建重组表达质粒,转染入BHK-21细胞,通过间接荧光免疫试验检测BPV2-L1蛋白的表达。本研究成功构建pcDNA3.1(+)-BPV2-L1重组真核表达质粒,证实能表达出BPV2-L1蛋白,为BPV2DNA疫苗的研发提供了前期基础。To express of L1 gene of bovine papillomavirus type 2 （BPV2-L1） in BHK-21 cells. BPV2 L1 gene was amplified from sample of skin lesions by PCR. Then the product was cloned into pMD18- Tvector, sequenced and then subcloned into eukaryotic expressing vector pcDNA3.1（＋）. The recombined vector was transfected into BHK-21 cells with lipofectin and the expression of L1 gene was detected by indirect immunofluorescence assay. The eukaryotic expressing vector encoding BPV2 L1 was successfully constructed and its expression in BHK-21 cells could be detected. The eukaryotic expressing vector encoding BPV2 L1 was constructed and it expressed L1 protein correctly in BHK-21 cells.

关 键 词：牛乳头瘤病毒2型 衣壳蛋白 L1基因 克隆 真核表达 

分 类 号：S852.653[农业科学—基础兽医学]