Highly efficient production of human interferon-α by transgenic cultured rice cells

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著者

    • KURITA Akihiro
    • Laboratory of Genetic Engineering, Graduate school of Agriculture, Kyoto Prefectural University
    • MORITA Shigeto
    • Laboratory of Genetic Engineering, Graduate school of Agriculture, Kyoto Prefectural University
    • KOGA Junichi
    • Advanced Medical Technology Research Center, Research Division, JCR Pharmaceuticals Co., Ltd.
    • TANAKA Kunisuke
    • Laboratory of Genetic Engineering, Graduate school of Agriculture, Kyoto Prefectural University
    • MASUMURA Takehiro
    • Laboratory of Genetic Engineering, Graduate school of Agriculture, Kyoto Prefectural University

抄録

Interferon-α (IFN-α) is an important antiviral pharmaceutical. A binary vector containing the first intron of the rice cytosolic SOD gene, the signal sequence of the 10 kDa rice prolamin, the amino-terminal region of β-glucuronidase, a thrombin recognition site, and the mature polypeptide region of human IFN-α was constructed, under the regulation of the cauliflower mosaic virus 35S promoter. Here, we report that transgenic rice cells transformed with this fusion protein vector produced a biologically active IFN-α. The vector was introduced into rice calli by <i>Agrobacterium</i>-mediated methods. Five lines of transgenic calli were obtained. IFN assay demonstrated that these calli expressed fusion proteins bearing biologically active IFN-α. Liquid-cultured cells exhibited stable growth and the production of active IFN-α during 10 successive generations, i.e. in 10 weeks. The expressed proteins were purified by immuno affinity chromatography and reverse-phase HPLC. Repeated selections of cultured cells that had been obtained by dividing calli into small cell aggregates considerably increased the production of IFN-α. Thrombin protease treatment of the fusion protein yielded the intact IFN-α polypeptide. Thus, transgenic suspension rice cells are expected to be useful for the production of large amounts of biologically active proteins at a low cost; moreover, such a system would be easier to employ than animal cell culture systems.

収録刊行物

  • Plant biotechnology  

    Plant biotechnology 23(3), 283-289, 2006-06-01 

    Japanese Society for Plant Cell and Molecular Biology

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各種コード

  • NII論文ID(NAID)
    10021908753
  • NII書誌ID(NCID)
    AA11250821
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    13424580
  • NDL 記事登録ID
    7943866
  • NDL 雑誌分類
    ZR3(科学技術--生物学--植物)
  • NDL 請求記号
    Z54-J126
  • データ提供元
    CJP書誌  NDL  J-STAGE 
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