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1、新疆農(nóng)業(yè)大學(xué)碩士學(xué)位論文扁桃花粉AcSFB基因的克隆及其表達(dá)分析姓名：郭長奎申請學(xué)位級(jí)別：碩士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)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

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