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Developmetn of a method for cloning of cDNAs by phenotypic complementation of mammalian or fission yeast cells

Research Project

Project/Area Number 01890009
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field 広領域
Research InstitutionOsaka University

Principal Investigator

OKAYAMA Hiroto  Osaka University, Research Institute for Microbial Diseases, Professor, 微生物病研究所, 教授 (40111950)

Co-Investigator(Kenkyū-buntansha) ONO Yasuko  Osaka University, Research Institute for Microbial Diseases, Technical Instructo, 微生物病研究所, 教務職員(文部技官) (70194602)
NAGATA Akihisa  Osaka University, Research Institute for Microbial Diseases, Instructor, 微生物病研究所, 助手 (50155933)
NOJIMA Hiroshi  Osaka University, Research Institute for Microbial Diseases, Associate Professor, 微生物病研究所, 助教授 (30156195)
Project Period (FY) 1989 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥23,400,000 (Direct Cost: ¥23,400,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1990: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1989: ¥19,400,000 (Direct Cost: ¥19,400,000)
KeywordscDNA / Expression Cloning / Schizosacchromyces Pombe / Heterologous Genetic Complementation / Cell Cycle / cdc25 / Cdc2 / Weel / Schizosaccharomyces pombe / 動物細胞 / 遺伝子相補 / 酵母ベクタ- / 酵母トランスフェクション法
Research Abstract

We have developed a method that permits cloning of CDNAs based on. phenotypic complementation of mammalian or fission yeast cells. This method is composed of 1)a high-efficiency full-length CDNA cloning method ; 2)the pcD2 expression vector ; 3)a high-frequency mammalian cell transfection method ; 4)a high-efficiency yeast transfection method ; 5)highly-stable yeast vectors for transducing CDNA libraries into fission yeast.
Using this method, we have isolated two distinct human and rat homologs of the cdc25^+ gene, a fission yeast mitotic inducer, and human and rat homologs of the weel^+ gene, a fission yeast mitotic inhibitor. In addition we have isolated several human genes that suppress temperal'ure-sensitive cdc2 mutants or/and a temperature-sensitive sexual differentiation-proficient patl mutant. Some of them encodes a protein with RNA-bindng domains and some others a protein which. might associate with cdc2 kinase.
This CDNA cloning system will be useful for the islation of a wide variety of mammalian genes.

Report

(4 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Toyama,R.: "Human chorionic gonadotropin alfa and human cytomegalovirus promoters are extremely active in the fission yeast S.pombe." FEBS Lett.268. 217-221 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tanaka,K.: "A cDNA sequence of xeroderma pigmentosum group A gene encoding a protein with zinc finger structure." Nature. 348. 73-76 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Inoue,H.: "High efficiency transformation of Escherichea coli with plasmids." Gene. 96. 23-28 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Okazaki,N.: "High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe." Nucl.Acid.Res.18. 6485-6489 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nagata,A.: "An additional homolog of the fission yeast cdc25^+ gene occurs in humans and is highly expressed in some cancer cells." New.Biol.3. 959-968 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Igarashi,M.: "Weel^+-like gene in human cells." Nature. 353. 80-83 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Eiden,M.: "Suppressionof the chemically transformed phenotype of BHK cells by a human cDNA." Mol.Cell.Biol.11. 5321-5329 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Okazaki,N.: "The ste4^+ gene,essential for sexual differentiation of Schizosaccharomyces pombe,encodes a protein with a leucine zipper motif." Nucl.Acid.Res.19. 7043-7047 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kimura,T.: "Structure and expression of a human oxytocin receptor." Nature.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Okayama,H.: "Recombinant DNA methodology III.Vectors or methods for gene cloning.(12) High-efficiency cloning of full-length cDNA;construction and screening of cDNA expression libraries." Academic press, 26(235-260) (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Okayama,H.: "Methods in molecular biology.Chapter 2.Calcium phosphate mediated gene transfer into established cell lines." The Human press Inc., 7(15-21) (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Toyama, R.: "Human chorionic gonadotropin alfa and human cytomegalovirus promoters are extremely active in the fission yeast S, pombe." FEBS Lett.268. 217-221 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tanaka, K.: "A cDNA sequence of xeroderma pigmentosum group A gene encoding a protein with zinc finger structure." Nature. 348. 73-76 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Inoue, H.: "High efficiency transformation of Escheriches coli with plasmids." Gene. 96. 23-28 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Okazaki, K.: "High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe." Nucl. Acid. Res.18. 6485-6489 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nagata, A.: "An additional homolog of the fission yeast cdc25^+ gene occurs in humans and is highly expressed in some cancer cells." New Biol.3. 959-968 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Igarashi, M.: "Weel^+-like gene in human cells." Nature. 353. 80-83 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Eiden, M.: "Suppression of the chemically transformed phenotype of BHK cells by a human cDNA." Mol. Cell. Biol.11. 5321-5329 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Okazaki, N.: "The ste4^+ gene, essential for sexual differentiation of Schizosaccharomyces pombe, encodes a protein with a leucine zipper motif." Nucl. Acid. Res.19. 7043-7047 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kimura, T.: "Structure and expression of a huma oxytocin receptor." Nature.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nagata,A.: "An additional homolog of the fission yeast cdc25^+ gene occurs in humans and is highly expressed in some cancer cells." New.Biol.3. 959-968 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Igarashi,M.: "wee1^+-like gene in human cells." Nature. 353. 80-83 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Eiden,M.: "Suppression of the chemically transformed phenotype of BHK cells by a human cDNA." Mol.Cell.Biol.11. 5321-5329 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Okazaki,N.: "The ste4^+ gene,essential for sexual differentiation of Schizosaccharomyces pombe,encodes a protein with a leucine zipper motif." Nucl.Acid.Res.19. 7043-7047 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kimura,T.: "Structure and expression of a human oxytocin receptor." Nature.

    • Related Report
      1991 Annual Research Report
  • [Publications] Toyama,R.,and Okayama,H.: "Human chorionic gonadotropin alfa and human cytomegalovirus promoters are extremely active in the fission yeast <S.>___ー <pombe>___ー." FEBS Lett. 268. 217-221 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Inoue,H.,Nojima,H.,and: "High efficiency transformation of <Escherichea>___ー <coli>___ー with plasmids." Gene. 96. 23-28 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Okayama,H.,and Chen,C.: "Calcium phosphateーmediated gene transfer into established cell lines." Method Mol.Biol.

    • Related Report
      1990 Annual Research Report
  • [Publications] Okazaki,K.,Okazaki,N.,Kume,K.,Jinno,S.,Tanaka,K.,and Okayama,H.: "Highーfrequency transーformation method and library transducing vectors for cloning mammalian cDNAs by transーcomplementation of <Schizosaccharomyces>___ー <pombe>___ー." Nucleic Acids Res.18. 6485-6489 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hiroto Okayama: "Efficient vector system for cloning mammalian cDNA by transcomplentation of Schizosaccharomyces pombe:isolation of human cDNAs that complement patl." Mol.Cell.Biol.in press.

    • Related Report
      1989 Annual Research Report
  • [Publications] Hiroto Okayama: "High efficiency calcium phosphate transfection of mammalian cells." Current Protocols. in press.

    • Related Report
      1989 Annual Research Report
  • [Publications] Hiroto Okayama: "An improved method for high-efficiency transformation of Echerichiacoli with plasmids."

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2019-02-15  

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