表达兔出血症病毒VP60蛋白重组牛疱疹病毒1型的构建及免疫原性分析  

Construction and immunogenicity analysis of recombinant bovine herpesvirus-1 expressing the VP60 protein of RHDV

作　　者：黄艳秋[1,2] 原冬伟[1] 刘行波[1,2] 刘家森[2] 陈淑慧[1,2] 单丽玲[2] 郭东春[2] 姜骞[2] 崔玉东[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163310 [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150001

出　　处：《中国预防兽医学报》2014年第8期597-601,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30800830);黑龙江省科技计划项目(PC10S02);黑龙江省教育厅面上项目(12541580)

摘　　要：为制备表达兔出血症病毒(RHDV)VP60蛋白的重组病毒疫苗,本研究将RHDV VP60基因插入到CMV启动子下,构建牛疱疹病毒1型(BoHV-1)gE基因缺失转移载体,利用磷酸钙介导转染法将该转移载体与gE基因缺失的亲本病毒基因组DNA共转染牛肺细胞,制备了表达RHDV VP60基因的重组BoHV-1(rBoHV-1/gE-/VP60)。通过PCR、western blot及间接免疫荧光鉴定表明VP60基因在rBoHV-1/gE-/VP60中获得表达。并且体内试验表明重组病毒能够有效诱导小鼠产生RHDV特异性抗体。本研究构建了表达RHDV VP60基因的rBoHV-1/gE-/VP60,为研制预防兔出血症的BoHV-1活载体疫苗奠定了基础。To construct the recombinant virus expressing VP60 gene of rabbit hemorrhagic disease virus （RHDV）, the cassette containing the VP60 gene under the control of the CMV promoter was inserted into the recombinant bovine hepervirus-1 （BoHV-1） of gE gene deletion transfer vector, and the resultant transfer vector was co-transfected into bovine lung cell line （BL） with the total DNA of parental virus （rBoHV-1/gE） by calcium phosphate to generate the recombinant BoHV-1 （rBoHV-1/ gE-/VP60）. In addition, the expression of RHDV VP60 gene from the cells infected with rBoHV-1/gE-/VP60 was confLrmed by PCR, western blot and immunofluorescence assay. Moreover, the tests showed that the recombinant virus was able to effectively stimulate the mice to produce specific antibody against RHDV. In conclusion, the construction of the rBoHV-1/gE-/VP60 provided a basis for development of BoHV-1 live vector vaccine against RHD.

关 键 词：兔出血症病毒 VP60基因 牛疱疹病毒1型 

分 类 号：S852.65[农业科学—基础兽医学]