| Project/Area Number |
24650266
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Fukushima Medical University |
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Principal Investigator |
WADA Ikuo 福島県立医科大学, 医学部, 教授 (40182969)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 蛍光タンパク質 / 分泌系 / 酸化的フォールディング / 蛍光相関分光法 / 小胞体 / 蛍光タンパク / FLIM / 一分子輝度計測 / 分泌 / 蛍光イメージング / 分子イメージング / 分泌過程 |
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| Outline of Final Research Achievements |
It was not clear if the fluorescent proteins, which are cytosolic nature, can function properly as an inert tag in the oxidative environment. Based on our finding that GFP forms disulfide-bonds in the ER and behaves anomalously, we aimed to develop small fluorescent proteins suitable for the extracellular space. Following the successful elimination of Cys from SGFP2 and TagRFP, we developed a series of fluorescent proteins with various photochemical properties including photo-chromism, which are not affected by the oxidative folding. Those properties can be applied for photoconversion-induced FRET measurement to monitor oligomerization of secretory cargo. To generate a practical smaller fluorescent tag, we focused on a FMN-dependent fluorescent protein and found a set of mutations to adapt the molecule for the oxidative folding. Those sets of improved fluorescent proteins should be useful to study proteins in the extracellular space.
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