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Why green fluorescent fusion proteins have not been observed in the vacuoles of higher plants
https://ir.soken.ac.jp/records/3923
https://ir.soken.ac.jp/records/392352aa8896-ca07-465c-9e99-92dce21967b8
Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2013-08-26 | |||||
タイトル | ||||||
タイトル | Why green fluorescent fusion proteins have not been observed in the vacuoles of higher plants | |||||
タイトル | ||||||
タイトル | Why green fluorescent fusion proteins have not been observed in the vacuoles of higher plants | |||||
言語 | en | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
アクセス権 | ||||||
アクセス権 | metadata only access | |||||
アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
著者 |
TAMURA, Kentaro
× TAMURA, Kentaro× SHIMADA, Tomoo× ONO, Eiichiro× TANAKA, Yoshikazu× NAGATANI, Akira× HIGASHI, Sho-ich× WATANABE, Masakatsu× NISHIMURA, Mikio× HARA-NISHIMURA, Ikuko |
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著者別名 |
渡辺, 正勝
× 渡辺, 正勝 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Green fluorescent protein (GFP) makes it possible for organelles and protein transport pathways to be visualized in living cells. However, GFP fluorescence has not yet been observed in the vacuoles of any organs of higher plants. We found that the fluorescence of a vacuole-targeted GFP was stably observed in the vacuoles of transgenic Arabidopsis plants under dark conditions, and that the fluorescence rapidly disappeared under light conditions. The vacuolar GFP was rapidly degraded within 1 h in the light, especially blue light. An inhibitor of vacuolar type H+-ATPase, concanamycin A, and an inhibitor of papain-type cysteine proteinase, E-64d, abolished both the light-dependent disappearance of GFP fluorescence and GFP degradation in the vacuoles. An in vitro assay showed that bacterially expressed GFP was degraded by extracts of Arabidopsis cultured-cell protoplasts at an acidic pH in the light. These results suggest that blue light induced a conformational change in GFP, and the resulting GFP in the vacuole was easily degraded by vacuolar papain-type cysteine proteinase(s) under the acidic pH. The light-dependent degradation accounts for the failure to observe GFP fluorescence in the vacuoles of plant organs. Our results show that stable GFP-fluoresced vacuoles are achieved by transferring the plants from the light into the dark before inspection with a fluorescent microscope. This might eliminate a large hurdle in studies of the vacuolar-targeting machinery and the organ- and stage-specific differentiation of endomembrane systems in plants. | |||||
書誌情報 |
Plant Journal en : Plant Journal 巻 35, 号 4, p. 545-555, 発行日 2003-08 |
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出版者 | ||||||
出版者 | Wiley-Blackwell | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 09607412 | |||||
PubMed番号 | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | PMID | |||||
関連識別子 | 12904216 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | https://doi.org/10.1046/j.1365-313X.2003.01822.x | |||||
関連名称 | 10.1046/j.1365-313X.2003.01822.x | |||||
権利 | ||||||
権利情報 | © Wiley-Blackwell | |||||
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識別子タイプ | URI | |||||
関連識別子 | http://olabout.wiley.com/WileyCDA/Section/id-406074.html | |||||
関連名称 | Copyright | |||||
フォーマット | ||||||
内容記述タイプ | Other | |||||
内容記述 | application/pdf | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 |