| 研究生: |
謝曜駿 Xie, Yao Jun |
|---|---|
| 論文名稱: |
以生物取像系統探討綠色螢光蛋白在不同培養條件下之基因重組大腸桿菌內涵體的形成 Study of Green Fluorescence Protein Inclusion Body Formation under Various Culture Conditions in Recombinant Escherichia coli with Bioimaging System |
| 指導教授: |
鄭智元
Zheng, ZiYuan |
| 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: | 2010 |
| 畢業學年度: | 98 |
| 語文別: | 中文 |
| 論文頁數: | 60 |
| 中文關鍵詞: | 綠色螢光蛋白 、大腸桿菌 、巨大原生質體 、生物取向系統 、內涵體 |
| 外文關鍵詞: | GFP, inclusion body, Escherichia coli |
| 相關次數: | 點閱:112 下載:1 |
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生物取象系統使用使用螢光蛋白當作報導蛋白,一般應用於真核生物,可即時精確地觀察到螢光蛋白在細胞或組織之行程及分布狀態;而原核生物細胞體積只有真核生物細胞的千分之一,通常無法使用生物取象系統精確地觀察螢光蛋白在細胞內之分佈狀態。本研究以特殊培養方法,將基因重組大腸桿菌BL21(DE3)/ pET21a-GFP 做成與酵母菌同等大小之巨大原生質體(giant protoplast) 。在IPTG誘導下,由於綠色螢光蛋白的大量表現,可即時觀察到內涵體之形成,藉此可評估基因重組大腸桿菌形成內涵體之培養條件。
實驗發現在不同培養條件下,藉由螢光顯微鏡觀察巨大原生質體螢光表現差異,可得到隨著誘導劑及培養溫度的增加或培養液PH值的下降,其內涵體的形成有增加的趨勢。
Bioimaging system usually utilize fluorescence protein as a reporter gene in eukaryotic system, and it gives a easy way for us to real-time monitor the distribution of fluorescence protein in bacteria or cell.Eukaryote is thousand times the size of prokaryote, so it is not suitable to this system.In our reserach, recombinant Escherichia coli BL21(DE3)/ pET21a-GFP is used to prepare as giant protoplast which size is similar to saccharomycete with particular method. The expression of GFP can be induced by IPTG to real-time monitor inclusion body formation.Using this approach, we can study inclusion body formation under various culture conditions in recombinant Escherichia coli.
By monitoring the fluorescence protein formation in the giant protoplast under various culture conditions, the result indicated that inclusion body formation would increase as increasing IPTG, increasing induction temperature or deceeasing ph value.
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