Department of Biology, C016, University of California San Diego, LaJolla, CA 92093.
Proc Natl Acad Sci U S A. 1987 Jul;84(14):4870-4. doi: 10.1073/pnas.84.14.4870.
The cauliflower mosaic virus (CaMV) 35S RNA promoter has been dissected and examined in a transient expression system using the firefly luciferase gene as a reporter of promoter activity. Deletion analysis has shown that the 35S RNA promoter is composed of at least three regions-distal, medial, and proximal-which are essential for activity. The distal region contains three smaller elements homologous to the simian virus 40 "core" enhancer element, the medial region possesses a CCAAT-like box, and the proximal region contains a TATA box. A DNA segment encompassing the distal region is capable of activating the CaMV 35S core promoter in an orientation-independent, but not position-independent, fashion. The distal region can also activate a heterologous weak promoter, the CaMV 19S RNA promoter, albeit not to the high levels of the 35S RNA promoter. Multimers of the distal region are able to activate the 35S RNA promoter core to even greater levels of expression than the native 35S promoter. These experiments demonstrate that elements outside the boundaries of the core promoter (composed of proximal and medial elements) are recognized in a plant cell transient expression system.
花椰菜花叶病毒 (CaMV) 35S RNA 启动子在瞬时表达系统中已被剖析和研究,该系统使用萤火虫荧光素酶基因为启动子活性的报告基因。缺失分析表明,35S RNA 启动子由至少三个区域组成 - 远端、中间和近端,这些区域对于活性是必需的。远端区域包含三个与猴病毒 40“核心”增强子元件同源的较小元件,中间区域具有 CCAAT 样框,近端区域包含 TATA 框。包含远端区域的 DNA 片段能够以不依赖于取向但不依赖于位置的方式激活 CaMV 35S 核心启动子。远端区域还可以激活异源弱启动子,即 CaMV 19S RNA 启动子,但不能达到 35S RNA 启动子的高水平。远端区域的多聚体能够将 35S RNA 启动子核心激活到比天然 35S 启动子更高的表达水平。这些实验表明,在植物细胞瞬时表达系统中,核心启动子(由近端和中间元件组成)之外的元件被识别。