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《番茄果實(shí)乙烯信號(hào)轉(zhuǎn)導(dǎo)組分EREBPs基因克隆和表達(dá)研究》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
1、中國(guó)農(nóng)業(yè)大學(xué)博士學(xué)位論文番茄果實(shí)乙烯信號(hào)轉(zhuǎn)導(dǎo)組分EREBPs基因克隆和表達(dá)研究姓名:蔚變?cè)粕暾?qǐng)學(xué)位級(jí)別:博士專(zhuān)業(yè):食品科學(xué)指導(dǎo)教師:羅云波20030501AbstractTheplanthormoneethylenehasaprofoundroleinplantgrowthanddevelopment.Anumberofethylanesignalingelementshavebeenisolatedandidentified.Althou【ghnuclearcascadesinethylenesignaltransductionpathwayfromE
2、IN3toERFltoPRproteinshasbeenestablishedinArabidopsisthaliana,asimilarroutefromEIN3toEREBPstofruitripeningandsenescencehasnotbeenelucidatedinLycopersiconesculentum(tomato).EREBPs(ethylene—responsiveelementbindingproteins)belongtoafamilyoftranscriptionfactors,whichregulateethylener
3、esponsesthroughbindingtothepromoterregionofethylene—regulatedtargetgenes.StudiesonEREBPsintomato,whichisamodelplantinfruitandvegetableripeningandsenescenceresearches,willmakeusbeRerunderstandthemechanismofripening-relatedgenesexpressionregulatedbyethyleneandfurtherelucidateethyle
4、netransductionpathway.Apairofdegeneraleprimersweredesignedintheconserveddomain(ERFdomain)whichbasedonthealignmentofEREBPsgeaefamilyintomatoandotherplantspecies.A14bpfragmentwasamplifiedbyRT—PCR(ReverseTranscriptionPolymeraseChainReaction),whichwasusedasaprobeforscreeningtomatofru
5、it(pinkstage)cDNAlibrary.Twopositiveclonescontainingentirecodingregionwereisolated,whichwereidentifiedasnewmembersofEREBPsfamilybyBI—ASTServer,andnamedasLeERFlandLeERF2.TheLeERFlcDNAis1041bpandthepredictedtranslationproductis204aminoacidsTheLeERF2cDNAis2898bpandthepredictedtransl
6、ationproductis210aminoacidsLeERFlwashomologonswithEREBP4andDDTFRl0A,with33%similarityattheaminoacidlevel.LeERF2washomologouswithEREBP3andpti5,with46%similarityattheaminoacidlevel.ThecDNAsequencesofLeERFlandLeERF2weredepositedinGenBank(GenBankAccessionNumberAY077626,AY275554respec
7、tively).NorthemblothybridizationshowedthatLeERFl,LeERF2andpti4(amemberofEREBPsintomato)expresseddifferentiallyindifl'erentpartsofwildtomatofruitduringripening,whichwasrelatedwithfruitripening.InbothNrfruitandantisenseACStransgenictomatofruit,expressionlevelsofLeERFI,LeERF2andpti4
8、weregenerallylowerthanthoseinwildfruitbu