| 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) |
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| 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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