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Development of molecular methods for epidemiology of insect viruses

Research Project

Project/Area Number 07556136
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 蚕糸・昆虫利用学
Research InstitutionThe University of Tokyo

Principal Investigator

KOBAYASHI Masahiko  The University of Tokyo, Graduate school of Agricultural and life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (60162020)

Co-Investigator(Kenkyū-buntansha) NOGUCHI Yoko  Saitama Sericultural Experiment Station, 主任研究員
KOBAYASHI Jun  Mie University, Faculty of Engineering, Assistant Professor, 工学部, 助手 (70242930)
SHIMADA Toru  The University of Tokyo, Graduate school of Agricultural and life Sciences, Asso, 大学院・農学生命科学研究科, 助教授 (20202111)
BANDO Hisanori  Hokkaido University, Faculty of Agriculture, Associated Professor, 農学部, 助教授 (20189731)
NAKAGAKI Masao  Shinshu University, Faculty of Textile Science and Technology, Associated Profes, 繊維学部, 助教授 (70135169)
阿部 広明  東京農工大学, 農学部, 助手 (80222660)
宮嶌 成壽  三重大学, 工学部, 教授 (80239409)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1997: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1996: ¥3,200,000 (Direct Cost: ¥3,200,000)
Keywordsinsect viruses / molecular epidemiology / Bombyx mori / PCR / NPV / CPV / DNV / retrotransposon / 養蚕 / レトロポゾン
Research Abstract

We developed a diagnosis system for virus diseases in silkworms and wild insects based on comparative studies on DNA sequences in the virus and host insects. A forecast method of virus disease expansion in insect populations was established by using molecular epidemiological techniques.
1.The polyhedrin genes and DNA helicase genes were cloned and sequences from Samia cynthia ricini nuclear polyhedrosis virus (SrNPV) and Antheraea yamamainuclear polyhedrosis virus (AyNPV).
2.An immunological sensor was developed using an monoclonal antibody against Bombyx moricytoplasmic polyhedrosis virus (BmCPV) and a quartz oscilator to detect BmCPV rapidly and easily.
3.Quantitative changes of two genomic DNAs in the Bombyx moridensonuclear virus type II (DNV2) were investigated in the midgut cells infected with DNV2, using the competitive PCR method.
4.We developed a very simple method to detect all of known viruses in Bombyx morisimultaneously in one reaction. The condition of PCR and primer designs were optimized.
5.We set a model rearing shelf containing healthy silkworm larvae and a few nuclear polyhedrosis ones. Expansion of the virus was monitored in the larvae, feces, diet, etc., on the shelf, using the PCR diagnosis system.
6.Random amplified polymorphic DNAs (RAPDs) were found to be closely linked to the nonsusceptibility gene to DNV2 (nsd-2). A linkage map was constructed based on recombination values among the RAPDs and nsd-2.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] 野口 洋子・小林 正彦・嶋田 透: "ポリメラ-セ連鎖反応によるカイコ核多角体病ウイルス検出技術の簡易化" 日本蚕糸学雑誌. 64,4. 352-359 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 安部 広明・嶋田 透・小林正 彦: "カイコ濃核病ウイルスI型感受性遺伝子に連関するランダム増幅多型DNA(RAPD)" 日本蚕糸学雑誌. 64,3. 262-264 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 安部 広明・嶋田 透・小林 正彦: "カイコ濃核病ウイルスI型感受性系統保存へのDNA診断法の応用" 日本蚕糸学雑誌. 65,3. 196-200 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 景安 聖士・伴戸 久徳 ら: "RPCを用いたカイコ疫病ウイルス遺伝子の検出" 日本蚕糸学雑誌. 66,6. 477-483 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Abe H., Shimada T., etal: "Nucleotide sequence of the random amplified poly-morphic DNA(RAPD) on the chromosame of the silkworm" Applied Ent. And Zool. 31. 633-637 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Isawa H., Bando H, et al: "Analysis of genetic, m formation of an insect picarna-like vrisus in fectilous feacherie uirees of silkworm." Archies of Virology. 143. 127-143 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Noguchi Y., Kobayashi M.and Shimada T.: "An improved method for PCR-based detection of nuclear polyhedrosis virus in Bombyx mori." Journal of Sericultural and Science of Japan. 64. 352-359 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Abe H., Shimada T., Kobayashi M,et al.: "Identification of random amplified polymorphic DNA linked to the densonucleosis virus type-1 susceptibility gene of the silkworm, Bombyx mori." Journal of Sericultural Science of Japan. 64. 262-264 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Abe H., Shimada T., Kobayashi M,et al.: "An application of DNA diagnosis method for preservation of the susceptible strain to densonucleosis virus type-1 of the silkworm, Bombyx mori." Journal of Sericultural Science of Japan. 65. 196-200 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kageyasu S., Hayakawa T., Isawa H., Asano K., Sahara K., Iizuka T.and Bndo H.: "Detection of the silkworm-pathogenic virus genomes by PCR." Journal of Sericultural Science of Japan. 66. 477-483 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Abe H,Shimada T et al.: "Nucleotide sequence of the random amplified polymorphic DNA (RAPD) on the Wchromosome of the silkworm, Bombyx mori (Lepidoptera : Bombycidae)." Applied Entomology and Zoology. 31. 633-637 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Isawa H., Asano S., Sahara K., Iizuka T.and Bando H.: "Analysis of genetic information of an insect picorna-like virus, infectious flacherie virus of silkworm : evidence for evolutionary relationships among insect, mammalian and plant picorna (-like) viruses" Archives of Virology. I 143. 127-143 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 後藤孝一郎・小林正彦: "家蚕のモザイク致死に関する遺伝学的解析" 日本蚕糸学雑誌. 66(2). 123-127 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 浜幸夫・小林正彦ら: "休眠性蚕卵の非休眠化に伴う卵内カルシウム量および存在状態の変化" 日本蚕糸学雑誌. 66(3). 219-221 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 洗幸夫・小林正彦ら: "家蚕雌蛹へのEGTA投与が産下卵の休眠性に及ぼす影響" 日本蚕糸学雑誌. 66(6). 403-408 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 板戸久徳ら: "PCRを用いたカイコ病原ウイルス遺伝子の検出" 日本蚕糸学雑誌. 66(6). 477-483 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Shimada, M.Kobayashi: "Absennce of dosage compen sation at the transcription level of a sex-linked gene in a female heterogametic insect, Bombix mori" Heredity. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] J.Kobayashi et al.: "Determination of the nuckeotide sequence of Bm CPV segment 9 and its wxpression in BmN4 cells" Journal of Virology. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 阿部広明: "カイコ濃核病ウイルス1型感受性系統保存へのDNA診断法の応用" 日本蚕糸学雑誌. 65・3. 196-200 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 大林富美: "限性形蚕,限性セ-ブルならびに限性黒色蚕のW染色体に存在する同一のRAPD" 日本蚕糸学雑誌. 65・5. 395-398 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hiroaki ABE: "Nucleotide Sequence of the Raindom Amplified Polymorphic DNA(RAPD)on the W Chromosome of the Silkworm,Bombyx mori(Lepidoptera:Bombycidae)" Applied Entomology and Zoology. 31・4. 633-637 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hajime TAKEDA: "Impaired yolk protein uptake by oocytes of a Bombyx mori mutant." Insect Biochemistry and Molecular Biology. 26. 607-616 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kozo TSUCHIDA: "Purification and properties of a lipid transfer particle from Bombyx mori." Biochem.Biophys.Acta. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takashi TOMITA: "Chromosomal localization of amplified esterase genes in insecticide resistant culex mosquitoes." Insect Biochemistry and Molecular Biology. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 前川 秀彰: "昆虫ゲノム解析の研究の現状(蚕糸・昆虫研資料)" 農林水産省蚕糸・昆虫農業技術研究所, 153 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 宮嶌 成壽: "ウイルス学" (株)朝倉書店(出版予定), (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 野口洋子・小林正彦・嶋田透: "ポリメラーゼ連鎖反応によるカイコ核多角体病イウルスの検出技術の簡易化" 日本蚕糸学雑誌. 64. 352-359 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] S. Hoshizaki and T. Shimada: "PCR-based detection of Wolbachia, cytoplasmic incompatibility microorganisms, infected in natural papulations of Laodelphax striatellus(Homoptera: Delphacidae)" Insect Molecular Biology. vol. 4.237-243 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K. Hashido, et al.: "Favorite site for recombination of a defective non LTR retrotransposon, BMC1." J. Cell. Biochem.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] H. Maekawa, et al.: "The Chromatin condensation could prevent expression of non-LTR retrotransposon,R1Bm. inserted into 28S rRNA region of rDNA." J. Cell. Biochem.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 阿部広明 その他: "カイコの系統保存へのDNA診断法の応用.-濃核病ウイルス1型感受性系統の保存-." 日本蚕糸学雑誌. 65巻(印刷中). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Jun Sasaki, et al.: "Insecticidal activity of the protein encoded by the cryV gene of Bacillus thuringiensis kurstaki INA-02" Current Microbiology. (in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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