1、運用NCBI數據庫查找目的蛋白序列或目的堿基序列第1種方法1.打開NCBI官網，并選擇Gene菜單，如下圖。%NCB1NAjnalCenterForBolechnciogvInformalionNCBI而色ResourceList(A-ZJAllResourcesChemicals&BioassBs流a&Sot出ergDNAMRNADcrnains&StrucesG日仆的&ExpfBSBiiQflGBnHtic&&MedcmeGannmes&MaosHorrwlngyGene*|AJIDataibases*一AssemblyOiQPno|

2、ec1BioSampletoNCBIBioSyslemsBoaks':印l卅teii把咱yhlbuiiMiM由出增訛h僮去xdlh«dJth幻沖力色的4值CmvarqgenomicmkirmalionCloneConservedDomaindbGaPdbVarESIGernneGEOData&elsGEOProfilesGSSGTRHamolcKj-eneM?<KnSearchLderalureProteirwSequence.niStysi4TaxnnaniyTrainingTutorialsVadaticnSubmitiormamj鈍沖值sDownloadT

3、ransferNCBId瀚atoyourcomputer±LftamFinddxu<rnt昆印dadas«orn就chatutorialPapularRescufePubMWBookshelfPubf/ledCefTbraiPubMMHullhBLASTMucleiOliile8nqrn.SNPGenePrgriinPubChemDevelopUseMCSI陰鼻andcodelibrariestobuidipplicalionsAnalyzeIri4ntrfy-anNCBItoolfuryourdatawmI彈is1askResearchExmoreICBIrwear

4、chandc4llatKiratr即用ctsNCElAnn-ouncendbVarqtnew1000Gefiomes由Tlh*h；W+i>hJhr?fl1IFAuguslSlatMCBir.2.在搜索欄內輸入想要查找的蛋白名稱或基因名稱，此處以基因RSK2為例，如圖。tiewsourcea&efTOftiit10Mlspe<rpage.Sflfl域RsHancg，Filters:MnnaiMHhfi0的超8知所孫孰M4巾即3IB日PrQteln<flolngSoqwncacMilsntCCO5r相聯R遍Sutil's,CurrentChrMiiDHffnckK

5、-aUonsrnoiB.口用融IWW"的血口用血6DidyoumeanRSK2asapenesyniWSeardnGanatarRSKJasasyrnlnlPane1trillKerf>Last»tiime>lGcneIDDescripliDnLoczrlianAiauesMIMRPSGKA3D6197rribci?CHialprvin舜kin#sA?|%tiq蕤”g(humanjChrommmflA.NCJWQ打11IK14991120K75T4，Mm"Eg叫CIS.HU-3.ISPK-1.CMPKAPK1S.MRX1$,RSK曄3SGK-alphs

6、J曲aSK2.pRSKa-WW5即血。©110651iribascmalpralainS5kinarMpclypogde3尼日eusclJuh4houMmouse)ChrDm&umaK,NC期10盹711592556961i936fl244jFMPKAPK.1tIMPK-SM2SK>alpha3p90fiSK3pO?SK2LIS6MI卻msiRitnis4malprc”i力爭5kinswIIl&MfiCiSi'fiE白n文曲酎iriuilt|ChrtrnwmeX,NCJ0043&M1529964325363T1)IMel_CG175»,

7、CG17596Dm白冠G仃5M.RSK.R$k,5K1dRSK碘.咕kiik2日儂效ilk(ibaa(xnalproleinS$kinase.90Mg.笛ypeptig3LhornCQ1Mrshek.rIhK一界r.,'ir：KErik冉SKI蛾.快船ii工幽.得心Seairhne-sullbsrum，；11to2QM如3G'S-hIb。1digetantm白dor忸miRecenl)時gQRSK2AH口間網£5HamosasHHEnbo3.在搜索結果中查找目的蛋白，欄會列出目的蛋白的各種別名。SearchresultsFindriii9lvcl日融才口削儂專gSel

8、ectSearchdeullBRStSUmFuldauSesrch注意Description欄會標明物種信息，以human為例，AliasesItems：1to20of203O'e電also1di-ccmtinueddrFiplaceditomnPag«1of11N&jd>Last>>flame/GeneIDDescriptionLocatiGnAliaisesMIM._RP號6KA3ID:6197ribosemalproteinSGkinaseA3Home?sapenshuman:ChromosomeX.NC_00002311(2014991120

9、267514complerrient)CLS.HU-3ISPKd.30007SMAFKAPK1B.MRX19,RSKRSK2SSK-alpha.p90.RSK2.pp90RSK24.在Searchresult頁面中點擊你要找的蛋白名稱(Name/GeneID),進入頁面并在頁面找到“NCBIReferenceSequences(RefSeq)”條目，并在其子條目中找到"mRNAandProtein(s)”，查找mRNA堿基序列點擊NM,查找蛋白氨基酸序列點擊NP,以下以查找mRNA堿基序列為例如下：-NCBIReferenceSequences(RefSeq)tRefSeqsmain

10、tainedindependentlyofAnnotatedGenomesThesereferencesequencesexistindependentlyofgenomebuilds.ExN的G&nomic1.riG_007488.1RefSeqGeneRange5001.121722DownloadG-n日anKFASIASgQgnceWgweHGra口hies)1.NM004516.21NP0D4577JribosomalproteinSBkinasealpha-3Seuid-ntic-IuQt®insdndttieirdnntxmtRdlecdtionsfoWNP0D

11、457L1N,ucleotideAdvanced點擊NM-004586.2進入頁面后如下：NucleotideNCBIisphasingoutsequenceGlnumbersinSeptember2016Pleaseuseaccession.version!ReadnrioreGenBankHomosapiensribosomalproteinS6kinaseA3(RPS6KA3),mRNATJCBlReferenceSequenceNMD045S62FA2TAGrmph記號Gt?to.©LOCUSWM_0C4E867723bpmRNAlinearPRI03-而R-2。16DEFI

12、NIIIONRomosapiensribosomalproteinS6kinaseA3(RPS6KA3>,mRNA.ACCESSI0W0.004536KM.939339KM.944112VERSI0F0.004586.2Gl;56243494KEYWORDSSOUBCEORGANISMRefSeq*Hbitosapietts(hman)Hguci3a口LeneEukaryota;Metazoa.Chordata;CramataVertebrataBEuteleostoairMaMialia;Eutheria;Euarchontoglires：Primates;Haplorrhim：Cat

13、arrhini:Honinidae;Homo在頁面下方會列出mRNA的序列:ORIGIN1611211812413013614214S1541601eei721761S41901961102110811141120112611321第2中方法1 .進入Uniplot數據庫，在搜索欄中直接輸入要查找的基因或蛋白名稱，如下圖:UniProtBLASTAJ>gnRjetnewe/IDmappingThemissionofUniProt論toprovidethecommunitywitihacomprehensiverhigh-qualityandfreelyaccessibleresource

14、ofpiUniProtKBniProtKnowledgebaseSwiss-Pnot(551r705Manuallyannotatedandreviewed.TrEMIBL(65.378749)iAutomsdcdlyannotatedandnotreviewed.UniRefSaqu6nceclustersUniParcSequencearchiv&LiteraturecitationsCross-ref.databasesSupportingdataTaxonomy中DiseasesXYXSubcellUarlocationsKeywordsu.-mtIIIniPrntHatafi

15、&ttiinna+artarl2 .進入以下頁面：.13招力ER那Filterby器HHtwH(30Sws-Prct*yr*電乜論師*($TrEMELPopularorganismsHuiunMouu舊Raving川RmI)5LfiogHi1U3.HP5LBI2UfilbiiMikialptrotoin5G-hmdoapqnE(Human'i740*fcinase-alpha-3MiSJPpPKIB.RSK2DidVMJmcBfi或273.J73<wult：s：fatnd加1DtfwirDrgvtsmsPl©爵4it§43_M0uSE*1HlboSOn

16、ilpvuEMiiNbAlphfi23*11國iRpsgka-rnijitak2Q15349!CEftA2_HUH4H為4ib»oma4pmteiviS&fcwia&e4lph«!2CPS4KA2M&PKmClC.K5K3-Homo-sapiEns(Human)733口Q9WUT3PC5ftft2_MOLHE斯fUboxinMproteinS&H>nasc-alpha-2Hps&lulRsk3MmeuhuLkJiMitxsa733o理&的N修iT心中XiAfJiViiil-HdpmMWiiUnacft1川Ml笄PAKM2

17、JfifiIP27341wos.MjwysN斯*tgE-acthfMedproHem1*4PUJEWCJ,FRKM3HwnnsjpiEns(Huntan)379PfiHDflSH<01_M0U5E囂pnttinkHiaxe1MapklErUrM叩匕PrkmlMjsmuscijlm；Mousa)1S8TuwW.ITME他il'lTADisplayEntryFeatureviewerFea&uretableEFunOWN臺m白看&Taxonomy3SubcelhilarlocationEPath&lpgy的tech0PTM/ProcessingExpressi

18、onImeractionQStructureEFamily&Domains日SequenceECross-rerfefencesNone3 .在搜索結果Entryname欄中會列出不同物種選項，且會有Proteinnames及Genenames，點擊所要選擇物種蛋白，進入以下頁面。UniProtKB-P51812(KS6A3_HUMAN)%blastQFormatAddtobasketGHistoryPi-y:u1|Riho&oinalprotein56kinasealpha-3GeneRP"KA3Organism也加sapiens(Humsn)StdftUS|筠Rv

19、iwd.AnrtgC?tiQnStore：.回.國-£即而已或31丫對口匚號MprQt&in帕卬回：FunctionSerine/threonine-proteinkinasethatactsdownEtreamofERK(MAPK1/ERK2andM4PK3/ERK1)signalingtranscoptionfa匚tursCREB1HETVl/ERSlandNR4A1/NUR77,regulatestranslationthroughRPS6andEIF4survival,anddifferentiationbymodulatingmTORsig巾曰lingandlrep

20、ressingpro-apoptotic：functionofBADphophoryla>tionofCRE01andhistoneH3at'S«r-W'fwhichresults.力thesubsequenttraciscriptionalacmitogenicstimulation(EGFandpm點)phosphorylatesaMactivatesMR4A1/NUR77andETvi/ERSltransderivedsignal,acsicdiicttyonthetranscriptionregruladonofseveralgenesbyptioaph

21、orylatingGSK3EresponsetoserumorEGFviaanrnTOR"independentmechanismandpramotestranslaticininitialtioriby匕tcii門與uliri消phosplvarylatesEIF4BrenhancingEIF4BaffinityfortheEIF3camplexandsbrnulatingcap-deppathwaybydrectlyphosplhoryiatingiTSC2at"Ser-US'jwhichpatentlyinhibitsTSC2abihtytosuppreswh

22、ichregulatesrtiiTORClactivityandmaypromoterapamytift-sensitivegignalingiindependntiyoftheFthepfo-ap&pt&ticpfotinsBAD.andDAPK1andsuppr<GS£ingtheirpro-ap-optotiefunction,Promotesmerasfionsetothehepatotoxincarbonttrachloricte(COM),isjnwoHviadincellcycleregulationbyphosphc*ssocistion叫i由14-3-3proteinsandpreventsitstranstocabontothenucleus日ndinhibitionofG1progicrhLq/4EnLrcfiECLi卜門七iuncr9LnE、nlcEnknllu1十匚-sc:CifFaL十crcFE!7mqHint*qrlftTnnc:FcFEntiGE-cin4 .點擊頁面頁面左側的Sequence欄，并找至USequencedatabases,其下面表格中有RefSeqi,其中包含NP及NMSequencedatabasesNoneliBIIMUIMIflselecttheimKdestinations:U083I6