『徳島大学 教育・研究者情報データベース (EDB)』---[学外] /
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登録内容 (EID=272381)

EID=272381EID:272381, Map:0, LastModified:2016年12月22日(木) 14:59:47, Operator:[三木 ちひろ], Avail:TRUE, Censor:0, Owner:[鬼塚 正義], 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):
共著種別 (推奨):
学究種別 (推奨):
組織 (推奨): 1.徳島大学.工学部.生物工学科.生物機能工学講座 [継承]
著者 (必須): 1. (英) Kyoung Ho Lee (日) (読)
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
2.鬼塚 正義 ([徳島大学.大学院社会産業理工学研究部.生物資源産業学域.応用生命系.生体分子機能学分野]/[徳島大学.生物資源産業学部.生物資源産業学科.応用生命講座])
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
3. (英) Honda Kohsuke (日) (読)
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
4. (英) Ohtake Hisao (日) (読)
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貢献度 (任意):
学籍番号 (推奨):
[継承]
5.大政 健史
役割 (任意):
貢献度 (任意):
学籍番号 (推奨):
[継承]
題名 (必須): (英) Rapid construction of transgene-amplified CHO cell lines by cell cycle checkpoint engineering  (日)    [継承]
副題 (任意):
要約 (任意): (英) The process of establishing high-producing cell lines for the manufacture of therapeutic proteins is usually both time-consuming and laborious due to the low probability of obtaining high-producing clones from a pool of transfected cells and slow cell growth under the strong selective pressure of screening to identify high-producing clones. We present a novel method to rapidly generate more high-producing cells by accelerating transgene amplification. A small interfering RNA (siRNA) expression vector against ataxia telangiectasia and Rad3 related (ATR), a cell cycle checkpoint kinase, was transfected into Chinese hamster ovary (CHO) cells. The influences of ATR downregulation on gene amplification and the productivity were investigated in CHO cells producing green fluorescent protein (GFP) and secreting monoclonal antibody (mAb). The ATR-downregulated cells showed up to a 6-fold higher ratio of GFP-positive cells than that of the control cell pool. Moreover, the downregulated mAb-producing cells had about a 4-fold higher specific production rate and a 3-fold higher volumetric productivity as compared with the mock cells. ATR-downregulated cells showed a much faster increase in transgene copy numbers during the gene amplification process via methotrexate (MTX) treatment in both GFP- and mAb-producing cells. Our results suggest that a pool of high-producing cells can be more rapidly generated by ATR downregulation as compared with conventional gene amplification by MTX treatment. This novel method may be a promising approach to reduce time and labor in the process of cell line development.  (日)    [継承]
キーワード (推奨): 1. (英) Animals (日) (読) [継承]
2.生物工学 (biotechnology) [継承]
3. (英) CHO Cells (日) (読) [継承]
4. (英) Cell Cycle Checkpoints (日) (読) [継承]
5. (英) Cell Cycle Proteins (日) (読) [継承]
6. (英) Cricetulus (日) (読) [継承]
7.遺伝子発現 (gene expression) [継承]
8. (英) Gene Silencing (日) (読) [継承]
9. (英) Genes, Reporter (日) (読) [継承]
10.遺伝子工学 (genetic engineering) [継承]
11. (英) Green Fluorescent Proteins (日) (読) [継承]
12. (英) Molecular Sequence Data (日) (読) [継承]
13. (英) Recombinant Proteins (日) (読) [継承]
14. (英) Sequence Analysis, DNA (日) (読) [継承]
15. (英) Transgenes (日) (読) [継承]
発行所 (推奨):
誌名 (必須): Applied Microbiology and Biotechnology ([Springer-Verlag])
(pISSN: 0175-7598, eISSN: 1432-0614)

ISSN (任意): 1432-0614
ISSN: 0175-7598 (pISSN: 0175-7598, eISSN: 1432-0614)
Title: Applied microbiology and biotechnology
Title(ISO): Appl Microbiol Biotechnol
Supplier: Springer Online Journal Archive
Publisher: Springer Berlin Heidelberg
 (NLM Catalog  (Scopus  (CrossRef (Scopus information is found. [need login])
[継承]
[継承]
(必須): 97 [継承]
(必須): 13 [継承]
(必須): 5731 5741 [継承]
都市 (任意):
年月日 (必須): 西暦 2013年 4月 25日 (平成 25年 4月 25日) [継承]
URL (任意):
DOI (任意): 10.1007/s00253-013-4923-9    (→Scopusで検索) [継承]
PMID (任意): 23615744    (→Scopusで検索) [継承]
NAID (任意):
WOS (任意): 000321121500008 [継承]
Scopus (任意): 2-s2.0-84879418366 [継承]
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: 10.1007/s00253-013-4923-9  (日)    [継承]
2.(英) Article.DataBankList.DataBank.DataBankName: GENBANK  (日)    [継承]
3.(英) Article.DataBankList.DataBank.AccessionNumberList.AccessionNumber: AB761036  (日)    [継承]
4.(英) Article.PublicationTypeList.PublicationType: Journal Article  (日)    [継承]
5.(英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't  (日)    [継承]

標準的な表示

和文冊子 ● Lee Ho Kyoung, Masayoshi Onitsuka, Kohsuke Honda, Hisao Ohtake and Takeshi Omasa : Rapid construction of transgene-amplified CHO cell lines by cell cycle checkpoint engineering, Applied Microbiology and Biotechnology, Vol.97, No.13, 5731-5741, 2013.
欧文冊子 ● Lee Ho Kyoung, Masayoshi Onitsuka, Kohsuke Honda, Hisao Ohtake and Takeshi Omasa : Rapid construction of transgene-amplified CHO cell lines by cell cycle checkpoint engineering, Applied Microbiology and Biotechnology, Vol.97, No.13, 5731-5741, 2013.

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