Construction of high-expression vector had on gene-amplified chromosomal location and its application to human cell
| Project/Area Number |
17560689
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Osaka University |
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Principal Investigator |
OMASA Takeshi Osaka University, Department of Biotechnology, Associate Professor (00252586)
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| Co-Investigator(Kenkyū-buntansha) |
OHTAKE Hisao Osaka University, Department of Biotechnology, Professor (10127483)
HONDA Kohsuke Osaka University, Department of Biotechnology, Assistant Professor (90403162)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,740,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥240,000)
Fiscal Year 2007: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | gene amplification / BAC library / Chinese hamster ovary cell / chromosome / MTX / CHO(Chinese hamster ovary) / CHO (Chinese hamster ovary) |
|---|
| Research Abstract |
Gene amplification means "the selective, repeated replication of a certain gene or genes without a proportional increase in other genes in the genome". Gene amplification is a widespread phenomenon in eukaryotes. It is an important process in the development of many organisms, the emergence of drug resistance in tumor cells and some human parasites and the maturation of tumors. From the early 1980s, this gene amplification was used in the industrial production of therapeutic proteins using Chinese hamster ovary (CHO) cells. The most pressing problem in such protein production in mammalian cells is the low productivity of the recombinant protein. To increase productivity, gene amplification cell engineering techniques for the transfected gene are extensively applied, whereby the amplifiable gene is used as a selectable marker for the transfected vector. Firstly the vector, which contains the cDNA of the objective protein and amplifiable marker gene, is introduced into the host cell line. When the selectable and amplifiable gene is amplified, genetically linked sequences containing the objective cDNA are co-amplified. Therefore, a high level of expression of the transfected DNA can be easily attained. In this study, we analyzed the gene amplification mechanism by construction of CHO bacterial artificial chromosome (BAC) library. We determined the structure of gene-amplified location and estimated the gene amplification mechanism.
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Report
(4 results)
Research Products
(32 results)
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[Journal Article] Development of cell-based simulator system for drug evaluation2005
Author(s)
Takeshi, Omasa, Tetsuji, Hayashi, Shinsuke, Onishi, Tadaaki, Hashimoto, Michimasa, Kishimoto, Tomohiro, Yoshikawa, Masato, Miyake, Hisao, Ohtake
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Journal Title
Proceeding book for Asia-Pacific Biochemical Engineering Conference ANI-O3
Pages: 1-4
Description
「研究成果報告書概要(欧文)」より
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[Presentation] BAC-FISHを用いたCHO-DG44株における染色体識別とその応用2008
Author(s)
曹 溢華, 矢野 秀法, Haghparast Seyed Mohammad Ali, 木村 修一, 高木 康弘, 本田 孝祐, 大政 健史, 淺川 修一, 清水 信義, 大竹 久夫
Organizer
日本農芸化学会2008年度大会
Place of Presentation
名古屋
Year and Date
2008-03-28
Description
「研究成果報告書概要(和文)」より
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[Presentation] CHO細胞における染色体の識別及び再配列解析2007
Author(s)
曹 溢華, 矢野 秀法, 木村 修一, 高木 康弘, 本田 孝祐, 大政 健史, 淺川 修一, 清水 信義, 大竹 久夫
Organizer
日本生物工学会、平成19年度大会
Place of Presentation
東広島
Year and Date
2007-09-26
Description
「研究成果報告書概要(和文)」より
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[Book] 抗体医薬の最前線2007
Author(s)
大政健史、奥村一夫
Total Pages
9
Publisher
シーエムシー出版
Description
「研究成果報告書概要(和文)」より
Related Report
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