拟南芥逆境诱导型启动子rd29A的克隆与活性检测.pdf

32 1 ( ) V o l 32, N o 1 200 8 1 Journa l o f N an jing Fo restry U n iversity( N atura l Sciences Ed ition) J an , 200 8 rd29A * , , , ( , , 2 10037 ) : 以拟南芥基因组 DNA 为模板, 通过特异 PCR 扩增, 克隆逆境诱导表达启动子 rd29A 序列分析表 , 该启 动子与NCBI上已报道 rd29A 的启动子有 9964% 的同源性, 其序列含有干旱诱导表达元件 DRE 和ABA 作用元 件ABR E 等顺式作用元件用 rd29A 启动子替换 pBI121载体上的 35S 启动子构建新的载体 pBrd - GU S, 并以农 杆菌介导法将其转入烟草转基因烟草的GU S 组织化学染色及 PCR 分析结果表 , 在低温干旱及高盐等胁迫 诱导下, rd29A 启动子可增强 GU S 基因表达, 因此, rd29A 启动子可应用于植物抗逆基因工程研究 : 逆境; rd29A 启动子; GUS 瞬时表达; 转基因烟草 : S722 : A : 1000 - 2006 ( 2008) 01- 0006- 05 Cloning of stressinducible promoter rd29A from A rab idop sis tha liana and its activity detection in transgenic tobacco * YANG Chunx ia, CHEN Y ing, HUANG M inren , L I H uog en ( N an jing F ores try U n iv ersity, T he K ey L aboratory of F orest G enet ics and G en e Eng ineering of th e S tate F ores try A dm in istration and Jiang su P rov in ce, N an j ing 2 10037, Ch ina) Abstract: Stress indu c ible prom oter rd29A w as cloned by PCR from A ra bidop sis thaliana genom ic DNA Sequ ence analysis show ed tha t th is prom ote r w as 9951% hom o logou sw ith reported prom oter rd29A in N CB I and conta ined severa l s isacting elem ents inc lu ding dehydration respon sive elem ent ( DRE) and A BA respon sive e lem ent ( ABRE ) P romo ter 35S o f vecto r pB I12 1 w as replaced by prom o ter rd29A, thu s vecto r pB rd - GU S w as con stru cted and transferred