3 4 205 7 CHINESEJOURNALOF VIROLOGY Vol.3 No.4 July 205 2, 2 2 * (., 02206;2., 50300) :R373.2 + 3 :205-06-04; :205-06-09 : (203ZX000460, 204ZX0004002,20ZX000400) :A :000-872(205)04-0333-08 DOI:0.3242/j.cnki.bingduxuebao.002726 (MERS-CoV), MERS-CoV, S β,sars-cov HCoV-HKU E M N ORF3 ORF4a [-4] [2] 202MERS-CoV, ORF4b ORF5 ORF8b (WHO), 205 6, 54 MERS-CoV, 43, : MERS-CoV, MERS-CoV ChinaGD0, S E M N ORF3 ORF4a ORF4b ORF5ORF8b S MERS-CoV,, MERS-CoV 5,N E M :ORF3 ORF4a ORF4b ORF5,ORF8b MERS- CoV MERS-CoV MERS-CoV : ; ; ; 37%,SARS-CoV, ( 2055 MERS- ) [5] CoV,205 5 28 : [4] 2055 26 MERS-CoV WHO MERS-CoV,5 28 MERS-CoV MERS-CoV,529 2 MERS-CoV, MERS- CoV, MERS-CoV MERS-CoV ChinaGD0 MERS-CoV, S E : (972-), * :,,E-mail:tanwj28@63.com;,kecw965@aliyun.com, M N ORF3 ORF4a ORF4b ORF5 ORF8b GenBank (KT00289-KT00294, KT003528-KT003530) MERS-CoV NCBI
334 3 MERS-CoV Joining Method, MERS-CoV 6 (), -6, 2 5 ; 4 6 205 6,GenBank, MERS-CoV MERS-CoV 86, MEGA5.0 Neighbour- MERS-CoV Figure PhylogenetictreeofcompletegenomesofMERS-CoV Theevolutionarytreewasconstructedfromnucleotidealignmentsusing MEGA5.0software,sequencesnamearede- rivedfrom GenBankaccessionnumberandvirusstrainname.
4 : 335 2 MERS-CoV S S MERS-CoV S MERS-CoV 5(2) RBD S Jeddah( )Riyadh( ) MERS- 86 S CoV, 98.9%-99.6% NC 09843.3 99.5%,, MERS-CoV Jed- S dahriyadh S ( RBD ) 2 S Figure2PhylogeneticanalysisofnucleotidesequenceinSgeneofMERS-CoV
336 3 3 N EM,N,ChinaGD0 MERS-CoV N E MERS-CoV E N 00% E Chi- M nagd0n MERS-CoV N Chi- 99.4%-99.9%,, nagd0m M ChinaGD0N 5 98.9% ~99.5%, ChinaGD0 (3) E M 5 (4) 3 N Figure3 PhylogeneticanalysisofnucleotidesequenceinNgeneofMERS-CoV
4 : 337 4 M Figure4 Phylogeneticanalysesofnucleotidesequencein M geneofmers-cov 4 ORF3 ORF4a ORF4b ORF5 98.7% E N,ORF3 F,86 P L ORF4a MERS- ORF8b CoV 98.5%~99.7%, ORF3 ORF4a ORF4b ORF5 ORF8b 2 MERS-CoV ORF4b,ChinaGD0 ORF3 86,7T C ORF3 97.3%-, 6 M T ORF5 2 : 2, 228 G T,56 C T 7 L ORF4bORF5
338 3 ChinaGD0 5 37% (5 6) ORF8b,MERS- CoV 99.% ~00%,,,MERS-CoV, MERS-CoV SAR-CoV 5 ORF4b Figure5 PhylogeneticanalysesofnucleotidesequenceinORF4bgeneofMERS-CoV
4 : 339 6 ORF5 Figure6 PhylogeneticanalysesofnucleotidesequenceinORF5geneofMERS-CoV, MERS-CoV, S N M E ORF4a ORF4b ORF5 ORF8b S M N E,ORF3 ORF4a ORF4b ORF5ORF8b,ORF3 MERS-CoV, ChinaGD0 ORF4a ORF4bORF5, Jeddah( )Riyadh( ) JeddahRiyadh S,N E M,MERS-CoV 5,ORF8b
340 3 solationofanovelcoronavirusfromamanwithpneumo- MERS-CoV niainsaudiarabia[j].n EnglJ Med,202,Nov8; 367(9):84-820. [3]vanBoheemenS,deGraafM,LauberC,etal.Genomic characterizationofanewlydiscoveredcoronavirusasso- MERS-CoV ciated withacuterespiratorydistresssyndromeinhu- MERS-CoV S mans[j].mbio,202,nov20;3(6).pi:e00473-2 5, [3]ChanJF,LauSK,ToK K,etal.MiddleEastrespira- N E M torysyndromecoronavirus:anotherzoonoticbetacoro- MERS- naviruscausingsars-likedisease[j].clin Microbiol [6-7] Rev,205,28(2):465-522. CoV [4]KayaliG,PeirisM.A moredetailedpictureoftheepi- demiologyofmiddleeastrespiratorysyndromecorona- virus[j].lancetinfectdis,205 May;5(5):495-497. : []degrootrj,bakersc,baricrs,etal.middleeast respiratorysyndromecoronavirus (MERS-CoV):an- nouncementofthecoronavirusstudygroup[j].jvir- ol,203,87:7790-7792. [2]ZakiA M,vanBoheemenS,BestebroerT M,etal.I- [5]Coten M,WatsonSJ,ZumlaAI,etal.Spread,circu- lation,andevolutionofthemiddleeastrespiratorysyn- dromecoronavirus[j].mbio,204,feb8;5().pi: e0062-3. [6]Coten M,WatsonSJ,Kelam P,etal.Transmission andevolutionofthe MiddleEastrespiratorysyndrome coronavirusinsaudiarabia:adescriptivegenomicstudy [J].Lancet,203,Dec4;382(9909):993-2002. SequencingandPhylogeneticAnalysesofStructuralandAccessoryProteinsof MiddleEastRespiratorySyndromeCoronavirusfromtheFirstImportedCasein China,205 LU Roujian,ZOU Lirong 2,WANG Yanqun,ZHAO Yanjie,ZHOU Weimin,WUJie 2, WANG Wenling,WU Guizhen,KEChangwen 2,TAN Wenjie (.NationalInstitutefor Viral DiseaseControl & Prevention,ChineseCenterfor DiseaseControland Prevention,China CDC,Beijing 00052,China;2.Guangdong ProvincialCenterfor DiseaseControland Prevention,Guangzhou50300,China) Abstract:ToinvestigatethegeneticcharacterandoriginofthefirstimportedinfectioncaseofmiddleEast respiratorysyndromecoronavirus(namedasmers-cov ChinaGD0),RNA wasextractedfromswabsof thispatientfolowedbyrt-pcramplification.alcodinggeneofstructural(s,e,m,e)andaccessory (ORF3,ORF4a,ORF4b,ORF5,ORF8b)proteinsweresequencedandanalyzed.Phylogeneticanalysesof structuralproteincodinggenesof MERS-CoV ChinaGD0indicatesthatseveralsubstitutesexistsinS codinggeneanditsoriginbelonggroup5ofmers-cov,whichwererecentcirculatedinsaudiarabiaare- a,whileotherthreestructuralgenes(n,e,m)wereveryconserved.phylogeneticanalysesofaccessory proteincodinggenesof MERS-CoV ChinaGD0indicatesthatseveralsubstitutesexistsamong ORF3, ORF4a,ORF4band ORF5,while ORF8b wasconserved.inconclusion,genomeof MERS-CoVChina GD0wasgeneralconservedalthoughseveralgeneticvariationswerefoundamongstructuralandaccessory proteincodinggenes.thisisthefirstreportonsequencingandphylogeneticanalysesofthefirstimported MERScaseinChina,whichmaypaythewayforpreventionandcontrolofimported MERS-CoVinfection. Keywords:middleEastrespiratorysyndrome;coronavirus;genome,sequencing;phylogeneticanalysis Corresponding authors:tan Wenjie,Email:tanwj28@63.com;KE Changwen,Email:kecw965@aliyun.com