亞洲Ⅰ型口蹄疫病毒VP1基因真核表達(dá)載體的構(gòu)建及其免疫活性初探.pdf

1、甘肅農(nóng)業(yè)大學(xué)碩士學(xué)位論文亞洲Ⅰ型口蹄疫病毒VP1基因真核表達(dá)載體的構(gòu)建及其免疫活性初探姓名：董金杰申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：王永錄伏小平20050620ConstructionofAsiaIFMDVVPlGeneRecombinantExpressionVectorandElementaryAnalysesofItsImmunologicalActivitySummaryFoot—and—mouthdiseaseVir

2、us(FMDV、causesahighlycontagiousdisease——foot—and—monthdisease(FMD)inclovenhoofedanimalsVaccineisoneofmajormethodsagainstFMDnotonlyatpresentbutalsointhefutureWiththedevelopmentofmolecularbiologyandfurtherunderstandingofst

3、ructureandfunctionofFMDVgenome，F(xiàn)MDvaccinehasgone“inactivevaccine”and“attenuatedvaccine”andhasalreadydevelopedtowardsnew—typedmolecularvacinneBecauseDNAvaccinehasmuchmerit，ithasbecomeimportantdirectionofnewtypedmolecularv

4、accineresearchInthisstudy，AsiaIserumTiletotalRNAwasextractedfromtypeofFMDVandtheeDNAfragmentofVP1wasamplifiedwithspecificprimersbyRTPCRthepurifiedeDNAfragmentwasinsertedintopGEMTeasyvectorTherecombinantplasmid‘wasidentif

5、iedbyrestrictionendonucleaseanalysisandPCRnwasprovedthattheacquiredrecombinantcontainscompleteVPlgenebyDNAsequencingAfterwardsthecompleteVPlgenefromtheidentifiedrecombinantwasamplifiedwithanotherpairofprimerscontainingBa

6、mHIandEcoRITheexpressionvectorpcDNA31()wasdigestedbyBamHIandEcoRIrespectivelyPositiveclonesnamedaspcDNAVwasidentifiedbyrestrictionendonucleaseanalysis，PCRandDNAsequencing；IL18wasinsertedintopcDNAVbythesarnemethod，itwasna

7、medpcDNAVIconstructingeukaryoticexpressionrecombinantplasmidspcDNAVI，300400gramguineapigswereimmunizedwithpcDNAVI，andaboostervaccinationwasgiventwoweekslaterKinetictestswerecarriedoutbyELISAResultsshowthathumoralimmunore

8、sponsewaselicitedbypcDNAVIDNAvaccinesandhadstrongerprotectioneffectThisencouragedfurtherworktowardsthedevelopmentofaDNAvaccineagainstFMDVandprovidedthebasisofresearchfurDNAvaccineKeywords：FMDV；AsiaIserumtype；expressionve