伪狂犬病病毒UL9基因突变株构建及其生物学特性分析  

Construction of the PRV UL9 Gene Mutant Strain and Analysis of Its Biological Characteristics

作　　者：阮可悦 桑国菊 童武[1] 许朝阳 于海[1] 单同领[1] 李丽薇[1] 高飞[1] 童光志[1] 郑浩[1] RUAN Keyue;SANG Guoju;TONG Wu;XU Zhaoyang;YU Hai;SHAN Tongling;LI Liwei;GAO Fei;TONG Guangzhi;ZHENG Hao(Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;College of Animal Science and Veterinary Medicine,Shandong Agricultural University,Taian 271018,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]山东农业大学动物科技学院,泰安271018

出　　处：《中国动物传染病学报》2023年第4期1-8,共8页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点基础研究发展计划(973计划)。

摘　　要：伪狂犬病病毒(PRV)是一种大的DNA病毒,编码蛋白众多,其中一些具有酶功能,涉及病毒复制、蛋白调控和核苷酸代谢等。本研究在实验室已有的PRV感染性克隆pBAC-PRV基础上,利用体外转座的方法将Tn5转座子随机插入伪狂犬病病毒基因组中,获得病毒复制相关功能基因UL9的突变体。将UL9基因突变体转染Vero细胞,获得重组病毒vPRV-Tn5-UL9。通过病毒空斑实验和多步生长曲线测定等方法对vPRV-Tn5-UL9的生物学特性进行研究分析,结果显示突变病毒可在体外传代,但病毒的增殖速度显著降低,这表明UL9基因为PRV复制非必需基因,但对病毒复制增殖存在重要影响。此外,动物试验显示,vPRV-Tn5-UL9失去对小鼠的致死能力。本研究成功利用体外转座技术获得了PRV的UL9突变病毒,为构建重组病毒的方法提供了新思路,同时也为研究PRV复制机制打下一定基础。Pseudorabies virus(PRV),a large DNA virus,encodes a large number of proteins.Some proteins are enzymes involving in virus replication,protein regulation and nucleotide metabolism.In this study,the Tn5 transposon was randomly inserted into the pseudorabies virus genome existing PRV infectious clone pBAC-PRV by vitro transposition to obtain a mutant of the functional gene UL9 related to virus replication.The UL9 gene mutant was transfected into Vero cells to obtain the recombinant virus vPRV-Tn5-UL9.The biological characteristics of vPRV-Tn5-UL9 were studied and analyzed by virus plaque experiments and multi-step growth curve determination.These results showed that the UL9 gene is a non-essential gene for PRV replication,and mutations do not affect the passage of the virus in vitro.However,the proliferation rate is significantly reduced.In addition,animal experiments show that vPRV-Tn5-UL9 loses its lethal ability to mice.In this study,the UL9 mutant virus of PRV was successfully obtained by technology of vitro transposition,which provided a new idea for the construction of recombinant virus,and also laid a certain foundation for studying the mechanism of PRV replication.

关 键 词：伪狂犬病病毒 UL9基因 Tn5转座子 BAC 

分 类 号：S852.65[农业科学—基础兽医学]