| Project/Area Number |
11794010
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| Research Category |
Grant-in-Aid for University and Society Collaboration
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied animal science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SAWASAKI Toru , Grad Sch of Agric Life Sci, Professor, 大学院・農学生命科学研究科, 教授 (00012047)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Satoshi , Grad Sch of Agric Life Sci, Associate Professor, 大学院・農学生命科学研究科, 助教授 (90242164)
IMAKAWA Kazuhiko , Grad Sch of Agric Life Sci, Associate Professor, 大学院・農学生命科学研究科, 助教授 (00291956)
SHIOTA Kunio , Grad Sch of Agric Life Sci, Professor, 大学院・農学生命科学研究科, 教授 (80196352)
HIYAMA Takashi Teikoku Hormone Mfg. Co. Ltd., Natural Products Research Dept., Researcher, 天然物研究部, 研究員
HIRATA Touichi Iwate Univ, Faculty of Agriculture, Assistant, 農学部, 助手 (20241490)
佐久間 秀樹 帝国臓器(株), 天然物研究部, 部長(研究職)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2001: ¥14,100,000 (Direct Cost: ¥14,100,000)
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| Keywords | Gonadotropin / Nuclear transfer / Livestock animals / Clone |
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| Research Abstract |
Aim of this study is to establish systems for stable production of safe chorionic gonadotropins (CGs) in E. coli and cloned livestock animals. CG consists of highly glycosylated α and β subunits. Correct conformation of the heterodimers as well as N-linked oligosaccharides on α subunit have been shown to be relevant for CG to elicit its biological activities, which make it difficult to produce large amount of active recombinant CG by using usual E. coli expression system. We have shown that FSH-like activity of equine CG (eCG) is resistant to the removal of N-linked oligosaccharides, and that α and β subunits of eCG can be synthesized as a single polypeptide chain (tethered-eCG) without any loss of biological activities. These findings have suggested that the recombinant eCG protein with a potent FSH-like activity can be produced by E. coli expression system in which very little of the glycosylation occurs. To establish such eCG producing system, we evaluated the conditions for efficient expression of tethered-eCG in E. coli and for the purification of the tethered-eCG protein froin crud E. coli extrancts. So far we have succeeded in obtaining the tethered-eCG protein as a single band on SDS-PAGE gel. By this achievement, it became possible to set up large-scale production system of a potent eCG protein. We also intended to establish production system of recombinant human CG (hCG) in tranagenic livestock animals. To this goal, we tried to construct a mammalian gland-specific hCG-expression vector. Once this vector is made, it should be possible to produce transgenic cows expressing hCG in their milk by so-called animal cloning technique which is now possible to perform at the sock farm of the University of Tokyo. To understand general abnonnalities in cloned animals, placental phenotypes and abnormalities in DNA methylation of cloned mice were also investigated.
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