| Project/Area Number |
13559007
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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 | 展開研究 |
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| Research Field |
広領域
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| Research Institution | Keio University |
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Principal Investigator |
YANAGAWA Hiroishi Fac. Sci. & Tech., Professor, 理工学部, 教授 (40327672)
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| Co-Investigator(Kenkyū-buntansha) |
DOI Nobuhide Fac. Sci. & Tech., Assistant Professor, 理工学部, 講師 (50327673)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 2002: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | Proteomics / Proteome / Micro array / Protein Chip / Fluorescence Labeling / Interaction / Puromycin |
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| Research Abstract |
Protein microarrays or proteome chips are potentially powerful tools for comprehensive analysis of protein-protein interactions. In interaction analysis, a set of immobilized proteins is arrayed on slides and each slide is probed with a set of fluorescently labeled proteins. Here we have developed and tested an 'in vitro' protein microarray, in which both arraying and probing proteins were prepared by cell-free translation. The in vitro synthesis of fluorescently labeled proteins was accomplished by a new method: a fluorophore-puromycin conjugate was incorporated into a protein at the C-terminus on the ribosome. The resulting fluorescently labeled proteins were confirmed to be useful for probing protein-protein interactions on protein microarrays in model experiments. We have used three pairs of model proteins that are known to interact: the classical oncogenes, c-Fos and c-Jun; Polycomb-related genes, RYBP and Ring1A; and calmodulin binding peptide (CBP) derived from skeletal muscle myosin light-chain kinase and calmodulin. The in vitro protein microarrays can easily be extended to a high-throughput format and thus should be useful for large-scale analysis of protein-protein interactions.
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