玉米小斑病菌糖蛋白激发子的纯化和性质研究.pdf

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2016-05-10 发布于安徽
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玉米小斑病菌糖蛋白激发子的纯化和性质研究.pdf

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10.70%，无规则弯曲占48.02%. 关键词:玉米玉米小斑病菌激发子 POD PAL ExtractionandCharacterizationofa GlycoproteinElicitorfromHaphalCellWallofBipolarismaydis WangLi (LaboratoryofPhysiologicalPlantPathology,SouthChinaAgricultural University,Guangzhou510640,PeoplesRepublicofChina) ABSTRACT Inthepresentstudy,experimentwascarriedoutfortheisolation,purificationand characterizationofelicitorrfomhyphalcellwallrfomBipolarisMaydis.Thestudyofthe elicitorwasmainlyperformedbySDSandMALDITOFMSetc. 1.Thecurdeelicitor(CEF)rfomBipolarisMaydiswaspurifiedprimarilyby deeprfozenofliquefiednitrogen，mechanicaldisintegration,highspeedcentrifugationand PM-10membraneultrafiltrationetc..Theactiviyttestshowedthatthecurdeelicitorhad elicitoractiviyt. 2.ThecrudeelicitorwaspurifiedutilizingHiTrap16/10DEAESepharoseFFion exchangerespectively.Fourabsorptionpeaksweredeterminedat280nmand220nmwith peakD4beingabletoincreasethecativiytofPODandPALinmaizeleaves,whichmeans ithadelicitoractiviyt.CollectpeakD4respectivelyaselicitorD4. 3.FractionD4waspurifiedutilizingSephacrylS-100gelfiltration.Onlyone absorptionpeakwasdeterminedat280nmand220nmwithitbeingabletoincreasethe activiytofPODandPALinmaizeleaves,whichmeansithadelicitoractiviyt.Collectpeak SaselicitorS. 4.SDSshowedthatCEF,D4andSweregraduallypurified.ElicitorShad onlyoneelectrophoresisband.Itsmolecularweight(Mw)was36.8kDa. 5.AlltheelicitorsCEF,D4andSwereabletoincreasetheactiviytofPODandPAL inmaizeleaves.Astheelicitorswereourifiedsteobystep,theirabiliytofthe theactiviytofPODandPALinmaizeplantswasheightened. 6.ItwasdeducedthattheelicitorSwashypotheticalproteinSNOG09682(accession no:gi111061827)whosemolecularweight(Mw)is35787Da,PIis6.04utilizingMALDI TOFMSmethod. 7.Itwaspredicted勿ExPASyproteinanalysistoolsonline,thatcontainedphysical andchemicalcharacters,primary structure,secondary structure,post-transla