| Project/Area Number |
09557014
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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 |
Pathological medical chemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
ABURATANI Hiroyuki The University of Tokyo, RCAST, Associate Professor, 先端科学技術研究センター, 助教授 (10202657)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMADA Masamitsu Takara shuzo, Biomedical Center, Researcher, バイオメディカルセンター, 研究員
KODAMA Tatsuhiko The University of Tokyo, RCAST, Professor, 先端科学技術研究センター, 教授 (90170266)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥9,200,000 (Direct Cost: ¥9,200,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1997: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | RDA / MICROARRAY / GENE EXPRESSION PROFILING / 染色体欠失 / トランスクリプトーム解析 / 組織特異性 / ゲノムスキャニング / サブトラクション / DNAアレイ / 遺伝子発現 |
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| Research Abstract |
We intended to develop a method to rapidly scan quantitative and qualitative variation throughout the genome. Representative Difference Analysis (RDA) was initially applied as an efficient subtraction method to isolate DNA fragments differentially represented between two genomes compared, especially to isolate tumor suppressor genes involved in multi-step carcinogenesis. In the meantime, the human genome project has progressed far more rapidly than expected, then has made possible the gene expression profiling analysis with microarray technology. We verified that Genechip, the oligonucleotide array made by photolithography was reliable in quantitation when compared to SAGE (Serial Analysis of Gene Expression) method. Expression profiling using Genechip was applied to identify the genes differentially expressed between stomach cancer cell lines with different metastatic potentials, OCUM-2M and OCUM-2MD3.
We also developed an oligonucleotide array system for single nucleotide polymorphism detection. Oligonuceotides were fixed on glass surface via biotin-avidin binding, where streptoavidin was bound on the glass slide by plasma polymerization technique to create hydrophobic surface, which resulted in higher signal-to-noise ratio. Discrimination of variant apo E alleles were successfully obtained.
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