小麥組蛋白修飾酶基因及DNA甲基轉移酶基因的分離及鑒定.pdf_第1頁
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1、中國農業(yè)大學博士學位論文小麥組蛋白修飾酶基因及DNA甲基轉移酶基因的分離及鑒定姓名:戴艷申請學位級別:博士專業(yè):作物遺傳育種指導教師:孫其信倪中福20050401AbstractEpigenelicregulationisamajoraspectofgenecontrolbywhichheritablechangesingeneexpressionOCCUrwithoutanalterationinDNAsequence,whichma

2、inlyincludeshistonecode,DNAmethylation,RNAinterferenceandgenomieimprintingThestudyofepigenticsinwheatlaggedbehindInthisarticle,genesencodinghistonemodificationenzymeandDNAmethltransferasewereisolatedusingdegenerateprimer

3、PCRand/nsilicocloningThemainresultsareasfollows:1)TwohistoneacetyltransferasegenesandfourhistonedeacetylasegeneswhichbelongedtodifferenttypeswereisolatedusinginsiHcocloningandRTPCRmethod,designatedTaHATI2,TaHDI一4respecti

4、velyAminoacidsequenceanalysissuggestedthattheyallcontainedtypicalstmctruecharacteristicsofhistoneacetyltransferaseanddeacetylaseSubeellularlocalizationindicatedthatTaHAT/proteinbecameconcentratedinthenucleusandTaHD3displ

5、ayednuclearcytoplasmicdistributionToourknowledge,nogenesencodinghistoneacetyltransferaseanddeacylasehasbeenreportedinwheat2)NineSETdomainproteingenefragmentswereobtainedbydegenerateprimerPCRPhylogeneticanalysissuggestedt

6、hattheybelongedtoSU(VAR)39classSETdomainproteinInaddition,twoSETdomainproteingeneswereobtainedusinginsilicocloningandRTPCRmethod,named砌齲刀2respectivelyPhylogeneticanalysisindicatedthattheybelongedtoASHandSU(VAR)39classSET

7、domainproteinrespectively3)WeisolatedfourwheateDNAfragmentsandoneeDNAwithopenreadingframeencodingDNAmethyltransferaseanddesignatedTaMETl,TaMET2a,TaMET2b,TaCMTandTaMET3,respectivelyBLASTsearchesandphylogeneticanalysissugg

8、estedthat5cDNAsbelongedtofourclasses(Dnmtl,Dnmt2,CMTandDnmt3)ofDNAmethyltransferasegenesTaMET2aencodedaproteinof376aaandcontainedeightoftenconservedmotifscharacteristicofDNAmethyltransferaseGenomicsequenceofTaMET2awasobt

9、ainedandfoundtocontaintenintronsandelevenexons4)Expressionanalysisofthesegenesobtainedinthisarticlerevealedthattheywereexpressedindevelopingseed,duringgerminationandvariousvegetativetissues,butinquitedifferentabundanceIt

10、wasinterestingtonotethatTaMETlandTaMET3involvedinmaintenancemethylationanddenOVOmethylationwereclearlydetectedindryseedsandothergeneshadnotbeendetectedMoreoverdifferentialexpressionpatternsofthesegenesWereobservedbetween

11、wheathybridanditsparentsinleafstemandrootofjointingstage,somewereupregulatedwhilesomeothersweredownregulatedinhybrid5)OverexpressionofTaHAT/andTaHD3genesinArabidopsissuggestedthattheywerebothassociatedwithvariousdevelopm

12、entalabnormalities,mainlyincludingleafandflowerdefectwhichsuggestedthat如J刪ZYandTaHD3maybeinvolvedinageneralregulatorymechanismresponsibleforplantplasticityandvariationinnatureThisisafirstreportaboutfunctionstudyofhistone

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