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

2010 Fiscal Year Final Research Report

• 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(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
• 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

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
  • 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
  • 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] 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] 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
  • 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
  • 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
  • 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
  • Year and Date: 20100921-20100922
• [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
  • Year and Date: 20100921-20100922
• [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
  • Year and Date: 20100918-20100919
• [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
  • Year and Date: 20091025-20091027
• [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
  • Year and Date: 20090912-20090913
• [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
  • Year and Date: 20090327-20090331
• [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
  • Year and Date: 20090327-20090331
• [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
  • Year and Date: 20080622-20080626
• [Remarks] ホームページ等