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

Basic and applied research on production of "double-muscle" cattle by use of a chimeric DNA/RNA oligo-nucleotide

Research Project

Project/Area Number 12660261
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied animal science
Research InstitutionKinki University

Principal Investigator

SAEKI Kazuhiro  School of Biology-Oriented Science and Technology, Kinki University, Department of Genetic Engineering, Professor, 生物理工学部, 教授 (10298937)

Co-Investigator(Kenkyū-buntansha) MATSUMOTO Kazuya  School of Biology-Oriented Science and Technology, Kinki University, Department of Genetic Engineering, Professor, 生物理工学部, 助教授 (20298938)
HOSOI Yoshihiko  School of Biology-Oriented Science and Technology, Kinki University, Department of Genetic Engineering, Professor, 生物理工学部, 教授 (70192739)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsTGP8 / Cattle / embryo / clone / transfection / somatic cells
Research Abstract

The following research was done for the production of transforming growth factor 8 gene (TGF8) knock-out cattle using a chimeric DNA/RNA oligo-nucleotide.
1) To investigate effects of cell cycle of the recipient cells, bovine fibroblasts and cumulus cells, on the gene expression, p-β-act/luc^+/IRES/EGFP/neo^r was injected into the cell nuclei. The luminescence was highly detected when cells were in a confluent state, but not after serum starvation. To examine the efficiency of gene transformation of the cells, gene introduction methods, DNA-microinjection, etectroporation and a transfection reagent were used for introduction of the p-β-act/luc^+/IRES/EGFP/neo^r into the bovine somatic cells. The gene was transfected most effectively by use of the transfection reagent by detecting EGFP fluorescence. Furthermore, in the transfected cells, the fluorescence was further detected after selection by G418, and in nuclear transplanted-embryos with the transfected. These indicated that the gene m … More ay be integrared in genome of the cells. From the data, the use of the transfection reagent was the most effective to introduce the exogenous gene into bovine somatic cells.
2) A point mutation of cyh2 encoding L29 which is a ribosomal protein in yeast induce cychloheximide tolerance. We examined whether the similar mutation of the homologous gene encoding L27, a mammalian ribosomal protein induced the cychloheximide tolerance during cell culture. We designed the chimeric DNA/RNA oligo-nucleotide which induced the mutation of L27 gene, and introduced the oligo-nucleotide into bovine and mouse fibroblasts by the transfection reagent The cells were cultured under cychloheximide for 10 days. All of the not-transfected and vecter-transfected cells were degenerated 10 days after culture with cychloxeximide, but bovine and mouse cells transfected with the oligo-nucleotide were survived even with cychloheximide for 10 days. The results showed the transfection of the oligo-nucleotide might induce the point-mutation of the L27 gene of the cells and consequently the cells acquired the cychloheximide tolerance.
3) We designed the the chimeric DNA/RNA oligo-nucleotide which induced point-mutation of bovine GDF8 gene. The use of the oligo-nucleotide may provide the alternative method to produce the KO animals. Less

Report

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

    (21 results)

All Other

All Publications (21 results)

  • [Publications] 佐伯和弘, 高塚秋光, 松本和也, 細井美彦, 加蒔博己, 入谷 明: "ウシ胚の体外発生に及ぼす培養液中の無機リン酸塩の影響"近畿大学生物理工学研究所紀要. 5. 25-29 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 保坂 卓, 谷口俊仁, 佐伯和弘子: "ウシ線維芽細胞を用いた核移植によるクローンウシの受胎"近畿大学生物理工学研究所紀要. 3. 45-51 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saeki K, Hosaka T, Maeda M, Minaminoto H, Okamoto C, Matsuba J, Matsumoto K, Hosoi Y, Iritani A: "Onset of gene expression in bovine embryos reconstructed with fibroblasts following reporter gene microinjection"Theriogenology. 55. 289 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 駒野徹, 佐伯和弘, 松本和也: "食資源動物の利用形質の遺伝子工学的制御-遺伝子工学がデザインする未来の食資源動物-"学術月報. 541. 447-450 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saeki K, Maeda M, Hori T, Okamoto C, Kasamatsu A, Taniguchi S, Matsumoto K, Hosoi Y, Nishimoto Y, Iritani A: "Early development and gene expression of reconstructed embryos following fusion of bovine enucleated oocytes with somatic cells from different species"Theriogenology. 57. 446 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Saeki, N.Sumitomo, Y.Nagata, Y.Hosoi, K.Matsumoto, A.Iritani: "Atomic force microscopy of bovine acrosome-intact and reacted spermatozoa ; their fine structure and numerical analysis"Theriogenology. 59. 468 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Saeki, N. Sumitomo, Y. Nagata, Y. Hosoi, K. Matsumoto and A. Iritani: "Atomic force microscopy of bovine acrosome-intact and reacted spermatozoa; their fine structure and numerical analysis."Theriogenology. 59. 468 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saeki K, Maeda M, Hori T, Okamoto C, Kasamatsu A, Taniguchi S, Matsumoto K, Hosoi Y, Nishimoto Y and Iritani A.: "Early development and gene expression of reconstructed embryos following fusion of bovine enucleated oocytes with somatic cells from different species."Theriogenology. 57. 446 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saeki K, Hosaka T, Maeda M, Minaminoto H, Okamoto C, Matsuba J, Matsumoto K, Hosoi Y and Iritani A.: "Onset of gene expression in bovine embryos reconstructed with fibroblasts following reporter gene microinjection."Theriogenology. 55. 289 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saeki K, Noichi T, Matsumoto K, Kaneko T, Harada M, Hosoi Y, Iritani A.: "Fate of DNA microinjected into bovine embryos."Theriogenology. 53. 520 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Saeki, N.Sumitomo, Y.Nagata, Y.Hosoi, K.Matsumoto, A.Iritani.: "Atomic force microscopy of bovine acrosome-intact and reacted spermatozoa ; their fine structure and numerical analysis"Theriogenology. No.59, vol.1. 468 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Y.Hosoi, M Takahashi, S Higashide, K.Matsumoto, K.Saeki, A.Iritani, T Miyano: "Development of rabbit preantral follicles isolated from ovaries of sexually matured doe in the SCID mice"Theriogenology. No.59, vol.1. 408 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Matsumoto, M Zhang, Y Yamamoto, K.Saeki, Y.Hosoi, K Lu, A.Iritani: "Molecular cloning and characterization of a novel gene specifically expressed in gonad"Theriogenology. No.59, vol.1. 428 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Y.Hosoi, R.Torii, N.Fujinami, K.Matsumoto, K.Saeki, A.Iritani: "Fertilization by intracytoplasmic sperm injection and subsequent embryo development in vitro to blastocysts in Japanese monkey (Macaca fuscata)"Journal of Mammalian Ova Research. No.20, vol.1(In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Saeki K et al.: "Early development and gene-expression of reconstructed embryos following fusion of bovine enucleated oocytes with somatic cells from different species"Theriogenology. 57. 446 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 佐伯和弘: "最近における胚の性判別および精子分別の研究開発の動向. I.PCR法による胚の性判別とその実用化"畜産技術. 11月号. 5-8 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 駒野徹, 佐伯和弘, 松本和也: "食資源動物の利用形質の遺伝子工学的制御-遺伝子工学がデザインする未来の食資源動物-"学術月報. 541. 447-450 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 佐伯和弘 他: "ウシ胚の体外発生に及ぼす培養液中の無機リン酸塩の影響"近畿大学生物理工学研究所紀要. 5. 25-29 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 佐伯和弘 他: "子ウシ線維芽細胞を用いた核移植によるクローンウシの受胎"近畿大学生物理工学研究所紀要. 3. 45-51 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Saeki,K et al.: "Onset of gene expression in bovine embryos reconstructed with fibroblasts following reporter gene microinjection"Theriogenology. 55. 289 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Saeki,K et al.: "Fate of DNA microinjected into bovine embryos"Theriogenology. 53. 520 (2000)

    • Related Report
      2000 Annual Research Report

URL: 

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

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi