Wu Tsung-Meng, Lin Ke-Chun, Liau Wei-Shiang, Chao Yun-Yang, Yang Ling-Hung, Chen Szu-Yun, Lu Chung-An, Hong Chwan-Yang
Department of Agricultural Chemistry, College of Bioresources and Agriculture, National Taiwan University, Taipei, 10617, Taiwan, ROC.
Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan, ROC.
Plant Mol Biol. 2016 Jan;90(1-2):107-15. doi: 10.1007/s11103-015-0397-8. Epub 2015 Oct 30.
In the post-genomic era, many useful tools have been developed to accelerate the investigation of gene functions. Fluorescent proteins have been widely used as protein tags for studying the subcellular localization of proteins in plants. Several fluorescent organelle marker lines have been generated in dicot plants; however, useful and reliable fluorescent organelle marker lines are lacking in the monocot model rice. Here, we developed eight different GFP-based organelle markers in transgenic rice and created a set of DsRed-based gateway vectors for combining with the marker lines. Two mitochondrial-localized rice ascorbate peroxidase genes fused to DsRed and successfully co-localized with mitochondrial-targeted marker lines verified the practical use of this system. The co-localization of GFP-fusion marker lines and DsRed-fusion proteins provide a convenient platform for in vivo or in vitro analysis of subcellular localization of rice proteins.
在后基因组时代，已经开发了许多有用的工具来加速基因功能的研究。荧光蛋白已被广泛用作蛋白质标签，用于研究植物中蛋白质的亚细胞定位。在双子叶植物中已经产生了几种荧光细胞器标记系；然而，单子叶模式植物水稻中缺乏有用且可靠的荧光细胞器标记系。在这里，我们在转基因水稻中开发了八种不同的基于绿色荧光蛋白（GFP）的细胞器标记，并创建了一组基于红色荧光蛋白（DsRed）的入门载体，用于与标记系结合。两个与DsRed融合的线粒体定位的水稻抗坏血酸过氧化物酶基因，并与线粒体靶向标记系成功共定位，验证了该系统的实际应用。GFP融合标记系和DsRed融合蛋白的共定位为水稻蛋白质亚细胞定位的体内或体外分析提供了一个方便的平台。
