Systematic screening of viral host factors using a transposon-tagging method.

• Project/Area Number: 20380111
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General fisheries
• Research Institution: Hiroshima University
• Principal Investigator: YASUSHI Okinaka Hiroshima University, 大学院・生物圏科学研究科, 准教授 (80363034)
• Co-Investigator(Kenkyū-buntansha): 中井 敏博 広島大学, 大学院・生物圏科学研究科, 教授 (60164117)
• Co-Investigator(Renkei-kenkyūsha): TOSHIHIRO Nakai 広島大学, 大学院・生物圏科学研究科, 教授 (60164117)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000); Fiscal Year 2010: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2009: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2008: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
• Keywords: ベータノダウイルス / 宿主因子 / トランスポゾンタギング / 遺伝子破壊系 / 魚類培養細胞 / メダカ培養細胞 / 遺伝子破壤系

Research Abstract

We cloned a medaka (Oryzias latipes) HNI-2 cell line, the chromosome number of which was 25. We then introduced a transposon into the cells for gene disruption and obtained 17 betanodavirus-resistant and GFP-expressed cell lines (GFP expression indicates transposon insertion). DNA fragments adjacent to the inserted transposon were isolated from the genomic DNA prepared from each of the virus-resistant cells and used for blastn searches against the Ensembl medaka genome database. Nine sequences identified from the 17 resistant cell lines hit database sequences and five of which were revealed to encode proteins.

Research Products

• [Journal Article] Susceptibilities of medaka (Oryzias latipes) cell lines to a betanodavirus. (2010)
  • Author(s): Adachi, K., Sumiyoshi, K., Ariyasu, R., Yamashita, K., Zenke, K., Okinaka, Y.
  • Journal Title: Virol.J. 7 Pages: 150-150
  • NAID: 120002577222
  • Peer Reviewed
• [Journal Article] Identification of RNA regions that determine temperature sensitivity in betanodaviruses. (2010)
  • Author(s): Hata N., Okinaka, Y., Iwamoto, T., Kawato, Y., Mori, K., Nakai, T.
  • Journal Title: Arch.Virol. 155 Pages: 1597-1606
  • NAID: 120002753670
  • Peer Reviewed
• [Journal Article] Trash fish can be a source of betanodaviruses for cultured marine fish. (2010)
  • Author(s): Gomez, D.K., Mori, K., Okinaka, Y., Nakai, T., Park, S.C.
  • Journal Title: Aquaculture. 302 Pages: 158-163
  • Peer Reviewed
• [Journal Article] Susceptibilities of medaka (Oryzias latipes) cell lines to a betanodavirus. (2010)
  • Author(s): Kei Adachi
  • Journal Title: Virol.J. Volume: 7 Pages: 150-150
  • Peer Reviewed
• [Journal Article] Identification of RNA regions that determine temperature sensitivity in betanodaviruses. (2010)
  • Author(s): Naomi Hata
  • Journal Title: Arch.Virol. Volume: 155 Pages: 1597-1606
  • Peer Reviewed
• [Journal Article] Production of biologically active Atlantic salmon interferon in transgenic potato and rice plants (2010)
  • Author(s): Noriho Fukuzawa
  • Journal Title: J.Biosci.Bioeng. (印刷中)
  • Peer Reviewed
• [Journal Article] Genetic heterogeneity of betanodaviruses in juvenile production trials of Pacific bluefin tuna, Thunnus orientalis (Temminck & Schlegel). (2009)
  • Author(s): Sugaya, T., Mori, K., Nishioka, T., Masuma, S., Oka, M., Mushiake, K., Okinaka Y., Nakai T.
  • Journal Title: J.Fish Dis. 32 Pages: 815-823
  • Peer Reviewed
• [Journal Article] Genetic analysis and pathogenicity of betanodavirus isolated from wild redspotted grouper Epinephelus akaara with clinical signs. (2009)
  • Author(s): Gomez, D.K., Matsuoka, S., Mori, K., Okinaka, Y., Park, S.C., Nakai, T.
  • Journal Title: Arch.Virol. 154 Pages: 343-346
  • Peer Reviewed
• [Journal Article] Genetic heterogeneity of betanodaviruses in juvenile production trials of Pacific bluefin tuna, Thunnus orientalis (Temminck & Schlegel) (2009)
  • Author(s): Takuma Sugaya
  • Journal Title: J.Fish Dis. 32 Pages: 815-823
  • Peer Reviewed
• [Journal Article] Genetic analysis and pathogenicity of betanodavirus isolated from wild redspotted grouper Epinephelus akaara with clinical signs (2009)
  • Author(s): Dennis K.Gomez
  • Journal Title: Arch.Virol. 154 Pages: 343-346
  • Peer Reviewed
• [Journal Article] Comparisons among the complete genomes of four betanodavirus genotypes. (2008)
  • Author(s): Okinaka, Y., Nakai, T.
  • Journal Title: Dis.Aquat.Org. 80 Pages: 113-121
  • NAID: 120001049000
  • Peer Reviewed
• [Journal Article] The variable region of RNA2 is sufficient to determine host specificity in betanodaviruses. (2008)
  • Author(s): Ito, Y., Okinaka, Y., Mori, K., Sugaya, T., Nishioka, T., Oka, M., Nakai, T.
  • Journal Title: Dis.Aquat.Org. 79 Pages: 199-205
  • NAID: 120006323780
  • Peer Reviewed
• [Journal Article] Phylogenetic analysis of betanodavirus RNA2 identified from wild marine fish in oceanic regions. (2008)
  • Author(s): Sakamoto, T., Okinaka, Y., Mori, K., Sugaya, T., Nishioka, T., Yamashita, H., Oka, M., Nakai, T.
  • Journal Title: Fish Pathol. 43 Pages: 19-27
  • NAID: 10021214757
  • Peer Reviewed
• [Journal Article] Comparisons among the complete genomes of four betanodavirus genotypes. (2008)
  • Author(s): Yasushi Okinaka
  • Journal Title: Diseases of Aquatic Organisms 80 Pages: 113-121
  • NAID: 120001049000
  • Peer Reviewed
• [Journal Article] The variable region of RNA2 is sufficient to determine host specificity in betanodaviruses. (2008)
  • Author(s): Yu Ito
  • Journal Title: Diseases of Aquatic Organisms 79 Pages: 199-205
  • Peer Reviewed
• [Journal Article] Phylogenetic analysis of betanodavirus RNA2 identified from wild marine fish in oceanic regions. (2008)
  • Author(s): Takahiro Sakamoto
  • Journal Title: Fish Pathology 43 Pages: 19-27
  • Peer Reviewed
• [Presentation] メダカ培養細胞におけるRNA干渉を利用した遺伝子ノックダウン法の最適化 (2010)
  • Author(s): 善家孝介、冲中泰
  • Organizer: 日本魚病学会、三重大学
  • Place of Presentation: 津
• [Presentation] ベータノダウイルスの温度感受性機構の解明?IIIRGNNV型近縁株を用いた研究の効率化 (2010)
  • Author(s): 有安亮、冲中泰
  • Organizer: 日本魚病学会
  • Place of Presentation: 三重大学、津
• [Presentation] メダカ培養細胞におけるRNA干渉誘導法の最適化 (2010)
  • Author(s): 善家孝介、冲中泰
  • Organizer: 小型魚類研究会、プラザイースト
  • Place of Presentation: さいたま
• [Presentation] メダカ培養細胞におけるRNA干渉を利用した遺伝子ノックダウン法の最適化 (2010)
  • Author(s): 善家孝介
  • Organizer: 日本魚病学会
  • Place of Presentation: 三重大学、津
• [Presentation] ベータノダウイルスの温度感受性機構の解明-III RGNNV型近縁株を用いた研究の効率化 (2010)
  • Author(s): 有安亮
  • Organizer: 日本魚病学会
  • Place of Presentation: 三重大学、津
• [Presentation] メダカ培養細胞におけるRNA干渉誘導法の最適化 (2010)
  • Author(s): 善家孝介
  • Organizer: 小型魚類研究会
  • Place of Presentation: プラザイースト、さいたま
• [Presentation] メダカ由来細胞株におけるベータノダウイルス増殖性の検討 (2009)
  • Author(s): 足立圭、有安亮、住吉洸介、冲中泰
  • Organizer: 日本ウイルス学会、都市センターホテル
  • Place of Presentation: 東京
• [Presentation] ベータノダウイルスの感染・増殖に適したメダカ培養細胞の同定 (2009)
  • Author(s): 冲中泰、足立圭、有安亮、住吉洸介
  • Organizer: 小型魚類研究会
  • Place of Presentation: 名古屋大学、名古屋
• [Presentation] ベータノダウイルスの温度感受性機構の解明-II 温度感受性に関わるゲノム領域の特定 (2009)
  • Author(s): 冲中泰・端直美・河東康彦・中井敏博
  • Organizer: 日本水産学会
  • Place of Presentation: 東京海洋大学、東京都
• [Presentation] ベータノダウイルスの宿主特異性決定機構の解明III 細胞膜上レセプター以外の決定機構の有無 (2009)
  • Author(s): 河東康彦・冲中泰・中井敏博・山下浩史
  • Organizer: 日本水産学会
  • Place of Presentation: 東京海洋大学、東京都
• [Presentation] Acute toxicity of polyphenols to young sea bass (Lates calcarifer) (2009)
  • Author(s): Tanasomwang Varin
  • Organizer: JSPS-NRCT Joint Seminar
  • Place of Presentation: Rayong, Thailand
• [Presentation] メダカ由来細胞株におけるベータノダウイルス増殖性の検討 (2009)
  • Author(s): 足立圭
  • Organizer: 日本ウイルス学会
  • Place of Presentation: 都市センターホテル、東京
• [Presentation] 各種ポリフェノールの抗ウイルス効果の検討 (2009)
  • Author(s): 山下佳澄
  • Organizer: 日本魚病学会
  • Place of Presentation: 東北大学、仙台
• [Presentation] 種々のメダカ由来細胞株におけるベータノダウイルス増殖性 (2009)
  • Author(s): 足立圭
  • Organizer: 日本魚病学会
  • Place of Presentation: 東北大学、仙台
• [Presentation] 柿渋を利用した抗ノロウイルス消毒剤の開発とオールマイティーな抗ウイルス薬への応用 (2009)
  • Author(s): 島本整
  • Organizer: 第21回微生物シンポジウム
  • Place of Presentation: 福山大学、福山
• [Presentation] ベータノダウイルスの宿主特異性決定機構の解明-III 細胞膜上レセー以外の決定機構の有無 (2009)
  • Author(s): 河東康彦
  • Organizer: 日本水産学会
  • Place of Presentation: 東京海洋大学、東京都
• [Presentation] Genotype-specific binding of betanodaviruses to sevenband grouper cultured cells, SGF. (2008)
  • Author(s): Kawato, Y., Okinaka Y., Yamashita H., Nakai T.
  • Organizer: In 7th Symposium on Diseases in Asian Aquaculture
  • Place of Presentation: Taipei, Taiwan
• [Presentation] 柿渋の抗ノロウイルス効果と他のウイルスに対する効果 (2008)
  • Author(s): 島本 整
  • Organizer: 日本食品微生物学会学術総会
  • Place of Presentation: 広島国際会議場、広島市
• [Presentation] Expression of Atlantic salmon interferon (IFN) in transgenic potato and rice plants. (2008)
  • Author(s): Noriho Fukuzawa
  • Organizer: Fifth International Symposium of The Japanese Society for Fish Pathology
  • Place of Presentation: Tokyo University, Japan
• [Presentation] E Genotype-specific binding of betanodaviruses to seve nband grouper cultured cells, SGF (2008)
  • Author(s): Yasuhiko Kawato
  • Organizer: 7th Symposium on Diseases in Asian Aquaculture
  • Place of Presentation: Taipei, Taiwan
• [Remarks] ホームページ等