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《番茄果實乙烯信號轉(zhuǎn)導組分EREBPs基因克隆和表達研究》由會員上傳分享，免費在線閱讀，更多相關內(nèi)容在學術論文-天天文庫。
1、中國農(nóng)業(yè)大學博士學位論文番茄果實乙烯信號轉(zhuǎn)導組分EREBPs基因克隆和表達研究姓名：蔚變云申請學位級別：博士專業(yè)：食品科學指導教師：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
