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《小黑楊ftlike基因的克隆、原核表達(dá)與植物超表達(dá)、rnai表達(dá)載體的構(gòu)建》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、AbstractAftervegetativegrowthinjuvenility，treesusuallyturnintotheperiodofreproductivegrowthofanthesisthroughtheresponsetoaseriesofintemalandexternalfactors(such舔vernalization，etc)，andvegetativereproductionoftreeswillslowdownandstagnate．Inplant，F(xiàn)Tgene，encodingaMADS-boxproteinwh
2、ichisactivatedbyCO，SOCl，AGL24andothergeneandsuppressedbyFLC,canpromotebyactivatingthefloralmeristemspecificgenesAPlandLFY．SointhecaseoftheexpressionlevelofFTgenecoulddeclineinpopulus，thefloweringofthepopuluswillbedelayedandmaintaininvegetativegrowthperiod(juvenilephase)togreat
3、lyincreasethetimberproduction．Onthecontrary，ifFTgenecouldoverexpressinpopulus，theanthesiswillbepromotedtoacceleratetheprocessandshortenthecycleofbreedingofpopulus．TotalRNAhasbeenextractedformtheleavesofPopulusXxiaohei，andthena535bpeDNAsequencehaSbeenclonedandcomparedthehomolog
4、ybetweenAtFTgeneofArabidopsisthaliana峨瓶a70％similarity．WenameditPnFTl．WeobtainedthePnFTI-pET-52brecombinantvectorbytheinsertionofPnFTlintotheMCSonpET-52bprokaryoticexpressionvector．BecauseofthepredictiveanalysisoftheE．colirarecodonsofPnFTlgeneshowing24．5％probability,weselectedt
5、heRosettaTMofE．coliasexpressstrains．AftertheinductionofTheRosettaTMofE．coliexpressstrainscarryingPnFTl一pET-52brecombinantvectorbyIPTC寫wehavefoundathe28KDPnFTlrecombinantinclusionproteinbytheSDS—PAGEdetectionanalysisoftheextractionproductsoftotalproteinofthallus．Thepurification
6、PnFTIrecombinantproteinhasrevertedtothesolvableandactivatedconditionthroughtheextractionofinclusion,thepurificationofelectroelution,dialysisandprecipitationbyacetone．TheproteinsamplewithalOug／ulconcentrationand90％puritydidn’tcontainthematerialswhichalenotbeneficialtotheprepara
7、tionofthepolyclonalantibodysuchaSSDS，urea,CBB(Coomassiebrilliantblue)andSOon．ThePnFTlproteinhasabsolutelyreachingthestandardofthesampleforthepreparationofthepolyclonalantibodyandbeensenttoAbmartcompanytofinishthepreparation．WehavetransformedtheplantoverexpressionandRNAiexpress
8、ionvectorsofPnFTlgenebyIn—fusionandtraditionalmethodtoresearc
