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Construction of new host-vector systems of the production of foreign proteins using a variety of protein targeting signals.

Research Project

Project/Area Number 12556061
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Applied molecular and cellular biology
Research InstitutionKagawa University

Principal Investigator

TAKEGAWA kaoru  Kagawa university, Faculty of Agriculture, Professor, 農学部, 教授 (50197282)

Co-Investigator(Kenkyū-buntansha) HAMA yuko  Asahi Glass Co. Ltd., Research Center, Project Leader, 中央研究所, 首席研究員
KIMURA yoshio  Kagawa University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (10243750)
SAKAI yasuyoshi  Kyoto University, Graduate School of Agriculture, Associate Professor, 農学研究科, 助教授 (60202082)
浜 祐子  旭硝子, 中央研究所, 首席研究員
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 2002: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2000: ¥4,700,000 (Direct Cost: ¥4,700,000)
KeywordsSchizosaccharomyces pombe / protein trafficking / sortiong signal / glycosylatopn / foreign gene expression / 物質生産
Research Abstract

1. We developed a new transformation system using multiple auxotrophic marker genes for the fission yeast Schizosaccharomyces pombe. This new transformation system would be an useful tool for molecular genetic studies of S. pombe where multiple recombinant modifications or multiple mutations are needed.
2. We have isolated many genes that are required for protein secretion and sorting in fission yeast. Using a variety of protein targeing signals and mutants, we constructed new types of host-vector systems to produce foreign proteins.
3. The glycoproteins of S. pombe contain a large amount of galactose in addition to mannose, indicating that S. pombe is equipped with mechanisms for galactosylation of glycoprotein, like mammalian cells. To elucidate the physiological role of galactosylation, we isolated several S. pombe mutants defective in protein galactosylation. We elucidate that the galactosylation of protein is required for the maintenance of normal cell shape, sexual agglutination, tolerance toward various drugs, and non-sexual flocculation in S. pombe.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Kaoru Takegawa 他: "Expression and characterization of Schizosaccharomyces pombe vacuolar carboxypeptidase Y in Saccharomyces cerevisiae"Curr.Genet.. 42. 252-259 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tomoko Iwaki 他: "Characterization of vps33+, a gene required for vacuolar biogenesis and protein sorting in Schizosaccharomyces pombe"Yeast. 20(In press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshio Kimura 他: "Myxococcus xanthus mokA encodes a histidine kinase-response regulator hybrid sensor required for development and osmotic tolerance"J.Bacteriol.. 183. 1140-1146 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Toshihiro Komeda 他: "Production of active bovine cathepsin C (dipeptidyl aminopeptidase I) in the methylotrophic yeast Candida boidinii"Appl.Microbiol.Biotechnol.. 59. 252-258 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Bumjun Lee 他: "Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii"Microbiology. 148. 2697-2704 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Atsushi Isoai 他: "Production of D-amino acid oxidase (DAO) of Trigonopsis variabilis in Schizosaccharomyces pombe and the characterization of biocatalysts prepared with recombinant cells"Biotechnol.Bioeng.. 80. 207-217 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tomoko Iwaki et al.: "Characterization of vps33+, a gene required for vacuolar biogenesis and protein sorting in Schizosaccharomyces pombe"Yeast. 20, in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kaoru Takegawa et al.: "Heterologous expression and characterization of Schizosaccharomyces pombe vacuolar carboxypeptidase Y in Saccharomyces cerevisiae"Curr. Genet.. 42. 252-259 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshi Kimura et al.: "Myxococcus xanthus mokA encodes a histidine kinase-response regulator hybrid sensor required for development and osmotic tolerance"J. Bacteriol. 183. 1140-1146 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Toshihiro Komeda et al.: "Production of active bovine cathepsin C (dipeptidyl aminopepetidase I) in the methylotrophic yeast Candida boidinii"Appl. Microbiol. Biotechnol.. 59. 252-258 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Bumjun Lee et al.: "Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii"Microbiology. 148. 2697-2704 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Atsushi Isoai et al.: "Production of D-amino acid oxidase (DAO) of Trigonopsis variabilis in Schizosaccharomyces pombe and the characterization of biocatalysts prepared with recombinant cells"Bioechnol. Bioeng.. 80. 207-217 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kaoru Takegawa他: "Expression and characterization of Schizosaccharomyces pombe vacuolar carboxypeptidase Y in Saccharomyces cerevisiae"Curr. Genet.. 42. 252-259 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masayuki Onishi他: "Role of phosphatidylinositol 3-phosphate in formation of forespore membrane in Schizosaccharomyces pombe"Yeast. 20. 193-206 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hong Cheng他: "Role of the Rab GTP-Binding Protein Ypt3 in the Fission Yeast Exocytic Pathway, and Its Connection to Calcineurin Function"Mol. Biol. Cell. 13. 2963-2976 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Toshihiro Komeda他: "Production of active bovine cathepsin C (dipeptidyl aminopeptidase I) in the methylotrophic yeast Candida boidinii"Appl. Microbiol. Biotechnol.. 59. 252-258 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Toshihiro Komeda他: "Construction of protease-deficient Candida boidinii strains useful for recombinant protein production : Cloning and disruption of proteinase A gene (PEP4) and proteinase B gene (PRB1)"Biosci. Biotechnol. Biochem.. 66. 628-631 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Bumjun Lee他: "Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii"Microbiology. 148. 2697-2704 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Naotaka Tanaka, 他: "Functional Characterization of Gms 1p/UDP-Galactose Transporter in Schizosaccharomyces pombe"Yeast. 18・8. 745-757 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Naotaka Tanaka, 他: "Characterization of a Schizosaccharomyces pombe Mutant Deficient in UDP-Galactose Transport Activity"Yeast. 18・10. 903-914 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Naotaka Tanaka, 他: "Biosynthetic pathway and physiological role of galactose-containing oligosaccharides in fission yeast Schizosaccharomyces pombe"Trends Glycosci. Glycotechnol. 13・73. 519-532 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hirofumi Horiguchi, 他: "Antioxidant system within yeast peroxisome : Biochemical and physiological characterization of CbPmp20 in the methylotrophic yeast Candida boidinii"J. Biol. Chem.. 276・17. 14279-14288 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hirofumi Horiguchi, 他: "Peroxisomal catalase in the methylotrophic yeast Candida boidinii. Transport efficiency and metabolic significance"J. Bacteriol. 183・21. 6372-6383 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hiroyuki Mukaiyama, 他: "Paz2 and 13 other PAZ gene products regulate vacuolar engulfment of peroxisomes during micropexophagy"Genes Cells. 7・1. 75-90 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hiroya Yurimoto 他: "Regulation and evaluation of five methanol-inducible promoters in the methylotrophic yeast Candida boidinii."Biochim.Biophys.Acta. 1493・(1-2). 56-63 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 阪井康能: "異種タンパク質生産のための"細胞内輸送工学""生物工学会誌. 78・9. 397-401 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yoshio Kimura 他: "Myxococcus xanthus mokA encodes a hybrid sensor required for development and osmotic torelance. "J.Bacteriol.. 183.4. 1140-1146 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hiroya Yurimoto 他: "Characterization and high-level production of D-amino acid oxidase in Candida boidinii."Biosci.Biotechnol.Biochem.. 65(in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Naotaka Tanaka 他: "Functional Characterization of Gms1p/UDP-Galactose Transporter in Schizosaccharomyces pombe."Yeast. 18(in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Naotaka Tanaka 他: "Characterization of a Schizosaccharomyces pombe Mutant Deficient in UDP-Galactose Transport Activity."Yeast. 18(in press). (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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