| Project/Area Number |
23651230
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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 |
Chemical biology
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| Research Institution | Tohoku University |
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Principal Investigator |
SAKAMOTO Seiji 東北大学, 多元物質科学研究所, 助教 (30335228)
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| Project Period (FY) |
2011 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2011: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | プロテアーゼ / 蛍光タンパク質 / 発光タンパク質 / バイオプローブ / バイオセンサー / 分割型タンパク質 / 分割型蛍光タンパク質 / 分割型インテイン / カスパーゼ / アポトーシス |
|---|
| Research Abstract |
We designed a dual-protease biosensor for two different protease activities using doubly inhibited split-GFP, which was fully activated only when two proteases were present at the same time. In the system, N- and C-terminal fragments of split-GFP of split-luciferase were cyclized via the distinct protease substrate sequences, respectively. The simultaneous addition of two proteases resulted in cleavage of both substrate linkers, leading to the functional recovery of split-GFP and split-luciferase. These results clearly demonstrate the successful construction of the dual-protease biosensor for two different caspase activities by using split-GFP and split-luciferase.
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