Skriver K, Olsen F L, Rogers J C, Mundy J
Carlsberg Research Laboratory Copenhagen, Denmark.
Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7266-70. doi: 10.1073/pnas.88.16.7266.
We have used a transient expression assay in aleurone protoplasts of barley to delineate hormone response elements of the abscisic acid (ABA)-responsive rice gene Rab16A and of the gibberellin A3 (GA3)-responsive barley alpha-amylase gene Amy 1/6-4. Our approach used transcriptional fusions between their 5' upstream sequences and a bacterial chloramphenicol acetyltransferase reporter gene. A chimeric promoter containing six copies of the -181 to -171 region of Rab 16A fused to a minimal promoter conferred ABA-responsive expression on the reporter gene. Transcription from this ABA response element (GTACGTGGCGC) was unaffected by GA3. A chimeric promoter containing six copies of the -148 to -128 sequence of Amy 1/6-4 fused to the minimal promoter conferred GA3-responsive expression on the reporter gene. Transcription from this GA3 response element (GGCCGATAACAAACTCCGGCC) was repressed by ABA. The effect on transcription from both hormone response elements was orientation-independent, indicating that they function as inducible enhancers in their native genes.
我们利用大麦糊粉层原生质体中的瞬时表达分析,来确定脱落酸(ABA)应答型水稻基因Rab16A和赤霉素A3(GA3)应答型大麦α-淀粉酶基因Amy 1/6 - 4的激素反应元件。我们的方法是利用它们5'上游序列与细菌氯霉素乙酰转移酶报告基因之间的转录融合。一个包含Rab 16A - 181至-171区域六个拷贝并与最小启动子融合的嵌合启动子,赋予了报告基因ABA应答型表达。来自这个ABA反应元件(GTACGTGGCGC)的转录不受GA3影响。一个包含Amy 1/6 - 4 - 148至-128序列六个拷贝并与最小启动子融合的嵌合启动子,赋予了报告基因GA3应答型表达。来自这个GA3反应元件(GGCCGATAACAAACTCCGGCC)的转录受到ABA的抑制。两种激素反应元件对转录的影响均与方向无关,表明它们在其天然基因中作为诱导型增强子发挥作用。