Abstract
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.
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Acknowledgments
We are grateful to the Joint Center for Instruments and Research of the College of Bioresources and Agriculture at National Taiwan University for confocal microscopy and technical support. This work was supported in part by the Ministry of Science and Technology (MOST) of the Republic of China (NSC 101-2313-B-002-008-MY3 to C. Y. Hong). T. M. Wu was supported by a postdoctoral fellowship from MOST (NSC 103-2811-B-002-142).
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Wu, TM., Lin, KC., Liau, WS. et al. A set of GFP-based organelle marker lines combined with DsRed-based gateway vectors for subcellular localization study in rice (Oryza sativa L.). Plant Mol Biol 90, 107–115 (2016). https://doi.org/10.1007/s11103-015-0397-8
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DOI: https://doi.org/10.1007/s11103-015-0397-8