伪狂犬病病毒gD、gE基因的克隆及转移载体的构建  

Cloning of the gD and gE of pseudorabies virus and construction of a universal transfer vector

作　　者：胡涛[1] 崔保安[1] 杨明凡[1] 王学斌[1] 张素梅[1] 王岩 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]郑州牧业高等专科学校,河南郑州450008

出　　处：《中国预防兽医学报》2004年第2期149-151,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：利用PCR扩增了伪狂犬病病毒Min＿A株gD、gE基因,扩增产物克隆于pGEM＿TEasy载体。将gD、gE基因连接于质粒pUC18获得pUgDgE,缺失质粒pUgDgE的BamHI和BstEII位点间391bp的片段。在此缺失位置插入来自质粒pcD NA3.1＿的一段含hCMV启动子、多克隆位点和Neo报告基因的片段,构建了通用转移载体pgD＿M＿gE。该转移载体缺失了伪狂犬病病毒gI基因和gE基因ORF5'端363bp的碱基,有11个酶切位点可供外源基因的插入,上下游侧翼分别为1.25bp和1.42bp。这为开发以伪狂犬病病毒为载体的多价基因工程疫苗提供了有力工具。The gD and gE gene of the pseudorabies virus Min-A strain were obtained by polymerase chain reaction(PCR),and then cloned into the pGEM-T Easy vector.The gD and gE gene is subcloned into pUC18,resulting in pUgDgE.The fragment from pcDNA3.1-including hCMV promoter/enhancer,MCS and neomycin resistance gene was inserted into the BamHI and BstEII sites of pUgDgE,resulting in the universal transfer vector pgD-M-gE.The universal transfer vector pgD-M-gE deleted the gI gene and 363 bp in the 5' end of the gE ORF of PRV.There were 11 restrication sites for insertion of the foreign gene.The upstream and downstream flanking sequences were up to 1.25 kb and 1.42 kb.It will be useful for developing the recombinant PRV expressing foreign gene(s).

关 键 词：伪狂犬病毒 GD gE 克隆 通用转移载体 

分 类 号：S852.65[农业科学—基础兽医学] Q785[农业科学—兽医学]