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1、VIROLOGY151.41-49(1986)SequenceAnalysisofthePorcineTransmissibleGastroenteritisCoronavirusNucleocapsidProteinGenePAULA.KAPKEANDDAVIDA.BRIAN2DepartmatofM~tiology,TheUniversityofTennessee,K?mxviU~,Tennessee$7996-0845ReceivedDecember12,1985;acceptedJanuary28,1986The3endofthe20-kbgeno

2、meofthePurduestrainofporcinetransmissiblegastroen-teritiscoronavirus(TGEV)wascopiedintocDNAafterprimingwitholigo(dT)andthedouble-strandedproductwasclonedintotheP&IsiteofthepUC9vector.Onecloneof2.0-kbcontainedpartofthepoly(A)tailandwassequencedinitsentiretyusingthechemicalmethodofM

3、axamandGilbert.Anothercloneof0.7kbalsocontainedpartofthepoly(A)tailandwassequencedinparttoconfirmtheprimarystructureofthemost3endofthegenome.Twopotential,nonoverlappinggeneswereidentifiedwithinthe3-terminal1663-basesequencefromanexaminationofopenreadingframes.Thefirstgeneencodesa3

4、82-aminoacidproteinof43,426molwt,thatistheapparentnucleocapsidproteinonthebasisofsize,chemicalproperties,andaminoacidsequencehomologywithothercoronavirusnucleocapsidproteins.Itisflankedonits5sidebyatleastpartofthematrixproteingene.Thesecondencodesahypothetical78.aminoacidproteinof

5、9101molwtthatishydrophobicatbothends.A3-proximalnoncodingsequenceof276baseswasalsodeterminedandaconservedstretchof9nucleotidesnearthepoly(A)tailwasfoundtobecommonamongTGEV,themousehepatitiscoronavirus,andtheavianinfectiousbronchitiscoronavirus.Q1986Academicpress,I~C.INTRODUCTIONwo

6、uldaccountforonlyapproximately8.4Thegenomeoftheporcinetransmissiblekbofcodinginformation.Inaddition,thegastroenteritiscoronavirus(TGEV)hasvirussynthesizesatleastonenonstructuralbeenshowntobeasingle-stranded,non-proteinduringitsreplication,anRNA-de-segmented,polyadenylated,infectio

7、usRNApendentRNApolymerase,thesizeofwhichmoleculeof6.8Xlo6molwtorapproxi-isnotyetknown(DennisandBrian,1982).mately20kbinlength(Brianetal,1980).Duringreplication,TGEVproducesnineThetotalnumberofgenesencodedbythespeciesofsubgenome-sizepolyadenylatedTGEVgenome,however,hasnotyetbeenRNA

8、moleculeseachofwhichmayfunctionde