PRVΔgE/TK/UL49.5基因缺失株的构建及相关特性研究  

Construction and characterization of PRVΔgE/TK/UL49.5three-gene-deleted strain

作　　者：丁晨梦 孙亚威 石蒙蒙 韩紫薇 许夕雅 吕晨哲 齐江坤 杨寒 于林洋 李永涛[1] 陈陆[1] DING Chenmeng;SUN Yawei;SHI Mengmeng;HAN Ziwei;XU Xiya;LYU Chenzhe;QI Jiangkun;YANG Han;YU Linyang;LI Yongtao;CHEN Lu(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学学院,河南郑州450046

出　　处：《中国兽医学报》2023年第9期1814-1819,1836,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31772781)。

摘　　要：为探究UL49.5基因缺失对伪狂犬病病毒(pseudorabies virus,PRV)变异株毒力的影响,本试验以PRVΔgE/TK株为亲本株,通过同源重组和Cre-loxP系统构建重组病毒PRVΔgE/TK/UL49.5,并对该毒株遗传稳定性、体外生长特性、干扰SLAⅠ抗原递呈能力及对小鼠的安全性进行初步研究。经PCR及测序确定PRVΔgE/TK/UL49.5株UL49.5基因缺失474bp。与PRV YY和PRVΔgE/TK株相比,PRVΔgE/TK/UL49.5在Vero细胞上增殖滴度相似,可稳定传代,具有良好的增殖能力。与PRV YY株相比,PRVΔgE/TK/UL49.5株感染PK-15细胞后下调SLAⅠ和TAP转录水平能力降低,并对小鼠具有安全性。本试验通过同源重组技术构建了PRVΔgE/TK/UL49.5株,为PRV基因缺失疫苗候选毒株筛选奠定了基础。To explore the effect of UL49.5gene-deleted on the virulence of the pseudorabies virus(PRV)variant,the PRVΔgE/TK/UL49.5strain was constructed by homologous recombination technology and Cre-loxP system using PRVΔgE/TK strain,and its genetic stability,in vitro growth characteristics,SLAI antigen presentation ability and safety to mice were studied.The deletion of 474bp UL49.5gene fragment of PRVΔgE/TK/UL49.5strain was confirmed by PCR and sequencing.Compared with PRV YY strain and PRVΔgE/TK strain,PRVΔgE/TK/UL49.5 strain replication titer in Vero cells was similar,and it could be passaged stably and had good proliferative ability.Compared with PRV YY strain,PRVΔgE/TK/UL49.5strain down-regulated SLAI and TAP transcriptional levels in PK-15cells and was safe for mice.In summary,PRVΔgE/TK/UL49.5strain was constructed by homologous recombination technique,which laid a foundation for screening PRV candidate gene-deleted vaccine strains.

关 键 词：伪狂犬病病毒 基因缺失 UL49.5基因 同源重组 

分 类 号：S855.3[农业科学—临床兽医学]