黄瓜转录因子cscbf3的克隆与其转化黄瓜的分析

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1、BioinformaticsanalysisshowedthatcScBF3proteinwasconsistedofAP2踟久bindingdommnwhichincludedaminoacidresidues，nuclearlocalizationsignalregionandacidicactivationdomain，andsomephosphorylationsitesandproteinsecondarystructuresmayplayimportantrolesinprocessofinteractionsbetweenproteinsandDNA．Qua

2、ntitativereal．timePCRresultsshowedthatexpressionofcscBF3couldbeinducedbylowtemperature，andthelevelreachedtopeakveryfastandthendecreased．TheseresultssuggestedthatCscBF3Wasarapidresponsegeneandplayedanimportantroleinresistancetolowtemperatureincucumber．2．InfluenceofacetosyringoneandpHofCO-c

3、ulturemediumoncucumbergenetictransformationefficiencyCucumbergenetictransformationefficiencyaffectedbyacetosyringoneconcentrationsandCO—culturepHwerestudiedwithco锣ledonarynodeofcucumber‘Changchunmici’．T11eresultsindicatedthatbothacetosyringoneaddedinbacterialiquidandCO-culturemediumcoulds

4、ignificantlyimproveresistancebudinductionrate，andtheoptimumASconcentrationWas100pmol／L．CucumberresistancebudinductionratewasalsoaffectedbydifferentpHofCO．culturemediumandthemostsuitablepHforcucumbergenetictransformationWas5．2．111eseresearchresultswerehelpfultooptimizesystemofcucumbergenet

5、ictransformationandincreasetransformationefficiency,aswellaspromptedtransgenicworkofcucumber．3．ConstructionofExpressionVectorofcscBF3andTransfelrmationtoCucumbercScBF3Wasclonedusingspecificprimersdesignedaccordingtoits0对sequenceandWasinsertedintopCAMBIAl304inforwardorientationbydoubleenzy

6、medigestion．TherecombinantplasmidWasthenintroducedintoAgrobacteriumtumefaciensstrainEHAl05byfreeze-thawmethod．ElectrophoresisresultssholwedthatrecombinantexpressionvectorpCAMBIAl304．cScBF3wassuccessfullyconstructedandintroducedintoEHAl05．Cotyledonnodeof‘Changchunmici’WastransformedbyAgrob

7、acteriumMediatedmethod，andhygromycinresistantplantswereobtainedafterselectionanddomestication．CscBF3WasprovedtobeintroducedintocucumbergenomesuccessfullybyPCRdetectionwithatransformationrateof4．8％．KEYWORDS：Cucumber；CBF3；Clone；Acetosyringone；VectorConstruction；Geneti