| Project/Area Number |
04454065
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
蚕糸学
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| Research Institution | MIE UNIVERSITY |
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Principal Investigator |
MIYAJIMA Shigetoshi Mie Univ., Fca.Eng., Prof, 工学部, 教授 (80239409)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Jun Mie Univ., Fca.Eng., Res.Ass, 工学部, 助手 (70242930)
TOMITA Masahiro Mie Univ., Fca.Eng., Ass.Prof, 工学部, 助教授 (20183494)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | Monoclonal Antibody / Silkworm / Virus / Diagnosis / Hybridoma / Crystal Bibrator / Immunosensor / Enhancing Factor / 細胞質多角体病ウイルス / CPV / バイオセンサー / PZセンサー / 共振周波数 / カイコウイルス / 迅速診断法 / 診断法の開発 / 細胞融合 |
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| Research Abstract |
In order to develop a simple ana quickly diagnosis for silkworm virus we made the monoclonal antibodies for cytoplasmic-polyhedrosis virus (CPV) and dnhancing factor of nuclear-polyhedrosis virus infection (EF) by hybridoma selection method.
1.Three-group clones of monoclonal antibody for CPV-I strain were obtained. Thst is, one group was remarkably reacted to CPV-I strain, the second reacted to CPV-H strain, and the third reacted in the intermediate degrees. These results suggested that both strains pessessed the common epitope between CPV-H and CPV-I strains. But some of them showed a little different reaction between two strains.
2. There are two types of inclusion body produced by Pseudaletia separata entomopoxvirus. One is called spheroid enclosed virus particles and the other is spindle without virus particle in it. Enhancing factor (EF) is known as the increasing the nuclear-polyhedrosis virus infection.
So we mede a monoclonal antibody for the enhancing factor protein. But this mo
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noclonal antibody did not react for spheroidin which was a main constructed protein of spheroid. On the contrary the another inclusion protein, spindle, reacted remarkably to this monoclonal antibody. From the result it was demonstrated that 49.5KDa protein, i.e.main constructed protein of spindle, had common epitope to enhancing factor.
We tried to develop the immunosensor constructed by monoclonal antibody and crystal bibrator to detect the cytoplasmic-polyhedrosis virus of the silkworm simply and quickly. At first we purified the IgG from monoclonal antibody for CPV and tested its reaction. The IgG showed the highly titers, i.e.1 x 10^<-5> g IgG could detect 1 ug/ml CPV by ELISA.In addition to this it was clear that this monoclonal antibody reacted both CPV core protein and CPV polyhedrin by western blotting analysis. The IgG conjugated the crystal bibrator smeared with protein A was showed the variation of co-frequency. By remove the non-specific adsorption we might develop the high sensitive immunosensor. Less
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