Project/Area Number |
15K14319
|
Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
Allocation Type | Multi-year Fund |
Research Field |
Neurophysiology / General neuroscience
|
Research Institution | Osaka University |
Principal Investigator |
Mori Yasutake 大阪大学, 医学系研究科, 准教授 (00343252)
|
Co-Investigator(Kenkyū-buntansha) |
岡 雄一郎 大阪大学, 連合小児発達学研究科, 講師 (30614432)
猪口 徳一 大阪大学, 医学系研究科, 助教 (60509305)
|
Co-Investigator(Renkei-kenkyūsha) |
SATO Makoto 大阪大学, 大学院医学系研究科, 教授 (10222019)
|
Project Period (FY) |
2015-04-01 – 2017-03-31
|
Project Status |
Completed (Fiscal Year 2016)
|
Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
|
Keywords | 局所翻訳 / 翻訳停止因子 / シナプス / 分子イメージング / 翻訳 / messenger RNA / リアルタイム解析 / 可視化 / 蛋白質翻訳 / イメージング / 脳・神経 |
Outline of Final Research Achievements |
To develop a novel detection method of protein synthesis on time at restricted region of the cells, we focused on molecular interaction between two translation factors at the last phase of ribosomal scanning on specific mRNA. Two different fluorophores CFP and YFP were fused with the interacting trasnlation termmination factors to produce a pair of molecular probe. We expressed these probed in the cells and anchored them around the stop codon of specific mRNA. In this preparation, the end of the mRNA scanning induced proximity of two fluorophores by interaction of the molecular probes, so that excitation of CFP emitted YFP fluorescence. We detected the fluorescence energy transfer in the cells that are activated by serum and growth factors. In conclusion, this energy tranfer from CFP to YFP well related to translation state od the cells. We successfully detected protein synthesis rate in living cells without destroying cell structure.
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