| Project/Area Number |
14370755
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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 | 一般 |
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| Research Field |
医薬分子機能学
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| Research Institution | Kyoto University |
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Principal Investigator |
SUGIURA Yukio Kyoto Univ., Inst.Chem.Res., Prof., 化学研究所, 教授 (40025698)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAOKA Makoto Kyoto Univ., Inst.Chem.Res., Assistant Prof., 化学研究所, 助手 (60314275)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2004: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2003: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥6,900,000 (Direct Cost: ¥6,900,000)
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| Keywords | Zinc finger / Multi finger / Molecular design / Transcription factor / DNA recognition / Architecture / Base specificity / Artificial protein / 遺伝子発現 / タンパク質工学 / タンパク質設計 |
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| Research Abstract |
In gene regulation, the positional finding of a unique target site in the genome by multi-zinc fingers has been widely studied. The custom sets of 6-zinc finger proteins are very practical in medicinal research for their adequate recognition length of DNA and manageability. To assess the possibility of multi-connection of finger domains for understanding of DNA binding mechanisms and gene regulation, the longest artificial zinc finger protein, Sp1ZF15, has been constructed. This zinc finger consists of 5 units of Sp1 zinc finger peptide connected by canonical short linker sequences (TGEKP). Recognition on the 50 successive base pairs of DNA by Sp1ZF15 was determined by gel mobility shift assays and DNase I footprinting analyses. Furthermore, capable bindings to shorter targets were also examined. Sequence alterations of the GCG triplet to ATA at a target site clearly showed that Sp1ZF15 alters its DNA binding mode depending on the target sequences. Of special interest is the fact that Sp1ZF15 controls the number of the finger domains active in DNA binding corresponding to the length and sequence of the target DNA. These results show the zinc finger protein to be an effective applicant for a binding module to long DNA sequence for the development of gone regulation. In addition, the present evidence suggest that a vast collection of distinct transcription factors by varying the choice, number and order, in multi zinc finger proteins, might be produced in a cell.
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