1、東北農業(yè)大學碩士學位論文野生大豆耐鹽相關基因的克隆與序列分析姓名：張春寶申請學位級別：碩士專業(yè)：作物遺傳育種指導教師：王鳳義董英山20070620IsolationandSequenceAnalysisofSalt—toleranceRelatedGeneinWildSoybeanAbstractSaltsUessisoneofthemainfactorswhichaffectthegrowthandoutputofcropsItise

2、stimatedthatupto4x1059x108hectarelandWaSsalinizeal，whichismorethanthreetimestopresentagriculturalland，SOhowtoimprovecropssalttaleranceisbecomingmoleandmoreimportantCultivatedsoybeanoaonsfromwildsoybean，wildsoybeanhasgrea

3、tresistanceandisallimportantgermplasmreSOllreOsforcultivatedsoybeanbreedingTherefore，itisveryurgenttostudysalttolerancemechanismandisolateexcellentgenesfromwildsoybeanforcultivatedsoybeanimprovementTheprolineisanimportan

4、tosmolyteNotonlylowerplantsbutalsohighel“plantsCanaccumulatefreeprolineinresponset0osmoticstressesTheprolinesynthesisenzymefP5CS)isakeyenzymeintheprolinebiosynthesiswhichlimitsthesynthesisrateofProlineIthasimportantsigni

5、ficancetoresearchonP5CSWildsoybeanZYD00659WaSusedinthisstudyTwopairsofprimersweredesignedaccordingtotheconservedregiomofthePSCSfromotherplantinGenhankThegenewasamplifiedbyRTPCRfromthetotalRNAwhichWasextractedfromwildsoyb

6、eanundersaltStressin200mMNaCIByhomologousbasedcloningweisolatedacDNAsequenceinwild∞ybearlwith1569bpinlengthItencodes523aminoacidsPredictedbysoftwarethemolecularweightofthispoptideis56514kDaitsisoeleetriepointis675Thededu

7、cedproteincontainsNADPHbindingdomainGlu—kinasedomainGSAdehydmgenasodomainandfeedbackinhibitionsiteNCBIdalabaseConservedDomainSearchithasthewholeProAdomainnamedGsPSCSlUsethe&anlemethod，wealsoisolatedanothereDNAsequenceinw

8、ildsoybeanwith560bpinlengthItencodesabout186aminoacids。PredictedbysoftwarethemolecularweightofthispeptidejS20，186kDaitsisoelectricpointis473NCBIdatabaseConservedDomainSearchithasthepartialProAdomainnamedGsP3CS2Wel螂GsPSCS

9、2eDNAsequencedesignedapairofprimeramplifiedaDNAsequencebyPCRingenomieDNAThelengthofthesequenceis1374bp，containsfourintronsandfiveexttonsAndbet、vcenintronsandextronwhichhavetypicalGT／AGstructurenamedgGsPSCS2Weblastthesequ