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Development of systems for cell culture and expression vector to generate transgenic marine invertegra

Research Project

Project/Area Number 08558080
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Developmental biology
Research InstitutionHIROSHIMA UNIVERSITY

Principal Investigator

SHIMADA Hiraku  Fuculty of Science, Hiroshima University, Professor, 理学部, 教授 (70011559)

Co-Investigator(Kenkyū-buntansha) MACHII Akira  Yamakatsu Pearl Co., Investigator, 養殖研究室, 顧問研究員
淡路 雅彦  水産庁中央水産研究所, 生物機能部・分子生物研究室, 室長
WADA Katsuhiko  National Research Institute of agriculture, Head Investigator, 遺伝育種部, 部長
NAKATSUBO Keiko  Fuculty of Science, Hiroshima University, Assistant, 理学部, 助手 (40192760)
AKASAKA Koji  Fuculty of Science, Hiroshima University, Associate Professor, 理学部, 助教授 (60150968)
MASAHIKO Awaji  National Research Institute of Fisheries Science, Investigator
小林 敬典  水産庁養殖研究所, 研究員
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥19,800,000 (Direct Cost: ¥19,800,000)
Fiscal Year 1998: ¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 1997: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1996: ¥10,500,000 (Direct Cost: ¥10,500,000)
Keywordssea urchin / pearl oyster / transgenic animal / insulator / expression vector / cell culture / 遺伝子導入 / 遺伝子組換え / バフンウニ / 遺伝子 / 転写調節 / 転写因子
Research Abstract

1. Identification of cis-active elements that regulate expression of sea urchin gene :
Several sequence elements that regulate transcription of the sea urchin Ars gene were detected by reporter assay around its transcription start site. Among them between Sox-binding sitebetween -168bp and -162bp is most prominent element. Around -186bp cis-element that represses the promoter activity was detected.
2. Validity of insulator element in gene transfer experiments :
An insulator element present at the upstream of the sea urchin Ars gene showed insulator activity also in cells of Drosophila, mouse and humane
3. Cell culture system for mantle epitermal cells of pearl oyster :
Primary cell culture system for mantle epitermal cells of pearl oyster was established, though they do not proliferate in vitro.
4. Culturing of sea urchin embryonic cells :
Though we succeeded in establishing in vitro culture of cells from sea urchin pluteus larvae, we failed to culture them beyond three days because of difficulty in preventing proliferation of contaminating fungi.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Ishii et al: "Hbox1 and Hbox7 are involved in the Pattern Formation of Sea urchin Embryo" Development Growth & Differentiation. 41印刷中. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kurokawa,et al: "HpEts,an ets-related transcription factor implicated in primary mesenchyme cell differentiation of the sea urchin embryo" Mechanism of Development. 80. 41-52 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawasaki,et al: "Lim1-related homeobox gene(HpLim1) expressed in sea urchin embryo" Development Growth&Differentiation. 41印刷中. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kiyama et al: "Structure and function of a sea urchin orthodenticle-related HpOtx gene" Developmental Biology. 193. 139-145 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Mitsunaga-Nakatsubo et al: "Differential expression of sea urchin Otx isoform (HpOtxE and HpOtxL) mRNA during early development" International Journal of Developmental Biology. 42. 645-651 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Mitsunaga-Nakatsubo et al: "Arylsulfatase exists as non-enzymatic cell surface protein in sea urchin embryos" Journal of Experimental Zoology. 280. 220-230 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Ishii, et al.: "Hbox1 and Hbox7 are involved in the Pattern Formation of Sea urchin Embryo" Dev.Growth & Differ.41(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] D.Kurokawa, et al.: "HpEts, an ets-related transcription factor implicated in primary mesenchyme cell differentiation of the sea urchin embryo." Mech.Dev.80. 41-52 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Kawasaki, et al.: "Liml-related homeobox gene (HpLim 1) expressed in sea urchin embryo" Dev.Growth & Differ.41(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Kiyama, et al.: "Structure and function of a sea urchin orthodenticle-relatedHpOtxgene." Dev.Biol.193. 139-145 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Mitsunaga-Nakatsubo, et al.: "Differential expression of sea urchin Otx isoform (HpOtxE and HpOtxL) mRNA during early development." Int.J.Dev.Biol.42. 645-651 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Mitsunaga-Nakatsubo, et al.: "Arylsulfatase exists as non-enzymatic cell surface protein in sea urchin embryos." J.Exp.Zool.280. 220-230 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ishii et al: "Hboxl and Hbox7 are involved in the Pattern Formation of Sea urchin Embryo" Development Growth & Differentiation. 41 (印刷中). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kurokawa,et al: "HpEts,an ets-related transcription factor implicated in primary mesenchyme cell differentiation of the sea urchin embryo" Mechanism of Development. 80. 41-52 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kawasaki,et al: "Liml-related homeobox gene (HpLiml) expressed in sea urchin embryo" Development Growth & Differentiation. 41 (印刷中). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kiyama et al: "Structure and function of a sea urchin orthodenticle-related HpOtx gene" Developmental Biology. 193. 139-145 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Mitsunaga-Nakatsubo et al: "Differential expression of sea urchin Otx isoform (HpOtxE and HpOtxL) mRNA during early development" International Journal of Developmental Biology. 42. 645-651 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Mitsunaga-Nakatsubo et al: "Arylsulfatase exists as non-enzymatic cell surface protein in sea urchin embryos" Journal of Expermintal Zoology. 280. 220-230 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] N.Sakamoto: "Two isoforms of orthodenticle-related proteins (HpOtx)bind to the enhancer element of sea urchin arylsulfatase gene." Developmental Biology. 181. 284-295 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Koji Akasaka: "Oral-aboral ectoderm differentiation of the sea urchin embryos is disrupted in response to calcium ionophore." Development Growth and Differentiation. 39. 373-379 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Junji Morokuma: "A cis-regulatory element within the 5′ flanking region of arylsulfatase gene of sea urchin,Hemicentrotus pulcherrimus." Development Growth and Differentiation. 39. 469-476 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Daisuke Kurokawa: "Cloning of cyclin E cDNA of the sea urchin,-Hemicentrotus pulcherrimus." Zoological Science. 14. 791-794 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Mitsunaga-Nakatsubo: "Arylsulfatase exists as non-enzymatic cell surface protein in sea urchin embryos." Journal of Experimental Zoology. 280. 220-230 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Kiyama: "Structure and function of a sea urchin orthodenticle-related HpOtx gene." Developmenttal Biology. 193. 139-145 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sakamoto et al.: "A triplex DNA configuration of the polypyrimidine : polypurine stretch in the 5' flanking region of the sea urchin arylsulfatase gene." Zoological Science. 13. 105-109 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Harada et al: "Spatial expression of a forkhead homologue in the sea urchin embryo." Mechanism of Development. 60. 163-173 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Mitsunaga-Nakatsubo et al: "cDNA cloning of Na+, K+-ATPase alpha subunit from embryos of the sea urchin, Hemicentrotus pulcherrimus." Zoological Science. 13. 833-841 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sakamoto et al: "Two isoforms of orthodenticle-related proteins (HpOtx) gind to the enhancer element of sea urchin arylsulfatase gene." Developmental Biology. 181. 284-295 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Akasaka et al: "Oral-aboral ectoderm differntiation of the sea urchin embryos is disrupted in response to calcium ionophore." Development Growth and Differentiation. 39 (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wada et al: "Color and weight of pearls produced by grafting the mantle tissue from a selected population for white shell color of the Japanese pearl oyster" Aquaculture. 142. 25-32 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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