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《大豆誘導(dǎo)型nac轉(zhuǎn)錄因子基因啟動(dòng)子克隆及功能研究》由會員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、萬方數(shù)據(jù)山東大學(xué)碩士學(xué)位論文對全長啟動(dòng)子轉(zhuǎn)基因擬南芥進(jìn)行了GUS染色分析，發(fā)現(xiàn)：(1)轉(zhuǎn)基因擬南芥子葉葉脈、初生真葉、主根根尖、萼片、花瓣、花絲、雌蕊柱頭以及角果的果皮和果柄中均有表達(dá)；(2)ABA處理24h條件下，GUS基因在子葉中受到顯著誘導(dǎo)卜調(diào)。確定了該啟動(dòng)子ABA誘導(dǎo)特性及表達(dá)部位。2．3GmS刀啟動(dòng)予ABA誘導(dǎo)關(guān)鍵片段的確定構(gòu)建了GmST2啟動(dòng)予不同5’缺失片段驅(qū)動(dòng)G淞報(bào)告基因的植物雙元表達(dá)載體，轉(zhuǎn)化擬南芥，狀得了片段P2一P7的T3代轉(zhuǎn)基因純系種子。GUS染色結(jié)果顯示，P2同啟動(dòng)子全長一樣具有啟動(dòng)轉(zhuǎn)錄活性
2、，P3．P5活性顯著降低，P7活性部分恢復(fù)，說明P2和P3之間的片段為轉(zhuǎn)錄增強(qiáng)的關(guān)鍵片段，且P7和P5之問(一1053bp～．350bp)存在轉(zhuǎn)錄活性抑制元件。ABA處理下，P7顯示出誘導(dǎo)活性，且誘導(dǎo)部位主要在子葉和真葉，其他片段未顯示誘導(dǎo)活性。以上結(jié)果說明P7(一350bp～一1bp)均對啟動(dòng)子在葉中響應(yīng)ABA起作用。關(guān)鍵詞：大豆，NAC轉(zhuǎn)錄因子，誘導(dǎo)型啟動(dòng)子，鹽脅迫，ABA萬方數(shù)據(jù)山東大學(xué)碩士學(xué)位論文CloningandfunctionalanalysisoftheinduciblesoybeanNACtrans
3、criptionfactorpromotersABSTRACTResearchontheresponsemechanisminenvironmentstressofplant，andimprovementofplantstresstolerancebygeneengineeringhasbecomeoneoftheefficientapproachesofobtainingstresstolerancecrops．Promoteristheswitchofgeneexpression，andregulatesgene
4、expressionpatternattranscriptionlevel．Stress-inducedpromoterisabletoavoidtheadverseeffectsonplantdevelopmentcausedbyectopicexpressionofexogenousgenes．Theirapplicationhasbecomeonethehotareaofstresstoleranceplantlransgenicengineering．Inourpreviousstudy，NACtranscr
5、iptionfactorgenesGmSTlandGmST2isolatedfromsalttolerancesoybean，ShengdouNo．9，havebeenprovedtobegenesresponsetoabioticstress．TheirexpressionswereinducedbysaltanddroughtetcOnthisbasis，inthisstudy，weclonedthepromotersofthetwogenes，analyzedtheirtranscriptionactivati
6、onactivity；determinatedthetheirinductioncharacteristicsbyabioticstressandtheirtissueexpressionpaaems；comfirmedthekeyfragmentinvolvedintheinductionbyanalyzingtheactivityofdifferentfragments；preliminarilyinvestigatedthefunctionofthekeycis—actingelement；providethe
7、fundamentalsfortheapplicationofinduciblepromoteranditskeycis．a(chǎn)ctingelementinstresstoleranceplanttransgenicengineering．Mainprocessesandresultsareasfollows：1FunctionstudyofthepromoterofNACtranscriptionfactorgeneGmST／fromShengdouNo．9．I．1SequceneofGmST／promoterandc
8、／s-actingelementsTheGmSTIpromoterwasclonedfromShengdouNo．9byusinghomology．basedcloning，anditssequencecontains2000bpnucleotidesupstreaminitiationcodonATG．Analysisofthepromote
