重组TMV的灭活与在烟草上诱导过敏性反应现象的初步分析

《重组TMV的灭活与在烟草上诱导过敏性反应现象的初步分析》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、上海大学硕士学位论文ABSTRACTByusingrecombinantplantvirusessuchastobaccomosaicvirus(TMV)asvectors，wecouldaddgenesequencesencodingantigenicdeterminantofvaccineorbiopharmaceuticalproteinsintoTMVgenome．Afterinfectingplants，thisrecombinantviruscouldmultiplyandresemblehi【ghlevelsinplants，leadingtohighlevelsofant

2、igengeneTherewereseveralpotentialadvantagesofusingthisexpressionsystemfortheproductionofvaccineandbiopharmaceuticalprotein．Theamountofrecombinantproductscouldbeproducedhighly,thepurificationrequirementwaseasy,andevencouldbeeliminated．Mostnotably,theproductioncostscouldbegreatlyreduced．Sotheseadva

3、ntagescaughtmanyscientists’eyes．Bynow,wehadstudiedverymuchabouttheexpressionofforeignpeptidesintobaccobyarecombinantTMV-basedvectorinthelaboratory．Theseforeignpeptidescamefromfoot-and-mouthdiseasevirusandsevereacuterespiratorysyndromecoronavirus．Recombinantvirusesthatwedescribedcouldinfectthetoba

4、ccosystematically,thoughtheirinfectionWaslowerthantheinfectionofwidetypeTMV，theystillengenderedsafetyconcerns．Inordertosolvetheproblem，weusedtwodifferentmethods：physicalinactivatedmethodandcloneconstructionofdefectiveTMVoVIII上海大学硕士学位论文Theformer，recombinantviralparticleWBSirradiatedusingdifferentd

5、osagesof60Coy．Throughthetest,wefoundthatTMVS6-1couldbetotallyinactivatedby60Coy．Thestudyhadsuggestedthatproductionspecificoptimizationofgammairradiationhadthepotentialtoprovideeffectivevirusinactivation，andthismethodhadlittleeffect011substantialantigenactivityforforeignpeptides．Soitcouldrelieveef

6、fectivelybiosafyhiddendanger．Thelatterutilizedthetheoryoffunctionalgenecomplementation,namely,thedefectiverecombinantviruswhichdeletedafunctionalproteincouldaccumulatetoacertainlevelintheplantcellwiththehelpoftransgenieplantsorhelpervirusesthatexpressedageneencodingthefunctionalTMVprotein．Forspec

7、ificstudiesofthefeasibilityoffunctionalcomplementaryintobacco，firstlyweconstructeddefectiveTMVmMPeDNAcloneencodingadysfunctionalMEThetobaccoseedlingsweTecoinoculatedwithmixtureofRNAanddRNAina1：1ratioofinvitrotranscript