| Project/Area Number |
05660046
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物保護
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
HOSOKAWA Daijiro Prof. Tokyo University of Agriculture and Technology, 農学部, 教授 (50014957)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Yuichiro Assoc. Prof. School of Science and Engineering Teikyo University, 理工学部, 助教授 (60183125)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | tobacco mosaic virus / potato virus X / cell-to-cell movement / movement protein / in situハイブリダイゼーション法 |
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| Research Abstract |
Cell-to-cell movement of tobacco mosaic virus (TMV) and potato virus X (PVX) were investigated using methods of molecular cell biology. The results are as follows.
1. The effects of deletion mutants in 30 k protein (movement protein) gene of TMV on cell-to-cell movement and localization of movement protein in tobacco protoplasts has been investigated. Cell-to-cell movement and localization of 30 k protein in protoplasts of 3 C-terminal amino acids deletion mutant indistiguishable from those of wild-type clone. When 33 C-terminal amino acids were deleted, virus moved slowly and lesser amounts of 30 k protein were found in the cytoplasms of protoplasts. Deletion of 77 C-terminal amino acids and residues from 9 to 142 at N-terminal abolish cell-to-cell movement and 30 k protein are not found in protoplasts, though virus particles aboundantly accumulated in protoplasts.
2. We have developed a in situ hybridization method using digoxygenin labelled-RNA probes to monitor the localization of TMV-RNA in tobacco plants and protoplasts. We are carring out studies to investigate behavior of virus RNA during cell-to-cell movement of TMV using this in situ hybridization method.
3. The 25 k protein which is part of the triple gene block of PVX are involved in cell-to-cell movement, In this experiment, we have used an E.coli expression system to produce significant amounts of 25 k protein. A polyclonal antiserum were produced using purified 25 k protein. An immunocytochemical analysis of infected tobacco leaves reveals the 25 k protein is exclusively associated with cytoplasmic inclusions in the cytoplasm of infected cells. These results indicate that the triple gene block proteins of PVX constitute a new class of movement protein.
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