1、甘肅農(nóng)業(yè)大學碩士學位論文極細鏈格孢菌66kDa激活蛋白的純化、理化性質和生物活性分析姓名：李杰申請學位級別：碩士專業(yè)：植物病理學指導教師：朱建蘭邱德文20080601甘菇農(nóng)業(yè)大學20儲屆碩士學位論文AbstractInthisreseal“ch，a66kDaactivatorproteinwasextractedandpurifiedfromtheAlternariatenuissimaThesilverstainingshowedas

2、ingleband，whichverifieditispurityThisproteinbiologicalactivityphysicalcharacterglycoproteinidentificationandMSanalysiswereexecutedtomaketheoreticalfoundationforfurtherresearchonthefunctionmechanismofthisprotein1Theextrac

3、tandpurificationof66kDaactivatorproteinfromAlternariatenuissimaThesuitablebufferforextractionandconservationofthisproteinw雒screenedfromGoodbuffersystemThemostnucleicacidintheextractwasremovedbyprotaminePigmentwaseffectua

4、llyabsorbedbyactivecarbonTheprimaryfractionationextractWasdonebyethanolprecipitationandthefollowingpurificationwasexecutedbyDEAEanionexchangechromatographyAfterdesaltingbyHiprepTMDesaltingcolumnandconcentratedbyultrafiRr

5、ation，asinglebandWasdetectedbySDSPAGEandsilverstaining2The66kDaactivatorproteinphysicalandchemicalcharactersTheapparentmolecularweightofthisproteinwas66kDaonSDS—PAGEThePIofthisproteinwasabout427on2D—electrophoresisThepro

6、teinwastreatedby100℃boilingbatheandthencooleddowntoroomtemperatureTheSDS—PAGEdetectionofthetreatedproteinverifiedthisproteinWasathermostableprotein3The66kDaactivatorproteinglyeoproteinidentificationTheglycoproteinspecifi

7、cstainingwasdonewitllhorseradishperoxidaseaspositivecontrolandsoybeantrypsinasnegativecontrol，andtheresultshowed66kDaproteinwasaglycoprotein4The66kDaactivatorproteinbiologicalactivitydeterminationTheTMVresistanceassaywas

8、respectivelydonebytreatingtobacumSamsun、Ⅳith41xg／mL，61xg／mL，8Itg／mLproteinsolutionandmechanicalinoculationafter72hourneresultWasthatthe8pg／mLproteinsolutionhadanecrosisinhibitrateat49％Wheatseedswerealsotreatedby8pg／mLand

9、thenculturedrespectivelyatroomtemperatureandlowtemperatureof15“CTheaverageplantheightdetectionwasdoneafter5daysandthetreatmentandcontrolatroomtemperaturewere987cmand982cm，whichhadnoobviousdifferenceButthetreatmentandcont