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研究生:陳怡秀
研究生(外文):Yi-Siou Chen
論文名稱:發展實驗方法用以探索專一結合於HIF-2α 或其下游基因Oct4 啟動子之蛋白質
論文名稱(外文):Methodological development to explore the proteins which specifically interact with HIF-2α or bind the promoter of Oct4
指導教授:周慰遠蕭嘉陽
指導教授(外文):Wei-Yuan ChouChia-Yang Shiau
口試委員:白壽雄周慰遠蕭嘉陽
口試委員(外文):Shou-Hsiung PaiWei-Yuan ChouChia-Yang Shiau
口試日期:2014-05-15
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:37
中文關鍵詞:缺氧誘導轉錄因子八聚體結合轉錄因子
外文關鍵詞:HIF-2αOct4
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Oct4 ( octamer-binding transcription factor 4 ) 是一類轉錄因子,與胚胎幹細胞維持多能性狀態以及胚胎早期分化有密切相關性,近年來已知許多腫瘤發展也和Oct4表現有關。 過去研究發現Oct4會受到HIF-2α ( Hypoxia Inducible Factor-2α ) 的調控,其結合位置在Oct4轉錄序列上游-2557/-2426及-1956/-1852處。 在我們的實驗中發現BEAS-2B細胞中也有Oct4及HIF-2α的蛋白質表現,但BEAS-2B是體細胞而非幹細胞,因此本實驗室嘗試發展一套實驗系統,主要是利用已知受HIF-2α調控之Oct4序列混合BEAS-2B細胞之核內蛋白質並分析與Oct4序列行交互作用之核內蛋白質,作為未來研究體細胞與幹細胞多能性差異之先驅。
Oct4 is a transcription factor involved in the maintenance of pluripotent embryonic stem cells, differentiation at the early stage of embryonic development and tumorigenesis. Recent reports indicated that Oct4 is directly regulated by HIF-2α. There are two HIF-2α binding sites at the upstream sequence -2557/-2426 and -1956/-1852 of Oct4. In our experiment data, BEAS-2B has Oct4 and HIF-2α proteins, but BEAS-2B is somatic cell but not stem cell. Therefore, we were trying to develop an experimental system by using the Oct4 sequence to mix with the nuclear proteins from BEAS-2B and analyze the captured proteins which interact with the sequence. This method is a pioneer for future research which investigates the difference in pluripotency between somatic cell and stem cell.
圖目錄 Ⅱ
中文摘要 Ⅲ
Abstract Ⅳ
第一章、緒論 1
第一節、Oct4 1
第二節、缺氧誘導轉錄因子 ( Hypoxia Inducible Factors, HIFs ) 2
第三節、HIF-2α 3
第四節、實驗目的 4
第二章、材料與方法 8
第一節、實驗材料與儀器 8
第二節、實驗用溶液配方 11
第三節、實驗方法 13
第三章、實驗結果 20
第一節、觀察BEAS-2B內HIF-2α及Oct4的蛋白質表現 20
第二節、尋找適當超音波實驗條件 20
第三節、穀胱甘肽轉移酶下拉實驗 21
第四節、發展實驗方法以探索結合於Oct4 啟動子之蛋白質 22
第四章、討論 24
附圖 28
參考文獻 36
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