安全选择标记和无选择标记转基因技术研究进展

:2008-10-01:863(2007AA10Z189);:(1984-),,,,:(1976-),,,,,1,2,1,1,1,2(11,130033;21,130118):,,,:;;:Q785:A:1000-7091(2008)-0096-07TheAdvancesinResearchoftheTransgenicPlantswithSafeorNoSelectionMarkersXUEJian1,2,GUODong2quan1,ZHAOGui2lan1,YANGXiang2dong1,WANGLi2jun2(11BiotechnologyResearchCenter,JilinAcademyofAgricultureSciences,Changchun130033,China;21CollegeofHorticulture,JilinAgriculturalUniversity,Changchun130118,China)Abstract:Thesafetyofthetransgenicplantswouldbeascribedtotheselectionmarkergenespresentintheirgenomestoagreatextent.Twostrategiesincludingutilizationofthesafemarkergenesandmarker2freetechniquesareusuallyexploitedfortheproductionofthetransgenicplants.Inthispaper,theadvancesintherecentresearchoftheuti2lizationofthesafemarkergenesandmarker2freetechniqueswerereviewedandthepossibleapplicationsofthesetech2niqueswerealsodiscussed.Keywords:Safemarker;Marker2free;Transgenicplants1983,,,1996,5177hm2,20061102hm2,7%,23,,,,21[1],,,,,(),,,,:,,,,;,,2008,23():962102,,1,,,,[2]:11111111ipt232IaaHiptT2DNA,(IAA),IAA,;,[2]IaaH,[3]11112pmi,xylArtlpmi626262,62,6262,,62,ATP[4],,62,62ATP,,,,,,[5]xylA,,D2D2D2,D2D2,xylA,D2D2,,,D2,,xylA,,xylA9[6]11113D2(DAAO)(AK),,,,AK,,akak,,AK,,[7];112,,,Daniell[8]BADH,,113(GFP),,,,GFP,,,GFP,,,GFP[9][10][11]2(gus),:97GUS,,,[12],gus62BA,Joersbo,gus,62BA,2[13],,ipt,,ipt,,;pmi,,pmi,,;GFP,,,,,GFP,,,,GFP,,,2,,,,,MAT,.;.;|.T2DNA.Targetgene;.Selectablemarkergene;|.T2DNAborder.1(,2005)Fig.1Theco2transformationsystem(Dengqiming,etal,2005)2112,,,,(1)McK2night[14],2,,Framond[15]T2DNATinosC58,2,63%,59%Matthews[16]2T2DNATi,367%,25%Lu[17]komari,2T2DNAT2DNAT2DNA,T2DNA,36%64%212DNA,9823,,2DNA,,2DNA,DNA,,1996FischerN[18](aaDA)2,,aaDA213(Multi2auto2transformationMAT)R/RS[19,20],,ipt(),[21,22]ipt,,iptipt,EbinumaACiptSugita[22],Z.rouxiiR/RSAC,R/RSR2s2(GST2227),lacZ,iptipt,SafenerR,,rolMAT,NIAES1724rolA,rolB,rolC,rol,rolMAT88%,14%Southern,86%GUS,rolMAT214,2DNA,DNA,(2)[23]Cre/loxpFLP/FRTR/RSGin/gixattp5Cre/loxP,2lox(34bp),lox,Cre,loxAndrewpART27pART542PCre21,pART54TDNAuidA,nptIIcodA,PCreCrehptZuoDNAPX62GFP,CreXVE,XVE2FLP/FRTsFRTsFLP,1989CreggJM[24]FRTSArg,FRTs2ARG+2FRTsArg-,Arg+,FLPArg21996LyznikLA[25]FTs2npt2FRTs,FLPnptR/RSR,RS,2RS[6]1995,Onouchi[26]RRS,Gin/gixGin,2gix,MaeserS[27]1991Gin,attP(attP)(attB)attP(int)(IHF)Zubko[20]2attP,INTIHF:992(Cre/loxP)(,2004)Fig.2Thesite2specificrecombinationsystem(Cre/loxP)(Dongwenqietal2004)215,,Ac/Ds[28],AcDs,,,,,(),Ebinuma[13]iptDs,MAT,GoldsbroughnptII,GUS,nptIIDs2GUS2Ds[33]213%616%Ds2GUS2DsnptII4,2,(1)1Tab.1Comparisonofdifferentmarker2freetechniquesTypesAdvantagesDisadvantageExamples,,,,,,,,,,,,(MAT),,,,,DNA,,,,,,,,,3,I2amtham[30],10023Haj2dukiewicz[31],2loxP,CRE,cre,CRE,CREloxP,cre,Lutz[32]HajdukiewiczCorneille,CRE,loxP,,,,Zuo[33]DNA,,,,,,,,4,,,,,,,,;,,,:[1]CliveJames,AnatoleFK.Globalreviewofthefieldtestingandcommercializationoftransgenicplants1986to1995[R].TheInternationalServicefortheAcquisitionofAgri2biotechApplications(ISAAA),1996.[2]EbinumaH,SugitaK,MatsunagaE,etal.Selectionofmark2er2freetransgenicplantsusingtheisopentenyltransferasegene[J].ProcNatlAcadSciUSA,1997,94:2117-2121.[3]EbinumaH,KomamineA.MAT(Multi2Auto2Transformation)vectorsystem,theoncogenesofAgrobacteriumaspositivemarkersforregenerationandselectionofmark2freetransgenicplants[J].InVitroCellDev2Pl,2001,37:103-113.[4]WeisserP,KramerR,SprengerGA.ExpressionoftheEs2cherichiacolipmigene,encodingphosphomannoseisomeraseinZymomonasmobilis,leadstoutilizationofmannoseasanovelgrowthsubstrate,whichcanbeusedasaselectivemarker[J].ApplEnvironMicrobiol,1996,62(11):4155-4161.[5]CorneilleS,LutzK,SvabZ,etal.Efficienteliminationofse2lectablemarkergenesfromtheplastidgenomebytheCRE2loxSite2specificrecombinationsystem[J].PlantJ,2001,27(2):171-178.[6]ReinerG.GenesforribitolandD2arabitolcatabolisminEs2cherichiacoli:theirlociinCstrainsandabsenceinK212andBstrains[J].JBacterial,1975,123:530-536.[7],,.[J].,2007,27(3):115-119.[8]DaniellH,MuthukumarB,LeeSB.Markerfreetransgenicplants:engineeringthechloroplastgenomewithouttheuseofantibioticselection[J].CurrGenet,2001,39:109-116.[9],,,.(GFP)[J].,2003,45(6):654-658.[10]ChungBC,KimJK,NahmBH.Inplantsvisualmonitoringofgreenfluorescentproteinintransgenicriceplants[J].MolCell,2000,10:411-414.[11]ZhangCL,ChenDF,McormaAC,etal.UseoftheGFPreporterasavitalmarkerforAgrobacterium2mediatedtrans2formationofsugarbeet(BetavulgarisL.)[J].MolBiotech2nol,2001,17(2):109-117.[12]OkkelsFT,WarsJ,JoersboM.Synthesisofcytokininglu2curonidesfortheselectionoftransgenicplantcells[J].Phy2tochem,1997,46:801-804.[13]EbinumaH,SugitaK,MatsunagaE,etal.Selectionofmark2er2freetransgenicplantsusingtheisopentenyltransferasegene[J].ProcNatlAcadSciUSA,1997,94:2117-2121.[14]McKnightTD,LillisMT,SimpsonRB.SegregationofgenestransferredtooneplantcellfromtwoseparateAgrobacteriumstrains[J].PlantMolBiol,1987,8:439-445.[15]DeFramondAJ,BackEW,ChilonWS,etal.Twoun2:101linkedT2DNAscantransformthesametobaccoplantcellandsegregateintheF1generation[J].MolGenGenet,1986,202:l25-l3l.[16]MatthewsPR,WangMB,WaterhousePM,etal.Markergeneeliminationfrom