| 研究生: |
林青璇 Lin, Ching-Shiuan |
|---|---|
| 論文名稱: |
綠色螢光蛋白應用於大腸桿菌生物取像
系統之研究 The Study of Green Fluorescent Protein Applications for E.coli Bioimaging System |
| 指導教授: |
鄭智元
Jeng, Jr-Yuan |
| 學位類別: |
碩士 Master |
| 系所名稱: |
工學院 - 化學工程學系 Department of Chemical Engineering |
| 論文出版年: | 2006 |
| 畢業學年度: | 94 |
| 語文別: | 中文 |
| 論文頁數: | 54 |
| 中文關鍵詞: | 巨大原生質體 、綠色螢光蛋白 |
| 外文關鍵詞: | GFP |
| 相關次數: | 點閱:112 下載:1 |
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本研究探討大腸桿菌綠色螢光蛋白基因(pGFP)在大腸桿菌Escherichia coli XL1-Blue 巨大原生質體之表現,確立以螢光顯微鏡及數位照相定量菌體內綠色螢光蛋白螢光強度之方法,並用此法來估算啟動(promoter)的效率。實驗發現pGFP 螢光基因可在大腸桿菌巨大原生質體表現,代表巨大原生質體具有正常細胞功能。比較空氣量對螢光亮度的影響,發現螢光強度會隨著空氣量的減少而延遲。本研究建立之數位影像系統可數值化螢光蛋白之螢光強度,具有快速、直接、簡便的優點。利用此結果,本研究之綠色螢光蛋白巨大原生質體之生物取像系統具有作為研究啟動子(promoter)效率及細胞微結構與生理反應工具的發展潛力。
To develop a rapid reporter system for estimating the promoter efficiency in Escherichia coli XL1B, the expression pattern of the green fluorescent protein(GFP) in the giant protoplast was investigated. In addition, the methods for quantifying the fluorescent intensity of GFP by fluorescent microscopy as well as digital camera were illustrated. The results showed that the protoplasts of E. coli was able to express GFP, indicating that giant protoplasts possess cell functionality. The expression level of GFP in E. coli protoplasts was affected by the air level, i.e. the fluorescence intensity increases as the air level does. The digital imaging method with the advantages of rapidity, directness, and easiness established in the work could digitalize the intensity of fluorescence emitted by GFP. The results suggested that the tested plasmids were stable in recombinant cells, no matter cells in normal situation or becoming the giant protoplasts. In conclusion, the bioimaging system of the giant protoplast of E. coli could be a suitable method to estimate the promoter efficiency and promise to be a tool for studying the protein expression and cellular activity.
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