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Establishment of recombinant gonadotropin-producing system by making use of E. coli ahd cloned aninals.

Research Project

Project/Area Number 11794010
Research Category

Grant-in-Aid for University and Society Collaboration

Allocation TypeSingle-year Grants
Research Field Applied animal science
Research InstitutionThe University of Tokyo

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)

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)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2001: ¥14,100,000 (Direct Cost: ¥14,100,000)
KeywordsGonadotropin / Nuclear transfer / Livestock animals / Clone
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.

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Li J.Y., et al: "Induction of short ovulatory cycle in Shiba-Goats by repeated treatments with prostaglandin F2α"Journal of Reproduction and Development. 47. 97-103 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saneyoshi T., et al: "Equine follicle-stimulating hormone : Molecular cloning of β subunit and biological role of the asparagines-linked oligosaccharide at asparagine^<56> of alpha subunit"Biology of Reproduction. 65. 1686-1690 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tanaka S., et al: "Placentomegaly in cloned mouse concepti caused by expansion of the spongiotrophoblast layer"Biology of Reproduction. 65. 1813-1821 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohgane J., et al: "DNA methylation variation in cloned mice"Genesis. 30. 45-50 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Li, J.Y. et al.: "Induction of short ovulatory cycle in Shiba-Goats by repeated treatments with prostaglandin F2α."J Reproduction and Development. 47. 97-103 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Saneyoshi, T. et al.: "Equine follicle-stimulating hormone : Molecular cloning of β subunit and biological role of the asparagines-linked oligosaccharide at asparagine^<56> of alpha subunit."Biology of Reproduction. 65. 1686-1690 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tanaka, S. et al.: "Placentomegaly in cloned mouse concepti caused by expansion of the spongiotrophoblast layer."Biology of Reproduction. 65. 1813-1821 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohgane, J. et al.: "DNA methylation variation in cloned mice."Genesis. 30. 45-50 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mogi K.et al.: "Characterization of growth hormone pulsatility in male Shiba goats : Effects of postpubertal castration and intravenous injection of KP10"Endocrine Journal. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Saneyoshi. et al.: "Equine follicle-stimulating hormone : Molecular cloning of β subunit and biological role of the asparagines-linked oligosaccharide at asparagine^<56> of alpha subunit"Biology of Reproduction. 65. 1686-1690 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tanaka S. et al.: "Placentomegaly in cloned mouse concepti caused by expansion of the spongiotrophoblast layer"Biology of Reproduction. 65. 1813-1821 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Ohgane J. et al.: "DNA methylation variation in cloned mice"Genesis. 30. 45-21 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Jun You Li et al.: "Induction of short ovulatory cycle in Shiba-goats by repeated treatments with prostaglandin F2a"Journal of Reproduction and Development. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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