表达兔瘟病毒VP60基因的重组病毒BHV-1的构建与鉴定  

Construction and Identification of Recombinant Virus BHV-1 Expression of VP60 Gene of RHDV

作　　者：原冬伟[1] 鲁文赓[1] 黄艳秋[1] 郭东华[1] 薛飞[2] 曲连东[2] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆 [2]中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室/自然疫源性人兽共患病研究团队, 黑龙江哈尔滨

出　　处：《免疫学研究》2017年第2期11-18,共8页Immunology Studies

基　　金：黑龙江省教育厅面上项目(12541580)。

摘　　要：为了构建表达兔瘟病毒VP60基因的牛疱疹病毒I型,首先构建含有兔瘟病毒VP60基因和CMV启动子的转移载体,然后将构建移载体与gE基因缺失的牛疱疹病毒I型基因组共转染牛肺细胞后收获增殖的病毒。通过蓝白斑筛选不含有LacZ基因的白色病毒蚀斑,反复纯化获得重组病毒BHV-1-VP60。PCR方法验证VP60基因的成功插入,间接免疫荧光试验和Western blot,结果证明了BHV-1-VP60中的VP60基因在易感细胞中获得了表达。本研究成功地构建了兔瘟病毒VP60基因的重组病毒BHV-1-VP60,为研制新型兔瘟活载体疫苗奠定了基础。In order to construct the recombinant bovine heper virus I which expressed RHDV VP60 gene, first, we constructed a transfer vector containing RHDV VP60 gene and CMV promoter, and then the mixtures of parental virus DNA and transfer vector were transfected into bovine lung cells. Then the propagated viruses were harvested. The recombinant virus was obtained by selection for white virus plaques which did not contain LacZ gene. PCR assay showed that VP60 gene had been inserted into the recombinant virus genome;the expression of VP60 in infected cells was proved by indirect immunofluorescence assay and Western blot. This study successfully constructed the recombinant virus BHV-1-VP60, which provided a basis for the development of a new type of RHDV vector vaccine.

关 键 词：兔瘟病毒 VP60基因 牛疱疹病毒1型 蓝白斑筛选 

分 类 号：S852[农业科学—基础兽医学]