The South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China.
Methods Mol Biol. 2023;2652:119-127. doi: 10.1007/978-1-0716-3147-8_6.
Fluorescent proteins (FPs) revolutionized the cell biology research by visualizing the dynamics of cellular events. In fusion with the targeted proteins, the FPs can be utilized to monitor the protein dynamics and localization in cells. Recently, FPs have been used as reporters for live cell imaging to study the protein localization or organelles dynamics in plants, allowing cell biologists to explore the plant cell function by obtaining tremendous details of cell structures and functions in combination with confocal imaging. To facilitate the usage of fluorescent proteins for protein localization and dynamic analysis in plant cell biology research, here we describe the updated protocol of Agrobacterium-mediated transformation of Arabidopsis thaliana using fluorescent proteins to generate the stable expression transgenic plants for protein trafficking and localization study. We further use the GFP-tagged SDP1 (sugar-dependent protein) lipase, mCherry-tagged peroxisome marker, and BODYPY or Nile Red (lipid droplet staining dye) as examples to introduce the method for the protein localization analysis in plants.
荧光蛋白（FPs）通过可视化细胞事件的动态，彻底改变了细胞生物学研究。与靶向蛋白融合后，FPs 可用于监测细胞内蛋白质的动态和定位。最近，FPs 已被用作活细胞成像的报告器，用于研究植物中蛋白质的定位或细胞器的动态，使细胞生物学家能够通过获得细胞结构和功能的大量细节来探索植物细胞功能，结合共聚焦成像。为了便于在植物细胞生物学研究中使用荧光蛋白进行蛋白质定位和动态分析，我们在此描述了使用荧光蛋白对拟南芥进行农杆菌介导的转化的最新方案，以生成用于蛋白质运输和定位研究的稳定表达的转基因植物。我们进一步以 GFP 标记的 SDP1（糖依赖性蛋白）脂肪酶、mCherry 标记的过氧化物酶体标记物以及 BODYPY 或尼罗红（脂滴染色染料）为例，介绍了植物中蛋白质定位分析的方法。
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