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Secretory production of a functional membrane protein

Research Project

Project/Area Number 24656505
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field Biofunction/Bioprocess
Research InstitutionKobe University

Principal Investigator

YAMAJI Hideki  神戸大学, 工学(系)研究科(研究院), 教授 (40283874)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords昆虫細胞 / 組み換えタンパク質生産 / 膜タンパク質 / 組換えタンパク質生産
Research Abstract

The DNA fragment encoding the bacteriorhodopsin gene from Halobacterium halobium was cloned into a plasmid vector that contained the Bombyx mori actin promoter downstream of the B. mori nucleopolyhedrovirus (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression. After transfection with the resultant plasmid into Trichoplusia ni BTI-TN-5B1-4 (High Five) cells, cellular extract was analyzed by western blot analysis. Specific protein bands were detected at electrophoretic mobility of approximately 20 kDa in the cell lysate, indicating successful expression of bacteriorhodopsin in insect cells. The plasmid vector was then transfected into recombinant High Five cells that expressed the precursor (prM) of the viral membrane protein (M) and the envelope glycoprotein (E) of Japanese encephalitis (JE) virus. Secretory expression of virus-like particles embedding bacteriorhodopsin has been examined.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (9 results)

All 2014 2013 2012

All Journal Article (5 results) (of which Peer Reviewed: 4 results) Presentation (4 results)

  • [Journal Article] Suitability and perspectives on using recombinant insect cells for the production of virus-like particles2014

    • Author(s)
      Hideki Yamaji
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 98 Issue: 5 Pages: 1963-1970

    • DOI

      10.1007/s00253-013-5474-9

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Production of Japanese encephalitis virus-like particles in insect cells2013

    • Author(s)
      Hideki Yamaji, Eiji Konishi
    • Journal Title

      Bioengineered

      Volume: (in press) Issue: 6 Pages: 438-442

    • DOI

      10.4161/bioe.24514

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Efficient production of Japanese encephalitis virus-like particles by recombinant lepidopteran insect cells2013

    • Author(s)
      Hideki Yamaji, Masataka Nakamura, Miwa Kuwahara, Yusuke Takahashi, Tomohisa Katsuda, Eiji Konishi
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 97 Issue: 3 Pages: 1071-1079

    • DOI

      10.1007/s00253-012-4371-y

    • NAID

      120005464744

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Production of Japanese encephalitis virus-like particles using the baculovirus-insect cell system2012

    • Author(s)
      Hideki Yamaji, Maiko Segawa, Masataka Nakamura, Tomohisa Katsuda, Miwa Kuwahara, Eiji Konishi
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 114 Issue: 6 Pages: 657-662

    • DOI

      10.1016/j.jbiosc.2012.06.012

    • NAID

      110009578162

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] 昆虫細胞を用いたウイルス様粒子の生産2012

    • Author(s)
      山地秀樹
    • Journal Title

      PHARM TECH JAPAN

      Volume: 28 Pages: 2503-2508

    • URL

      http://www.jiho.co.jp/shop/list/detail/tabid/272/catid/75/pdid/92354/Default.aspx

    • Related Report
      2013 Final Research Report
  • [Presentation] Efficient production of viral envelope proteins in recombinant insect cells2013

    • Author(s)
      Hideki Yamaji, Miwa Kuwahara, Eiji Konishi
    • Organizer
      23rd ESACT (European Society for Animal Cell Technology) Meeting
    • Place of Presentation
      Lille, France
    • Year and Date
      2013-06-24
    • Related Report
      2013 Final Research Report
  • [Presentation] Efficient production of viral envelope proteins in recombinant insect cells2013

    • Author(s)
      Hideki Yamaji, Miwa Kuwahara, Eiji Konishi
    • Organizer
      23rd ESACT Meeting
    • Place of Presentation
      Lille Grand Palais, Lille, France
    • Related Report
      2013 Annual Research Report
  • [Presentation] Production and purification of Japanese encephalitis virus-like particles from recombinant insect cells2012

    • Author(s)
      Yuri Hotta, Azusa Minamitani, Kazuma Narita, Tomohisa Katsuda, Hideki Yamaji
    • Organizer
      The 25th Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2012)
    • Place of Presentation
      名古屋
    • Year and Date
      2012-11-30
    • Related Report
      2013 Final Research Report
  • [Presentation] Secretory production of virus-like particles by recombinant insect cells2012

    • Author(s)
      Hideki Yamaji, Miwa Kuwahara, Eiji Konishi
    • Organizer
      The 15th European Congress on Biotechnology of the European Federation of Biotechnology (ECB15)
    • Place of Presentation
      Istanbul, Turkey
    • Year and Date
      2012-09-26
    • Related Report
      2013 Final Research Report

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Published: 2013-05-31   Modified: 2019-07-29  

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