狂犬病毒重组致病株Mu11的定点回复突变及生长特性的研究  

Site-directed Reverse Mutation and Studies on Growth Characteristics of Rabies Virus Recombinant Pathogenic Strain Mul1

作　　者：金蓉 梁晶晶 张传亮 韦显凯 于冬玲 刘俊丹 李晓宁[1,2,3] 罗廷荣 JIN Rong;LIANG Jingjing;ZHANG Chuanliang;WEI Xiankai;YU Dongling;LIU Jundan;LI Liaoning;LUO Tingrong(College of Animal Science and Technology,Guangxi University,Nanning,530004;Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics,Nanning,530004;Guangxi Colleges and Universities Key Laboratory of Prevention and Control for Animal Disease,Nanning,530004;Department of Pathobiology Laboratory of Infectious Diseases&Immunology,School of Veterinary Medicine,University of Pennsylvania,Philadelphia,PA19104,U.S.A;Guangxi Center for Animal Disease Control and Prevention,Nanning,530000)

机构地区：[1]广西大学动物科学技术学院,南宁530004 [2]广西壮族自治区兽用生物制品工程研究中心,南宁530004 [3]广西高校动物疫病预防与控制重点实验室,南宁530004 [4]美国宾夕法尼亚大学兽医学院病理生理学系传染病与免疫学实验室,费城PA19104 [5]广西壮族自治区动物疫病预防控制中心,南宁530004

出　　处：《基因组学与应用生物学》2023年第6期620-628,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31902311);广西自然科学基金项目(202100192)共同资助。

摘　　要：狂犬病毒GX074株于2003年自广西健康犬脑分离得到,将其G蛋白第253、263、273位氨基酸通过反向遗传学技术替换到弱毒疫苗株rRC-HL对应位点得到重组毒株Mu11。为验证G蛋白第253、263、273位氨基酸对狂犬病毒的生长特性的影响,本研究以Mu11株的感染性cDNA克隆质粒为基础,将Mu11株G蛋白第253、263、273位氨基酸回复突变为亲本弱毒株rRC-HL的对应氨基酸,得到突变病毒株Mu11^(R253.263.273)。应用间接免疫荧光实验及对拯救病毒基因测序,证明Mu11^(R253.263.273)毒株突变位点正确。通过对其病毒滴度和多步生长曲线的测定,证明突变病毒株Mu11^(R253.263.273)基因组稳定,在细胞中的复制能力与亲本弱毒株rRC-HL一致。研究表明狂犬病毒G蛋白253、263、273位氨基酸对Mu11和亲本强毒GX074的生长特性具有重要影响,为进一步对其致病性功能的研究提供科学依据。The rabies virus strain GX074 was isolated from the brain of a healthy dog in the Guangxi Province in 2003,and the amino acids at positions 253,263,and 273 of its G protein were replaced to the corresponding sites of the attenuated vaccine strain rRC-HL by reverse genetic technology to obtain the recombinant strain Mull.To verify the effects of amino acids at positions 253,263 and 273 of G protein on the growth characteristics of rabies virus,this study used the infectious cDNA cloning plasmid of Mull strain as the basis,and restored the amino acid mutations at positions 253,263,and 273 of strain Mul1 G protein to the corresponding amino acids sites of the parental attenuated strain rRC-HL,and the mutant virus strain Mu11^(R253.263.273)was obtained.It was confirmed that the mutant sites of the strain Mu11^(R253.263.273)were correct by indirect immunofluorescence assay and gene sequencing experiment.By measuring itsvial te ad muli-sep growh curve,it was demonstrate tht the mutant vrustran Mu11^(R253.263.273)had stablegenomeand isreplication ability was consistent with the parental attenuated strain rRC-HL in cells.These studies indicate that amino acids 253,263,and 273 of rabies virus G protein have an important effect on the growth characteristics of the strain Mull and the parental virulent strain GX074,which provides scientific basis for further research on its pathogenicity.

关 键 词：狂犬病毒 G蛋白 反向遗传技术 病毒拯救 生长特性 

分 类 号：S852.65[农业科学—基础兽医学]