| Project/Area Number |
16380010
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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 | 一般 |
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| Research Field |
Breeding science
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| Research Institution | YOKOHAMA CITY UNIVERSITY |
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Principal Investigator |
HIRANO Hisashi YOKOHAMA CITY UNIVERSITY, International Graduate School of Arts and Sciences, Professor, 大学院国際総合科学研究科生体超分子科学専攻, 教授 (00275075)
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| Co-Investigator(Kenkyū-buntansha) |
KAWASAKI Hiroshi YOKOHAMA CITY UNIVERSITY, International Graduate School of Arts and Sciences, Associate Professor, 国際総合科学研究科, 准教授 (70169704)
SASSA Hidenori Chiba University, Department of Horticulture, Associate Professor, 園芸学部, 准教授 (50295507)
TANGA Michifumi Toyo Kohan Co., Director, 部長
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,900,000 (Direct Cost: ¥15,900,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥11,400,000 (Direct Cost: ¥11,400,000)
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| Keywords | proteome / protein interaction / DLC plate / protein chip / glycoprotein / rice / soybean / プロテオーム / 蛋白質間相互作用 / タンデムアフィニティー精製 |
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| Research Abstract |
Protein chip has been one of the most promising tools for identification of expressed proteins and study of protein-protein interaction in proteome analysis. We developed a novel protein chip plate, which is made of diamond-like carbon-coated stainless steel (DLC), and modified with N-hydroxysuccinimide ester on the surface of plate. Proteins separated by SDS gel electrophoresis or two-dimensional electrophoresis (2-DE) were successfully electroblotted and immobilized covalently onto the DLC plate to produce easily and rapidly a high-density protein chip. High blotting efficiency (25-70 %) of proteins from the gels onto the DLC plate was obtained by improvement of electrophoresis and electroblotting techniques. Using the DLC plate, we established the techniques for identification of proteins immobilized on the DLC plate, for detection of proteins or peptides interacted with proteins immobilized on the plate using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry, and also we developed the technique for identification of post-translational modification of proteins on the plate. In this study, we applied these techniques to identification of a number of proteins and N-linked glycoproteins of the rice calli and soybean seeds, which were separated by 2-DE. The developed techniques have a great potential to make possible the high-throughput proteome analysis in plants.
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