豬偽狂犬病毒（PRV）糖蛋白gD基因的植物轉(zhuǎn)化與表達研究.pdf

1、河南師范大學碩士學位論文豬偽狂犬病毒（PRV）糖蛋白gD基因的植物轉(zhuǎn)化與表達研究姓名：祁喜濤申請學位級別：碩士專業(yè)：植物學指導教師：劉萍胡建廣20060601AbstractTakjngtobaccoasmetypeplant，thepresentthesisstudies血eexpressionofpseudorabiesvirus(PRV)glyc叩roteingDgeneintraIlsgeilicplantsandsetsupa

2、setofmethodsforplallttr孤sfonnin島transgenicplamscreemn＆】eafprote砸extractingandotherS；allthesemethodsh“elaidthegroundworkfortransgenicmaize0nsuchabasiswetransformhypersweetmaizeimmatureembryowiththepGLB—gDplaSmidwhichwas叭1

3、tbypGLBpla鋤id，GlobulpromoterandPRV出ycoproteingDgenethoughmemediationofAgrobacteriumtumefaciens卸dsuccess刪lyobtainthemmsgenjcp】a工lts，Thein丑uenceofAgrobact“umnunefaciensdenSity，t11e1eng血ofco—cul“Vationpefioda11dtempemtureon

4、mmsfonnationratehasbeenstudiediIltheexperimemThefindingsoftheexperimemalstudyareasfollows：1Bymeansof1eafdisc廿卸sfomationme也od，tobaccoistransformedbyAgrobacteriumt啪efaciens，success如Ilyinducingca工ius；thecailusthrou曲secondar

5、ycuItivation靜owsintoanindependentplanLPCRd或ection，PCR—sollthernhy城dizationandWestem坶蹦dizalionallprove恤tPIⅣ西ycoproteingDgenehasbeenintegratedintothetobaccogenomeandexpressedin也eri曲tway；thjsshowsthatPRVglycoproteingDgeneca

6、nberightlyexpressediIlplaIlts2Whenthetemperattlrestaysbetween26℃to28℃a11dtheOD600valueofthebact麗umsolutionarerespectjVelyO，3，O6a11d1O，themaizeimmatureembryoafter5minutesinfectionuIldergoesco—cultiVation，restorativecultiV

7、ationarldresistalltcalIusscreenjngat22℃nisfoundt11at廿1emmsfornlationrateisthehighestandfomulationrateOfresistantcallusreaches237％、vhentheOD600Valueofmeoriginalbacteri啪solutionisO5andinfectiondensityis033The0D600Valueforo

8、riginalbacteriumsolutiondensityandinfectiondensitybeingO5and03，theresistantcaIlusfonnulationratcofimmatureembryoisbe洫gstudi。datt11etemperatllreof22℃，25℃and28℃forone，twoandthreedayseachht啪souttllatthetmsfonnationemciencyr

9、ategoeshighestat22℃forthreedays、vitlla246％f0咖ulationrateofresistantcallus4TransgenicmaizeplantSareobtainedwi也maizeimmatureembryothrought11emediatbnofAgmbacteriumtuIllef如iensAninvestigationofthetransgenicmaizeⅡlolcculesth