| Project/Area Number |
05454653
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Developmental biology
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
SHIMADA Hiraku Hiroshima University Faculty of Science, Professor, 理学部, 教授 (70011559)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Kazumi Hiroshima University, Faculty of Science, Instructor, 理学部, 助手 (80220367)
AKASAKA Koji Hiroshima University, Faculty of Science, Associate Professor, 理学部, 助教授 (60150968)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1994: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥4,600,000 (Direct Cost: ¥4,600,000)
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| Keywords | sea urchin / development / gene expression / arylsufatase / cis-element / DNA binding protein / G-string / luciferase assay / ウニ発生 / アリールスルファターゼ / 転写調節 / 遺伝子発現 / 転写因子 / インビトロ転写 |
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| Research Abstract |
This research project has been proposed to obtain clues to understand a mechanism underlying temporal and spatial expression of genes during early development of sea urchin. For this purpose we selected a gene for arylsulfatase (Ars) that expresses in the temporally and spatially regulated manner in sea urchin embryo. The sea urchin Ars gene contains a TATA box in its basal promoter, several G-string sequences and a long polypyrimidine region in its 5' flanking region. Following results were obtained in this study. (1) The long polypyrimidine region can form a triplex DNA configurationthrough Hoogsten base parings, (2) four cDNA clones for G-string DNA-binding proteins were obtained by south-western blotting using a synthetic poly (dG)-poly(dC) as the probe. All these cDNAs contain GA-rich sequence in its ORF.(3) Temporal expression of the Ars gene is primarily determined by a short region between -1 and -100b positions. Two cis-elements between -100 and -252 position seem to be related to temporal regulation of the Ars gene expression. (4) A 229nts sequence in the first intron strongly enhances the activity of these elements. all these results were obtained by luciferase assays. (5) A nuclear extract system to effectively transcribe the Ars gene in vitro was constructed.
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