MOA Key Laboratory on Safety Assessment (Molecular) of Agri-GMO, Institute of Biotechnology, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
College of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China.
Int J Mol Sci. 2018 Jul 10;19(7):2009. doi: 10.3390/ijms19072009.
Promoters play a very important role in the initiation and regulation of gene transcription. Green-tissue promoter is of great significance to the development of genetically modified crops. Based on RNA-seq data and RT-PCR expression analysis, this study screened a gene, (GREEN SPECIAL EXPRESS), which is expressed specifically in green tissues. The study also isolated the promoter of the gene (OrGSEp), and predicted many cis-acting elements, such as the CAAT-Box and TATA-Box, and light-responding elements, including circadian, G-BOX and GT1 CONSENSUS. Histochemical analysis and quantification of GUS activity in transgenic plants expressing GUS under the control of OrGSEp revealed that this promoter is not only green tissue-specific, but also light-inducible. The ability of a series of 5’-deletion fragments of OrGSEp to drive GUS expression in was also evaluated. We found that the promoter region from −54 to −114 is critical for the promoter function, and the region from −374 to −114 may contain core cis-elements involved in light response. In transgenic rice expressing GUS under the control of OrGSEp, visualization and quantification of GUS activity showed that GUS was preferentially expressed in green tissues and not in endosperm. OrGSEp is a useful regulatory element for breeding pest-resistant crops.
启动子在基因转录的起始和调控中起着非常重要的作用。绿色组织启动子对转基因作物的发展具有重要意义。本研究基于 RNA-seq 数据和 RT-PCR 表达分析，筛选出一个在绿色组织中特异性表达的基因 (GREEN SPECIAL EXPRESS)。本研究还分离了该基因 (OrGSEp) 的启动子，并预测了许多顺式作用元件，如 CAAT-Box 和 TATA-Box，以及光响应元件，包括circadian、G-BOX 和 GT1 CONSENSUS。在转基因植物中，通过 OrGSEp 控制下表达 GUS 的组织化学分析和 GUS 活性的定量分析表明，该启动子不仅具有绿色组织特异性，而且具有光诱导性。还评估了一系列 OrGSEp 5’-缺失片段驱动 GUS 表达的能力。我们发现，−54 到−114 的启动子区域对于启动子功能至关重要，而−374 到−114 的区域可能包含参与光反应的核心顺式元件。在受 OrGSEp 控制表达 GUS 的转基因水稻中，GUS 活性的可视化和定量分析表明，GUS 优先在绿色组织中表达，而不在胚乳中表达。OrGSEp 是一种用于培育抗虫害作物的有用调控元件。
