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動物細胞を用いたDNA複製/チェックポイント因子Claspinの発現、精製と機能解析
https://doi.org/10.15083/00005585
https://doi.org/10.15083/000055852cdeadc3-5826-43a7-a96f-5268a3d2502a
名前 / ファイル | ライセンス | アクション |
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K-03492.pdf (2.7 MB)
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Item type | 学位論文 / Thesis or Dissertation(1) | |||||
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公開日 | 2014-02-24 | |||||
タイトル | ||||||
タイトル | 動物細胞を用いたDNA複製/チェックポイント因子Claspinの発現、精製と機能解析 | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題 | DNA replication | |||||
主題Scheme | Other | |||||
キーワード | ||||||
主題 | checkpoint | |||||
主題Scheme | Other | |||||
キーワード | ||||||
主題 | Claspin | |||||
主題Scheme | Other | |||||
キーワード | ||||||
主題 | expression | |||||
主題Scheme | Other | |||||
キーワード | ||||||
主題 | purification | |||||
主題Scheme | Other | |||||
資源タイプ | ||||||
資源 | http://purl.org/coar/resource_type/c_46ec | |||||
タイプ | thesis | |||||
ID登録 | ||||||
ID登録 | 10.15083/00005585 | |||||
ID登録タイプ | JaLC | |||||
著者 |
Uno, Syuzi
× Uno, Syuzi |
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著者別名 | ||||||
識別子 | 11646 | |||||
識別子Scheme | WEKO | |||||
姓名 | 宇野, 修司 | |||||
著者所属 | ||||||
著者所属 | 東京大学大学院新領域創成科学研究科生命科学研究系メディカルゲノム専攻 | |||||
著者所属 | ||||||
著者所属 | Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo | |||||
Abstract | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Mrc1/Claspin are evolutionally conserved and has been known to play a major role in replication checkpoint regulation. Furthermore, checkpoint-independent roles at a replication fork have been suggested in yeasts. However, the information on biochemical functions of Claspin has been limited, due to difficulties associated with purification of this protein. I have improved expression and purification procedures for human Claspin as well as other high molecular weight replication proteins. I used a CSII-EF-MCS expression vector and Polyethyleneimine (PEI) as a transfection reagent for high-level expression in 293T cells. This method could be generally applicable to purification of large-sized proteins which have been difficult to handle. Purified Claspin stimulated the helicase activity of the MCM4-6-7 complex in vitro. This finding can at least partially explain a stimulatory role of Claspin at a replication fork. I also identified interactions of Claspin with replication and checkpoint factors, which may bear physiological significance. | |||||
書誌情報 | 発行日 2011-06-27 | |||||
日本十進分類法 | ||||||
主題 | 467 | |||||
主題Scheme | NDC | |||||
学位名 | ||||||
学位名 | 博士(生命科学) | |||||
学位 | ||||||
値 | doctoral | |||||
学位分野 | ||||||
Integrated Biosciences (生命科学) | ||||||
学位授与機関 | ||||||
学位授与機関名 | University of Tokyo (東京大学) | |||||
研究科・専攻 | ||||||
Department of Medical Genome Sciences, Graduate School of Frontier Sciences (新領域創成科学研究科メディカルゲノム専攻) | ||||||
学位授与年月日 | ||||||
学位授与年月日 | 2011-06-27 | |||||
学位授与番号 | ||||||
学位授与番号 | 甲第27368号 | |||||
学位記番号 | ||||||
博創域第711号 |