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研究生: 林政源
Lin, Cheng-Yuan
論文名稱: The Design and Engineering of a Molecular Probe for Dopamine Detection in vivo
活體多巴胺探針之設計及開發
指導教授: 桑自剛
Sang, Tzu-Kang
口試委員:
學位類別: 碩士
Master
系所名稱: 生命科學暨醫學院 - 生物科技研究所
Biotechnology
論文出版年: 2009
畢業學年度: 97
語文別: 英文
論文頁數: 55
中文關鍵詞: 多巴胺活體
外文關鍵詞: dopamine, in vivo
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  • Abstract.........................................................................................................................ii 中文摘要……………………………………………………………………………...iii Acknowledgement……………………………………………………………………iv Introduction…………………………………………………………………………...1 Materials and Methods……………………………………………………………….8 Plasmids and constructs…………………………………………………………....8 Over-expressions of BL21 and analysis of SDS-PAGE with coomassie blue staining……………………………………………………………………………..9 PC12 cell culture…………………………………………………………………...9 Transfect and selection……………………………………………………………10 Living cells confocal imaging…………………………………………………….10 Western Blot……………………………………………………………………....11 PC12 cells immunohistochemistry………………………………………………..12 Flow cytometry measurement…………………………………………………….13 Cell microinjection………………………………………………………………..13 Drosophila stocks and genetics……………………………………………………14 Results………………………………………………………………………………..14 MAO-GFP fusion proteins could block the emission fluorescence of the GFP in living bacteria……………………………………………………………………..15 The shield effect of MAO-GFP in living mammalian cells………………………16 The expression of dopamine probes in cytoplasm of PC12 cell………………….18 PC12 cells express dopamine transporter that could uptake exogenous dopamine 19 The short form probe could detect endogenous and exogenous dopamine through shield effect……………………………………………………………………….20 Short form probe expressing cell could detect dopamine when microinjecting cell with dopamine directly…………………………………………………………...22 Discussion……………………………………………………………………………25 Dopamine involved in neurological disorders……………………………………25 A new approach for detecting dopamine in living organism……………………..26 Future expansions of the present study…………………………………………...28 Figures……………………………………………………………………………….30 References…………………………………………………………………………...48

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