WEKO3
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In vitroプリオン増殖機構解析：PrP遺伝子欠損細胞を用いた改良PMCA法
http://hdl.handle.net/20.500.12000/18127
http://hdl.handle.net/20.500.12000/181277a14a7b1-9561-497f-869e-8c55f83ad0cc
| 名前 / ファイル | ライセンス | アクション | |
|---|---|---|---|
20780219seika.pdf
|
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| Item type | デフォルトアイテムタイプ（フル）(1) | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| 公開日 | 2010-09-29 | |||||||||
| タイトル | ||||||||||
| タイトル | In vitroプリオン増殖機構解析：PrP遺伝子欠損細胞を用いた改良PMCA法 | |||||||||
| 言語 | ja | |||||||||
| 作成者 |
作道, 章一
× 作道, 章一
× Sakudo, Akikazu
|
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| アクセス権 | ||||||||||
| アクセス権 | open access | |||||||||
| アクセス権URI | http://purl.org/coar/access_right/c_abf2 | |||||||||
| 主題 | ||||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | 微生物 | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | 獣医学 | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | 感染症 | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | ウイルス | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | 神経科学 | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | プリオン | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | 人獣共通感染症 | |||||||||
| 言語 | ja | |||||||||
| 主題Scheme | Other | |||||||||
| 主題 | 人畜共通感染症 | |||||||||
| 内容記述 | ||||||||||
| 内容記述タイプ | Other | |||||||||
| 内容記述 | 科研費番号: 20780219 | |||||||||
| 内容記述タイプ | Other | |||||||||
| 内容記述 | 2008年度～2009年度科学研究費補助金（若手研究(Ｂ)）研究成果報告書 | |||||||||
| 内容記述タイプ | Other | |||||||||
| 内容記述 | 研究概要（和文）：本研究では、プリオン蛋白質(PrP)遺伝子欠損マウス神経細胞株(HpL)にマウスPrP遺伝子もしくは異種PrP(ハムスター、ウシ)を導入した細胞のライゼートを正常型PrP(PrP^C)供給源としてprotein misfolding cyclic amplification (PMCA)を行うとともに、マウススクレイピープリオンもしくは異種プリオン（それぞれハムスタースクレイピープリオン、Bovine spongiform encephalopathy (BSE)プリオン）存在下で、異常型PrP(PrP^<Sc>)増幅の有無からPrP^<Sc>増幅に必要な条件についての基礎的データを得ることを目的とした。PMCA法によるハムスター263K株、マウスObihiro株、マウスChandler株の試験管内増幅による比較を行った結果、263K株とChandler株は脳ホモジネートをPrP^C供給源とした場合、既存の条件（1% TritonX-100, 4mM EDTA含PBS、40 cycle）でPrP^<res>（蛋白質分解酵素抵抗性PrP）の増幅が可能であったが、Obihiro株では同条件では増幅がされなかった。さらに、HpLにマウスPrP遺伝子やハムスターPrPを導入した細胞の細胞ライゼートをPrP^C供給源として試験管内増幅を行ったところ、同条件では263K株とchandler株のいずれも増幅がされなかった。これらのことから、プリオン株ごとに条件設定が必要であるとともに、細胞ライゼートを用いた場合、脳ホモジネートの条件とは異なる増幅条件が必要であることが明らかとなった。本研究の結果から、PMCA法によるPrP^<res>の増幅条件はプリオン株やPrP^C供給源の性状ごとに最適化する必要があることが明らかになったとともに、動物を用いないプリオン検出系として、広範なプリオン株に適応可能なPMCA条件の構築が必要であることが示唆された。 | |||||||||
| 内容記述タイプ | Other | |||||||||
| 内容記述 | 研究概要（英文） : 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, PrP^<res> 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 PrP^<res> amplification compared to brain homogenate. Therefore, we concluded that the adaptation of PMCA conditions were required for efficient PrP^<res> proliferation depending on prion strains and PrP^C sources. | |||||||||
| 内容記述タイプ | Other | |||||||||
| 内容記述 | 研究報告書 | |||||||||
| 出版者 | ||||||||||
| 言語 | ja | |||||||||
| 出版者 | 作道章一 | |||||||||
| 言語 | ||||||||||
| 言語 | jpn | |||||||||
| 資源タイプ | ||||||||||
| 資源タイプ | research report | |||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_18ws | |||||||||
| 出版タイプ | ||||||||||
| 出版タイプ | NA | |||||||||
| 出版タイプResource | http://purl.org/coar/version/c_be7fb7dd8ff6fe43 | |||||||||
| 識別子 | ||||||||||
| 識別子 | http://hdl.handle.net/20.500.12000/18127 | |||||||||
| 識別子タイプ | HDL | |||||||||
| 書誌情報 |
発行日 2010-03-31 |
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