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An establishment of cell line with high sensitivity to rotavirus by transfection of receptor gene and the application for diagnosis in rotavirus infection

Research Project

Project/Area Number 07556121
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Applied veterinary science
Research InstitutionGifu University

Principal Investigator

MINAMOTO Nobuyuki  Gifu University, Faculty of Agriculture, Professor, 農学部, 教授 (10144007)

Co-Investigator(Kenkyū-buntansha) SUGIYAMA Makoto  Gifu University, Faculty of Agriculture, Assisatnt professor, 農学部, 助教授 (80196774)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥2,500,000 (Direct Cost: ¥2,500,000)
KeywordsRotavirus / Sialic acid / Mucin / Molecular cloning / Anti-receptor antibody / Expression of gene / VP4 / Ganglioside / 欠損VP4 / 牛乳ムチン / MA-104細胞 / 発現 / レセプター遺伝子 / クローニング / モノクローナル抗体 / 腸管上皮細胞
Research Abstract

In this study, to establish cell lines with high sensitivity to rotavirus, we determined complete sequence of all genomic segments of avian rotavirus, and characterization of a receptor for avian rotavirus on MA-104 cells. Moreover, we attempted to produce monoclonal antibodies directed to receptor for rotavirus. The ressults run as follows :
(1) The complete nucleotide sequence of 11 genomic segments of avian rotavirus, P0-13 strain, which was isolated from a pigeon in our laboratory, was determined to elicit binding protein of rotavirus to cellular receptor. Then, the cloning genes encoding several proteins of rotavirus were expressed at a high level in simian COS7 cells and the expressed proteins were examined in the function of receptor binding.
(Arch.Virol., 1995. Virology, 1995. Vet.Microbiol., 1996. Arch.Virol., 1996. Virus Genes, 1997. Virus Res., 1997)
(2) By the treatment of MA-104 cell monolayrs with several enzymes, and by binding activity of rotaviruses to gangliosides, we suggested that sialic acid served as the surface molecule used by the virus for binding and entry into host cells.
(Arch.Virol., in preparation)
(3) It was indicated that ovomucin in hen egg white and mucin in cow milk bind to avian rotavirus, and these mucins inhibit infection of the virus to MA-104 cells. Although we produced many of monoclonal antibodies to these mucins, all of the resulting antibodies failed to inhibit infection of rotavirus to MA-104 cells so far.
In the future, we will make an attempt to map receptor binding region in VP8 of rotavirus and to produce monoclonal antibodies against the synthetic peptides corresponding to the amino acid region

Report

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

    (38 results)

All Other

All Publications (38 results)

  • [Publications] Ito, H.,: "Sequence analysis of cDNA for the VP6 protein of group Aavian rotavirus : a comparison with group Amammalian rotaviruses." Archives of Virology. 140. 605-612 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rohwedder, A.,: "Sequnce analysis of pigeon,turkey,and chicken rotavirus VP8 identifies rotavirus 993/83,isolated from calf feces,as a pigeon rotavirus." Virology. 210. 231-235 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.,: "Expression of the majorinner capsid protein,VP6,of avian rotavirus in mammalian cells." Veterinary Microbiology. 49. 257-265 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.,: "Mapping of antigenic sites on the major capsid protein of avian rotavirus using Escherichia coli expression system." Archives of Virology. 141. 2129-2138 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rohwedder, A.,: "Bovine rotavirus 993/83 shows athird subtype of avian VP7 protein." Virus genes. 14(2). 147-151 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.,: "Sequence analysis of the VP6 gene in group Aturkey and chicken rotaviruses." Virus Research. 47. 79-83 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.: "Sequence analysis of cDNA for the VP6 protein of group A avian rotavirus : a comparison with group A mammalian rotaviruses" Archives of Virology. 140. 605-612 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rohwedder, A.: "Sequnce analysis of pigeon, turkey, and chicken rotavirus VP8 identifies rotavirus 993/83, isolated from calf feces, as a pigeon rotavirus" Virology. 210. 231-235 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.: "Expression of the major inner capsid protein, VP6, of avian rotavirus in mammalian cells" Veterinary Microbiology. 49. 257-265 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.: "Mapping of antigenic sites on the major capsid protein of avian rotavirus using Escherichia coli expression system" Archives of Virology. 141. 2129-2138 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Rohwedder, A.: "Bovine rotavirus 993/83 shows a third subtype of avian VP7 protein" Virus genes. 14. 147-151 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito, H.: "Sequence analysis of the VP6 gene in group A turkey and chicken rotaviruses" Virus Research. 47. 79-83 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ito,H.: "Sequence analysis of cDNA for the VP6 protein of group A avian rotavirus:a comparison with group A mammalian rotaviruses." Archives of Virology. 140. 605-612 (1995)

    • Related Report
      1997 Annual Research Report
  • [Publications] Rohwedder,A.: "Sequnce analysis of pigeon,turkey,and chicken rotavirus VP8 identifies rotavirus 993/83,isolated from calf feces,as a pigeon rotavirus." Virology. 210. 231-235 (1995)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ito,H.: "Expression of the major inner capsid protein,VP6,of avian rotavirus in mammalian cells." Veterinary Microbiology. 49. 257-265 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ito,H.: "Mapping of antigenic sites on the major capsid protein of avian rotavirus using Escherichia coli expression system." Archives of Virology. 141. 2129-2138 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Rohwedder,A.: "Bovine rotavirus 993/83 shows a third subtype of avian VP7 protein." Virus genes. 14(2). 147-151 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ito,H.: "Sequence analysis of the VP6 gene in group A turkey and chicken rotaviruses." Virus Research. 47. 79-83 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 伊藤啓史: "トリロタウイルスPO-13株ゲノム遺伝子の全塩基配列の決定" 第45回日本ウイルス学会総会. (1997年9月発表).

    • Related Report
      1997 Annual Research Report
  • [Publications] Sugiyama,M.: "Characterization of the receptor on MA104 cells for avian rotaviruses" Archives of Virology. (発表予定).

    • Related Report
      1997 Annual Research Report
  • [Publications] 川村尚良、: "トリロタウイルスレセプターに対するモノクローナル抗体の作出" 第126回日本獣医学会. (1998年8月発表予定).

    • Related Report
      1997 Annual Research Report
  • [Publications] Song,X,G.: "Antiviral activities in bovine colostrum that prevent human rotavirus infection in vivo." Milchwissenschaft. (発表予定).

    • Related Report
      1997 Annual Research Report
  • [Publications] Kanamaru,Y.: "Potent inhibitory activity against human rotavirus infection of high-Mr glycoproteins in cow milk." Archives of Virology. (発表予定).

    • Related Report
      1997 Annual Research Report
  • [Publications] Kanamaru,Y.: "Inhibition of human rotavirus infection by aglycoprotein in cow milk." Archives of Virology. (発表予定).

    • Related Report
      1997 Annual Research Report
  • [Publications] Ito,H.,: "Sequence analysis of cDNA for the VP6 protein of group A avian rotavirus : a comparison with group A mammalian rotaviruses." Archives of Virology. 140. 605-612 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Rohwedder,A.,: "Sequnce analysis of pigeon,turkey,and chicken rotavirus VP8 identifies rotavirus 993/83,isolated from calf feces,as a pigeon rotavirus." Virology. 210. 231-235 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ito,H.,: "Expression of the major inner capsid protein,VP6,of avian rotavirus in mammalian cells." Veterinary Microbiology. 49. 257-265 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ito,H.,: "Mapping of antigenic sites on the major capsid protein of avian rotavirus using Escherichia coli expression system." Archives of Virology. 141. 2129-2138 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Rohwedder,A.,: "Bovine rotavirus 993/83 shows a third subtype of avian VP7 protein." Virus genes. (in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Ito,H.,: "Sequence analysis of the VP6 gene in group A turkey and chicken rotaviruses." Virus Research. (in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Sugiyama,M.,: "Characterization of the receptor on MA104 cells for avian rotaviruses" Archives of Virology. (発表予定).

    • Related Report
      1996 Annual Research Report
  • [Publications] 川村尚良、: "トリロタウイルスレセプターに対するモノクローナル抗体の作出" 第124回日本獣医学会(1997年10月). (発表予定).

    • Related Report
      1996 Annual Research Report
  • [Publications] Ito H.,: "Sequence analysis of cDNA for the VP6 protein of group A avian rotavirus: a comparison with group A mammalian rotaviruses." Archives of Virology. 140. 605-612 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Rohwedder A.,: "Sequnce analysis of pigeon, turkey, and chicken rotavirus VP8 identifies rotavirus 993/83, isolated from calf feces, as a pigeon rotavirus." Virology. 210. 231-235 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Ito H.,: "Expression of the major inner capsid protein, VP6, of avian rotavirus in mammalian cells." Veterinary Microbiology. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Ito H.,: "Mapping of antigenic sites on the major capsid protein of avian rotavirus using Escherichia coli expression system." Archives of Virology. (発表予定).

    • Related Report
      1995 Annual Research Report
  • [Publications] Rohwedder A.,: "VP7 from rotavirus 993/83 isolated from a calf with diarrhea is closely related to VP7 of pigeon rotavirus PO-13." Virology. (発表予定).

    • Related Report
      1995 Annual Research Report
  • [Publications] 後藤和男: "トリロタウイルスの細胞レセプターに対する抗体の作出。" 第44回日本ウイルス学会(1996年11月). (発表予定).

    • Related Report
      1995 Annual Research Report

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

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