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1、JournalofGeneralVirology(1994),75,1789-1794.PrintedinGreatBritain1789Nucleotidesequenceandexpressionofthespike(S)geneofcaninecoronavirusandcomparisonwiththeSproteinsoffelineandporcinecoronavirusesJohnG.Wesseling,'~HarryVennema,Gert-JanGodeke,MarianC.HorzinekandPeterJ
2、.M.Rottier*VirologyDivision,DepartmentofInfectiousDiseasesandImmunology,VeterinaryFaculty,UtrechtUniversity,P.O.Box80.165,3508TDUtrecht,TheNetherlandsWehavecloned,sequencedandexpressedthespike(S)twofelinecoronaviruses.Theporcinesequencesaregeneofcaninecoronavirus(CCV
3、;strainK378).Itsclearlymoredivergentmainlyduetothelargedifferencesdeducedaminoacidsequencehasrevealedfeaturesinintheamino-terminal(residues1to300)domainsofthecommonwithothercoronavirusSproteins:astretchofproteins;whenonlythecarboxy-terminalparts(residueshydrophobicam
4、inoacidsattheaminoterminus(the301andon)areconsideredthehomologiesbetweentheputativesignalsequence),anotherhydrophobicregioncanine,felineandporcineSpolypeptidesaregenerallyatthecarboxyterminus(themembraneanchor),heptadquitehigh,withidentitiesrangingfrom90.8%to96-8%.re
5、peatsprecedingtheanchor,andacysteine-richregionThehumancoronavirusislessrelatedtotheotherlocatedjustdownstreamfromit.Likeotherrepresent-membersoftheantigenicgroup.Aphylogenetictreeativesofthesameantigeniccluster(CCV-Insavc-1constructedonthebasisoftheSsequencesshowedt
6、hatstrain,felineinfectiousperitonitisandentericcorona-thetwoCCVsareevolutionarilymorerelatedtotheviruses,porcinetransmissiblegastroenteritisandres-felinethantotheporcineviruses.ExpressionoftheCCVpiratorycoronaviruses,andthehumancoronavirusSgeneusingthevacciniavirusT7
7、RNApolymeraseHCV229E),theCCVSpolypeptidelacksaproteolyticsystemyieldedaproteinoftheexpectedMr(approxi-cleavagesitepresentinmanyothercoronavirusSmately200K)whichcouldbeimmunoprecipitatedwithproteins.PairwisecomparisonsoftheSaminoacidananti-felineinfectiousperitonitisv
8、iruspolyelonalsequenceswithintheantigenicclusterdemonstratedthatserumandwhichwasindistinguishablefromtheSthetwoCCVstrains(K378andIn
