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| 研究生： |
張兆毅 Chang, Zhao-Yi |
|---|---|
| 論文名稱： |
綠色螢光蛋白發光團對位吡啶類似物合成、光/熱異構化反應、酸性水解及與DNA嵌合反應之探討 para-Pyridinium Chromophore of Green Fluorescence Protein Analogue： Synthesis, Photo/Thermal Isomerization, Acidic Hydrolysis and Discussion the Mechanism of Intercalation with DNA |
| 指導教授： |
宋光生
Sung, Kuang-Sen |
| 學位類別： |
碩士 Master |
| 系所名稱： |
理學院 - 化學系 Department of Chemistry |
| 論文出版年： | 2017 |
| 畢業學年度： | 105 |
| 語文別： | 中文 |
| 論文頁數： | 80 |
| 中文關鍵詞： | 綠色螢光蛋白發光團類似物 |
| 外文關鍵詞： | GFP, protonation, acidic condition, hydrolysis |
| 相關次數： | 點閱：148 下載：0 |
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近年來已有許多綠色螢光蛋白的相關研究，部分研究表示綠色螢光蛋白在酸性環境下會變性，但都是取自天然的GFP模型直接與酸進行作用，且顯少有研究提出在酸性作用下，致使其變性的機制及變性結果。我們的方法則是合成出綠色螢光蛋白發光團類似物，利用甲基化的方法替代氫的質子化，以進行模擬綠色螢光蛋白發光團在酸性環境下受到質子化後的效果。藉由比較我們合成出的綠色螢光蛋白類似物p3-Bn和p4-Bn，發現甲基化後五元雜環上7號碳的位置(C7)變得較缺電子，更易受到親核基的攻擊而導致水解。此外，化合物照射波長350 nm之光源後，藉由追蹤核磁共振儀積分值比例得知光異構化轉換率，並搭配sigma-plot軟體計算出熱異構化速率常數，以及測量吸收、螢光放射光來探討其光物理性質。
在我們合成出我們需要的目標產物之後，我們利用了可見光-紫外分光光譜儀以及螢光光譜儀來檢測綠色螢光蛋白類似物與DNA行反應之後會產生的光譜變化並對其進行探討確定是否有進行反應。
To simulate other GFP chromophoric analogues under the acidic condition, we used methylation method to synthesize GFP chromophoric analogues p4-Bn and planned to imitate protonated type of GFP simultaneously. In this way, we found that water could easily affect compound p4-Bn. Due to the positively charged nitrogen on p4-Bn, the methyl carbon on five-membered heterocyclic ring became electron deficient and was easily attacked by nucleophile such as H2O. We monitored the integration value of 1H NMR and calculated the rate of photo-isomerization and thermal-isomerization with software. Finally, the result of single-crystal X-ray diffraction clarified that p4-Bn would ultimately hydrolyze in water. These results could offer an evidence to previous reports that protonation of GFP derivatives will change its conformation or denaturation as a result of the hydrolysis reaction. Moreover, we used GFP analogue m4-Bn to intercalate with E. coli plasmid DNA. By using UV and FL detection to observe the change of wavelength.
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校內：2022-01-01公開校外：不公開