日本囊對蝦凝集素的分離及其功能研究.pdf

1、分類號——UDC廈門大學(xué)博士后研究工作報告密級編號——日本囊對蝦凝集素的分離及其功能研究楊海杰合作導(dǎo)師：徐洵教授李少菁教授工作完成時間：2003年1月2005年9月報告提交時間：2005年12月廈門大學(xué)海洋與環(huán)境學(xué)院2005年12月AbstractAbstractCrustaceansassumeaparticulareconomicimportanceasafoodsourceandtheiruseinaquaculture，espe

2、ciallypenaeidshrimps，iscurrentlyexpandingTheSHCCeSSofshrimpcultivationis，howeverseriouslyendangeredbydiseasesLectinshavebeenregardedasanimportantroleinnon—selfrecognizationandpathogenclearanceininvertebrateimmunityThisdi

3、ssertationaims砒isolatingandcharacterizingthelectinsinshrimpinordertohelpthepreventionoftheshrimpdiseasesByusingaffinitychromatographywithfetuinSepharose，anaturallectin(nominatedPjLec)wasisolatedfromthehemolymphoftheshrim

4、pMarsupenaeusjaponicusSDSPAGEshowedthatthepurifiedPjLecproteinconsistedof37kDasubunitsThenativePjLecbehavedasa452kDaproteiningelfiltrationchromatographywithSuperdex200ThosedatastronglysuggestthatPjLecisbuiltupby12subunit

5、sofsimilarmolecularweightPjLeccouldagglutinateallthemammalianerythrocytestested，andthehemagglutinationiscalciumindependentTheagglutinationactivitywasstablebetweenpH6andpH8，andwastemperature—dependentThehemagglutinationof

6、PjLecproteinwasinhibitedbyManNAc，Neu5Aandlipopolysaccharide(LPS)Bovinesubmaxillarymuein(BSM)，whichcontainsmainlyNeu5A，isthemostpotentinhibitorofPjLec謝tllaMICofO0006medmLFurthermore，PjLechasabroadagglutinationactivityonGr

7、amnegativeandpositivebacteriaand4of5strainsofpathogeneticbacteriaofshrimp(Vibrioparahaemolyticus，Aeromomashydrophilic，VibrioalginolyticusandMicrococcuslysodeikticus)couldbeagglutinatedbyPjLeeItsuggestsPjLecmightplayanimp

8、ortantroleindefenseagainstbacterialinfectioninPenaeusjaponicusInpreviousstudies，weisolatedanovelCtypelikelectin(nominatedPjLec2)fromtheSSHlibraryofhepatopancreasinMarsupenaeusjaponicusTheexpressionlevelofPjLec2mRNAinviru

9、sresistantshrimpsisobviouslyhigherthanthatinnormalshrimpsInthisstudylectinPjLec2wasexpressedinEcoliandpartialfimctionofPjLec2wasanalyzedItcouldbehighlyexpressedinEcoliwhenfusedwithGSTHoweverhemagglutinationassayshowedtha

10、tpurifiedPjLee2waslackofhemagglutinatingactivitytoall7testedmammalRBCsincludingmouse，miceandrabbit，Inaddition，9721eccanagglutinatetwoGramnegativebacteriaAeromomashydrophilicandHbrioalginolyticusbothofwhicharemayorpathoge