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Studies on the post-mortem fish muscle tenderization -Analysis using the transgenic red seabream

Research Project

Project/Area Number 15380142
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Fisheries chemistry
Research InstitutionKyoto University

Principal Investigator

TOYOHARA Haruhiko  KYOTO UNIVERSITY, GRADUATE SCHOOL OF AGRICULTURE, ASSOCIATE PROFESSOR, 農学研究科, 助教授 (90183079)

Co-Investigator(Kenkyū-buntansha) KINOSHITA Masato  KYOTO UNIVERSITY, GRADUATE SCHOOL OF AGRICULTURE, ASSISTANT PROFESSOR, 農学研究科, 助手 (60263125)
KATO Keitaro  KINKI UNIVERSITY, FISHERY RESEACH INSTITUTE, 水産研究所, 講師 (90330240)
Project Period (FY) 2003 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥15,200,000 (Direct Cost: ¥15,200,000)
Fiscal Year 2005: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 2004: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2003: ¥6,200,000 (Direct Cost: ¥6,200,000)
KeywordsRED SEABREAM / TRANSGENIC FISH / MEAT QUALITY / CULTURED FISH / TENDERIZATION / MUSCLE / AUTOLYSIS / CONNECTIVE TISSUE / 発現ベクター / トランスジェニックフィッシュ / アクチン / MMP / TIMP / 鮮度 / コラーゲン
Research Abstract

By the injection of 1, 10-phenanthroline, a specific inhibitor for metalloproteinase, meat tenderization of red seabream was significantly suppressed, suggesting that matrix metalloproteinase is involved in the post-mortem meat tenderization of red seabream. Then, Genes encoding matrix metalloproteinases (MMP) and tissue inhibitor of metalloproteinases (TIMP) were isolated from red sea bream. Actually, recombinant MMP degraded collagen, a main component of the muscle connective tissue, and the activity was inhibited by the addition of TIMP. These facts suggested MMP-TIMP system plays a very important role in the post-mortem tenderization of red seabream muscle.
These findings suggested the transgenic expression of TIMP might suppress the post-mortem meat tenderization. We made a transgenic medaka overexpressing TIMP. Histological analysis demonstrated the effectiveness of the transgenic expression of TIMP for the suppression of collagen breamdown.
To apply this technology to cultured fish, we adopted red seabream, an important species for fish culture. For the construction of the expression vector, three distinct α-actin genes (two skeletal muscle types and one cardiac muscle type) were also isolated. These actin genes are composed of 8 exons and having E Box and CArG Box in the up-stream region, but the expression pattern in the fish was clearly distinct. We made the expression vectors harboring the promoter regions of these genes with GFP as a reporter gene. As a result, all vectors were revealed to be useful for making transgenic red seabream.
By using these vectors, we introduced TIMP gene into red seabream. We are now keeping 200 fish possibly having TIMP gene.

Report

(4 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (12 results)

All 2006 2005 2004 2003

All Journal Article (9 results) Book (1 results) Patent(Industrial Property Rights) (2 results)

  • [Journal Article] Molecular cloning and characterization of a tissue inhibitor of metalloproteinase, TIMP-2, from red seabream cultured cells.2006

    • Author(s)
      K.Touhata
    • Journal Title

      Fisheries Science (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Molecular cloning and characterization of a tissue inhibitor of metalloproteinase, TIMP-2, from red seabream cultured cells2006

    • Author(s)
      K.Touhata, Y.Touhata, M.Yamashita, H.Toyohara
    • Journal Title

      Fisheries Science (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Establishment of a transgenic medaka line expressing Japanese flounder tissue inhibitor of metalloproteinase for the suppression of post-mortem meat tenderization.2004

    • Author(s)
      H.Toyohara
    • Journal Title

      Marine Biotechnol. 6

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Establishment of a transgenic medaka line expressing Japanese flounder tissue inhibitor of metalloproteinase for the suppression of post-mortem meat tenderization.2004

    • Author(s)
      H.Toyohara
    • Journal Title

      Marine Biotechnol. (6)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Establishment of a transgenic medaka line expressing Japanese flounder tissue inhibitor of metalloproteinase_for the suppression of post-mortem meat tenderization2004

    • Author(s)
      Haruhiko Toyohara
    • Journal Title

      Marine Biotechnology 6

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Occurrence of two distinct types of tissue inhibitor of metalloproeinase-2 (TIMP-2) in teleost fish.2003

    • Author(s)
      S.Kubota
    • Journal Title

      Biochim. Biophys. Acta. 1629

      Pages: 102-108

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Solubilization of Type I collagen from fish muscle connective tissue by matrixmetalloproteinase-9 at chilled temperature.2003

    • Author(s)
      M.Kubota
    • Journal Title

      Fisheries Science 69

      Pages: 1053-1059

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Occurrence of two distinct types of tissue inhibitor of metalloproeinase-2 (TIMP-2) in teleost fish. Biochim.2003

    • Author(s)
      S.Kubota
    • Journal Title

      Biochim. Biophys. Acta. (1629)

      Pages: 102-108

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Solubilization of Type I collagen from fish muscle connective tissue by matrixmetalloproteinase-9 at chilled temperature. Fisheries Science2003

    • Author(s)
      S.Kubota
    • Journal Title

      Fisheries Science (69)

      Pages: 1053-1059

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] バイオサイエンスの新戦略-魚類生産とバイオテクノロジー2004

    • Author(s)
      豊原治彦
    • Publisher
      丸善
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Patent(Industrial Property Rights)] α-アクチンプロモーター-遺伝子,発現ベクター及び遺伝子導入魚類2005

    • Inventor(s)
      家戸敬太郎, 豊原治彦
    • Industrial Property Rights Holder
      独立行政法人科学技術振興機構, 学校法人近畿大学
    • Filing Date
      2005-03-22
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Patent(Industrial Property Rights)] α-アクチンプロモーター遺伝子,発現ベクター及び遺伝子導入魚類2005

    • Inventor(s)
      家戸敬太郎, 豊原治彦
    • Industrial Property Rights Holder
      近畿大学, 京都大学
    • Industrial Property Number
      2005-081280
    • Filing Date
      2005-03-22
    • Related Report
      2005 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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