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1、P1:OTA/XYZP2:ABCJWST211-c02JWST211-BridgenAugust23,201214:57PrinterName:YettoComeTrim:244mm×168mmPartIPositivesenseRNAvirusesP1:OTA/XYZP2:ABCJWST211-c02JWST211-BridgenAugust23,201214:57PrinterName:YettoComeTrim:244mm×168mm2CoronavirusreversegeneticsMariaArmesto,Kirste
2、nBentley,EricaBickerton,SarahKeepandPaulBrittonAvianViralDiseases,InstituteforAnimalHealth,ComptonLaboratory,Newbury,Berkshire2.1TheCoronavirinaeTheCoronaviridaeformpartoftheorderNidovirales,whichcomprisestwosub-families,theCoronavirinaeandTorovirinae.Therearethreegen
3、eraofcoron-aviruses,alpha-,beta-andgammacoronaviruses(Carstens,2010),whichweresonamedfortheirvisualresemblancetothecoronaofthesuninnegativelystainedpreparations(Figure2.1)(Tyrrelletal.,1968).RepresentativemembersofeachofthecoronavirusgeneraareshowninTable2.1.Possiblyt
4、hemostpublicisedcoronavirusofrecentyearshasbeenthehumancoronavirusSARS-CoV,whichemergedinChinain2002causingthesevereacuterespiratorysyndromeepidemic(Drostenetal.,2003;Ksiazeketal.,2003;Marraetal.,2003;Rotaetal.,2003).Coronaviruseshave,however,beenisolatedfrommanyverte
5、bratesandcausesev-eraleconomicallyimportantdiseasesinlivestockspecies,includingpigs,cowsandchickens,anddomesticspecies,suchasdogsandcats.Coronavirusesareenvelopedviruseswithasingle-strandedpositive-senseRNAgenomeof26–32kb,thelargestgenomesofallRNAvirusescurrentlyknown
6、,thatreplicateinthecytoplasmofinfectedcells.Thegenomeassociateswiththenucle-oprotein(N),formingahelicalnucleocapsidwithinthevirusparticles.Althoughcommonamongnegative-senseRNAviruses,coronavirusesaretheonlypositive-senseRNAvirusestopossesshelicalnucleocapsids,whichare
7、enclosedwithinlipidenvelopescontainingthespike(S)glycoprotein,membrane(M)proteinanden-velope(E)protein(Figure2.1).Forgeneralreviews,see(Siddell,1995;LaiandCavanagh,1997;Enjuanes,2005;Siddelletal.,2005;Enjuanesetal.,2006;Gor-balenyaetal.,2006;Masters,2006;Thiel,2007;Br
8、ittonandCavanagh,2008;Perl-manetal.,2008;Norkin,2010).ReverseGeneticsofRNAViruses:ApplicationsandPerspectives,FirstEdition.E