雞傳染性法氏囊病毒河南H-1株VP2基因的克隆及原核表達(dá).pdf

1、中國(guó)農(nóng)業(yè)大學(xué)碩士學(xué)位論文雞傳染性法氏囊病毒河南H1株VP2基因的克隆及原核表達(dá)姓名：薛協(xié)超申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：預(yù)防獸醫(yī)指導(dǎo)教師：劉金華李文剛20051101AbstractInfectiousbursaldisease(IBD)isahightangentinfectiousdiseasecausedbyinfectiousbursaldiseasevirus(IBDV)PreBlymphoidcellsorimmatureBlym

2、phoidcells，thetargetofIBDV、oftenappeardegenerationandnecrosisSincethebursaofFabficiusisimportantorgantothematurationofB—lymphoidcells，amountofBcellsinthelymphoidfollicleofbursaFabriciusislytic，atrophyofbursacausingimmuno

3、suppression，leadingtoincreasedsusceptibilitytootherpathogensanddecreasedimmunologicalresponsetovaccine(newcastlediseaseandavianinfluenza)，producingconsiderableeconomiclossinpoultryindustriesVP2proteinisthemainstructurepr

4、oteinandprotectiveantigenofIBDV，containingtheBcellepitopesinducingtheneutralizationantibodiesThemaincontentsareasfollows：reversetranscriptionpolymerasechainreaction(RTPCR)wasusedtoamplifyVP2genefromidentifiedIBDVfieldiso

5、lateH1isolatedfromapoultryfarminHenanTheamplifiedVP2genewasclonedintoprokaryotic(Esghedchiacoti)expressionsystemtoproduceVP2proteins，InthisstudyRTPCRwasconductedafterisolatingtotalRNAofIBDVandthe1461bpVP2genewasamplified

6、TheVP2genewasthenclonedintothepGEM—TvectorRecombinantplasmidpTVP2wasconstructedsuccessfullySequenceanalysisshowedthatthehighhomologyoftargetgeneandanimoacidwiththevvlBDVsfromJapanandEuropeThesimilarvariationofcharacteris

7、ticaminoacidimpliedstrainH1wasprobablywlBDV，PhylogeneticanalysisshowedH1wasinthesamelineagetowlBDVUK661andOKYMisolatedfromEuropeandJapanrespectivelyimplyingtl—ImightbewlBDVTheVP2geneinpTVP2vectorwassubclonedintoprokaryot

8、icexpressionvectorpET28aAfterdoubleenzymedigestionandPCR，thevectorpETVP2wassuccessfullyconstructedThe537Kdrecombinantproteinwasexpressedwithhi曲efficiencyinEcoilafterinductionwithIPTGTheexpressedproteininformofinclusionbo

9、dyshowedspecificimmunoreactivitywithchickenpolyclonalantiserumtoIBDVinwesternblotassaysTheresearchaboveprovidedabasisforstudyingontheroleofVP2proteinandestablishingrelateddiagnosissystemKeywords：Chicken；Infectiousbursald