系統識別號 | U0002-2706200515450200 |
---|---|
DOI | 10.6846/TKU.2005.00641 |
論文名稱(中文) | 重組蛋白OraSE及GlmE最適化之生產條件探討 |
論文名稱(英文) | Optimization of the production conditions for recombinant proteins OraSE and GlmE |
第三語言論文名稱 | |
校院名稱 | 淡江大學 |
系所名稱(中文) | 化學學系碩士班 |
系所名稱(英文) | Department of Chemistry |
外國學位學校名稱 | |
外國學位學院名稱 | |
外國學位研究所名稱 | |
學年度 | 93 |
學期 | 2 |
出版年 | 94 |
研究生(中文) | 王立婷 |
研究生(英文) | Li-Ting Wang |
學號 | 692170144 |
學位類別 | 碩士 |
語言別 | 繁體中文 |
第二語言別 | |
口試日期 | 2005-04-29 |
論文頁數 | 32頁 |
口試委員 |
指導教授
-
鄭建中(cccheng@mail.tku.edu.tw)
共同指導教授 - 陳灝平(hpchen@mail.ntut.edu.tw) 委員 - 陳銘凱(mkchem@mail.tku.edu.tw) |
關鍵字(中) |
純化 濃度 管住層析 |
關鍵字(英) |
OraSE GlmE |
第三語言關鍵字 | |
學科別分類 | |
中文摘要 |
本研究藉由改變溫度的培養或使用不同的誘導因子,找出誘導生產蛋白質的最適化條件,以Eschericha coli合成的重組蛋白質OraSE及GlmE為探討對象。OraSE蛋白質可先後經由疏水性管柱及金屬螯合親和層析管柱分離,得到純的蛋白。另一蛋白質GlmE,也可經由疏水性管柱及陰離子交換管柱分離,達到純化的目的。並比較在不同溫度方式及誘導因子的培養下,所獲得蛋白質產量的差異。 |
英文摘要 |
Optimization of the production for recombinant proteins, OraSE and GlmE, was investigated by using several defferent growth conditions in this stusy . OraSE protein was purified by Phenyl – Sepharose HP and Ni2+ - NTA column ; GlmE protein was purified by Phenyl – Sepharose HP and Q – Sepharose FF. Significant difference in protein yield was observed when induction temperature and inducer were changed . |
第三語言摘要 | |
論文目次 |
中文摘要.................................................I 英文摘要................................................II 目錄..................................................III 圖目錄...................................................IV 表目錄.....................................................V 第一章 序論............................................1 1.1 前言.................................................1 1.2 T7基因表系統.........................................1 1.3 誘導因子乳糖及IPTG...................................2 1.4 乳糖操縱組 ( The lac operon )..................................3 第二章 實驗材料.....................................5 2.1 實驗藥品.........................................5 2.2 各式溶液組成......................................6 2.3 儀器設備..............................................7 第三章 OraSE蛋白質最適化生產條件探討................8 3.1 OraSE介紹.....................................................8 3.2 實驗方法.......................................................8 3.2.1菌種培養....................................................8 3.2.2 OraSE蛋白質的純化..........................................9 3.3 結果與討論....................................................12 3.3.1 管柱層析...................................................12 3.3.2 不同條件下OraSE蛋白質濃度比較.............................15 第四章 GlmE蛋白質最適化生產條件探討.............................18 4.1 GlmE蛋白質介紹...............................................18 4.2 實驗方法......................................................18 4.2.1 菌種培養...................................................18 4.2.2 GlmE蛋白質的純化...........................................19 4.3 結果與討論....................................................23 4.3.1 管柱層析...................................................23 4.3.2 不同條件下GlmE蛋白質濃度比較..............................26 第五章 結論.......................................................29 參考資料...........................................................31 圖目錄 圖 1-1乳糖結構式……………………………………………………..2 圖 1-2 IPTG結構式…………………………………………………….2 圖 1-3 乳糖操縱組示意圖……………………………………………..4 圖 3-1 OraSE蛋白質過完疏水性管柱Phenyl-Sepharose HP後層析.圖………………………………………………………………12 圖 3-2 OraSE蛋白質過完Ni2+-NTA column所得之層析圖………..13 圖 3-3 OraSE蛋白質純化過程的電泳膠片………………………….14 圖 3-4 不同OD值加入誘導因子所得濕菌重 ( 2 L )……………….17 圖3-5 不同OD值加入誘導因子單位細胞OraSE蛋白質的量……..17 圖4-1 GlmE蛋白質過完疏水性管柱Phenyl-Sepharose HP後之層析圖………………………………………………………………23 圖 4-2 GlmE蛋白質過完陰離子交換管柱所得之層析圖…………..24 圖 4-3 GlmE蛋白質純化過程的電泳膠片……………………………25 圖 4-4 不同條件下所得GlmE濕菌重………………………………27 圖 4-5 不同條件下所得GlmE蛋白質產量………………………….28 表目錄 表3-1 OD值0.7時不同誘導因子的蛋白質表現量………………..15 表3-2 OD值0.8時不同誘導因子的蛋白質表現量…………………16 表4-1 以37℃培養所得GlmE蛋白質產量.........................................26 表4-2 以降溫方式培養所得的GlmE蛋白質產量…………………..27 |
參考文獻 |
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