| Project/Area Number |
15K14461
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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 |
Structural biochemistry
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| Research Institution | Nagasaki University |
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Principal Investigator |
MASUMOTO Hiroshi 長崎大学, 医歯薬学総合研究科(医学系), 講師 (80423151)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | 質量分析 / タンパク同定 / タンパクの安定同位体標識 / タンパクのビオチン化修飾 / タンパク間相互作用 / タンパク定量 / タンパクの翻訳後修飾 / 同位体解析 / ゲノム編集技術 / タンパク修飾 / ビオチン化修飾 |
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| Outline of Final Research Achievements |
We tried to explore novel protein analysis combined with SILAC (Stable Isotope Labeling by Amino acids in cell culture) and chemical crosslinking to identify the components in protein complex named as SILAC-X-linking-protein purification-Mass spectrometry (SXP-MS). However, SXP-MS did not work well to identify proteins because of a huge number of contaminated proteins caused by cross-linking.
To substitute SXP-MS analysis, we applied BioID method in budding yeast to identify the interacting factors by biotiylation. This method was applicable for protein identification. During this research, we have developed the novel genome editing technology named as CRIPSR/Transposon gene integration (CRITGI) to achieve the protein overexpression as a by-product.
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