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1、VIROLOGY185,732-740(1991)ProcessingandAntigenicityofEntireandAnchor-FreeSpikeGlycoproteinSofCoronavirusTGEVExpressedbyRecombinantBaculovirusMURIELLEGODET,DENISRASSCHAERT,ANDHUBERTLAUDE’LaboratoiredeVirologieetImmunologicMokxlaires,InstituteNationaldelaR
2、echercheAgronomique,78350Jouy-en-Josas,FranceReceivedJune3,1991:acceptedAugust23,1991TheSgeneoftransmissiblegastroenteritisvirus(TGEV)wasinsertedintothegenomeofAutographacalifornicanuclearpolyhedrosisvirus(AcNPV)usingthetransferplasmidpVL941.Infectionof
3、Sf9insectcellswiththerecombi-nantvirusresultedinthesynthesisofa175KpolypeptidewhichwasabletotrimerizeandwastransportedtothecellsurfaceasistheauthenticTGEVSprotein.Despitethelackofcompletecarbohydrateprocessing,therecombinantSproteinexhibitedantigenicpro
4、pertiessimilartoTGEVSandinducedhighlevelsofneutralizingantibodiesinimmunizedrats.Engineeringadeletion(70aminoacids)intothecarboxy-terminuscontainingthemembraneanchorofthepolypep-tidealloweditssecretion.Theoligomerizationprocessandtheantigenicprofileofth
5、eanchor-freeSproteinwereshowntobepartiallyaltered.oI&IAcademic&s,kc.INTRODUCTIONgatedstructurepredictedfromthesequence,neartheC-terminalquarterofthepolypeptidechain(DeGrootTransmissiblegastroenteritisvirus(TGEV)causesaneta/.,1987;RasschaertandLaude,1987
6、).acuteenteritiswhichisoftenfatalinnewbornpigs.ItTheSproteinofTGEVistheonlyantigentoelicitthebelongstothecoronaviridae,afamilyofenvelopedproductionofhighlyneutralizingantibodies(Laudeetpositive-strandedRNAviruses.Virionsarecomposedal.,1986;Jimenezetal.,
7、1986)assumedtomediatetheofanucleocapsidproteinand2or3envelopeglyco-passiveprotectionconferredtosucklingpigletsproteins.Thespike,orpeplomerproteinS(formerly(Garwesetal.,1978/79).TheantigenicstructureofSnamedE2),formslargesurfaceprojectionscharacter-prote
8、inhasbeenstudiedusingmonoclonalantibodies.isticofcoronaviruses.ItexhibitsatypicalclassIglyco-AtleastfivemajorgroupsofepitopesorsiteshaveproteinprofilewithacleavedN-terminalsequenceandbeenidentified(Delmasetal.,1986;Correaeta/.,at
