大豆诱导型nac转录因子基因启动子克隆及功能研究

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1、万方数据山东大学硕士学位论文对全长启动子转基因拟南芥进行了GUS染色分析，发现：(1)转基因拟南芥子叶叶脉、初生真叶、主根根尖、萼片、花瓣、花丝、雌蕊柱头以及角果的果皮和果柄中均有表达；(2)ABA处理24h条件下，GUS基因在子叶中受到显著诱导卜调。确定了该启动子ABA诱导特性及表达部位。2．3GmS刀启动予ABA诱导关键片段的确定构建了GmST2启动予不同5’缺失片段驱动G淞报告基因的植物双元表达载体，转化拟南芥，状得了片段P2一P7的T3代转基因纯系种子。GUS染色结果显示，P2同启动子全长一样具有启动转录活性

2、，P3．P5活性显著降低，P7活性部分恢复，说明P2和P3之间的片段为转录增强的关键片段，且P7和P5之问(一1053bp～．350bp)存在转录活性抑制元件。ABA处理下，P7显示出诱导活性，且诱导部位主要在子叶和真叶，其他片段未显示诱导活性。以上结果说明P7(一350bp～一1bp)均对启动子在叶中响应ABA起作用。关键词：大豆，NAC转录因子，诱导型启动子，盐胁迫，ABA万方数据山东大学硕士学位论文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．actingelementinstresstoleranceplanttransgenicengineering．Mainprocessesandresultsareasfollows：1FunctionstudyofthepromoterofNACtranscriptionfactorgeneGmST／fromShengdouNo．9．I．1SequceneofGmST／promoterandc

8、／s-actingelementsTheGmSTIpromoterwasclonedfromShengdouNo．9byusinghomology．basedcloning，anditssequencecontains2000bpnucleotidesupstreaminitiationcodonATG．Analysisofthepromote