1999 Fiscal Year Final Research Report Summary
Development of a rapid scanning method for quantitative and qualitative variation throughout the genome.
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 |
Section | 展開研究 |
Research Field |
Pathological medical chemistry
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Research Institution | The University of Tokyo |
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)
|
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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Keywords | RDA / MICROARRAY / GENE EXPRESSION PROFILING |
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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[Publications] Murakami T, Mataki C, Nagao C, Umetani M, Wada Y, Ishii M, Tsutsumi S, Kohro T, Saiura A, Aburatani H, Hamakubo T and Kodama T.: "The gene expression profile of human umbilical vein endothelial cells stimulated by tumor necrosis factor a using DNA microarray analysis."Journal of Atherosclerosis and Thrombosis. in press. (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Ishida, T., Takashima, R., Fukayama, M., Hamada, C., Hippo, Y., Fujii, T., Moriyama, S., Matsuba, C., Nakahori, Y., Morita, H., Yazaki, Y., Kodama, T., Nishimura, S., and Aburatani, H.: "New DNA polymorphisms of human MMH/OGG1 gene : prevalence of one polymorphism among lung-adenocarcinoma patients in Japanese."Int J Cancer. 80. 18-21 (1999)
Description
「研究成果報告書概要(欧文)」より
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