Bang Seung Woon, Park Su-Hyun, Kim Youn Shic, Choi Yang Do, Kim Ju-Kon
Crop Biotechnology Institute, GreenBio Science and Technology, Seoul National University, Pyeongchang, 232-916, Korea,
Planta. 2015 Jun;241(6):1529-41. doi: 10.1007/s00425-015-2278-4. Epub 2015 Mar 26.
We have characterized four novel constitutive promoters ARP1, H3F3, HSP and H2BF3 that are active in all tissues/stages of transgenic plants and stable over two homozygous generations. Gene promoters that are active and stable over several generations in transgenic plants are valuable tools for plant research and biotechnology. In this study, we characterized four putative constitutive promoters (ARP1, H3F3, HSP and H2BF3) in transgenic rice plants. Promoter regions were fused to the green fluorescence protein (GFP) reporter gene and transformed into rice. Single-copy transgenic lines were then selected and promoter activity was analyzed in various organs and tissues of two successive homozygous generations. All four promoters showed a broad expression profile in most tissues and developmental stages, and indeed the expression of the ARP1 and H3F3 promoters was even greater than that of the PGD1 promoter, a previously described constitutive promoter that has been used in transgenic rice. This observation was based on expression levels in leaves, roots, dry seeds and flowers in both the T2 and T3 generations. Each promoter exhibited comparable levels of activity over two homozygous generations with no sign of transgene silencing, which is an important characteristic of promoters to be used in crop biotechnology applications. These promoters therefore have considerable potential value for the stable and constitutive expression of transgenes in monocotyledonous crops.
我们已经鉴定出四个新的组成型启动子ARP1、H3F3、HSP和H2BF3，它们在转基因植物的所有组织/阶段均有活性，并且在两个纯合世代中保持稳定。在转基因植物中几代都具有活性且稳定的基因启动子是植物研究和生物技术中的宝贵工具。在本研究中，我们在转基因水稻植株中鉴定了四个假定的组成型启动子（ARP1、H3F3、HSP和H2BF3）。将启动子区域与绿色荧光蛋白（GFP）报告基因融合并转化到水稻中。然后选择单拷贝转基因株系，并在两个连续纯合世代的各种器官和组织中分析启动子活性。所有四个启动子在大多数组织和发育阶段均表现出广泛的表达谱，实际上ARP1和H3F3启动子的表达甚至高于PGD1启动子，PGD1是先前描述的已用于转基因水稻的组成型启动子。这一观察结果基于T2和T3代叶片、根、干种子和花中的表达水平。每个启动子在两个纯合世代中均表现出相当的活性水平，没有转基因沉默的迹象，这是用于作物生物技术应用的启动子的一个重要特征。因此，这些启动子对于单子叶作物中转基因的稳定组成型表达具有相当大的潜在价值。
