基因Ⅰ型日本脑炎病毒GX毒株感染性克隆的构建  

Construction of infectious clone of genotypeⅠJapanese encephalitis virus GX strain

作　　者：颜梦学 叶静[1] 曹胜波[1] 熊俊垚 YAN Mengxue;YE Jing;CAO Shengbo;XIONG Junyao(State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学农业微生物国家重点实验室,湖北武汉430070

出　　处：《中国兽医学报》2025年第3期482-488,共7页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2022YFD1801500,2022YFD1800105);国家自然科学基金重点资助项目(32030107)。

摘　　要：通过RT-PCR方法从基因Ⅰ型(GⅠ)日本脑炎病毒(Japanese encephalitis virus,JEV)GX株中扩增覆盖全长cDNA的3段基因片段,将3个基因片段依次克隆到pBR322载体上以获得感染性克隆质粒pGX,测序正确后将该感染性克隆质粒与pCAGGS-T_(7)聚合酶真核表达质粒共转染至BHK-21细胞中进行病毒的拯救。间接免疫荧光和噬斑试验结果显示,成功拯救GⅠJEV rGX;噬斑试验和小鼠感染试验结果显示,拯救的病毒rGX与野生型病毒GX具有相似的复制能力与致病性。结果表明,成功构建了GⅠJEV GX毒株感染性克隆。The infectious clone plasmid of genotypeⅠJapanese encephalitis virus GX strain was successfully obtained by reverse transcription-polymerase chain reaction,where the cDNA of GX strain is divided into three fragments for amplification and the three fragments were sequentially cloned into pBR322 vector.After the infectious clone plasmid pGX was sequenced correctly,the pGX and pCAGGS-T_(7)eukaryotic plasmid were co-transfected into BHK-21 cells for virus rescue.The experimentalresults indicated that the Japanese encephalitis virus could be successfully rescuedfrom BHK-21 cells.The plaque experiment and mouse experiment indicated that the rescued virus had similar replication ability and pathogenicity with wild type virus.It was confirmed that the infectious clone of genotypeⅠJapanese encephalitis virus GX strain was successfully constructed in this study.

关 键 词：基因Ⅰ型 乙型脑炎病毒 感染性克隆 复制 致病力 

分 类 号：S852.65[农业科学—基础兽医学]