功能基因的序列比對方法

功能基因的序列比對<1>.切除載體和（或）引物打開所有的原始引物序列于一個EditSeq的窗口中exportallasone始UN門Mr"曰j,File]EdilSeardn-SperchFeffturesGoDidiesWindermNetSearchHelpODen...SaveAs...PrintSetup,,-Print..PrintSelection.,sRecentDocumentsCtri+P*hc?口凸wn-iunn匚aj?匚匚nil匚2廠匚4N.hAITFdTf"17ie?始UN門Mr"曰j,File]EdilSeardn-SperchFeffturesGoDidiesWindermNetSearchHelpODen...SaveAs...PrintSetup,,-Print..PrintSelection.,sRecentDocumentsCtri+P*hc?口凸wn-iunn匚aj?匚匚nil匚2廠匚4N.hAITFdTf"17ie?￡F「!I忡MfFHH「口應(yīng)E_Fu1Tn匚STU11UUL?CUCIIC~M廣UHj-R8!!R-rvnn匚ATIT1~7■:-z，EMl~ILIUIM廠IF4-;"irn"們tfghe?《mimF匚LIT勺umc?板匚I°ilI匚0廣匚xal口n11」hJH3匚Hiririohc.euciiEhiruUTiri-WMfJL1W-IM1KEmunn,kECII匚M廣居Mf~SlJL"Id-RJlN匚HTE01zc,虹「FH.I匚IN「匚”了n”waeILWIF&TElum?EFrHlFWF色"TEW-：!7廠W.1R-hA1：!匚ATl~tri『rar.匚匚i"ilI匚％[廠匚m廠林n匚at食noc-n?匚urni區(qū)Mf~E婿ihjl.noiri17^-2KAr-Sffi.-Hi3FA7m哼匚e.CTCIIUkJLW容MD91D.1759-3_MCRA._22-M13F_4T.FD2.seq:SEQUENCE?[>暮0I%跖「：[mport...OpenEntrez如q“.CloseExpert..45erne...Ctri4SL「口％』i3」xjik匚d~runjke?匚匚rtii性Mf~uW：M廣QhQ-fLHTW匚HT■&「1u旦c，匚匚Cll匚2f~UTd■了-#"!"：?￡.1門-函/1#匚ATPiFHv-n?匚Hflll缶^?i_h/T|CiMl~K穗WLEW■蚣◎GAACGTTTffTGSC心心CTGATOnGC海TCG/CHAA皿T皿ACTCTGO：C€AI'2OICCnGT<^?ZEI^ZAIGGAGA&^DZA&；CTGAGjl-GACCGGC&TTI&ZGIIT<^GTICC<^I'3CZiCJiTGC^C2LCC<^ri<^-mGC-^CiCGGTT&Zl'ZTCT-3?MLTg湖I海neC&gTC€TmCT心TC&AGGGITGCmrrMEGTTGT?河ITCC&AGGTTC砌gC&mST缸TITgSnGMGmSTlSLWm湖AmCA湖TEGTCGEGGAIGKAIZEGLHiIGCA&HnMRLmCHGIGTgmHmmmHCtrl+OCtrl4-R口「l+W您心i〕_i占■、秘；MPK感iIiZfK保存打開這個保存的文件，開始切除載體和引物選擇載體插入點兩側(cè)的序列（10-15個的樣子）搜索注意：不存在正反向的問題，都是一個方向，因為測序的時候是選擇兩個載體上的引物其中的一條來往后測序的!DataEditSearch]AlignmentWebSequencer1Find.MotifCtrl十F□NASequer郭我2白w疝1.\H09LQ-13.M09L0.1蛛Fin如泓F3部FindPireviousShift4-F3神FindMarkedSiteBH*由叩?口,|TiarmWMlFroMnSmimws:cX，!》LE.BCU上4?:F5a.??iJ:cX，!》LE.L'E匿-5BCU上4?:三5a.??ujlSU.「PI技L~=HLH47jam-■mg■mg■i。L1.MMIClLM■:■no.L:Li.明“。cesiyjiIIIHIIIMIIIHIIIUIIIHIIIIIIIHIIIMIIIHIIIHIIIHIIIIllllllll■fllll■IIIMIII■lllll一一一—蛆成方云mm石lh而訝?Il面荷iiiuiiiaLihBiiiiUiiiiiiiiiuaiiiiiiiLiiiiikiiaiiiiii:°j-C&4A.TCTTCCHE-AG-L1WG-takTCTTaCAt^4A.[I心ElMMTTWPmMl耳:?口耳：瓦&耳：?盤耳耳:■口JimMN,ri-ji-fcMfri*gLt-MLW■i*gLULAF■:i：hZM■EHL?XJrJ營?！鰻I?！?偵:iMl?JEM■AHi心7■iiii.L"!U-3女UL￡-ML^FNDll.aan；ICMEiJ立F47Ml；kZZJL4：30iTWE口ATIll|cJ-EBAAEETTfiC■EaC^USCTTCCLCkJLEi^TIIIIN|cncaaATCTT：EaaAiKrlMlHMt&T>：ETCd^lSLEilTlIIcfiC'JTCCAlUfiXTHIH|k：}-C-BAA；TC7TCCPifiAGh1|em<ietcttcc?°i-?ibatIUAIIIBIII心網(wǎng)K-ATCTTC￡!AD3J.E!ATfllllllBIIIl||:acaaA.TETT|AliET|||||||cEiCAaXTCTTAaiiMiiBkis&uiiiikiiiiiKiiikiiiLaiULiiiiiiB.iiiLififiiiiiKiaiK^SLidaiii^SiLiiii&iLiaBkaiiikgiu&iiiiuaiiEimiiillklllilk.imh.llll￡lfllHllailk.l&IHBll>.flailllliaik.Blaik.lliiiiaiiiiii&HiiiiiiiiiiM.iiiiiiiiiLaiiuiiiiHiiiEiaaiiiiigiiiiiiLici^iiuiLimiiiiiiiiiiuiiiiiiiiiiiiiiaiiEiiLiiixiBiiiiBimuiaiiiiiLiBiiiiiiKiiiiuiiiiiiLiakiuLiiiiidI曜盅黯皚盛酰部噓臨嵯噓雎恐IIIIUl|li||t||BJLI.|L|||||L|l.|L|||li|BJLB||||||U?|U|1l|MiliHiaiEig|?￡iiaB￡iiiHiaimi5SHiBgikg|kHex|||i|gi.UI!ll^ilxlIIHIIIiLlIi:::aIIIHIIllllllUlllllllIlLlI^'：3SA7CETCCA3曲”■麗IIH頁幣“亙1￡麗|兩@麗11"|而,麗*"丞|■頁“而|兩“"■麗而?IIIBIIIIHIIIMIIIMIIIMIII>：3：suAriT7.::KSAGAiIIMIIIIIIIIMIIIIIIIIMIIIBIIIIMIIIIIIIIHIIIMIIIMIIII1IIIMIIII1|IIII1￡|S|||||||?1||^3:5?。簎e-：A弭以i:|||HIIIUIIillimiimil|LL|B||HlB|L|g||Bk||Ll￡|||||HIU||iHlllLllIISL_._i■■■■■■■■?iin■■■■■■■■h■■■■■■■■—.sa*jAiIIBHH1M'HIBl111UL■■■，■一」■■■■■■■■h-■■■■一■■■■■■■■■=一，■■■■■h■IIIHII|iLfll|iliH|l|?|IIIIUI|C^diuA[CT7CL^AuA.|||||||||E||M||llin|Bk|||iU|8||B||LBl|||Mli||iL|li||||||ilillt|iliik||l?filIIHIIIMIIlllllllllllr^llk^BfiATziTCCA^uAclllUIIIIIIIIIIIIIIIIIIIIIIEHIIIUIIIIIIIIMIIIHIIIHIIIHIIIIlLllAllA：I—，弘EllllHIIIHIIIMIIIUIIIMIIUlilMIIIUlilMIIIMIIIHIIIHIIIMIIIMIll■llll■llll■IIIHIIIHIII■lll=□=i-at=3t.=.=a,…rIIIUIIIHIIIHIIIHIIIHIIIMIIIMIIIHIIIMIIMIIIHIIIMIIIHIIIHIIIHIIIHHIMIIH=-^^TCTCi=CADx?K7MIIHIIIHIIIIUIII4IIIMIIIMIIIfllllHIIIHIIIHIIIUIIIHIIIHIIIHIIIHIIIimiHIIIHIIIHIIIUII<?>=虹aemums■草l||■lll■IIIIHIIIMIIIHIII■llll■lll■IIIIHIII■fllllHIII■Mll■lIIBIgB?B|l?M|ilBllllHIIIIIIII^=口A"-IIIHIillllKgll>i<|B|Th1|CTIIHIIIMIII&IxI&HIIIHIIIMII=3=r=AT=TT.=.=AESrIIIHIIIHIIIHIIIMIIIHIIIMIIIHIIIHIIIMIIMIIIHIIIMIIIHIll■lll■llll■IIIHIIIHIII■lll=o=w=山rlll■IIIMIIIHIII■IIIIHIII■llll■lll■IIIIHIII■IIIIMIIIMIII■lcTc^^aA^rHIIIHIIIHIIIMIIIHIIIIHIIIMIIHIIIMHIHHIHlIIHIIIHHMBIIHI切完之后另存為

口CreateMewOpenReo-penCloseiSequencesPhylogenetirAnaly^isIHISaveSessionft|ExportAlignment/DNASequencesProteinSequencesCtrl4-S47Flz.geqrcCI47S12.seqrcGd47C12-3eqrcGd47H04.3EqidlCMEGAFormatFASTAformatPAUP^orinsttipTranslate/Untrans\ate而SelectGen&tieCodeTableR.BverseComplementR.everseCornplementExitAlignmentExpl&rerIff-，crr>.rr.rjiitn,47C04.3?qEM47BO4.seiq:^c|ca|47AO4.aeqIlliHII47HOS.seq47GDS.seq^c|ca|47TO3.seq^|ca|47SO3.seQllllll47BO3-misqt^|ca|=gt|||ttcc|ca|47COS.與g147BO3-seqhf.重新打開這個文件，開始切除引物方法同切載體，但是要注意正反向的問題。比如mcrA基因，其引物為Forward:5'-GGTGGTGTMGGATTCACACARTAYGCWACAGC-3'Reverse:5'-TTCATTGCRTAGTTWGGRTAGTT-3'先找Forward5’端，此時只找到的部分序列。切去5’端?！鮊ASequences|TranslatedProteinSequences|MC313C04M弓DOtM海光-M13FtrirCTQTCGGATTC^C?lC：?l^rkrTGGtlGtlGIAGEAIIC&CJiSA^rRrtCAACAGGTECAlAJA^GGJLTGllllllllllllllIIIIIIIIIIIHiTllATACACCIml.”5皿加?IGdAlAldllUllTll^TACACclirlIIIIIIIIIIIHIt||Atacacc|^t|catacaIII^tIlllllAcd^rlIIIIIIIIIIUIIIIIaccI^tI然后再切這些切掉5’端序列的3’端的序列，此時其3’端序列應(yīng)該是Reverse的反向互補序列。SearchSpeechFeaturesCreated:21:17[Gciodi乾]Windows(NetSearchHelp籍Untitled&eq#1:SEQUENCE囹D11<]|Selectkin:-S3=23fl亙甚ReverseComplementTranslateDNAReverseTranslateProteirGeneticCodesEditSelectedCodeDNAStatisticsProteinStatisticsR&verseSequenceIIu2±IIcC^IA-GIIWGGRIASTI切去這個反向互補序列，這樣一來這個些序列就已經(jīng)被切去兩端的引物了。但此時還剩下另一部分未切除任何引物的序列，此時記下這些序列的編號，先切去Reverse5’端。□NASequencesTranslatedProteinSequences|Sc■三亡土白目JAbZiru****||||||*|Illi「MDMCkl7Sm?3MCSAWAM13F47網(wǎng)八非習MD91D.1T5日一3MCBAW：L-M：13F47MD91D.1T5直-3MCBA47CD芝MD91D.1754-3MGaAJ■，一HJ.3F弓7&D芝.meqMQ9J,Q.：LT53-3MGaAJ■有一M13E47HOl.m已qMQSilD.1T5￡-：BMESAL5-1C13：F耳7GOL.mmq"MOSilD.1.751-3MC心且7FDl.muqLMD91t).l"7GO-：5NtSA47民01■與目q9.MDSlO.l^耳日一3MURAH-M：13F厘1?何lt)MU91K：L7心-3:MCBALD-UUF皆RDl.mg1：L_MD9：LD.：L746-：3HCSA9—M13F城7M)」，gIN.:^。910.]7冬日?3IMCBA8?M13F47H12.mEQ[13_:MU9：LD.：L744-3:MCKA，一MJ.3F471W_HUSID.11743-3:MCRA臼一H13F哼7Fl芝.megt-MD3；LQ?374/一3MCKAS-MJ.3E47El/.mmq1=5-MD3；L0?IT弓口一3劉匚您3-MJ.3E街Cl芝.m巳口1T『M0S?18.。宅■日一3ECSA5D-M1：BF耳，H04.misq;IS.M&91S.0^47-3MCSA43-M13F47C04.seqrcc■IIrlr192021232425Z電27箜箜3031323334353-537_Ma918.0€4￡-3MCRA48-M13F47F04.seq一MD3旺一賓駐-3且＜?規(guī)47-Ml，F(xiàn)野一MQ31白一06,4一3:MC口4S-M13F47。口4一旦白口一一13丘43-：3:MC；U145-M10F4F匚口4一日日q一MU51日一0電4衛(wèi)一3也匚既43-M13E4，日口冬一耳巨叫一MQ918一喝口42：-M13F方一MQ918一購弓口-3MC*4-M1郭野皿3一睥[.110318節(jié)35—3耳匚您40-M13E4F—MDSIE；.叮枝3日一3HCSA39-M：13E霸FOm.misq;-轉(zhuǎn)091占.七在37—：3HCaama-Ml：BFED5.m三理_右一之理亡云47Bg*耳:MCRA32-M1WF47CDZ.m目口MU91日.0634-3:MCRAai-MlWF5RD3.與色耳超U9：L8.t)Emm?8:MC3AmU-MIEF4，JLD3.日白巧MD9：L8.t)6mN?m苴匚口ZT?M1：3F47底口芝.日￡巧.宜31—3WCSAZ4-M13F47?與皂qHU9：L8.0630-3:MCa&ZD-M13E弓，F(xiàn)OZ.meq-MD3；L8?g關(guān)-3職您LE；-M],3EVE"m”??M03ia.0e2a-3HCSALZ-MISEi7DOZ.ssqccGIncc■■ccc

...cc

II"--Gc3FA.AATTIT是mA，nrlrGHini■■■■■IHIHI—In■ic:a|?瓦kat孔kAtAtAta|?aI?llllllllllllt~nIHIHI—l_

n_

—l_

n_llllllHHHIcccccccccc——————————cccccccccc■InsSI—llllllHHHIHHHIHillIMillllllllllllllHHHIUllllIlliImlAAAA

TTTT

AAAAcccsiTill

AAAA

SI—

AA.A.A.SI—IHIHIllllllllllllHIHIIIlliIIAAAAAAAAA—AAAAAAAAAAAA—AAAIAIAAAA—AA再用Forward的反向互補序列切去3’端，這樣剩下的序列也都被切除兩端的引物了。<2>將所有序列調(diào)整為同向序列：a.選擇前面記錄編號的序列，將這些序列一個個都轉(zhuǎn)換為其反向互補序列。這樣一來所有的序列都成為同向序列了，即在DNA兩條反向互補鏈的其中一條上的比較了。b.保存該文件卜1SequencerPhylogeneticAnalysisMEGAFormatPAUPF口4*7已凸4一mwq47忠04一月g47HCI3一目g5仁口巴一與白可Ccunplement47三口3一目白q47口皿一米口ExitAlignmentExplorer47C03.aaqBL-_QE34-3MGRA3【一M：L3F47巳口3一metj4*72C)4-meqjDNASequencesProteinSequencesReverseCo-mpletne-ntReverseDCreateNew?OpenReopenCloseupTranslate/Untranslate亟SelectGenetic(ZodeTable47GUmeg：47F2LW一會q：ExportAlignmentFASTAfgTGT1IICtrl-I-S47GIZ.asq：<3>生成OTUsGoogle搜索”FastgroupII”或/fg_tools.htm

FastGroupllFasrGroupHisanonlinetooltoinvestigatethebiodiversityofniicrobialconimijniries.FastGrouplltrims±eDNA/RNAsequenceswiththeuserspecifiedparameters.Tienitdeprdicatestheseqiaetucesandgroupsthe'similar1sequencestogetiief.AttheenciyoucangetthedeTEpEcErtingandgioupinginformationofyoursequencesas.wellaastatistiz-alcsthnatonofthec:ommunityolderstody.ForhelponhowtousetUstoolpleasegotofasthelp.Uploaddata?Uploadyourfastsfile|選擇苴件亍芭擇丈件.PasteyourPastadataThesequenceswillbereadinrmdotnorder.Setparameters匚TrimS'已ndwithoutMindiefirst10basspairsTrim3'endwithoutNinthefirst10basepairs匚Tfim5'endUserdefined||^|of7DJ%similarity□nimS'endv^ifliBact(S17to634}Q|dCAGCAGCDGCGGTAAT]of70]%similar^[ZSequencesshouldbeatIsast300EasepairslongtliMsemetliodPSI(TcrccntSequenceIdainty)of97%similaTityaPST(PeroefitSequenceIden^lywithgaps)of97%simihri^:5<qurticcMarchof%BimilirkyTrimofftheprmersonlyFormalouipnt區(qū)GeneralgrouprifomiatiQnI/Rjepresmotivesequences(F.4.STAformat^VDetailedsreuprifbnnatiflnIZGroupstatisticsIZRiclmess(ChaoLiIZEHvti'sih-fSliaiaioii-WiaLerindex)IZS.ank-abujidancecurvr[ZRarcfacdcncalcula^ciis厘textformat,'■plotl_Alltimmedsequences(TASTAfennit；Calculate!Clearfields（Onlinegrouping--注意勾選的選項）Choosemethod里面相似度可以選97%或98%提交之后出現(xiàn)的窗口如3套口含十71曰jfwh土」」DereplicationresultsNumberofvalidsequences:50Numberofgroups:10G已門已「日1泛口infcHLTDM~ti「nR己rxr己日己ntetiv己m己口u己口口已，。巳t日il巳dEECiur)infuirn日tiuiiGrciLipstatisticsDiversityandstructureestimatesChaol:18.0000ribotypesShannon-Wienerindex!1.7260nalsRank-abundancecurve:GetcurveRar己factiun己mDownstreamapplicationsRDFCl占mmifi己rBl日mtGeneralgroupinfoxmation-NumberofGroupnamesequencesTOC\o"1-5"\h\zM0819.0S76-AJCRA_39-“M13F_47_BH.SEQ"N0819.0881-AJCRA_44-〔M13F_47_G11.SEQ1Mnxiqn辯r-afcra亦-可以看到被分為了10個OUT每個OUT都自動選擇了一個代表序列。全選將其復制到word中，備用。并把其中的那些代表序列都復制下來粘貼到TXT保存。<4>尋找嵌合體：一般是對16SrRNA來說的兩個網(wǎng)站：/FindChimerasOutputs.html(或搜decipherchimera).au/bellerophon/bellerophon.pl(或搜bellerophonchimeracheck)<5>翻譯網(wǎng)站：http://pfam.sanger.ac.uk/在保存有OTUs的TXT文件中，一個一個翻譯成蛋白質(zhì)序列。最后保存。JOTU-IODNA-Pro-記事本1=〔□I—文件舊掘舊格式〔0）晏（Vl幫助［HJ>M0819.0885-A_MCRA_48-M13F_47_C12.SEQAGCCTACACCAACGATGTGCTGGACGACTTCTCCTACTACGGCGTGGACTATGCCGTGGACAAGTTCGGAGGATTTGCCAAGGCCCCCAAGACCATCGATGTCGCCAAGGAGCTGGCAACTGA>CACGCTGTACGGCATTGAGCAGTACGAGGCCTTCCCGACATTGCTCGAGGMCACTTCGGCGGATCCCAGAGGGCAGCCGTCCTGGCAGCCGCATCCGGTATCACCTCTGCCATCGCCTCTGGCCACAGCCAGGTAG&CCTTGCCGGATGGTATCTGTCCATGCTCCTGCACMGGAAGCCTGGGGCAGACTGGGATTCTTCGGCTACGACCTGCAGGATCAGTGCGGTCCAACCAACGTGTTCTCCTJCCAGTCAGACGAGGGCAACCCGCTGGAGCTGAGGGGCGCC^YTNDVLDDFSYYGVDYAVDKFGGFAKAPKTIDVAKELATEVTLYGIEQYEAFPTLLEDHFGG￡QRAAVLAAASGITSAlASGHSQVGLAGWYLSMLLHKEAWGRLGFFGYDLQDQCGFTNVF￡YQSDEGNPLELRGA—^，，fe，M涕，?,強筆?連費爹■鎧'■■農(nóng)宮I>M0819.0861-A_MCRA_24-M13F_47_C09.SEQGGCATACACCGACATCATCCTGGATGAGTTCACATACTACGGTATGGACTACATCAAGGACAAATATGGCGTAGACTGGAAGAACCCGAGCCCGAAGGACCGGGTCAAGCCAACCCAGGAAGTCGTTAACGACATCACAACCGAGGTGGCCCTCAACGCCATGGAGCAGTACGAGCAGTACCCGACCATGATG&AAGACCACTTCGGCGGCTCACAGCGTGCTGGTGTCATCGCCGCTGCCTGCGGTCTCTCGACCTCGATTGCAACCGGGMCTCCAACGCTGGTCTGAACGGATGGTATCTCTCCATGCTCCTGCACAAGGACGGCTGGTCACGTCTCGGGTTCTTCGGCTACGACCTCCAGGACCAGTGTGGTTCCGCGAACTCGCTCTCCATCGAACCC&ATCGCGGTCTGATGGGAGAACTGCGTGGGCCAAYTDHLDEFTYYGMDYIKDKYGWKNPSPKDRVKPTQEWNDITTEVALNMEQYEQYPTMMEDHPGGSQRAGVIAAACGLSTSIATGNSNAGLMGWYLSMLLHKDGWSRL&FFGYDLQDQCGSANSLSIEPDRGLMGELRGP>M0819.0859-A_MCRA_22-M13F_47_A09.SEQCTACACCAACGATGTGCTGGACGACTTCTCCTACTACGGCGTGGACTATGCCGTGGACAAGTTCGGAGGATTTGCCAAGGCCCCCAAGACCATCGATGTCGCCAAGGAGCTGGCAACTGAGGTCACGCTGTACGGCATTGAGCAGTACGAGGCCTTCCCGACATTGCTCGAGGATCACTTCGGCGGATCCCAGAGGGCAGCCGTCCTGGCAGCCGCATCCGGTATCACCTCTGCCATCGCCTCTGGCCACAGCCAGATAGGCCTTGCCGGATGGTATCTG/在用Expasy翻譯的時候選擇第二個選項TranslatetoolTranslateisatoolwhichallowsthetranslationofanucl&oti-de(DNA/RNA)sequelPleaseenteraDNAorRNAsequenceintheboxbelow(numbersandblanksI.AC4C.CAACSAI&IG.C.T.或叫￡0觥一1.1匚1C.GI4QIAC&5C.G.IS5AQIAI就偵叫.G偵恩0ftGT.TC.GE.姬.GA以聯(lián)威艘藪/MC堿版成溟弱溟威&懿堿&.必瓢溥&以受齦偵項好泣［:§邛￡裁就皿做氨麒皿匚叫姬依項匚羔恩C.A.UG.或CGAG.叫攻姬：Q.CG.觥成.皿工球四麟G.C0J.或K.GC.A氨。成AHXGG.IAIC.ACC.I.C.T.GC.GAIC.SC.C.T.CI^&CCAQ^&CC.A&G.TA&GC.G.T.I務(wù)匚偵GGA.T.G&IAIC.IS.IC.GAI敏XS式恕堿.◎姍冬寒您威.蜘或甄爆.A.T.KN.g冬.1版網(wǎng)&以阪服叫甄敲耳依以CQUC尊郵&照丁黑霽叫.如器峨瞬&抵丑夠京霧螂舄帔L4隔叫￡■#.Outputformat：CormpactOutputformat：Cormpact["M","-",nospaces).GeneticcodeVerboseTM^\"Stop^\space^betweenresiduesCompact("M",nospatesResetor|IncludenucIwLdP崩quinceTRANSLATESEQUENCE點擊翻譯53Frame1SLHQRCAGELLLLRRGLCRGQVRRIC^GPQDHRCRQGAGN-GHA面feGLPDIARGSLRRIPEGSEPGSRIRYHLCHRLl^PQPGRPCRJffVSVJUPAQGSLGQTGILRLRPAGSVRSNQRVLLPVKRGQPA&AEGK53Frame2A￥TNDVLDI)F5?VI)YAVDKF&&FMJiPKTIDVAXELATEVTLYGIEQYEAFPTLLEDHFGGSQRAAyLMASGITSAIASGHS如GLAG皿SMLHKEATCELGFFGYDLQDQCGPTNVFSTQSDEGWFLELRGA53Frame3PIPrMCWTTSPTWTMP^TSSEDLPEPPRPSMSPRSWQLRSRCTALSSTRPSEHCSRirSADPRGQPSWHPVSPLPSPLATIR-ALPDGICPCSCTRKPGADVDSSAnCKISAVQPTCSPTSQmiRVS-GA35Frame1G泌LQRYJ1V-LVGEHVGWT虬ILQWAEESQSAPGFLgEHGQIPSGKAYLJW折GgRGDrCCGC^DGCPLGSAEVILEQCREGLVLLNAVQEDLSCQLLGDIDGLGG-LGXSSELVHGIVHATCEWQHIVGVC35Frame2APLSSSGLPSSDV-EmVGFH-SCRS-FKNPSLFQASLCRSORYHFARFTV/LWEAlUEVIFDAAAETAALTOFFX-SSSWGKASYCSMPYSVTSVASSLATSJWLGALAUFFULSTA-STP—EESSSTSLV-^理想的情況是這段序列中應(yīng)該是沒有終止序列的即”-”符號，因此先選擇閱讀框較長，整段序列也沒有終止子的那些，如圖，先選擇第二個。復制紅色的區(qū)域，在blast上比對，看是否是需要的序列，如果是。那么就選擇此結(jié)果，如果不是，再一一比對其他的羅列結(jié)果?；蛘咧苯訉NA序列提交到sanger上，出現(xiàn)如下結(jié)果UFIA?quienc?searchresultsThispageshmstheresdtsafsearchingyourDNLAsequenceFarPFam-Amatdies.Tddothiswcperform占six-frametranslation!:□generate占setofpnotenissthenormalPfem-AWMsmndGAcut-offe.削gwrh?dstaiifddMHpSofinigre^ulHpage*Wehisrve柘Lird1si^iificaithit-ardninsgniFic-anthitsn1frame..Mid史theDNAar?dMid史theDNAar?dprotein尖quent耳andFrame2中有一段綠色，顯示就是mcrA的保守家族。那么Frame2即為正確的翻譯方法。另存為，只保留pro的序列的TXTOTU-IOonlyPro-1己事本文件⑴漏^〔日格式（OJ辭（Vl卷助〔H）|>M0819.0S85-A_MCRA_4S-M13P_47_C12.SEQAYTNBVLDDFSYYGVDYAVDKFGGFAKAPKTIDVAKELATEVTLYGIEQYEAFPTLLEDHFGGSQRAAVLAAASGITSAIASGHSQVGLAGWYLSMLLHKEAWGRLGFFGYDLQDQCGPTNVFSYQSDEGNFLELRGA>M0819.0861^_MCRA_24-M13F_47_C09.SEQAYTDIILDEFTY-ZGMDYIKDKYGVDWKNPSPKDRVKPTQEWNDITTEVALNAMEQYEQYPTMMEDHFGGSQRAGVIAAACGLSTSIMGNSNAGLNGWYLSMLLHKDGWSRLGFFGYDLQDQCGSANSLSIEPDRGLMGELRGP>M0819.0859-A_MCRA_22-M13F_47_A09.SEQYTNDVLDDFSYYGVDYAVDKFGGFAKAPKTIDVAKELATEVTLYGIEQYEAFPTLLEDHFGGSQRAAVLAAASGITSAIASGHSQIGLAGWYLSMLLHKEAWGRLGFFGYDLQDQCGPTNVFSYQSDEGNPLELRGA>M0819.0S52-A_MCRA_l5-M13P_47_B0g.SEQAYTNBVLDDFSYYGVDYAVEKYGGFAKAPANLEWKDLATEVTLYALEQYESFPTLLEDHFGGSQRAGVTAAASGITCAIATGNSQAGLAGWYLSQLLHKEAHGRLGFFGYDLQDQCGPTNVFSYQSDEGNFLELRGA>M0819.0S54-A_MCRA_l7-M13F_47_D08.SEQAYTDNILDEFTYYGMDCINDKYGVDWKNPSPKDRVKPTQEWNDITTEVALNAMEQYEQYPTMMEDHFGGSQRAGVIAAACGLSTSIATGNSNAGLNAWYLAfflLLHKEGWSRLGFFGYDLQDQCGSANSLSIRGDEGAIGEIRGP>M0819.0849-A_MCRA_12-M13F_47_G07.SEQAYTDDILDNNVYYDVDYINDKYNGAANLGTDNKVKATLDWKDIATESTLYGIETYEKFPTALEDHFGGSQRATVLAAAAGVACALGTANANAGLSGWYLSMYLHKEAWGRLGFFGFDLQDQCGATNVLSYQGDEGLPDELRGP>M0819.0S57-A_MCRA_2O-M13?_47_G08.SEQPYTNPVLDDFCYYGVDYAVDKFGGFAKAPKTIDVAKELATEVTLYGIEQYESFPTLLEDHFGGSQRAGVTAAASGITCAIATGNSQAGLAGWYLSQLLHKEAHGRLGFFGYDLQDQCGPTNVFSYQSDEGNFLELRGA>M0819.0S42-A_MCRA_5-M13F_47_HO6.SEQAYTNBVLDDFSYYGVDYAVEKYGGFAKAPANLEWKDLATEVTLYALEQYESFPTLLEDH改名為.FAST格式<6>尋找最相似序列打開這個FAST文件，開始一個個找最相似序列了。IllyIllllllllllHillv||=IIIatIItIIfaIaIIIlI/^II^II/IlyIiIII-j||vII|x|￡|l|rJu||Li||v|lt￡l||IlIIIyIvIIIIIIIIHzIIIIItvIIzIII^l|||v||^|L||||ir|L|7||1^||||^7||E||TA|^|||l|vA||LA||v|Lr|=||AvHf||eJJBIItaIIlaIItIle|t|||『IIIIVtllllllllllvlllllVL||:IIIJvlrlrrlnkrlr11~11|~1』」叩18<1匚|1.偵|7?{DataEditSearchAlignmentWebSequenDBirDisplayHelpProlainS^quonDDSM同，明叩WI■XDeletrRe^'eirseCom^pleme-nitSite#哪M5:alignmentExplorerfOlU-LDonlyPro.FASTAJDeleteGapsCtrl+Del3.:比踞色王15.L了■t.KDei9.D24S-AMCDi11-MC3AQSKiDeiHMCCli1-日-丑心9一。3穴*幻10MOS19.OSS1-AMCBA4UrdoCtrl+Z如亡/Ahbsrv*2.KDei9.D3€l-AMCBA￡4LM」￡19.DMCDA1=BL^^TSequence匚utCtirlfCCtd+XEditSequenceNameShewFullInfairmationiOpenSequencerFile-RedfertoGenEank在這個窗口，開始blast。找到一個序列后復制其DNA的編號Navigate