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2002 Fiscal Year Final Research Report Summary

Production of Transgenic Silkworms Resistant to BmNPV

Research Project

Project/Area Number 12306002
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field 蚕糸・昆虫利用学
Research InstitutionHokkaido University

Principal Investigator

BANDO Hisanori  Hokkaido Univ., Grad.School of Agr., Prof., 大学院・農学研究科, 教授 (20189731)

Co-Investigator(Kenkyū-buntansha) OKANO Kazuhiro  Riken, Special Postdoctoral Researcher, 微生物制御研究室, 基礎科学特別研究員 (70322691)
MATSUMOTO Tsuguo  Kyoto Institute of Technology, Faculty of Textile Science, Prof., 繊維学部, 教授 (40107355)
TAMURA Toshiki  National Institute of Agrobiological Sciences, Insect Gene Engineering Laboratory, Chief, 遺伝子工学研究室, 室長
Project Period (FY) 2000 – 2002
Keywordssilkworm / RNAi / NPV / virus resistance / piggy Bac / DNA Microarray / primase / helicase
Research Abstract

The nuclear polyhedorosis disease caused by BmNPV infection is a major problem in suriculture around the world. The inhibition of the functional expression of these essential genes in cells infected with NPV should be an effective strategy for preventing mortal infections of BmNPV in the silkworm.
An immediate-early gene product of baculovirus, IE1, is essential for viral gene expression and for viral DNA replication. We constructed an antagonistic IE1 (IE1TN) of Bombyx mori nuclear polyhedrosis virus (BmNPV)s and demonstrated markedly reduced virus production in BmN cells expressing IE1TN. On the other hand, recent study demonstrated that dsRNA is a powerful tool for gene-specific silencing in plants and animals. Then, we constructed recombinant plasmids designed for expressing dsRNAs with the sequences of the essential viral gene, such as Ief-1 and dnahel and demonstrated that viral production was effectively inhibited in BmN cells expressing dsRNAs. Finally, using a transposon piggyback system, we confirmed the moderate BmNPV-resistance caused by the transgenesis of the IE1TN-or dsRNA-expressing gene in the transgenic silkworms.

  • Research Products

    (82 results)

All Other

All Publications (82 results)

  • [Publications] K.Sahara: "W-derived BAC probes as a new tool for identification of the W chromosome and its aberrations in Bombyx mori"Chromosoma. 112. 48-55 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Tomita: "Transgenic silkworms produce recombinant human type III procollagen in cocoons"Nature Biotechnology. 21. 52-56 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Matsuyama: "Functional Analysis of an Immediate Early Gene, ie1, of Bombyx mori Nucleopolyhedrovirus in Mammalian Cells"Journal of Insect Biotechnology and Sericology. 72. 87-94 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] V.S.Mikhailov: "Baculovirus Alkaline Nuclease Possesses a 5'→3'Exonuclease Activity and Associates with the DNA-Binding Protein LEF-3"Journal of Virology. Feb,2003. 2436-2444 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Yamagishi: "Differential Expression of Viral Genes in Two Susceptible Insect Cell Lines"Archives of Virology. 148-3. 587-597 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Luque: "Characterization of a novel silkworm (Bombyx mori) phenol UDP-glucosyltransferase"European Journal of Biochemistry. 269. 819-825 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] GX.Quan: "Induction of the white egg 3 mutant phenotype by injection of the double-stranded RNA of the silkworm white gene"Insect Molecular Biology. 11. 217-222 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Li: "Genome organization of the densovirus from Bombyx mori (BmDNV-1) and enzyme activity of its capsid"Journal of General Virology. 82. 2821-2825 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] R.Isobe: "Antisense and Double-Strand RNA Interference in a Silkworm Ovarian Cell Line"Journal of Insect Biotechnology and Sericology. 71. 43-49 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ito: "Cloning and Expression of Novel Crystal Protein Genes cry39A and 39orf2 from Bacillus thuringiensis subsp. aizawai Bun1-14 Encoding Mosquitocidal Proteins"Journal of Insect Biotechnology and Sericology. 71. 123-128 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] GX.Quan: "Characterization of the kynurenine 3-monooxygenase gene corresponding to the white egg 1 mutant in the silkworm Bombyx mori"Molecular Genetics and Genomics. 267. 1-9 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Yamada: "Use of an N-terminal half truncated IE1 as an antagonist of IE1, an essential regulatory protein in baculovirus"Virus Research. 90. 253-261 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Kojima: "Tandem repetition of baculovirus ie1 promoter results in upregulation of transcription"Archives of Virology. 146. 1-8 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] F.Okabayashi: "A homologue of the Drosophila doublesex gene is transcribed into sex-specific mRNA isoforms in the silkworm, Bombyx mori"Comparative Biochemistry and Physiology Part B. 128. 145-158 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Matsumoto: "Characterization of acyl-CoA-Binding protein (ACBP) in the pheromone gland of the silkworm, Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 603-609 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] GX.Quan: "Isolation and expression of the ecdysteroid-inducible angiotensin-converting enzyme-related gene in wing discs of Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 97-103 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Takeda: "Mass isolation of cuticle protein cDNAs from wing discs of Bombyx mori and their characterizations"Insect Biochemistry and Molecular Biology. 31. 1019-1028 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] X.Zhao: "Isolation and expression of an ecdysteroid-inducible neutral endopeptidase 24.11-like gene in wing discs of Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 1213-1219 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Masaki: "Terminal stability and enzymatic activity of a smaller lysozyme originated from silkworm moth (Bombyx mori)"Journal of Protein Chemistry. 20. 107-113 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] N.Koganesawa: "Construction of an expression system of insect lysozyme lacking thermal stability : the effect of selection of signal sequence on level of expression in the Pichia pastoris expression system"Protein engineering. 14. 705-710 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 原 和二郎: "カイコにおけるEST化したcDNAクローンのRFLPによる連関検索"The Journal of Sericultural Science of Japan. 70. 135-143 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Okano: "Comparative Expressed-Sequence-Tag Analysis of Differential Gene Expression Profiles in BmNPV-Infected BmN Cells"Virology. 282. 348-356 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 礒部良子: "カイコ培養細胞におけるアンチセンスRNA及び日本鎖RNA遺伝子発現抑制効果"東北蚕糸・昆虫利用研究報告. 26. 11 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 伊藤 岳: "蚊に殺虫活性を持つB. thuringiensisのcry遺伝子の構造解析"東北蚕糸・昆虫利用研究報告. 26. 27 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 松山高央: "昆虫細胞と哺乳動物細胞における転写機構の比較解析"東北蚕糸・昆虫利用研究報告. 26. 7 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 吉戸敦生: "カイコ染色体の分裂様式に関する研究"東北蚕糸・昆虫利用研究報告. 26. 26 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐原 健: "天蚕人工飼料育に用いる試料樹葉特性の検討"日本応用動物昆虫学会誌. 45. 94-98 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 小島 桂: "家蚕濃核病ウイルス(BmDNV-2)特異的DNAポリメラーゼ活性と末端配列認識タンパク質の解析"日本蚕糸学雑誌. 70. 103-108 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Hayakawa: "Enhancement of baculovirus infection in Spodoptera exigua (Lepidoptera : Noctuidae) larvae with Autographa calofornica nucleopolyhedrovirus of Nicotiana tabacum engineered with a granulovirus enhancin gene"Applied Entomology and Zoology. 35(1). 163-170 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Hashimoto: "Physical map of Plutella xylostella granulovirus genome"Applied Entomology and Zoology. 35. 45-51 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Shimizu: "Extrachromosomal transposition of the transposable element Minos occur embryos of the silkworm Bombyx mori"Insect Molecular Biology. 9. 277-281 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Hagiwara: "Nucleotide sequences of genome segments 6 and 7 of Bombix mori cypovirus 1, encoding the viral structural proteins V4 and V5, respectively"Journal of General Virology. 81. 1143-1147 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] R.Ko: "Structural and Functional Analysis of the Xestia c-nigrum Granulovirus Matrix Metalloproteinase"Journal of Virology. Dec.2000. 11240-11246 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Komada: "N-Glycosylation contributes to the limited cross-reactivity between hemagglutinin neuraminidase proteins of human parainfluenza virus type 4A and 4B"Medical Microbiology and Immunology. 189. 1-6 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E.Q.Abraham: "Identification and characterization of a silkworm ABC transporter gene homologous to Drosophila white"Molecular and General Genetics. 264. 11-19 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Hashimoto: "Sequence Analysis of the Plutella xylostella Granulovirus Genome"Virology. 275. 358-372 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Hayakawa: "Analysis of proteins encoded in the bipartite genome of an new type of parvo-like virus isolated from silkworm -structural protein with DNA polymerase motifs-"Virus Research. 66. 101-108 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山多洋司: "NPV必須制御タンパク質(IE1)の機能阻害によるNPV増殖抑制"東北蚕糸研究報告. 25. (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 阿部 巧: "NPVによるCaMV35Sプロモーターの転写活性化"東北蚕糸研究報告. 25. (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山岸潤也: "PfDNV遺伝子産物によるウイルスRNAスプライシングの抑制"日本蚕糸学雑誌. 69. 271-276 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Tamura: "Transgenic silkworm research in Japan : Recent progress and future"Proceeding of Joint International Symposium of Insect COE Research Program and Insect Factory Research Project. 77-82

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Sahara: "W-derived BAC probes as a new tool for identification of the W chromosome and its aberrations in Bombyx mori"Chromosoma. 112. 48-55 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Tomita: "Transgenic silkworms produce recombinant human type III procollagen in cocoons"Nature Biotechnology. 21. 52-56 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Matsuyama: "Functional Analysis of an Immediate Early Gene, ie1, of Bombyx mori Nucleopolyhedrovirus in Mammalian Cells"Journal of Insect Biotechnology and Sericology. 72. 87-94 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] V.S.Mikahailov: "Baculovirus Alkaline Nuclease Possesses a 5' → 3' Exonuclease Activity and Associates with DNA-Binding Protein LEF-3"Journal of Virology. Feb, 2003. 2436-2444 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Yamagishi: "Differential Expression of Viral Genes in Two Susceptible Insect Cell Lines"Archives of Virology. 148-3. 587-597 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Luque: "Characterization of a novel silkworm (Bombyx mori) phenol UDP-glucosyltransferase"European Journal of Biochemistry. 269. 819-825 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] GX.Quan: "Induction of the white egg 3 mutant phenotype by injection of the double-stranded RNA of the silkworm white gene"Insect Molecular Biologyk. 11. 217-222 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Li: "Genome organization of the densovirus from Bombyx mori (BmDNV-1) and enzyme activity of its capsid"Journal of General Virology. 82. 2821-2825 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] R.Isobe: "Antisense and Double-Strand RNA Interference in a Silkworm Ovarian Cell Line"Journal of Insect Biotechnology and Sericology. 71. 473-479 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ito: "Cloning and Expression of Novel Crystal Protein Genes cry39A and 39orf2 from Bacillus thuringiensis subsp aizawai Bun1-14 Encoding Mosquitocidal Proteins"Journal of Insect Biotechnology and Sericology. 71. 123-128 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] GX.Quan: "Characterization of the kynurenine 3-monooxygenase gene corresponding to the white egg I mutant in the silkworm Bombyx mori"Molecular Genetics and Genomics. 267. 1-9 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Yamada: "Use of an N-terminal half truncated IE1 as an antagonist of IE1, an essential regulatory protein in baculovirus"Virus Research. 90. 253-261 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Kojima: "Tandem repetition of baculovirus iel promoter results in upregulation of transcription"Archives of Virology. 146. 1-8 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] F.Okabayashi: "A homologue of the Drosophila doublesex gene is transcribed into sex-specific mRNA isoforms in the silkworm, Bombyx mori"Comparative Biochamistry and Physiology Part B. 128. 145-158 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Matsumoto: "Characterization of acyl-CoA-Binding protein (ACBP) in the pheromone gland of the silkworm, Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 603-609 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] GX.Quan: "Isolation and expression of the ecdysteriod-inducible angiotensin-converting enzyme-related gene in wing discs of Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 97-103 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Takeda: "Mass isolation of cuticle protein cDNAs from wing discs of Bombyx mori and their characterization"Insect Biochemistry and Molecular Biology. 31. 1019-1028 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] X.Zhao: "Isolation and expression of an ecdysteroid-inducible neutral endopeptidase 24 11-like gene in wing discs of Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 1213-1219 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Masaki: "Terminal stability and enzymatic activity of a smaller lysozyme originated from silkworm moth (Bombyx mori)"Journal of Protein Chemistry. 20. 107-113 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N.Koganesawa: "Construction of an expression system of insect lysozyme lacking thermal stability the effect of selection of signal sequence on level of expression in the Pichia pastoris expression system"Protein engineering. 14. 705-710 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] W.Hara: "Linkage analysis of EST cDNA clones by using RFLP in the silkworm"The Journal of Sericultural Science of Japan. 70. 135-143 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Okano: "Comparative Expressed-Sequence-Tag Analysis of Differential Gene Expression Profiles in BmNPV-Infected BmN Cells"Virology. 282. 348-356 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] R.Isobe: "Inihibition of specific gene expression by antisense-or ds-RNAs in BmN Cells"Touhoku Sanshi Kenkyu Houkoku. 26. 11 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ito: "Structure of a cry gene coding for a mosquitocidal protein"Touhoku Sanshi Kenkyu Houkoku. 26. 27 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Matsuyama: "Analysis of transcription using insect and mammalian cell lines"Touhoku Sanshi Kenkyu Houkoku. 26. 7 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Yoshido: "Chromosomal division in the silkworm, Bombyx mori"Touhoku Sanshi Kenkyu Houkoku. 26. 26 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Sahara: "Study on the novel forage tree for Antheraea yamamai Guerin"Applied Entomology and Zoology. 45. 94-98 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Kojima: "Analysis of the BmDNV-2 specific DNA polymerase and the common terminal sequence binding proteins"The Journal of Sericultural Science of Japan. 70. 103-108 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Hayakawa: "Enchancement of baculovirus infection in Spodoptera exigua (Lepidoptera-Noctuidae) larvae with Autographa calofornica nucleopolyhedrovirus of Nicotiana tabacum engineered with a granulovirus enhancin gene"Applied Entomology and Zoology. 35(1). 163-170 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Hashimoto: "Physical map of Plutella xylostella granulovirus genome"Applied Entomology and Zoology. 35. 45-51 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Shimizu: "Extrachromosomal transposition of the transposable element Minos occur embryos of the silkworm Bombyx mori"Insect Molecular Biology. 9. 277-281 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Hagiwara: "Nucleotide sequences of genome segments 6 and 7 of Bombix mori cypovirus 1, encoding the viral structural proteins V4 and V5, respectively"Journal of General Virology. 81. 1143-1147 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] R.Ko: "Structural and Functional Analysis of the Xestia c-nigrum Granulovirus Matrix Metalloproteninase"Journal of Virology. Dec.2000. 11240-11246 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Komada: "N-Glycosylation contributes to the limited cross-reactivity between hemagglutinin neuraminidase proteins of human parainfluenza virus type 4A and 4B"Medical Microbiology and Immunology. 189. 1-6 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] E.G.Abraham: "Identification and characterization of a silkworm ABC transporter gene homologous to Drosophila white"Molecular and General Genetics. 264. 11-19 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Hashimoto: "Sequence Analysis of the Plutella xylostella Granulovirus Genome"Virology. 275. 358-372 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Hayakawa: "Analysis of proteins encoded in the bipartite genome of an new type of parvo-like virus isolated from silkworm -structural protein with DNA polymerase motifs-"Virus Research. 66. 101-108 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Yamada: "Effect of an IE1-antagoniat on NPV-infection"Touhoku Sanshi Kenkyu Houkoku. 25. (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Abe: "Activation of 35S promoter in the insect cells infected with NPV"Touhoku Sanshi Kenkyu Houkoku. 25. (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Yamagishi: "PfDNV nonstructural proteins suppress viral RNA splicing"The Journal of Sericultural Science of Japan. 69. 271-276 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Tamura: "Transgenic silkworm research in Japan Recent progress and future"Proceeding of Joint International Symposium of Insect COE Research Program and Insect Factory Research Project. 77-82

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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