[Visualization of actin cytoskeleton in living cells of Torenia fourineri using GFP-mTn fusion protein].

Shi yan sheng wu xue bao Pub Date : 2005-04-01
Yi Bo Qi, Jun Bai, Yu Zhen Han
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Abstract

By transformation mediated by Agrobacterium tumefacien, we successfully transferred the chimeric gene of GFP-mTn ( mTn is the binding domain of microfilament binding protein talin from mouse, which can show the microfilament in living cell ) into Torenia fournieri. Using confocal laser scanning microscopy (CLSM), the distribution of fusion protein in different kinds of tissues and cell in transgenic Torenia fournieri was observed. GFP fluorescence was found in leaf epidermal cell, stomatal guard cell and root epidermal cell. Actin filaments can be visualized clearly only in guard cells. In the guard cells of open stomata under light, actin filaments arrange reticularly and randomly in cortical cytoplasm. In the guard cells of closed stomata under darkness, actin filaments arrange curly along the longitude of guard cell, and some helix and ring structures were found. GFP fluorescence was not found in other cell types, including stem epidermal cell, root hair cell and reproductive organs. The transgenic Torenia fournieri we got provides a suitable material to study dynamics of actin filament in stomatal guard cell.

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[利用GFP-mTn融合蛋白可视化富氏托雷尼亚活细胞中的肌动蛋白细胞骨架]。
通过农杆菌介导的转化,我们成功地将小鼠体内的GFP-mTn (mTn是微丝结合蛋白talin的结合域,可以显示活细胞中的微丝)嵌合基因转移到富氏托伦菌中。利用激光共聚焦扫描显微镜(CLSM)观察了转基因富氏托伦虫不同组织和细胞中融合蛋白的分布。叶片表皮细胞、气孔保护细胞和根表皮细胞均可见到荧光。肌动蛋白丝仅在保护细胞中可见。在光照下开放气孔的保护细胞中,肌动蛋白丝在皮层细胞质中呈网状和随机排列。在黑暗条件下封闭气孔的保护细胞中,肌动蛋白丝沿保护细胞的经度卷曲排列,并有螺旋和环状结构。在茎表皮细胞、根毛细胞和生殖器官等其他类型的细胞中均未发现GFP荧光。所获得的转基因富氏托氏菌为研究气孔保护细胞中肌动蛋白丝的动态提供了合适的材料。
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