2009 Fiscal Year Final Research Report
In vitro analysis of prion proliferation : Modified PMCA method using PrP-gene deficient cell line
| Project/Area Number |
20780219
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied veterinary science
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| Research Institution | Osaka University |
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Principal Investigator |
SAKUDO Akikazu Osaka University, 医学部, 准教授 (10397672)
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| Project Period (FY) |
2008 – 2009
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| Keywords | 微生物 / 獣医学 / 感染症 / ウイルス / 神経科学 / プリオン / 人獣共通感染症 |
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| Research Abstract |
In this study, protein misfolding cyclic amplification (PMCA) was performed using prion protein (PrP)-gene deficient cell line as the source of cellular PrP. Mouse and hamster scrapie and bovine spongiform encephalopathy (BSE) prion were used for seed. In 2008, cell lysate was prepared from prion protein (PrP)-gene deficient cell re-introduced with mouse, hamster, and bovine PrP-gene. In 2009, we compared the in vitro amplification efficiency in PMCA among hamster scrapie 263K, mouse scrapie Obihiro, mouse Chandler scrapie prions. As the results, PrPres could be amplified from brain homogenates infected with 263K and Chandler prion under the condition of PBS containing 1% TritonX-100, 4mM EDTA (40 cycles), whereas Obihiro prion could not be amplified. In addition, PMCA using cell lysate required different conditions for PrPres amplification compared to brain homogenate. Therefore, we concluded that the adaptation of PMCA conditions were required for efficient PrPres proliferation depending on prion strains and PrPC sources.
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Research Products
(28 results)
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[Journal Article] Distinct immunohistochemical localization in Kuru plaques using novel anti-prion protein antibodies.2008
Author(s)
Hosokawa T, Ono F, Tsuchiya K, Sato I, Takeyama N, Ueda S, Zanusso G, Takahashi H, Sata T, Sakudo A, Suguira K, Baj A, Toniolo A, Yoshikawa Y, Onodera T.
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Journal Title
Microbiol Immunol. 52
Pages: 25-29
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[Journal Article] A monoclonal antibody (1D12) defines novel distribution patterns of prion protein (PrP) as granules in nucleus2008
Author(s)
Hosokawa T, Tsuchiya K, Sato I, Takeyama N, Ueda S, Tagawa Y, Kimura KM, Nakamura I, Wu G, Sakudo A, Casalone C, Mazza M, Caramelli M, Takahashi H, Sata T, Sugiura K, Baj A, Toniolo A, Onodera T.
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Journal Title
Biochem Biophys Res Commun 366
Pages: 657-663
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