National Engineering Laboratory for Tree Breeding (NDRC), Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants (MOE), the Tree and Ornamental Plant Breeding and Biotechnology Laboratory (SFA), College of Biological Science and Biotechnology, Beijing Forestry University, Qinghua Eastern Road No.35, Haidian District, Beijing 100083, China.
Int J Mol Sci. 2013 Mar 18;14(3):6187-204. doi: 10.3390/ijms14036187.
A novel sequence that functions as a promoter element for moderate constitutive expression of transgenes, designated as the PtMCP promoter, was isolated from the woody perennial Populus tomentosa. The PtMCP promoter was fused to the GUS reporter gene to characterize its expression pattern in different species. In stable Arabidopsis transformants, transcripts of the GUS reporter gene could be detected by RT-PCR in the root, stem, leaf, flower and silique. Further histochemical and fluorometric GUS activity assays demonstrated that the promoter could direct transgene expression in all tissues and organs, including roots, stems, rosette leaves, cauline leaves and flowers of seedlings and maturing plants. Its constitutive expression pattern was similar to that of the CaMV35S promoter, but the level of GUS activity was significantly lower than in CaMV35S promoter::GUS plants. We also characterized the promoter through transient expression in transgenic tobacco and observed similar expression patterns. Histochemical GUS staining and quantitative analysis detected GUS activity in all tissues and organs of tobacco, including roots, stems, leaves, flower buds and flowers, but GUS activity in PtMCP promoter::GUS plants was significantly lower than in CaMV35S promoter::GUS plants. Our results suggested that the PtMCP promoter from poplar is a constitutive promoter with moderate activity and that its function is presumably conserved in different species. Therefore, the PtMCP promoter may provide a practical choice to direct moderate level constitutive expression of transgenes and could be a valuable new tool in plant genetic engineering.
从木本多年生植物毛白杨中分离到一个新的序列,该序列可作为转基因中度组成型表达的启动子元件,命名为 PtMCP 启动子。PtMCP 启动子与 GUS 报告基因融合,以表征其在不同物种中的表达模式。在稳定的拟南芥转化体中,通过 RT-PCR 可以在根、茎、叶、花和蒴果中检测到 GUS 报告基因的转录物。进一步的组织化学和荧光 GUS 活性测定表明,该启动子可以在包括根、茎、莲座叶、茎生叶和花的幼苗和成熟植物的所有组织和器官中指导转基因表达。其组成型表达模式与 CaMV35S 启动子相似,但 GUS 活性水平明显低于 CaMV35S 启动子::GUS 植物。我们还通过在转基因烟草中的瞬时表达来表征该启动子,并观察到类似的表达模式。组织化学 GUS 染色和定量分析检测到烟草的所有组织和器官中都有 GUS 活性,包括根、茎、叶、花蕾和花,但 PtMCP 启动子::GUS 植物中的 GUS 活性明显低于 CaMV35S 启动子::GUS 植物。我们的结果表明,杨树的 PtMCP 启动子是一个具有中度活性的组成型启动子,其功能在不同物种中可能是保守的。因此,PtMCP 启动子可能为指导转基因的适度组成型表达提供了一个实用的选择,并且可以成为植物遗传工程中的一个有价值的新工具。