| Project/Area Number |
07454221
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物生理
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| Research Institution | Himeji Institute of Technology |
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Principal Investigator |
SONOBE Seiji Himeji Institute of Technology, Faculty of Science, associate professor, 理学部, 助教授 (30197024)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1995: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | actin filament / microtubules / Tobacco BY-2 Cells / microtubule-associated protein / actin-bindring protein / Hydrocharis / root hair / Cytoplasmic streaming / 植物細胞 / アクチン結合タンパク質 / タバコ培養細胞BY-2 / 微小管 / タンパク質精製 / 架橋タンパク質 |
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| Research Abstract |
In order to clarify the function of actin filaments in the organization of cortical microtubules, we tried to identify a protein (s) which could bind to both microtubules and actin filaments. Crude cytoplasmic extract was prepared from evacuolated protoplasts of tobaco BY-2 cells and tubulin and actin were polymerized upon addition of taxol and phalloidin and the polymers were collected. After depolymerization of microtubules followed by repolymerization, microtubule-associated protein fraction were obtained by salt extraction and the fraction was further fractionated by MonoQ column. On the other hand, we also tried to purify tubulin and atin and succeeded in their purification. An monoclonal antibody specific to plant actin could be prepared. Cosedimentation experiments using purified microtubules and actin filaments demonstrated that 120 and 90 kDa polypeptides could bind to both microtubules and actin filaments. These polypeptides induced bundle formation of actin filaments. Now, we are trying to prepare antibodies against these polypeptides to investigate colocalization of them with cortical microtubules. On the other hand, organization of actin filaments in Hydorcharis root hair cells was studied. In the cortical region, actin filaments colocalized with microtubules and reconstruction of actin filaments in ordered array was inhibited in the absence of microtubules, suggesting that arrangement of actin filaments was regulated by microtubules in Hydrocharis root hair.
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