Dynamic analysis of cellulose synthases in xylem cells

木质部细胞纤维素合酶的动态分析

• 批准号: 12306009
• 负责人: ITOH Takao
• 金额: $ 21.75万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12306009/
• 关键词: Cellulose synthases; GFP; GhCesA::GFP proteins; セルロース; セルロース生合成; TCの移動; ロゼット; 蛍光標識; ターミナルコンプレックス; レーザー顕微鏡

(1) It was demonstrated that part of cellulose synthase, c-di-GMP binding domain, is localized in the specific particle rows in cell membrane.(2) GhCesA::GFP was prepared for the visualization of TC movement(3) Three different techniques that is microinjection, electro-polation, and particle-gun was examined to introduce GhCesA::GFP proteins into tobacco BY2 cells. The particle-gun method was the best to introduce the proteins in the cells with a little damages.(4) The network structure was confirmed as endoplasmic reticulum (ER) by comparing the fluorescence pattern of GFP-fused proteins and DiOC6 staining images specific to ER.(5) It was shown that the partial CesA3 gene can be expressed in ER.(6) It was suggested that ChCesA::GFP proteins were localized in Golgi bodies as well by the distribution pattern of sphere-like fluorescence and their diameter similar to Golgi bodies.

(1)纤维素合酶的一部分c-di-GMP结合域定位于细胞膜上的特定颗粒行。(2)（3）采用显微注射、电脉冲和粒子枪三种不同的方法将GhCesA：：GFP蛋白导入烟草BY2细胞。粒子枪法对细胞损伤小，蛋白质导入效果最好。(4)通过比较GFP融合蛋白的荧光模式和内质网特异性DiOC 6染色图像，证实网络结构为内质网（ER）。(5)结果表明，CesA 3基因的部分片段可在ER中表达。(6)ChCesA：：GFP蛋白的荧光分布呈球形，其直径与高尔基体相似，表明ChCesA：：GFP蛋白也定位于高尔基体。

项目成果

伊東隆夫: "セルロース合成酵素はターミナルコンプレックスに局在する-フリーズ・フラクチャ法と免疫金標識法を組み合わせての実証-"植物の生長調節. 37(印刷中). (2002)

Takao Ito：“纤维素合酶位于末端复合物中 - 通过结合冷冻断裂法和免疫金标记法进行演示 -”植物生长调节 37（出版中）。

Brown RM Jr.: "The biosynthesis of cellulose"Pure Appl. Chem.. 10. 1345-1373 (1996)

Brown RM Jr.：“纤维素的生物合成”Pure Appl。

Karlson D.T., S.Kimura, T.Fujino, T.Itoh, E.N.Ashworth: "Elimination of artifactual immunogold labeling from secondary walls of woody stem sections"Plant Cell Rep.. 20. 786-790 (2002)

Karlson D.T.、S.Kimura、T.Fujino、T.Itoh、E.N.Ashworth：“消除木质茎切片次生壁中的人工免疫金标记”Plant Cell Rep.. 20. 786-790 (2002)

Itoh T.: "Deep-etching Electron Microscopy and 3-dimensional Cell Wall Architecture, In,"Wood Fomation in Trees", Nigel Chaffey, Ed."Taylor & Francis, London and New York. 83-98 (2002)

Itoh T.：“深蚀刻电子显微镜和 3 维细胞壁结构，见“树木中的木材形成”，Nigel Chaffey，Ed.“泰勒

Karlson, D.T. et al.: "Elimination of artifactual immunogold labeling from secondary walls of woody stem sections"Plant Cell Rep.. 20. 786-790 (2002)

Karlson, D.T. 等人：“消除木质茎切片次生壁中的人工免疫金标记”Plant Cell Rep.. 20. 786-790 (2002)