| Project/Area Number |
12206001
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | University of Tsukuba |
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Principal Investigator |
URUSHIHARA Hideko (2003-2004) University of Tsukuba, Graduate school of Life and Environmental sciences, Professor, 大学院生命環境科学研究科, 教授 (00150087)
田仲 可昌 (2000-2002) 筑波大学, 生物科学系, 教授 (80091908)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAKA Yoshimasa University of Tsukuba, Graduate school of Life and Environmental sciences, Professor, 大学院生命環境科学研究科, 教授 (80091908)
MORIO Takahiro University of Tsukuba, Graduate school of Life and Environmental sciences, Assistant Professor, 大学院生命環境科学研究科, 助手 (10292509)
MAEDA Mineko Osaka University, Graduate School of Sciences, Associate Professor, 大学院理学研究科, 助教授 (70029700)
OCHIAI Hiroshi Hokkaido University, Graduate School of Sciences, Professor, 大学院理学研究科, 教授 (10002122)
漆原 秀子 筑波大学, 生物科学系, 助教授 (00150087)
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| Project Period (FY) |
2000 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥136,800,000 (Direct Cost: ¥136,800,000)
Fiscal Year 2004: ¥30,000,000 (Direct Cost: ¥30,000,000)
Fiscal Year 2003: ¥30,000,000 (Direct Cost: ¥30,000,000)
Fiscal Year 2002: ¥33,800,000 (Direct Cost: ¥33,800,000)
Fiscal Year 2001: ¥43,000,000 (Direct Cost: ¥43,000,000)
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| Keywords | Dictyostelium discoideum / EST analysis / Cell differentiation / Gene expression / Microarray analysis / transcription factors / Upstream sequences / Cis-elements / 遺伝子発現 / 遺伝子ネットワーク / 脱分化 / 転写制御 / 配列アセンブル / DNAマイクロアレイ / cDNA解析 / ゲノム / 細胞分化 / マイクロアレイ / in situハイブリダイゼーション / 転写制御因子 / 完全長cDNA / ポリAトラップ法 |
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| Research Abstract |
The cellular slime mold, Dictyostelium discoideum, was used to understand the genetic control of development and differentiation. This organism normally proliferates as unicellular amoebae but gathers and constructs multicellular structures upon starvation. First, large scale analysis of cDNAs was performed to establish the genetic resources. Fourteen cDNA libraries were constructed from the cells at 5 developmental stages, and the nucleotide sequences of 95,884 clones, including 72,703 full-length ones, were determined. The sequences were assembled into 6,790 independent genes, which corresponded to 54% of the predicted 12,500 genes. These resources were then used for reverse-genetic and bioinformatic analyses. The following results were obtained.
1. Gene expression patterns during normal development were obtained showing occurrence of the overall drastic changes in expressed genes after cell aggregation. Whole mount in situ hybridization revealed the fine pattern formation in the prestalk region.
2. Two hundred and sixty genes were extracted to encode probable transcription factors, and precise expression patterns were obtained by real-time PCR for 40 of them. Thirteen genes were disrupted.
3. The Genetic network among 9 probable transcription factors was obtained using the S-system calculation based on the gene expression patterns during development in the wild type and disruption mutants.
4. Upstream promoter sequences were obtained by aligning the full-length cDNA and genome sequences, and candidate cis regulatory elements were extracted referring to the expression specificities.
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