對(duì)蝦白斑綜合癥病毒特異性卵黃免疫球蛋白的制備及性能研究.pdf

1、大連理工大學(xué)碩士學(xué)位論文對(duì)蝦白斑綜合癥病毒特異性卵黃免疫球蛋白的制備及性能研究姓名：劉君君申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：金禮吉20071221對(duì)蝦白斑綜合癥病毒特異性卵黃免疫球蛋白的制備及性能研究PreparationofEggYolkImmunoglobulin(IgY)SpecifictoWhiteSpotSyndromeVirus(wssv)andEvaluationofitsPropertiesAbstr

2、actWhitespotsyndromevirus(wssv)isoneofthemostvirulentpathogens，causinghighmortalityanddramaticeconomiclossesinculturedshrimpworldwideUptodatenoeffectiveme目lsnerisavailabletopercentthespreadingofWSSVTherefore，itisargentto

3、fredaneffectivemethodtOcontrolthispathogenChickeneggyolkimmunoglobulin(IgY)providesaninexpensiveandeffectivesourceofantibodiesforthepassiveimmunizationofanimalssufferingfromdifferentkindsofdiseasesItisapromisingimmunityp

4、reparationforthetreatmentandpreventionofaquaticdiseasesInthisstudy，WSSVwereseparatedfrominfectedcrayfishThemajorantigenregiongeneVP28，VPl9，VPl5ofWSSVwereamplifiedbyPCRandthenlinkedtogetherbyoverlapextensionPCRwitllthedes

5、ignedlinkersasoverlapregionsThefusiongeneWaSclonedintotheeukaryoticexpressionplasmidpCl，andthenpCI28—1915DNAvaccineWassucessfulyconstructedTheHeLacellsweretransfectedbyDNAvaccineandtheexpressionoffusiongeneWasdetectedbyi

6、ndirectimmunofiuoreseeneeassay(／FA)TheresultsshowedthatthefusiongeneobtainedfunctionalexpressioninHeLacellsSubsequently，theDNAvaccineandformalinkilledWSSVwereusedasantigenstoimmunizelayinghensfortheproductionofIgYspecifi

7、ctoWSSVAndtheactivityofIgYWasdetectedbyindirectenzyme—linkedimmunosorbentassay(ELISA)Theresultsdemonstratedthattheactivitiesofwatersolublefraction(wsr3ofIgYspecifictoinactivatedWSSV(wssvqgY)andIgYspecifictoDNAvaccinewith

8、cpoODNsasadjuvent(DNAIgY)showedasignificantdifferencecomparedwiththecontrolgroupFurthermore，thecpGoligodeoxynucleotides(CpGODNs)enhancedtheilnnluneresponseofDNAvaccinesignificantlyTheprotectiveeffectsofIgYpowderandeggyol

9、kpowderagainstWSSVinthecrayfish(Procambaruselarki0wereevaluatedInviralneutralizationexperimentbyintramuscularinjeetioncrayfishtreatedwithWSSVIgYandDNAIgYpowerexhibitedmortalitiesof20％and80％atday9postchallengewithWSSV，res

10、pectivelyInoraladministrationexperimentthosefedadietcontaining1％WSSVIgYpowerand10％eggyolkpowdershowedmortalitiesof533％and67弼，respectivelyTheWSSV—IgYimmersiongroupshowedamortalityof267％Theseresultshavesignificantlydiffere