蛋白(多肽)的DS模擬流程

1、本文以INGAP為例，闡述使用DS對某個蛋白從建模到動力學模擬，到結(jié)果分析的流程示意圖。一同源建模1.導入目標序列2.搜尋模板AnJI.RunX-廿二7?DplySLylB.?frIKtWQPiltsBuildUL>iEdittTuclticAcid.UntitkedlBu.ildul-1Eds1FtE-iaiaGLHDP5HGTLPFJG勺BLASTS-archNCENELPVCKFKY4-arahj4ijQAnc4-;'t>yuiriLlAriLyfSI-IUfil亂ifcbL.PretardNme吐tuw匚MILClJ-”BLA$TSerdiNa+JBLASTSear

2、ch*AnAkrHTrAic：ftnKijSimulationKeceiht4-LigandlataiactionsPhaEBacGfihcreStalllolftculsItcrq.iltc-SiailvriI7Se-arcb日'丄丁FOalLTiHDI.LhlsXHl再曰也H4ToqLlQBuildHon-iolagyhNa+1AlighE-equenceto|QjMq騎河芮也3lhw詒!MMVS.5.0J.215S可AnalyzeIrajectOK回詫&+Xl.ndly-zeiraje-d:口>回Vts玖GuildHcmolngyN口No亠.AlignSequenc

3、eLeONo羽B(yǎng)LA£TSejrdiNorrotejnBLfrortttrwpu-ijfrDtaja No NaBLASTS«-u-diCHE肛Sorva).J-昭曲創(chuàng)丫S：jc：i.35C:eilFile-EdrtView匸he-mi-rtrpStructureE-equente匚hrtScriptsTddI-sWiridev/Help1詢12丁l弓M14fl|i訓iMLPMTL匚P.M彳Ui'MLL呂匚L朗F卜三WYE石EE呂門耳號LP£勺R.IT匚F心£呂Y4TG呂Ti；¥ELL】P口i屈；諒；孑丨-五|ildT1LUUI.1U1

4、Ur*1CLILiUULLJI.1UIIUT廠ETT1UL*UUtAlignSeque-nEEto*'AlignSequenctito3.模板選取PfiMmeterName：1PfirdmeterlilueInputSequcncKNGAP:UntidedlTnputDjtabjEEPDB_nr95OrganismE-u'lueCutoff10MamumHits1QQ可Adv心riced_Shcfli-F«twtcrKtlpSuCCE-5550hits:UntHedlOiDDzSSH三月？16:17Ioc£lho5t441IDmodBl?:Unti'

5、0:D8S7雋一三冃ai5s斯io“iih加凹列肖cce-s-s7B.9%identrtuUD:DDSTH三冃m1513CIo»lho5h£i943Succaas50hit穿UrYtrtiedl0:00;B8處一三月3151loelhomt槨朋mSuoceis7H.9%identityUO:DDBH三冃315i4loE£lhD5t943Success70,9%dent.txU0:0106)S目?tlactlh.)stSJM3Sijccess-50hits;UrrthJedli0;DDJ50H二弓320S7locAlho5L9943fiiirrng3nDfrrtiT

6、iHC!TNGD-DlriT母穴二曰1l5-1ClnrfllhhO<MASuccessSuccess-Gucce-55SuccessDi-5ca«rySludia2.SClienthiltEditLlieirirttr；DTdtryTudi口2.5LinnrfTVU*in>-31lf'T-?tlJ£i3riLIUrd.il«xd.14ILIU:r±iL&LQ.Lkittbd<jT：.NAEL5COL£0MLFMTL<：H.H5WHLL3CLFITLjWVtGLEjQKKLPd."nP.IP-AT

7、ialyxi£fxudAriBlyxv.FxIild.C'u.ifn-iqn口firtndSlxartariTE4-it*inIroitLDtkjahiF丫爼崖OriaUlhEtihcti險呻nW5ibySiijLiTLl.yRlhamaau-din»FriL»in尊Lhrpl吁S+rl»BUnhtledl0ProtocdINane'z-tfr'frd'itdtLBDfft也ikElapsedTimsitirtDjte，StrverLucatim-+：Mg中aTrajpG曰ry隔EliCC&iEG1£IO

8、rris-fdlNG.DiflK月二三月衛(wèi)15JSIZha曲!Mm+At*la1-26"TrajpG曰rp回仙EliCC&iCiElfllIrfriwlNGHJCrtTU二三目A15dJIZha曲!MM.-if.：.亠mn匸2rThi廠n.gyiahna-Id4站上一.込XQilalcylOdilL5tUiIluUi-Ql-UX£iI*fckbliodrIndexNameVifUe-1.IUHK0T<rIfadtledLQLAittkdIUWAt-abwhariiProtocolNrn?OetjikEhpred亍吋StartDate"ServerLo

9、cation亠+.AjulyltTfbj亡ClCXFy血100FrifrewNG01DO55雋二三月4M聖lx川a購站？+ArwIyH亡T“i亡clmp0*sID!FrirrewlNG.01DO57雋二三月41514Idc川a理1站21T1J、*El“s.r'l-lk、4e亠4L.4.序列比對I5jkTkoTAndlyzflTrtjertoryFrflrnt5；N6.lrDscciverystudlci3.5ClientioniLece-ptar-LifamidlInternetianaPhiarBiflCdpStOEe-sSbaIIciIL«：4ziu1亡丑Verifyl&

10、#171;d.«ls-F2E-drl:dmChQfriijdrySwquwriCbChrt:Script-;TooleuUndaivHalpProtocolN-smpSjv?dDetail!：EljpedTfpStatus-No+AlignSpquenceto口Nu+BlastSEirth£ervErcii：all-iDs-lS9J3®y3.5c0.12158:口TeEpZ巴-;nSequenrs2«4'ihcuc«&bySLtilttTLjStartD-ateEeri.irLocatiori423%idcntilUDd0;56

11、b9hrt:UntitledL0!fiOd9HE四月9&遠hcalhortflaag罔三陰9UilCI口胡J罔二三月415Clo"lhMiW3厝QFrapwr-tPrEt«3：nSUCCP55Succe-ss回伽UCCB39101tf令母窯&X>:iSfrciia.'Dlc-cuiles7*rif7Fr七d*IlKHiILERjVAfLfjLfaUjJiiPrtLliE-ai).悍r*P呂IF7&ilQFil<tQuryOrJ-LMthuei-etbOtuLU££J4b't辱燉陥必酣51711Iz*Li

12、nniiD畫EhCLtlJLQAlsTiS«qaw.-=BiulJStruc-i-nr-iTAniHyE-i£：iFt扌:iaCanx»rvil3diPaht«rni*05i!iiProtsinsDuil>iJkfulC'gyMdd415.hb-riHASTfi.ilxLa£mdIwiflili-ux»FSI-EJAST2aruhla3JmiLiJyr-nDklsrr-tla-l»1L»plati-iiEaaoh£id*Ch<iLiMinanit«u*rlRi-Ein.

13、71;F'r-aLbstiUodlAnlLt-j>iyAflilyc-iTruwiH-ilrui-Fro-t»ihtUrttkdJ汁LLMDAKatschFC6Lh-itttadl嚴LLMjAHuaidtnJI-li1HtcIalrAaadBkDld-Uld£>111HL4LJiS'Wfili加伽K«|Ui;n"真皿中l(wèi)oljaDG.x.Dackudkriilya»0t5JCkFlPt亡i=TLP5HGTKWP.DFMn.KDyNEraktaHfdIv審I-aJallPinJlertplilesxiswciLAiK$

14、亡ud.«rHAJT5亡arrb.OOIEexyfr)Stltetctntaract亡kiisft也tk亡prrtpntmdImJEtTUJtTirm血”AliffwmiUnctfeai嚴LuKatHrtiarFffi"NC5eIeGVilt：W555E555AT.¥1W1G5¥"¥UWIGLHDLHDalndl*；SLUaLsili.CacLCtlParjrnpbrrNampParamr'trir'i.i'iBlue-InputRnlodel站quinceUntrtJedldUntdedl】nput"

15、;femphtetrudurMlU70ATv1ffbfeSCreateSequenceProFileRaise上ilinStrudturefTrueShtvFxrm-iLarK«lpTFV-lVF5.構(gòu)建模型Sludio寸5BulIJ.«ndEJltHutL-aicMidBuild,xndEditPr&t<tnCtliA*!Hh-kIc-cFlc-ihlsniIdHibdIoyNdLiIx<血*PratucolName：Tsri£jr岸口LrtttdLLTAMLrrtiedDbli0PxaLblti(luwyOtrlin亡Ibl百h&s

16、亡三Fre-Ivsji.E«pwh.Prsj-irtTtoleini3ia.ulajtiQn|KcceM0r-Li®aid1strxitici皿斷FIlaj：atGe著孫uullIclBEUleEFTaaliQiUlpiaridn«l4£4>IU4Tf：4Enhly-EBJ'fqIi-ld.Ccm-irvaiLiSuperinbpiystFrot亡LiraTii>ltemla'i»uiluVttifyJfotftiniWDELai.VtrifvThNta.LD.iFT-aialii-JD).Search.i-Ldt-

17、i-J-fiinHajLUXLmdJrifani.FroWi_alodelDoiJciuviLyzi打心寸匸mplu述glyzTf-knsr-Jiibro>a際血Lhttkdlrl心Urtked?*LLWM.PiraiTiete-rPdidfri±t±rVluehr>jtSequenceAligrrnentUntitiedl.tfiriirsJiite&lhf>jtlecriplatStructuresiuwOAhputMadalSfiqucncAhtidvdl団CjoppFromTeinplilesE9NurnherofModels5Ciptim

18、izjtiDnLn-ielHigh日AckntedCutOvErhdKigsTue»DifuHid*Drklge;£y?4Q-rCysSljCCyp&fl-<y51Tl1TZy?14GKyCis-ProlintsAdditionalFtrjints-®RjpfneLoops-Tut同ParjHtlPrQopjjingFall?B-uildHoimologyModQ-口罰Ii5h5L汁：YALIc：LHDF1<；LH0P1uLOliRDFHCEKDNtND*£tdtu$et£isEkpstdTime=tdrtDateTEeru

19、itrL口丘血11QNoSuccess42.3feidtntitY.U0iDM7KJI7!：:localhD5ti9943No'luCCA-Sfi47.3%汩抽現(xiàn)UEDfiSiS雋三四月口“工lacdlhoEt943NoSuccessB9hrte=UrtftiipdljgiD&4.9*.=!»：:：locjlhDsU994審上dignSequence+AlignSwqufrncft由BLAST5tard-i動力學模擬1、選擇ser36-val175部分日0*INAP.rewWI麗1皿0"白gjiMINGAP&0/SER.36tEH/mFAR.G37&

20、#177;回WILE期FF0/THR39&0/ME"&EJ«PCMl莊0/MFGLN2E回沖GIY恥FT回沖EER44&VAL45&0ALA46tE底imnwTE回"GD華FF05ER49&EJaIRSO&0/CVS51空H/MFTYR52匡回"ER52FT回舛LEUS4.I.-hLL屮丨HI"f!IFGAF_P*_1ej1stssQ6dNGAPj-uIQCQfi1LHAMGAP_nwXTJGAF_m;.Mi：PJGAP_nwM.NGAPjreMINaF_new.沖NGAPjre-.wINGFWW

21、.HIU,JGAP_nw.>1.dMLPMTLCn.M5WM顯尺畠IkIH"aIaHte門MMLPMTMMLPMTMMLPMTMMLPMTMMLPMTMMLPMTMMLPMTMMLPMTMMLPMTLCR.MSMJMLLUH.M5WMLLCR.MSMJMLLCRMSWMLLCH.MLLCRMSWMlLUH.MSWMLLCR.MSMJMLLUH.M5WML|DM4MlUISCLMFLSWVESCLMFL5WVESCLMFLSWMESCLMFL5WVESCLMFL5W¥ESCLMFL5WVESCLMFL5ESCLMFLSWVESCLMFL5WVErIMFIClUFERRR

22、R&»"A432、prepareprotein（對模型進行prepare,處理蛋白并加力場）|AaQNhighr亡*l.iwtHGAF_rerfJJKAeuifrciteiriShirClRwcrt即0宅曲十*IM口LtpTMTS-'flUhFCMTdL口總±£-*tXapxviacLvltbLuo.Cd-cal-steJr&teLn.Iwiiibiy-n4iilSesLdiiep£r'T-it-ias.r；ipcr,lf'x-ipuifrt-iaxHjEkO:emo*enol£15B.ecff

23、ptor-LiEanjfiIaiterarijtis-fftsphora-jsSmallMdlecnil>EisS-raylyToalji|窩：StVl-孝審客&X：iPfllcilR.jfcIKkSiP匚!TrnllLn.1<E3TI!L-luLLXyr>iTci-n*horrEudmalhi.-pr-ia¥L-n*«HUHBcy£Lh«aLructuii.ARG-57:LE3BFHR39W.041GLN42GLY43筑gVAL45ALA4j5TR4?GLY4BPiaR50-LEU54帀i-ErtmBSterunrprVski

24、BInputPrrairJNGAP3fi-175iNGAP用BuildLOOpGTfdd卬ProkiriflieTrueHAdviariredRn>.DpLiciTTCUiE«lKilp3、minimizationoffiEJidroihiliEi£d-lJjwwG»mii*TrnxivrSvgumoathirl3»liJjSndowjj*b檢*0£打但=311說：£4站.問n円m&cdNameSiwetlDeldkRacfwii!*幗ueUCC«ll*>jWHciHCjioModsAccess+.Mmn

25、Saqyno*冶TampxiJ27*討ml卸1忖3andWffltideH：DINSiP.m用用isTempos&JEM55+：PtbbeFraoiin*EkildHurwlowMcdiiFnpvKi:JDnwdili：INGAPHChiiMnkiLRradixtiMilSuCwflEEMDL1-33Mkcn.j'mfltPM>N血JlJwESlP»PWJ95HJWECwrirdre也rrfetrdTF*too«iidii54A5T5b-eKiTSCDI£«ivwr；i+.免朋T£rrh»5J.AST5«

26、;HShiTM：Bl&e(wr；i*&I.ASTSuch£U££<&UCC4II5ThfeicMAF.iHbtUtz嚀誣E左除idanlit2i»«qu«iEvWWi5T4PP.IimMISIMPRT田wlygahiTinryGsk疋曲ctiib&o幻iE電flyuIRodEdl£ucc«ai5U£«5SS1C«55Euhkk£0LFnnn:AWMHA£00TrnruAW-NPAa5U33n&lEm"】

27、4;«MZWZMkjAwidUIMVWSIf.AruaT-df*rlaryrAnauVdyKlory2(UDn*MTMi:a>rrb£ucc«aiIriA爐*dMaliciivMrirTisBb'SnLTM£lAPj3-ipirMijAPSmanMNniSnrOil>Enangy匚hingvSorefirwi呂frcqucE3ImpSck虧出EiMcrfel圈NanbarLkxFLadiM&UK4II&1CM55SuccessAigorttimHM5C-rjidkn口占edri注PrepareProteinnIn忡El

28、郵Vru3itt|-LodeadjSuhihiaryHeiiniibtxndfhn!ZCfE5driiMm叭imfroiuihvafim-rhrt>VKh>Hpr*»i出i2frwni.d>w*FpraiisvilMi«dGfaDetail*”,屁”|汕¥申仝住忙乂，|;卜P、I二陽fE3血址jMp口H田尹IM2P-*:；蠱+QM-MM-&milMo-KuhlsAQMET圧口fMfiOEWiiw*?+OvniriljurpowS*l_JLibniryJkid-iia+口Ubr-ary口占小BoQ5郵E口儷Tida口rrsfcix0Til.

29、口曰DktiMrpEhdb*OMatfQfflrfiKuies*l_J1*jrTn«-=phsm+口RKipiorUandInKr-adanfE*SmMjOPE»-l_J»OChTralc?EQEiedradiarii：5AEn«wmdMrirrisilKn滬CdricddMErMry滬CJcweEhwDRl_滬CakdafcabriTBcripnEnirgy護MrniEUiBnETt¥*e治QhtdiphU$MIU«Bmc-wmMwrpt.rtii5Un-BHai12.1.|'1=»PPMituoiB-AI3ID9

30、CBcnlVcfwcti1-J9.I.IZ1M-海H血VCiaHJiXMtiilPitfMgu(W£=TimaIUHfMK:3ft«rriiJiTithUlW2550EiichwiTirmHOIHiSlanvVriibbTagad”亠taiirrMLciidtal=bAfirXli«i=B時處Inj帀茁看Kimr:callchQ|RaramcttirNameferaiiTieterValueInputTypedMoleculeINGAP_prep:ING/lP-MinimizafiiernAlgoirithmiSmarlIMinimizerMas璽mpt5000R

31、MSGiradienliD.lEnergyChangeD.OSaveResultsFreque-ncy10003ImplicitSolwnlMcrcH凹NonbcndLi?tRadiusDi'stancrDupmiduritDidretries14,0HElectrostEfticsSphericalCutoffI-AdvancedMinimizationConstraintsApplySHAKEConEtraintFalseINumberofPcs匸essoirsBUameForcefieldInitialPotentialEnergy(kcal/mol)PotentialEnerg

32、y(kcal/mol)VanderWaalsEnergy(kcal/mol)ElectrostaticEnergykcal/mol)InitialRMSGradient(kcal''(niolkA,FinalRMSGradient(kcaVrnolkAMinimizationCriteriaINGAPCHARMm545.S8767-2745.17219-1034.53S06-2367.49SS414.7Q785Q.09677RMS<0.1C0NJUG>Minimizationexitingwithgradienttol&rance(0.1000000)sat

33、isfiedl.4、heatingNiuinizatioiL0DynamicsQte-ilingwCooling)Q£3RarameterNameRaram&t&irValueInputTypedMoleculeINGAPifNGAPSHeatingSseps|SOODTimeStepO.QOJInitialTemperature50.0Tisrget"Femperaiiiurc-3G0.0AdjuiEtVelocityFrequency50SaveRjesultiFrequency|00D創(chuàng)ImplicitSok'entMedelDistance-

34、DependentDidectirics用NonbandListRadius14.0+EleclnostaticsSphericalCutoff日AdvancedHeatinqConstraintsApplySHAKE匚口nslnraint|thj&1HeatingSaveFfestartFileTrue1HeatinqRandomNumber£.31415931415&3141593141S9NumberofPro匚實町口itsNameForcefieldStartTime(ps)EndTime(ps)InitialPotentialEnergy(kcal/mol)

35、TotalEnergy(kcal/mol)PotentialEnergy(kcal/mol)KineticEnergy(kcal/mol)Temperature(K)VanderWaalsEnergy(kcal/mol)ElectrostaticEnergy(kcal/mol)INGACHARMm0.000010.0000-2739.314424.2269-1248.3261672.553307.79560-881.2306-2452.87736P003123023T在300左右4mmnJedEQJ5、平衡Dynairiics(XeatingorCooling)DyitimLceCEquili

36、brationj*ParameterNamePsram&t&rValueInputTypedMoleculeliNGAP-flJINGAP-EquilibrationSteps5<W0TimeStep0.002Targetlemperature300.0AdjustVelocityFrequency50SaveResultsFrequency10000ConstantPressureFalseEImplicitSolventModdDistance-DependenitDielectrics3INonbondListRadius14.00ElectrostaticsSph

37、ericalCuEoffSAdvancedDynamicsIntegratorLeapfrogVerletEquilibratianConstraintsAdplySHAKEConEtraintTrueEquilibrationSaveRestartFileTrueEquilibrationRhstartFileF:TNGAPMDDynamicslrleatingorCooling2QEquilibrationRandomINum1415fl314159314159314159NumberofProcessorsCO-1.125_=-irnnnnMonnnclnECuooooGoG0-GOQG

38、01C：=lhcelHnlpChoosePilotAxesDataRowIndexIndexNameVisibleAngleEnergyElectrostaticEnergyPotentialEnergyVanderV/aalsEnergyBondEnergyStepTimeTemperatureTotalEnergyKineticEnergyReferenceNameColcrbyDdt.赴Series-t875-1.5LILI-t250-1.375-1.625-1.75U£03040IndexPlotsel>=ctedrmsBilly6取樣（這里以10ns為例）Plarai

39、TieterNameParameterValueInputTypedMoleculeINGAP-2iINGAP'.ingl.aIipmiiiriicsi'Equi1ibration)-J1DynamiC5(Froduction);|-Production5000000TimeStepRestartFile-F:NGAP_MDDy3mic5Equilibration_2014J&1.,TargetlemperatLrm300.0TemperatureCouplingDeca,.5,0SaveResultsFrequency100&0NVTDistance-Depe

40、ndentDielectrics+NonbondListRadius14,0+Productionlyps+ImplicitSolventModelElectrostaticsSphericsICutof?AdvancedLeapfrogVerletDynamicsIntegratorProductionConstraintsApplySHAKECo-nstraintProductionRandomIMurrbw”.”True314159314159314155314159TrueProductionSaveRe-startFileNumberofPro-cesaors三.MD軌跡分析i.勢能

41、分析DataRowIndexIndexNameVisibleAngleEnergyElectrostaticEnergyPo-tenii自丨EnergyVanderV/aalsEnergyBondEnergyStepTimeTemperatureTotalEnergy_=-irnnnnMonnnclnE_(:'§'uoc二uoooo(_uo_u_|二01C：=lhcelHnlp2.RMSD分析KineticEnergy血ferenceNameAxLialyzeTr=1jectory|ParameterNameParameterValueInputMoleculeING

42、AP:NGAPPropertiesTorsionAngleProperties-RMGDRefe-rence-1ScopeMoleculeLabelRMSDto-confAtom<3blipEackb口ngAtomGrouptoFitBackboneSaveFittedTrajectoryTrue一TrajectoryFileEnergyFileFrameStartFrameStopFrame1ncr&ment3.RMSF分析（氨基酸殘基的柔性）先根據(jù)RMSD等，選取平穩(wěn)后的構(gòu)象這里假設取最后100個構(gòu)象。VisibleReferenceNTreNameTimelbLalEner

43、gyKineticEnergyC«nEorwraCanf-aiC->TitQtTia"3W.iCanf-ara1C>XritQtTi"raIIPotAntii-I.TTE*"Ia.t；£a>>oiILec;iLleirE.hceptoi-LieandIntsrticin思疋ctphoT耳吁SmjbI1Hol洋匚©lei鑫JUL-dyiti：血鈕叔唧Clui卒r-3h-a*fl«1-3FileEdit¥irvChemisbStructure15equ<«nctChartSicn

44、ptFTools則ndawHp'爭些會喚xX'+.AlignSequencetoMo1-lMi-.VBjLwe口m*=e4、*”衿話8*£昏白霸I鳳m寸5眄i.”PlBuildl-lofriDlDgQjhQNqSuccessServtrilQCdlliAEbffiJBv35jO.1215Q甌plhTF甸？?IoFFPratDEDlNametfh'edDetdikElpfedTim,madek-UndoiODIdentityUQ：D0j+7j1:>dliIce曲TixaSiri-alit3dti>SaI.UpChlSU'ALUUCr-tah&

45、#187;Fln-ITrjtctoryTiatipJ.CompciDitni：Jut-ilyiis.ClientZll-rtV-Khtij3EVclx曲la廿；nohtidnohiJClioid.JlIeBond為爐£Cn&rvvtLmStatusSuccessStdrtD血StrvtrLoiZiSTtiunffl=四月9L5品Icicfilh口"朋朗HE四月9L咄申珂恤曲艸田二ffflFicIh:山E*amauLilyiuT±*«jct<UF.Ctlaallit-bVij-al*HM«nlCUcoliA電EpliustriGyr

46、aLi沁匚lIodlIi.t4Txit-irbQhijdEtht歹.DL-L<b£iLDusULbiiUDJiPuhlLqil.Ql'tlcI1|ibgaf_EDI2,ODD3.000i.ODD5.000113.L百5412.5EB.J32T.1L6T9.WIHCAPIH5XF-IW5KF-L.655.2LPrw'tstopm-i»Lncrw-inlCmcil_;H:ty：y-二拉z>：茁;；*:jA：y-ZAZA:_；:;；_攔二竝二嵐Cor'i'fcfiTiatiOilCor,ifOr,friitic>nCQrrf&am

47、p;fTiJtiDri匚口rTfarmatiDnCarrformatiDn亡口nfcrmatiDn匚口nfcrmatiDnCorTForrrjfltisnCOr'ifcXiTlfltiDilCor'rfGfiTifltiDilCariferfTiatiDnShrFwmdtiDn匚口ri+wmatiDn匚口nfcrmatiDn匚口nfarmatiDnCdnormadisnCor'i'fcffTiflilsnCor'iftxfriatiDrjCar-rfer-rriiatiDnCarrferfTidtiDn匚口rrformatiDnCahrFofmjtiDn匚口nfcrmatiDn匚anfcsrmatiDnCarYfcrmaticin|Cor-ifGrrnflcion取平均構(gòu)象DMCFbirildSifffGjTiltfcflUJ.!FXii!TrvJiiEtHriFIHG世*一曰-»Q口-J.525T目iwgMk囪ftp購HaHTiOfikEfiitfllfD&rweiySsudb5口耐LEUL34PF.OH5-AiNue-Hzrf*1LT朋LM曰&.科4mlyiiiH曰TZ:他曲宓祖r©西hm二Ti_l軸佃血<i曲p喚mM鼻Dkositfy矗皿。3