MODULAR ORGANIZATION AND DEVELOPMENTAL ACTIVITY OF AN ARABIDOPSIS-THALIANA EF-1-ALPHA GENE PROMOTER
MODULAR ORGANIZATION AND DEVELOPMENTAL ACTIVITY OF AN ARABIDOPSIS-THALIANA EF-1-ALPHA GENE PROMOTER
复制标题
DOI：
10.1007/bf00292002
复制
发表时间：
1993-04-01
期刊：
影响因子：
--
通讯作者：
LESCURE, B
中科院分区：
文献类型：
--
作者：
CURIE, C;AXELOS, M;LESCURE, B
在转基因拟南芥植物中分析拟南芥A1 EF-1 α基因启动子的活性。将A1基因的5'上游序列和几个启动子缺失融合到β-葡萄糖醛酸酶（GUS）编码区。通过GUS活性的定量和组织化学测定来监测启动子活性。结果表明，A1启动子表现出模块化的组织。相对于转录起始位点的上游和下游序列参与营养生长期间的定量和组织特异性表达。一个上游元件可能参与激活分生组织中的表达;下游区域，对应于5'非编码区（5' IVS）内的内含子，对于根中的表达是重要的;上游和下游序列都是叶中表达所必需的，表明EF-1 α顺式调控元件的组合特性。在转染的拟南芥原生质体中的瞬时表达实验的结果加强了这种特定的组合调控的概念。当启动子活性在A1 EF-1 α上游序列的控制下时，5 ′ IVS的缺失对表达的影响比这些上游序列被35 S增强子取代时大得多。类似地，对应于A1 EF-1 α上游顺式作用元件（TEF 1盒）的合成寡核苷酸在与不含TEF 1的EF 1-α启动子融合时能够部分恢复原始活性，但在与不含增强子的35 S启动子融合时没有显著效果。
The activity of the Arabidopsis thalana A1 EF-1alpha gene promoter was analyzed in transgenic Arabidopsis plants. The 5' upstream sequence of the A1 gene and several promoter deletions were fused to the beta-glucuronidase (GUS) coding region. Promoter activity was monitored by quantitative and histochemical assays of GUS activity. The results show that the A1 promoter exhibits a modular organization. Sequences both upstream and downstream relative to the transcription initiation site are involved in quantitative and tissue-specific expression during vegetative growth. One upstream element may be involved in the activation of expression in meristematic tissues; the downstream region, corresponding to an intron within the 5' non-coding region (5'IVS), is important for expression in roots; both upstream and downstream sequences are required for expression in leaves, suggesting combinatorial properties of EF-1alpha cis-regulatory elements. This notion of specific combinatorial regulation is reinforced by the results of transient expression experiments in transfected Arabidopsis protoplasts. The deletion of the 5'IVS has much more effect on expression when the promoter activity is under the control of A1 EF-1alpha upstream sequences than when these upstream sequences were replaced by the 35S enhancer. Similarly, a synthetic oligonucleotide corresponding to an A1 EF-1alpha upstream cis-acting element (the TEF1 box), is able to restore partially the original activity when fused to a TEF1-less EF1-alpha promoter but has no significant effect when fused to an enhancer-less 35S promoter.