美洲商陸抗病毒蛋白基因的表達(dá)和遺傳轉(zhuǎn)化的研究.pdf

1、中國農(nóng)業(yè)科學(xué)研究院碩士學(xué)位論文美洲商陸抗病毒蛋白基因的表達(dá)和遺傳轉(zhuǎn)化的研究姓名：韓政申請學(xué)位級別：碩士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：蘇金王錫鋒20040601ABSTRACTIthasbeenrevealedthatPokeweedAntiviralProtein(PAP)caninhibittheinfectionofmanyplantvirusesSoitcanbeusedtocontrolplantviraldiseaseT

2、hestudiesincludetwopansInthefirstpart，cultivationconditionssuchascarbonsources，inoculationration，methanolconcentration，pH，Dbiotinconcentration，shakingspeedandinducedtimewerestudiedInfurtherresearch，thefermentationstrateg

3、ywasoptimizedbyorthogonaloperationandabetterstrategytoproducePAPwasobtainedSDS—PAGEresultsshowedthatthefinalPAPconcentrationswere45mg／LWesternblottingshowedthattheproteincouldspeciallyreactwithPAPantiserumActivitytestsre

4、vealedthattheproteincouldinhibitTMVinfectionwithhi曲efficiencyInthesecondpart，PAPgenewastransferredintoPetuniahybridainordertoobtaintransgenicplantsthatcancontroltheinfectionofsomeplantvirusesAtfirsttheplantexpressionvect

5、orpBll21withPAPgenewasclonedintoAgrobacteriumstrainLBA4404byfreeze—thawmethod，andtheresuldngAgrobacteriumtransformantswereverifiedbyPCRamplificationThenthePAPgenewastransferredintoPetuniahybridabyleafdiscmethodmediatedby

6、theAgrobacteriamtransformantsTransgenieplantswereobminedthroughthescreeningwithkanamyeinPCRamplificationconfirmedtheresultsThispartofstudiesaccumulatedthefoundationfortheadvancesofplantgeneengineeringofPhybridaKeywords：P