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系統識別號 U0002-2706200515450200
中文論文名稱 重組蛋白OraSE及GlmE最適化之生產條件探討
英文論文名稱 Optimization of the production conditions for recombinant proteins OraSE and GlmE
校院名稱 淡江大學
系所名稱(中) 化學學系碩士班
系所名稱(英) Department of Chemistry
學年度 93
學期 2
出版年 94
研究生中文姓名 王立婷
研究生英文姓名 Li-Ting Wang
學號 692170144
學位類別 碩士
語文別 中文
口試日期 2005-04-29
論文頁數 32頁
口試委員 指導教授-鄭建中
共同指導教授-陳灝平
委員-陳銘凱
中文關鍵字 純化  濃度  管住層析 
英文關鍵字 OraSE  GlmE 
學科別分類 學科別自然科學化學
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 .

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


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