| Project/Area Number |
23510266
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Living organism molecular science
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| Research Institution | Doshisha Women's College of Liberal Arts |
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Principal Investigator |
NEGI SHIGERU 同志社女子大学, 薬学部, 助教 (50378866)
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| Project Period (FY) |
2011 – 2013
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2011: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | 亜鉛フィンガー / 転写因子 / 金属結合 / DAN結合 / レドックス / SELEX法 / タンパク質工学 / 蛋白質 / DNA / 人工蛋白質 / 人工ヌクレアーゼ / 酸化還元 / 金属イオン / DNA結合 / シグナル伝達 / レドックスセンサー / 金属置換 / 細胞移行 / 分子進化実験 / レドックス挙動 / ＤＮＡ結合 / 無細胞系タンパク質発現 |
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| Research Abstract |
In this study, we tried to create metallo fingerwith Co(II) and Ni(II) to investigate its metal-binding and secondary structure, DNA-binding property and oxidation reactivity using GAGA zinc finger protein (GAGA Zf). From UV and CD data,metallo fingers have almost similar coordinate and secondary structure as wild-type GAGA Zf. In DNA-binding assay using gel shift method, metallo finger can bind to cognate DNA sequence (GAGAG). From SELEX experiment, metal-substitution has no effect on the DNA sequence selectivity. Finally, we tried to study the oxidation reactivity metallo finger. As a result of an oxidation reaction with the diamide, reactivity became the following order, that is, apo>Ni(II)>>Co(II)>Zn(II) indicating that reactivity strongly depended on the metal ion. Probably, this reactive difference caused by the difference in stability of coordinate bond and domain structure among metallo fingers.
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