2007 Fiscal Year Final Research Report Summary
Proteome Analysis of Disease-related Genes based on Alternative Splicing
| Project/Area Number |
16201042
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied genomics
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| Research Institution | Keio University |
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Principal Investigator |
YANAGAWA Hiroshi Keio University, Faculty of Science and Technology, Professor (40327672)
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| Co-Investigator(Kenkyū-buntansha) |
DOI Nobuhide Keio University, Faculty of Science and Technology, Associate Professor (50327673)
WASHIO Takanori Nara Institute of Science and Technology, Graduate School of Information Science, Research Associate (50338266)
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| Project Period (FY) |
2004 – 2007
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| Keywords | Alternative splicing / Disease-related genes / Proteome / In vitro virus / Protein-protein interactions / Gene network / Bioinformatics / High-throughput screening |
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| Research Abstract |
(1) Computational analysis of disease-related genes with splice variants: We searched genes containing both ASDB (database of alternatively spliced genes) and OMIM (Online Mendelian Inheritance in Man) databases to extract alternatively spliced genes that related to diseases such as cancer and Alzheimer-disease.
(2) Construction of a parallel IVV screening system: We optimized the conditions of IVV screening for protein-protein interactions (PPIs) in 96-well microplates with a QIAGEN BioRobot 8000. We also improved the performance of computer systems for analysis of the huge sequence data (from 1000 sequences per day to 16000 sequences per day).
(3) A large-scale analysis of PPIs : As a result of the parallel IVV screening against 68 distinct bait proteins, total of -1000 PPIs were successfully detected. When arbitrary chosen PPIs were tested by pull-down assays, -70% of PPIs were verified.
(4) Confirmation of PPIs with splice variants : Among the above PPI data, 24 gene fragments contain their protein-binding regions within variable regions derived from alternative splicing. We confirmed that the in vitro binding ability of these genes was certainly affected by the change of splicing patterns.
(5) Construction of a PPI database : We developed a database containing not only the binding region from the PPI data but also the variable region of alternative splicing and SNP, the crystal structure of protein complexes, gene localization, etc. The database is useful for analyzing the effect of alternative splicing on disease-related gene networks.
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Research Products
(38 results)
