Ha S B, An G
Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.
Proc Natl Acad Sci U S A. 1988 Nov;85(21):8017-21. doi: 10.1073/pnas.85.21.8017.
We studied cis regulatory elements controlling the light-dependent organ-specific expression of Arabidopsis thaliana chlorophyll a/b binding protein gene (cab1) by stably transforming tobacco plants using a tumor-inducing (Ti) plasmid vector system. The results from the 5' and internal deletion analyses indicate that there are at least three cis-acting elements that are involved in the light-dependent developmental expression of cab1 gene. Two such elements are located at the immediate upstream regulatory region and the other element is located at the further upstream region. The 1120-base-pair (bp) DNA fragment containing the immediate and far upstream region can confer light-inducible organ specificity on the truncated nos promoter. However, deletion of the 39-bp DNA fragment at the immediate upstream regulatory region from this hybrid promoter resulted in a nonfunctional promoter, revealing that the 39-bp region is important for the cab promoter specificity. Further analyses of this region suggest that a potential Z-DNA-forming sequence (ATACGTGT) is involved in light-dependent developmental expression of the cab1 gene. Two additional Z-DNA-forming sequences (ACACATAT) that are inverted repeats of this sequence are also found in the upstream region where the additional regulatory elements are expected.
我们利用肿瘤诱导(Ti)质粒载体系统稳定转化烟草植株,研究了控制拟南芥叶绿素a/b结合蛋白基因(cab1)光依赖型器官特异性表达的顺式调控元件。5'端和内部缺失分析结果表明,至少有三个顺式作用元件参与了cab1基因的光依赖型发育表达。其中两个元件位于紧邻上游调控区,另一个元件位于更上游区域。包含紧邻上游和远上游区域的1120碱基对(bp)DNA片段可赋予截短的胭脂碱合成酶(nos)启动子光诱导的器官特异性。然而,从该杂合启动子中删除紧邻上游调控区的39 bp DNA片段会导致启动子失去功能,这表明该39 bp区域对cab启动子特异性很重要。对该区域的进一步分析表明,一个潜在的Z-DNA形成序列(ATACGTGT)参与了cab1基因的光依赖型发育表达。在预期存在其他调控元件的上游区域还发现了另外两个该序列的反向重复Z-DNA形成序列(ACACATAT)。