『徳島大学 教育・研究者情報データベース (EDB)』---[学外] /
ID: Pass:

登録内容 (EID=296077)

EID=296077EID:296077, Map:0, LastModified:2015年11月11日(水) 20:24:04, Operator:[大家 隆弘], Avail:TRUE, Censor:0, Owner:[山﨑 尚志], Read:継承, Write:継承, Delete:継承.
種別 (必須): 学術論文 (審査論文) [継承]
言語 (必須): 英語 [継承]
招待 (推奨):
審査 (推奨):
カテゴリ (推奨): 研究 [継承]
共著種別 (推奨):
学究種別 (推奨):
組織 (推奨):
著者 (必須): 1. (英) Ishido Tomomi (日) (読)
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
2.山﨑 尚志 ([徳島大学.大学院医歯薬学研究部.薬学域.薬科学部門.生命薬学系.薬物治療学])
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
3.石川 満 ([独立行政法人 産業技術総合研究所])
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
4. (英) Hirano Ken (日) (読)
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
題名 (必須): (英) Characterization of DNA polymerase from Danio rerio by overexpression in E. coli using the in vivo/in vitro compatible pIVEX plasmid.  (日)    [継承]
副題 (任意):
要約 (任意): (英) Eukaryotic DNA polymerase β (pol β), the polymerase thought to be responsible for DNA repair synthesis, has been extensively characterized in rats and humans. However, pol β has not been purified or enzymatically characterized from the model fish species Danio rerio (zebrafish). We used the in vitro/in vivo dual expression system plasmid, pIVEX, to express Danio rerio pol β (Danio pol β) for biochemical characterization. Danio pol β encoded by the in vitro/in vivo-compatible pIVEX plasmid was expressed in E. coli BL21(DE3), BL21(DE3)pLysS, and KRX, and in vitro as a C-terminal His-tagged protein. Danio pol β expressed in vitro was subject to proteolysis; therefore, bacterial overexpression was used to produce the protein for kinetic analyses. KRX cells were preferred because of their reduced propensity for leaky expression of pol β. The cDNA of Danio rerio pol β encodes a protein of 337 amino acids, which is 2-3 amino acids longer than other pol β proteins, and contains a P63D amino acid substitution, unlike mammalian pol βs. This substitution lies in a hairpin sequence within an 8-kDa domain, likely to be important in DNA binding. We performed extensive biochemical characterization of Danio pol β in comparison with rat pol β, which revealed its sensitivity to metal ion activators (Mn2+ and Mg2+), its optimum salt concentration (10 mM KCl and 50 mM NaCl), alkaline pH optimum (pH 9.0), and low temperature optimum (30°C). Substituting Mn2+ for Mg2+ resulted in 8.6-fold higher catalytic efficiency (kcat/Km). Our characterization of pol β from a model fish organism contributes to the study of the function and evolution of DNA polymerases, which are emerging as important cellular targets for chemical intervention in the development of anticancer agents.  (日)    [継承]
キーワード (推奨): 1. (英) Amino Acid Sequence (日) (読) [継承]
2. (英) Animals (日) (読) [継承]
3. (英) DNA Polymerase beta (日) (読) [継承]
4. (英) Enzyme Stability (日) (読) [継承]
5. (英) Escherichia coli (日) (読) [継承]
6. (英) Fish Proteins (日) (読) [継承]
7. (英) Gene Expression (日) (読) [継承]
8. (英) Kinetics (日) (読) [継承]
9. (英) Molecular Sequence Data (日) (読) [継承]
10. (英) Plasmids (日) (読) [継承]
11. (英) Protein Structure, Tertiary (日) (読) [継承]
12. (英) Rats (日) (読) [継承]
13. (英) Recombinant Fusion Proteins (日) (読) [継承]
14. (英) Sequence Homology, Amino Acid (日) (読) [継承]
15. (英) Zebrafish (日) (読) [継承]
発行所 (推奨): BioMed Central Ltd. [継承]
誌名 (必須): Microbial Cell Factories ([BioMed Central Ltd.])
(pISSN: 1475-2859)

ISSN (任意): 1475-2859
ISSN: 1475-2859 (eISSN: 1475-2859)
Title: Microbial cell factories
Title(ISO): Microb Cell Fact
Publisher: BioMed Central
 (NLM Catalog  (Scopus  (CrossRef (Scopus information is found. [need login])
[継承]
[継承]
(必須): 10 [継承]
(必須): [継承]
(必須): 84 84 [継承]
都市 (任意):
年月日 (必須): 西暦 2011年 10月 21日 (平成 23年 10月 21日) [継承]
URL (任意):
DOI (任意): 10.1186/1475-2859-10-84    (→Scopusで検索) [継承]
PMID (任意): 22018137    (→Scopusで検索) [継承]
NAID (任意):
WOS (任意):
Scopus (任意):
評価値 (任意):
被引用数 (任意):
指導教員 (推奨):
備考 (任意): 1.(英) Article.ELocationID: 10.1186/1475-2859-10-84  (日)    [継承]
2.(英) Article.PublicationTypeList.PublicationType: Journal Article  (日)    [継承]
3.(英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't  (日)    [継承]
4.(英) OtherID: PMC3214136  (日)    [継承]

標準的な表示

和文冊子 ● Tomomi Ishido, Naoshi Yamazaki, Mitsuru Ishikawa and Ken Hirano : Characterization of DNA polymerase from Danio rerio by overexpression in E. coli using the in vivo/in vitro compatible pIVEX plasmid., Microbial Cell Factories, Vol.10, 84, 2011.
欧文冊子 ● Tomomi Ishido, Naoshi Yamazaki, Mitsuru Ishikawa and Ken Hirano : Characterization of DNA polymerase from Danio rerio by overexpression in E. coli using the in vivo/in vitro compatible pIVEX plasmid., Microbial Cell Factories, Vol.10, 84, 2011.

関連情報

Number of session users = 0, LA = 0.38, Max(EID) = 375659, Max(EOID) = 1006582.