| Project/Area Number |
16380040
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied entomology
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| Research Institution | Nagoya University |
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Principal Investigator |
IKEDA Motoko Nagoya University, Graduate School of Bioagricultural Sciences, Asscciate prefessor, 大学院生命農学研究科, 助教授 (20262892)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Michihiro Nagoya University, Graduate School of Bioagricultural Sciences, Profossor, 大学院生命農学研究科, 教授 (60111837)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2006: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2005: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2004: ¥7,700,000 (Direct Cost: ¥7,700,000)
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| Keywords | nucleopolyhedrovirus / host DNA replication arrest / PCNA / apoptosis / アポトーシス制御 / Tn368細胞 / Tn-caspase-1 / アポトーシス誘導因子 / 初期遺伝子 / 相同繰り返し配列 / プロモーター / gp64 / IE1 / バキュロウイルス / DNA複製 / ウイルス増殖 / 宿主特異性 |
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| Research Abstract |
A homologue of proliferating cell nuclear antigen (pcna) gene is identified in Autographa californica nucleopolyhedrovirus genome, but the function in viral replication has not been elucidated. In the present study, AcMNPV mutants defective in the pcna gene (vAcApcnas) were generated and the function of the pcna gene was analyzed. Biological characterization revealed that vAcApcnas were viable in the Sf9 cells and the kinetics of viral DNA replication, viral structural protein synthesis, budded virus production and polyhedron production in the vAcApcna-infected cells were comparable to those in the cells infected with wt AcMNPV. It was also demonstrated that the amount of cellular PCNA in the nuclear fraction of Sf9 cells infected with vAcApcna was significantly higher than that of wt AcMNPV-infected cells. Further analysis by DNase I treatment and chromatin immunoprecipitation assay revealed that the cellular PCNA in the nucleus of Sf9 cells bound AcMNPV DNA. The amount of cellular PCNA associated with viral DNA replication sites was greater in cells infected with vAcApcna than in cells infected with wt AcMNPV. These results suggest that both cellular and viral PCNAs are involved in AcMNPV DNA replication and that vAcApcna is able to substitute cellular PCNA for viral PCNA. This deprivation of cellular PCNA may cause host DNA synthesis shut down.
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