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Study on the production of N-Glycolylneuraminic acid-deficient animals using gene targeting techniques

Research Project

Project/Area Number 07557155
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Biological pharmacy
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KAWASAKI Toshisuke  Department of Biological Chemistry, Kyoto University, Professor, 薬学研究科, 教授 (50025706)

Co-Investigator(Kenkyū-buntansha) TAKEUCHI Makoto  Central Laboratory, Kirin Brewery Co., Ltd., 主任研究員
SUZUKI Akemi  Tokyo Metropolitan Institute of Medical Science, 部長 (70134533)
KOZUTSUMI Yasunori  Dept.of Biol.Chem., Kyoto University, Assistant Professor, 薬学研究科, 助教授 (70205425)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥19,800,000 (Direct Cost: ¥19,800,000)
Fiscal Year 1997: ¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1996: ¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1995: ¥8,300,000 (Direct Cost: ¥8,300,000)
Keywordscarbohydrate antigen / CMP-NeuAc hydroxylase / N-Glycolylneuraminic acid / HD antigen / gene targeting / 遺伝子クローニング / ノックアウトマウス / クローニング
Research Abstract

N-Glycolylneuraminic acid (NeuGc) is derived from NeuAc by hydroxylation of a methyl moiety in the N-acetyl residue. This hydroxylation occurs by the conversion of CMP-N-acetylneuraminic acid (CMP-NeuAc) to CMP-N-glycolylCMP-NeuGc, which is carried out by an electron transport system involving NADH-dependent cytochrome b5 reductase, cytochrome b5, and CMP-NeuAc hydroxylase. CMP-NeuAc hydroxylase is the key enzyme in the hydroxylation of CMP-NeuAc, and functions at the final step of the electron transport system. We have previously purified CMP-NeuAc hydroxylase from the mouse liver, and isolated a cDNA for this enzyme which contains an ORF encoding a protein of 577 amino acids. All mammals examined except human have NeuGc and therefore NeuGc is known to be an epitope of one of major carbohydrate antigens in human. Recently, it is reported that NeuGc is detected in the sialic acid-containing medicines such as human erythropoietin produced in CHO cells.
In this study, we plan to make NeuGc-deficient animals using gene-targeting technique in order to produce NeuGc-free tsialic acid-containing medicines.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] T.Kawano et al: "Molecular cloning of CMP-N-acetylneuraminic acid hydroxylase from mouse liver" J.Biol.Chem.270(27). 16458-16463 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Takematsu et al: "Isolation and characterization of the rabbit cytochrome b 5 gene" J.Sequencing and Mapping. 6. 123-125 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Koyama et al: "A naturally occurring 46-amino acid deletion of cytidine monophospho-N-acetylneuraminic acid hydroxylase leads to a change in the intracellular distribution of the protein" Glycoconjugate J.13. 353-358 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Kawano, S.Koyama, H.Takematsu, Y.Kozutsumi, H.Kawasaki, S.Kawashimai, T.Kawasaki, & A.Suzuki: "Molecular cloning of CMP-N-acetylneuraminic acid hydroxylase from mouse liver" J.Biol.Chem.270(27). 16458-16463 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Takematsu, Y.Kozutsumi, A.Suzuki, & T.Kawasaki: "Isolation and characterization of the rabbit cytochrome b5 gene" J.Sequencing and Mapping. 6. 123-125 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Koyama, T.Yamaji, H.Takematsu, T.Kawano, Y.Kozutsumi, A.Suzuki & T.Kawasaki: "A naturally occurring 46-amino acid deletion of cytidine monophospho-N-acetylneuraminic acid hydroxylase leads to a change in the intracellulardistribution of the protein" Glycoconjugate J.13. 353-358 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yamaji et al: "Neutral glycosphingolipids induce cell-cell aggregation of a variety ofhematopoietic cell lines." Eur.J.Biochem.247(1). 21-29 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Koyama et al.: "A naturally occurring 46-amino acid deletion of cytidine monophospho-Nacetylneuraminic acid hydroxylase leads to a change in the intracellular distribution of the protein" Glycoconjugate J.13. 353-358 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Takematsu et al.: "Isolation and characterization of the rabbit cytochrome b 5 gene" J.Sequencing and Mapping. 6(2). 123-125 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kawano, S. Koyama, H. Takematsu, Y. Kozutsumi et al.: "Molecular Cloning of CMP-N-cetylneuraminic Acid Hydroxylase from Mouse Liver." J. Biol. Chem.270 (27). 16458-16463 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] H. Takematsu, Y. Kozutsumi, A. Suzuki. and T. Kawasaki: "Isolation and Characterization of the Rabbit Cytochrome β Gene." J. Sequencing and Mapping,. 6. 123-125 (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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