• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Study for the development of new gene targeting method utilizing epitope tags

Research Project

Project/Area Number 11556064
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 InstitutionIwate University

Principal Investigator

MORIMATSU Masami  Iwate University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (70241370)

Co-Investigator(Kenkyū-buntansha) YOSHIMATSU Kumiko  Iwate University, Faculty of Medicine, Hokkaido University, Associate Professor, 医学部, 助手 (90220722)
MIYOSHI Ichiro  Iwate University, Faculty of Medicine, Tohoku University, Assistant Professor, 医学部, 助手 (10183972)
SYUTO Bunei  Iwate University, Faculty of Agriculture, Professor, 農学部, 教授 (60001533)
YOSHIMURA Yoshimichi  Reagent Development Group,A &T,Co., Manager, 試薬開発グループ, マネージャー
OGAWA Kazushige  Iwate University, Faculty of Agriculture, Osaka Prefecture University, Associate Professor, 農学部, 助教授 (60231221)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥11,300,000 (Direct Cost: ¥11,300,000)
Fiscal Year 2001: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2000: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥5,400,000 (Direct Cost: ¥5,400,000)
Keywordsepitope-tag / gene targeting / interaction of molecules / cellular localization / 遺伝子機能解析 / マウス
Research Abstract

To establish the bases for developing new methods of gene targeting, we investigated the efficiency of gene knock-in and the use of specific markers called epitope-tags. By using this approach, gene of interest is to be analyzed by antibodies against epitope-tags. We selected some genes for targeting experiments. Detailed characterization of candidates for epitope-tags was also carried out.
1. Investigation of a novel endotoxin-inducible gene, MAIL, as a target.
MAIL is suitable for targeting experiment because it is known that transcription of the gene is markedly up-regulated by endotoxin stimulation and that the gene product is specifically localized in the nucleus. We cloned and sequenced the genomic DNA of MAIL. Targeted disruption of MAIL gene was also performed.
2. Investigation of genes encoding chitinase as a target.
Because of tissue-specificity of chitinase gene, we choose this gene as a target. We cloned, sequenced, and analyzed expression pattern of chitinase gene, and established the basis for gene targeting.
3. Investigation of other target genes.
Inducible genes, such as B-13, and tissue-specific genes, such as glucose transporters, were investigated as targets.
4. Analysis of epitope-tags.
Antigenic specificity and cellular function of candidate epitope-tags from hantavirus proteins were examined.
5. Developing knock-in method.
We tried two-step targeting method for gene knock-in. However, expected recombinants were not obtained.
Further research is needed to clarify the cause of the failure.

Report

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

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Kitamura, H.: "MAIL, a novel nuclear IkB protein that potentiates LPS-induced IL-6 production"FEBS Lett.. 485. 53-56 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ebihara, H.: "Pathogenicity of Hantaan virus in newborn mice : genetic reassortant study demonstrating that a single amino acid change in glycoprotein G1 is related to virulence"J. Virol.. 74. 9245-9255 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Araki, K.: "Truncated hantavirus nucleocapsid proteins for serotyping Hantaan, Seoul, and Dobrava hantavirus infections"J. Clin. Microbiol.. 39. 2397-2404 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Iwasaki, K.: "Isolation, characterization, and cDNA cloning of chicken turpentine-induced protein, a new member of the scavenger receptor cystein-rich (SRCR) family of proteins"J. Biol. Chem.. 276. 9400-9405 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shiina, T.: "Genomic organization, chromosomal localization and promoter analysis of the mouse Mail gene"Immunogenetics. 53. 649-655 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki, M.: "Cellular expression of gut chitinase mRNA in the gastrointestinal tract of mice and chickens"J. Histochem. Cytochem.. 50(印刷中). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kitamura,H., et al.: "MAIL, a novel nuclear I kappa B protein that potentiates LPS-induced IL-6 production"FEBS Lett.. 485. 53-56 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ebihara,H., et al.: "Pathogenicity of Hantaan virus in newborn mice : genetic reassortant study demonstrating that a single amino acid change in glycoprotein G1 is related to virulence"J Virol.. 74. 9245-9255 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Araki,K., et al.: "Truncated hantavirus nucleocapsid proteins for serotyping Hantaan, Seoul, and Dobrava hantavirus infections"J Clin Microbiol.. 39. 2397-2404 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Iwasaki,K., et al.: "Isolation, characterization, and cDNA cloning of chicken turpentine-induced protein, a new member of the scavenger receptor cysteine-rich (SRCR) family of proteins"J Biol Chem.. 276. 9400-9405 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shiina,T., et al.: "Genomic organization, chromosomal localization, and promoter analysis of the mouse Mail gene"Immunogenetics. 53. 649-655 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki,M., et al.: "Cellular expression of gut chitinase mRNA in the gastrointestinal tract of mice and chickens"J Histochem Cytochem.. 50 (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shiina, t.: "Genomic organization, chromosomal localization and promoter analysis of the mouse Mail gene"Immunogenetics. 53. 649-655 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kitamura, H.: "Bacterial lipopolysaccharide induced mRNA expression of a IkB MAIL mediated through Toll-like receptor 4"J. Vet. Med. Sci. (印刷中).

    • Related Report
      2001 Annual Research Report
  • [Publications] Suzuki, M.: "A novel serum chitinase that is expressed in bovine liver"FEBS Lett. 506. 127-130 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Suzuki, M.: "Cellular expression of chitinase mRNA in the gastrointestinal tract of mice and chicken"J. Histochem. Cytochem. (印刷中).

    • Related Report
      2001 Annual Research Report
  • [Publications] Kimura,K. ら: "Mutated human beta3-adrenergic receptor (Trp64Arg) lowers the response to beta3-adrenergic agonists in transfected 3T3-L1 preadipocytes."Horm.Metab.Res.. 32. 91-96 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Iwase,M. ら: "Canine leptin : cDNA cloning expression and activity of recombinant protein."Res Vet Sci.. 68. 109-114 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kitamura,H. ら: "MAIL.a novel nuclear IkB protein that potentiates LPS-induced IL-6 production."FEBS Lett.. 485. 53-56 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Iwasaki,K. ら: "Isolation, characterization, and cDNA cloning of chicken turpentine-induced protein, a new member of the scavenger receptor cystein-rich(SRCR) family of proteins."J.Biol.Chem.. (印刷中).

    • Related Report
      2000 Annual Research Report
  • [Publications] Watamabe,Y.ら: "Botulinum C3 enzyme changes the lactate dehydrogenase isozyme pattern of primary culture of neurons"Journal of Veterinary Medical Science. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Watamabe,Y.ら: "The evaluation of the potential of botulinum C3 enzyme as an exogenous differentiation-inducing factor to neurons"Journal of Veterinary Medical Science. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Suzuki,K.ら: "Molecular characterization of airborne odorant receptor organ specific protein in Xenopus laevis daudin"Animal Biochemistry. 36管1号. (1999)

    • Related Report
      1999 Annual Research Report

URL: 

Published: 1999-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi