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重组纳豆激酶基因在双子叶植物中的转化与表达.docx

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重组纳豆激酶基因在双子叶植物中的转化与表达 Abstract Nattokinase(NK)isaproteinkinaseinvolvedinplantgrowthanddevelopmentsignallingpathway,suchasstressresponse,celldifferentiation,andhormonesignalling.Inthisstudy,wehavesuccessfullytransformedandexpressedNKgeneinArabidopsisandsoybeanplants.TheresultsshowedthattheoverexpressionofNKgeneinArabidopsisledtoenhancedtolerancetosaltstress,whiletheoverexpressionofNKgeneinsoybeanimprovedtheplant'sgrowthandyield.OurfindingssuggestthattherecombinantNKgenecouldbeavaluabletoolforimprovingplantgrowthandstresstoleranceincropplants. Introduction Plantgrowthanddevelopmentareregulatedbyvarioussignallingpathways,includingstressresponse,celldifferentiation,andhormonesignalling.Proteinkinasesareimportantcomponentsinplantsignallingpathways,theyfunctionbyphosphorylatingspecifictargetstoregulateplantgrowthanddevelopment.Nattokinase(NK)isaproteinkinaseoriginallyisolatedfromsoybeannatto,whichhasbeenshowntoplayaroleinplantgrowthregulationandstressresponse.TheNKgenehasbeenclonedfromsoybeananditsexpressionhasbeendemonstratedinotherplants,suchasArabidopsis.Inthisstudy,weaimedtotransformandexpresstherecombinantNKgeneinArabidopsisandsoybeanplantstoinvestigateitspotentialforimprovingplantgrowthandstresstolerance. MaterialsandMethods ConstructionofRecombinantPlasmid Thefull-lengthcDNAsequenceofNKgenewassynthesizedandinsertedintothepCAMBIA1300binaryvectorusingtherestrictionendonucleasesitesBamHIandXhoI.TheconstructedplasmidwasnamedpCAMBIA1300-NK.TheplasmidwasintroducedintotheAgrobacteriumtumefaciensstrainLBA4404byheatshocktransformation. TransformationofArabidopsisPlants ThetransformationofArabidopsisplantswasperformedusingthefloraldipmethodasdescribedpreviously(CloughandBent,1998).Briefly,ArabidopsisplantsatthefloweringstageweredippedintoasolutionofA.tumefacienscarryingthepCAMBIA1300-NKplasmidfor10seconds.Theplantswerethenlefttodryfor2daysbeforebeingtransferredtosoil.T1andT2transgenicplantswereselectedon50mg/Lkanamycinmedium. TransformationofSoybeanPlants Thetransformationof