冠状病毒Coronavirusl论文-2004 Cloning, Expression, and Purification of the Nucleocapsid Protein of SARS Coronavirus.pdf

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1、DokladyBiochemistryandBiophysics,Vol.397,2004,pp.239–241.TranslatedfromDokladyAkademiiNauk,Vol.397,No.4,2004,pp.546–548.OriginalRussianTextCopyright©2004byNetesova,Belavin,Seregina,Ignat’ev,Sandakhchiev.BIOCHEMISTRY,BIOPHYSICS,ANDMOLECULARBIOLOGYCloning,Expression,andP

2、urificationoftheNucleocapsidProteinofSARSCoronavirusN.A.Netesova,P.A.Belavin,E.V.Seregina,G.M.Ignat’ev,andAcademicianL.S.SandakhchievReceivedJanuary12,2004AnoutbreakofsevereacuterespiratorysyndromeAccordingtothepublisheddata,thenucleocapsid(SARS)wasforthefirsttimedescri

3、bedintheGuang-proteinN,animportantstructuralprotein,canbeuseddongprovince(China);then,thisdiseaseexpandedtoasanantigenfordetectingantibodiestocoronavirusessomeothercountries[1].AsearlyasinApril2003,[10,11].closecooperationofresearchersfrommanylaborato-Althoughtheextent

4、ofhomologyoftheaminoacidriesworldwideenabledtheWHOtoannouncethatthesequenceoftheSARS-CoVnucleocapsidproteinNetiologicalagentofthediseasewasanewpathogen,withothercoronavirusesis22–30%,ashortsequenceSARS-coronavirus(SARS-CoV).AlthoughSARS-enrichedwithlysineresidues(KTFPP

5、TEPKKD-CoVbelongstothefamilyofcoronaviruses,itisrelatedKKKKTDEAQ)isuniquetotheSARSvirus[9].Prob-tononeofthestrainsofthefamilydescribedsofar[2].Apurifiedpreparationoftherecombinantnucleo-capsidproteinoftheSARS-CoVstrainFrankfurtwas1234forthefirsttimeobtainedinthiswork.The

6、yieldoftheproteinwas80mgperliterofculture.ThisproteincanbeusedasanantigenfordetectingantibodiestoSARS-CoV.Inaddition,recombinantproteincanbeusedinfurtherstructuralandfunctionalstudiesofSARSvirusandconstructionofmolecularmodelforscreeninganti-SARSdrugs.66kDaAccordingtot

7、heliterature,thesizeoftheSARSpathogengenomerangesfrom29.797to29.751bp[3,4].Analysisofcompletegenomesofcoronaviruses45kDafailedtorevealthegenomeclosesttoSARS-CoV.ThegreatestdegreeofhomologywasfoundbetweenSARS-CoVandtype2bovinecoronavirus[3–5].Thefunctionsofviralproteins

8、shouldbestudiedtoprovideadeeperinsightintothemechanismsofSARSinfectionitself,improvediagnostictests,andrapidly24kDade