Super-high Efficient Foreign Protein Expression System in Plants

• Project/Area Number: 16580282
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied molecular and cellular biology
• Research Institution: ISHIKAWA PREFECTURAL UNIVERSITY
• Principal Investigator: MORI Masashi Ishikawa Prefectural University, Faculty of Bioresources and Environmental Sciences, Associate Professor, 生物資源環境学部, 助教授 (00320911)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: Plant Virus Vector / Silencing Suppressor / Transgenic Plants / Foreign Gene Expression / ウイルスベクター

Research Abstract

The aim of this study was to establish super-high efficient foreign protein expression system in plants by controlled expression of a plant virus based vector and silencing suppressors, which prevents specific degradation of viral vector RNA in a later stage of the infection. Genes of three silencing suppressors, cucumber mosaic virus 2b, tomato bushy stunt virus p19 and tomato spotted wilt virus. NSs, which suppressed gene silencing by possibly different mechanisms, were introduced to estradiol-and dexamethasone-inducible expression vectors, and transformed into Nicotiana benthamiana plants. Transgenic plants that inducibly expressed one of these suppressors were successfully produced. However, the expression of the suppressors in the cross-progenies that had two or three different suppressors were often significantly suppressed, suggesting a synergistic adverse effect of these suppressors. Next, transgenic plants that inducibly expressed one of the silencing suppressors and a replicable brome mosaic virus (BMV)-based vector by treatment of dexamethasone were successfully produced by crossing of the transgenic plants. Significantly, in the transgenic plants that inducibly expressed NSs and the BMV vector, the decreasing of the viral RNA and the foreign protein encoded in the vector, which was usually observed in a later stage of the infection, was suppressed. Various useful proteins may be produced in transgenic plants that inducibly express NSs and a BMV vector.

Research Products

• [Journal Article] Membrane-bound tomato mosaic virus replication proteins participate in RNA synthesis and are associated with host proteins in a pattern distinct from that observed for non-membrane-bound ones (2006)
  • Author(s): Nishikiori, M., Dohi, K., Mori, M., Meshi, T., Naito S., Ishikawa,M.
  • Journal Title: Journal of Virology 80 Pages: 8459-8468
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of an enhancin gene from Trichoplusia ni granulosis virus confers the resistance to lepidopterous insect pests to rice (2006)
  • Author(s): Mori, M., Kitamura, H., Kondo, A., Dohi, K., Mori, M., Kaido, M., Mise, K., Shimojyo, E., Hashimoto,Y.
  • Journal Title: Plant Biotechnology 23 Pages: 55-61
  • NAID: 10021907868
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inducible virus-mediated expression of a foreign protein in suspension-cultured plant cells (2006)
  • Author(s): Dohi, K., Nishikiori, M., Tamai, A., Ishikawa, M., Meshi, T., Mori,M
  • Journal Title: Archives of Virology 151 Pages: 1075-1084
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Membrane-bound tomato mosaic virus replication proteins participate in RNA synthesis and are associated with host proteins in a pattern distinct from that observed for non-membrane-bound ones. (2006)
  • Author(s): Nishikiori, M., Dohi, K., Mori, M., Meshi, T., Naito S., Ishikawa, M.
  • Journal Title: Journal of Virology 80 Pages: 8459-8468
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression of an enhancin gene from Trichoplusia ni granulosis virus confers the resistance to lepidopterous insect pests to rice. (2006)
  • Author(s): Mori, M.Kitamura, H., Kondo, A., Dohi, K., Mori, M., Kaido, M., Mise, K., Shimojyo, E., Hashimoto, Y.
  • Journal Title: Plant Biotechnology 23 Pages: 55-61
  • NAID: 10021907868
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inducible virus-mediated expression of a foreign protein in suspension-cultured plant cells. (2006)
  • Author(s): Dohi, K., Nishikiori, M., Tamai, A., Ishikawa, M., Meshi, T., Mori, M.
  • Journal Title: Archives of Virology 151 Pages: 1075-1084
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The "resurrection method" for modification of specific proteins in higher plants (2005)
  • Author(s): Mori, M., Dohi, K.
  • Journal Title: FEBS Letters 579 Pages: 6210-6216
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genetic Differences in Nitrate Uptake in Two Clones of the Common Reed, Phragmites australis (2005)
  • Author(s): Araki, R., Mori, M., Mori, M., Hasegawa,H.
  • Journal Title: Breeding Science 55 Pages: 297-302
  • NAID: 110001828197
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The "resurrection method" for modification of specific proteins in higher plants. (2005)
  • Author(s): Mori, M., Dohi, K.
  • Journal Title: FEBS Letters 579 Pages: 6210-6216
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic Differences in Nitrate Uptake in Two Clones of the Common Reed; Phragmites australis. (2005)
  • Author(s): Araki, R., Mori, M., Mori, M., Hasegawa, H.
  • Journal Title: Breeding Science 55 Pages: 297-302
  • NAID: 110001828197
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Downregulation of the NbNACal gene encoding a movement-protein-interacting protein reduces cell-to-cell movement of Brome mosaic virus in Nicotiana benthamiana.
  • Author(s): Kaido, M., Inoue, Y., Takeda, Y., Sugiyama, K., Takeda, A., Mori, M., Tamai, A., Meshi, T., Okuno, T., Mise,K.
  • Journal Title: Molecular Plant-Microbe Interactions (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Downregulation of the NbNACal gene encoding a movement-protein-interacting protein reduces cell-to-cell movement of Brome mosaic virus in Nicotiana benthamiana.
  • Author(s): Kaido, M., Inoue, Y., Takeda, Y., Sugiyama, K., Takeda, A., Mori, M., Tamai, A., Meshi, T., Okuno, T., Mise, K.
  • Journal Title: Molecular Plant-Microbe Interactions (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Patent(Industrial Property Rights)] 形質転換植物、形質転換細胞、タンパク質生産キットおよびタンパク質の生産方法 (2007)
  • Inventor(s): 森正之, 土肥浩二
  • Industrial Property Rights Holder: (独)科学技術振興機構
  • Industrial Property Number: 2007-082289
  • Filing Date: 2007-03-27
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] 新規遺伝子発現方法およびその利用 (2005)
  • Inventor(s): 森正之, 土肥浩二
  • Industrial Property Rights Holder: (独)科学技術振興機構
  • Industrial Property Number: 2005-020455
  • Filing Date: 2005-01-27
  • Description: 「研究成果報告書概要(和文)」より