Getah virus(GETV)is a mosquito-borne virus of the genus Alphavirus in the family Togaviridae and,in recent years,it has caused several outbreaks in animals.The molecular basis for GETV pathogenicity is not well un-derstood.Therefore,a reverse genetic system of GETV is needed to produce genetically modified viruses for the study of the viral replication and its pathogenic mechanism.Here,we generated a CMV-driven infectious cDNA clone based on a previously isolated GETV strain,GX201808(pGETV-GX).Transfection of pGETV-GX into BHK-21 cells resulted in the recovery of a recombinant virus(rGETV-GX)which showed similar growth characteristics to its parental virus.Then three-day-old mice were experimentally infected with either the parental or recom-binant virus.The recombinant virus showed milder pathogenicity than the parental virus in the mice.Based on the established CMV-driven cDNA clone,subgenomic promoter and two restriction enzyme sites(BamHI and EcoRI)were introduced into the region between E1 protein and 3'UTR.Then the green fluorescent protein(GFP),red fluorescent protein(RFP)and improved light-oxygen-voltage(iLOV)genes were inserted into the restriction enzyme sites.Transfection of the constructs carrying the reporter genes into BHK-21 cells proved the rescue of the recombinant reporter viruses.Taken together,the establishment of a reverse genetic system for GETV provides a valuable tool for the study of the virus life cycle,and to aid the development of genetically engineered GETVs as vectors for foreign gene expression.

Tongwei Ren;Xiangling Min;Qingrong Mo;Yuxu Wang;Hao Wang;Ying Chen;Kang Ouyang;Weijian Huang;Zuzhang Wei

Laboratory of Animal Infectious Diseases and Molecular Immunology,College of Animal Science and Technology,Guangxi University,Nanning 530005,China

中国病毒学

[1]Tongwei Ren;Xiangling Min;Qingrong Mo;Yuxu Wang;Hao Wang;Ying Chen;Kang Ouyang;Weijian Huang;Zuzhang Wei-.Construction and characterization of a full-length infectious clone of Getah virus in vivo)[J].中国病毒学,2022(03):348-357

Alphavirus,Togaviridae,derstood,GX201808,pGETV,rGETV,binant,subgenomic,iLOV,GETVs

Construction,characterization,full,length,infectious,clone,Getah,vivo,mosquito,borne,genus,family,recent,years,has,caused,several,outbreaks,animals,molecular,basis,pathogenicity,not,well,un,Therefore,reverse,system,needed,produce,genetically,modified,viruses,study,viral,replication,its,mechanism,Here,generated,CMV,driven,cDNA,previously,isolated,strain,Transfection,into,BHK,cells,resulted,recovery,recombinant,which,showed,similar,growth,characteristics,parental,Then,three,day,old,mice,were,experimentally,infected,either,milder,than,Based,established,promoter,two,restriction,enzyme,sites,BamHI,EcoRI,introduced,region,between,E1,protein,UTR,green,fluorescent,GFP,RFP,improved,light,oxygen,voltage,genes,inserted,constructs,carrying,reporter,rescue,Taken,together,establishment,provides,valuable,tool,life,cycle,aid,development,engineered,vectors,foreign,expression

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