图位克隆基因原理与方法

oftheW0rkshOPonRrCh&loPm.tofBio10Hainan,31lul1AugU2)6,Sya,Cina27318l,,221,,57202,;2,,571158,[e-mail],,.,,,,,TheoryandMethodinMaPasedCloningofGenessongChenghiao.wangRulpinDuDaolinzLIDi,BaiYananl.IHDaD11.ofTiRux,5725;2ofBiolo,HanNOn.IColl,571158.dmgauIOr,[e-mail]915conductedwhenthesttucandctinnOfgenproduCtSwereunclear.lyknownandthectio.geneswetheirpaxtiauargenelocioncrom~es,withtheraPiddevel-Pmentofmulecularmtertechnologiesandconstructingtecbnologiesofhighalltygenomiclibra,mabasedclonmghasalleadybecomeagenebiotecbJqueforgeneisolation5palr~dthe~ProcessesofthemaPasedcloDJ9includingPrelLinmaPping,scalemaPPingofcandi-dategene,bulldinggenetiom,chromosomewal9orlandj9and6nallyctioncolrlPlementexln-menttoidenticandidategene.Theprobl~anddeveloPmenttrendsofmaseddoningarealso~1dinspaper.KeyMaP-bddog,MOlecularmter,FinemaPPing,Genomiclibrary,:,,;,,,,,,,(maPbaseddoning)sitionalclonin,1986Coulson,,DNA,(),,1.oftheWorkshoPonRh&v.loPm.tofBiotecbnologyinHainan,31ul1Aug2006,Sanya,Cina267318l,,11992ABI3(Giraudatatal.,1992)FADndeletai.,1992),,,DNA(CdYAcBAc),,,(l)1FigureIKste5inmbasedelo9Pnx552DNADNA,,2.1,COsndCOsmid,,25SKb,,,DNA,YAC1983MSzosta55KbYAC1987BteDNAYACYACC)(cEN)(L)(ARs),YAC,:(l)(chnericPhenope)(40%60%)YAC,,;(2)(stability),(sequencere~gement)(sertdeletion);(3)DNA;(4)YAcP1,BAC,PAC(MC)C).ZJ,DNA,,oftheWorkshoPonR.rCh&DeloPmentofBiothaologyinHai,3ljulIAugt,20062(X)67318l,,(restrictionentlengthpolsm,RFLp)DNA(randomlyaJplifiedpolymorphioDNA,RApD)(seeneetaggedsite,sTs)(eressedseneetag,EsT)(singlenucleotidepolymorphissN)(sPlosequcucelengthpOIrphism,SSLP)DNA(sPlesequencerepeat,ssR)(amPlifiedgmentlenpolyrnorphism,AFLP),,,,,,,,,,HajrLha(l998)2275,1521.6cM:20013000,.lian(1997)RP,3.6/0.IcM,BAC,20Kb33.1,,,(nearisogeaiclines,Nll)usegregantanalysis,BsA),,,,,Liang(1994)s,,,,Ns,sA)Michelinore(l991)BC,DH)(),DNA,DNA(),510cM,3J0.scM,,30004000,IcM1oooKb,,,eoftheWorkshoPonRah&LveloPmentofBiotechnoogyinHai,31July--lAugt,26,Sanya,ma2(X)67318l,,,,,,,;,,DNA(ssRterPrimerPreer5.0);,,DNA,Chujrchill(1993)DNADNA()(520),DNA,DNA,DNA,,DNA,,,,,,DNA,DNA,;,.,,iber.FIS,FlsHIKb(0.14DNA),Fiber-FIsH,,33(chromosomewang)DNA,,RFLPRAPD,DNA,,,,,,,,,,DNA,;,,,,,oftheWOrksbOPonRrCh&veloPmentofBIOtecbDologymHainan,31JulIAu2006,Sanya,CbIa2673181,,,.,,,14ORF(nreingfraJe),ORF,BAcYAccDNA,,,:(1)cDNA;(2)cD-NA;(3)cDNA,,;(4),DNA;(5),RNA)4,,,,.,,,Kashjmir--1()Conbin(),,,,,;,(C0nhaveretal.,1998),,,1%10(K,1%1X2sooK,(Janderetal.,2002).521,,,,,(),,,,(titativetraitlocis,Q),,(,).,,QTLRoftheworkshoPonRh&loPmentofBinteCbnologyinHainan,31JulIAut,2(X)6,Sanya,Chma267318l,,QTL,QTLQTL,(2005032)ArondelV,LmieuxB.,Hwang1.,bson5.,GaledmanH.M.,dSomenleC.R.,1992,MaP-bdeloningofageneeon-llingomega3tlyaciddesatorationinArab:,Seience,258:13531355B.ten.T.,1957,Clo9ofeseentsofexogeneousDNAintoyeastmeansof6cialehromosomeveetors,sei-enee,236:806812ChmhillG.A.,GiovannomJ.J.,dTankslS.D.,1993,led-nPlingmakeshighsolutionIrlaPPingPraetiealwithDNAmar,Proe.NaU.Acad.Sci.,91):16-20CopenhaverG.P.,BrowneW.E.,dP~5D.,1998,AssaLnggenome.widembinadonandcentromerectionswithArabidz,:tetrads,Gties,95(l):247252CoulsonA.,SulstonJ.,Be5.,dKamJ.,1986,Genetiesto,wardaPhysicallrPofthegenomeofthenematCe(norhabe,ROe.N.ad.Sei.,83:78217825GiraudatJ.,HaugeB.M.,valonC.,smalleJ.,pyF.,dGOCanH.M.,1992,IsolationoftheArabid,s.ABgenepositionaleloning,PlantCell,4(l0):12511261Hsha,YanoM.,ShoroujraA.,SatoM.,ShimanoT.,Kki,Y....otoT.,Lins.,AnioB.A.,ParcoA.,Ka-jiH.,HUangN.,YamamotoK.,Nag.ra,KrataN.,KushG.s.,dsasT,1998,Ahighensityrieegenetie1kagemwith2275mtersusingasingleatiGOetics,148:47994JanderG.,N5S.R.,Rounsleys.D.,BushD.F.,LM.,d5R.L,2(X)2,Arab:maPasedcloninginthepost-genomeera,PlantPhysiol.,129:050IianA.,ChenJ.,HanF.,SteffensonB.,deinhoA.,1997,TowmbaefoningofthebarleystemrustresistaneegeneRPglandRPusingriceasaninteenoeel~9vchicle,plantMol.Biol.,35:187LiangC.Z.,GuM.H.,PanX.B.,LiangG.H.,dZbuL.H.,1994,Pta9ofanewsedwhaggeneinriee,Teor.APPI.Genet,88:89800MIChelmoreW.,Paran1.,dKesseleliR.V.,1991,dentificationofrkersledtodiseascsistacegenesbulkedsegregantanaysis:araPidmethodtodetectmtersinscificgenomicrensusingsegatingpoPulations,Proc.Nati.Aead.Sei.,88(21):98289832MunyAW,dSzostakJ.W.,1983,Constlucdonofficiaiemo,OmesinyeaSNaIre,305:189193105