扁桃花粉AcSFB基因的克隆及其表達分析.pdf

1、新疆農業(yè)大學碩士學位論文扁桃花粉AcSFB基因的克隆及其表達分析姓名：郭長奎申請學位級別：碩士專業(yè)：生物化學與分子生物學指導教師：羅淑萍20090601TheCloningandExpressionAnalysisofPollenAcSFBGenesfromAlmond似mygdaluscommunisL)AbstractAlmond似mygdaluscommunis工)isthediploidmemberofRosaceaeandis

2、crosspollinatdandselfincompatibleplantStyleS—determinantshavebeendeterminedtobeSRNaseandtheSFB(Shaplotype—specificFboxproteingene)allelesareconsideredtobethefirstcandidatesofthepollenSdeterminantsNowthereisnotanyresearch

3、ofalmondpollenGSIinXinjiangInthisstudy，theinitialaimWaStoisolateandcharacteriseAcSFBallelegenesfromalmondcultivarinXinjiangChinatogainfurtherunderstandingoftheevolutionaryrelationshipsandphylogenyofSFBalleleswithinPrunus

4、ExpressionanalysisandsemiquantitativeRTPCRwereusedtofindwhichtissuecouldexpressSFBproteinandwhichfloweringperiodexpressedmore5’UTRand3’UTRanalysisandbioinformaticsanalysisstillhadstudiedinthisexpedmentThestudyofalmondsel

5、fincompatiblemechanismusingmolecularbiologicaltechnologywouldfastpromotetheindustryofalmondAllresultsweresummarizedasfollows：1TotalRNAfromalmondantherwasextractedwiththeimprovedCTABmethodThealmondantherRNAobtainedWas900d

6、integrityandwithoutstainofpolysaccharideandprotein，whoseyieldwasveryhi曲2Inthisstudy，fifteennovelAcSFBgenes(GenBankaccessionnumbers：EU909685，F(xiàn)J362524FJ362529，F(xiàn)J231204，F(xiàn)J415321andFJ514935～FJ514940)wereidentificatedusingRT—

7、PeRandRACEtechnolgyinAcommunismainlyplantedinChinaforthefirsttime而eidenffiedAcSFBgenesshowedShaplotypespecificpolymorphismTheseallelesencodedproteinswiththeFboxmotifintheN—aminoterminus，andwithfour(hyper)variableregions，

8、ie，V1，V2，HvaandHvb，consideredtoberecognitionsitebetweenSFBandSRNaseanddeterminedGSIWiththebioinformaticsanalysis，theAcSFBproteinswereinstable，hydrophilicityandnonsecretoryproteinandhadligaseorlyaseenzymeactivity3Usingthe

9、DNAMANsoft，phyletieevolutiontreewasconstructedTheresultshowedtheinterspeciesaminoacididentitiesobtainedbycomparisonwithotherPrunusSFBswereoftenhigherthanintraspeciesidentitiesTheevolutionrelationshipofpollenSgenesbetween

10、ChineseandAmericanalmondsWasreciprocalchiasmataandtherewerenoplacesdifferences4AcSFBgeneWasexpressedonlyinpollenandnotinotherorgan，whichshowedAcSFBexpressionhadorganspecificityTheAcSFBWasexpressedmoreinballstagethaninoth

11、erstage，whichwouldgiveareferenceforarrayingpollenizerStudyonexpresspaRemofAcSFBmaycontributetoconfirmationthattheAcSFBgeneisthepollenSdeterminantStillweprovedtherewasnointronintheAcSFBgenesandasingleintronhadbeenfoundint

12、hes’untranslatedregionoftheAcsFBgenesofalmond5ThevectorofpETAcSFBWasconstructed，aftertransformedtoEscherichiacoliBL21andinducedbyisopropyl—pD—thiogalactopyranoside(IPTG)，recombinantproteinabout60kDwasexpressedinpETAcSFBs