Development of live cell imaging techniques for cell wall polymers

细胞壁聚合物活细胞成像技术的发展

• 批准号: 24657034
• 负责人: HASHIMOTO Takashi
• 金额: $ 2.66万
• 依托单位: Nara Institute of Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24657034/
• 关键词: セルロース; 可視化; 蛍光標識; 植物細胞; 植物

Fluorescent probes that track and monitor dynamic status of biopolymers, such as cellulose microfibrils, in living plant cells are essential for next-generation dynamic studies of plant cell wall. In this study, we fused in tandem a signal peptide from cell wall protein (expansin), a crystal cellulose-binding module from cellulose degrading enzyme (cellobiohydrolase from Trichoderma reesei), a membrane-spanning domain of plasma membrane receptor (BRI1), and green fluorescent protein (GFP), and transiently expressed this fusion protein in onion epidermal cells. GFP fluorescence was detected in cortical regions as dotts but not in linear filaments. Detailed observation using transgenic plants is necessary.

荧光探针用于跟踪和监测活植物细胞中生物聚合物（如纤维素微原纤维）的动态状态，对于下一代植物细胞壁的动态研究至关重要。在这项研究中，我们串联融合了细胞壁蛋白（expansin）的信号肽、纤维素降解酶（里氏木霉的纤维素生物水解酶）的晶体纤维素结合模块、质膜受体的跨膜结构域（BRI1）和绿色荧光蛋白（GFP），并在洋葱表皮细胞中短暂表达了这种融合蛋白。GFP荧光在皮质区呈点状，但在线状细丝中未见。利用转基因植物进行详细观察是必要的。