臺灣博碩士論文加值系統

指導教授推薦書
論文口試委員審定書
誌謝 iii
中文摘要 iv
英文摘要 v
目錄 vi
圖目錄 viii
第一章 緒論 1
1.1 RNA 編輯 1
1.2 ADAR 簡介 1
1.3 TMEM63B 簡介 3
1.4 ADAR2 與 TMEM63B 之關聯 4
1.5 研究動機 5
第二章 實驗方法與材料 6
2.1 細胞株及細胞培養 6
2.2 細胞轉染 7
2.3 RNA 萃取 8
2.4 反轉錄聚合酶連鎖反應 8
2.5 即時定量聚合酶連鎖反應 9
2.6 蛋白質分析 9
2.7 聚合酶連鎖反應 10
2.8 表現載體之建構 11
2.9 CRISPR/Cas9 系統 12
2.10 小鼠組織 Genomic DNA 之萃取 12
2.11 免疫螢光染色法 13
2.12 膜蛋白萃取 13
第三章 實驗結果 15
3.1 ADAR2 是否造成 TMEM63B 之影響與 TMEM63B 之交互作用關係 15
3.2 藉由 knockdown方式分析 Tmem63b 之功能 16
3.3 TMEM63B 客製化抗體之測試與分析 17
3.4 藉由過量表現方式分析 Tmem63b 編輯前後之差異 18
3.5 探討 Tmem63b 在體內 (in vivo) 之表現及 Tmem63b 編輯事件之影響 19
第四章 討論 21
4.1 TMEM63B 之功能與機制探討 21
4.2 利用實驗動物進行 TMEM63B 生理功能之研究 22
參考文獻 23
圖表 27
附錄 46


圖目錄
Figure 1. ADAR2 之 RNA 編輯與 TMEM63B 表現量之關係……..27
Figure 2. 透過 Tmem63b knockdown 分析 Tmem63b 的功能及神經
細胞分化型態之影響..............................……………………………….29
Figure 3. 藉由 RNA-Seq 技術分析與 Tmem63b 可能相關之基因..33
Figure 4. 客製化 TMEM63B 抗體之測試…………………………...35
Figure 5. TMEM63B 蛋白編輯前後於細胞不同區域表現量之差異..38
Figure 6. TMEM63B 過量表現與 Na+
/K+
-ATPase 之交互作用.…39
Figure 7. Tmem63b 在體內之表現及 Tmem63b 編輯事件之影響….40
Figure 8. CRISPR/Cas9 系統建立 Tmem63b 之基因剔除鼠及 RNA
編輯前後之 knock-in 小鼠………............……………………..……..42
Figure 9. Tmem63b knockout 小鼠之蛋白結構預測與外表型之影
響..............................................................................................44

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