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| 研究生： |
蔡俊廷 Tsai, Jun-Ting |
|---|---|
| 論文名稱： |
過度表現Atg5-RNAi影響果蠅翅膀成蟲盤之發育 Overexpression of Atg5-RNAi Influence development of drosophila wing disc |
| 指導教授： |
徐瑞洲
Hsu, Jui-Chou |
| 口試委員: |
桑自剛
Sang, Tzu-Kang 張慧雲 Chang, Hui-yun |
| 學位類別： |
碩士 Master |
| 系所名稱： |
生命科學暨醫學院 - 分子醫學研究所 Institute of Molecular Medicine |
| 論文出版年： | 2019 |
| 畢業學年度： | 107 |
| 語文別： | 英文 |
| 論文頁數： | 36 |
| 中文關鍵詞： | 發育學 、黑腹果蠅 、成蟲盤 、自噬作用 |
| 外文關鍵詞： | Flip-Out system, wing disc |
| 相關次數： | 點閱：2 下載：0 |
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Flip-Out系統是一種基因工具，常用來產生表達特定基因的clone細胞群，並且可以藉由標誌蛋白來追溯細胞分裂後子細胞間的關係。將果蠅幼蟲經過短時間的熱休克處理後，幼蟲體內細胞會開始生產基因重組酶Flipase，它會結合在Flip-Out cassette中標誌基因兩側的FRT序列進行基因重組，將原先不斷表達的標誌基因剪出基因體並環化,啟動子轉而表達Gal4，Gal4會結合染色體上的UAS序列並表達綠螢光蛋白和其他特定基因，藉由綠螢光蛋白可以知道在翅膀成蟲盤上追蹤細胞分裂後屬於異種細胞的位置。我們發現過度表達Atg5 RNAi的clone細胞面積遠大於過度表達LacZ的clone細胞面積。這有可能是Atg5 RNAi會使得clone細胞更容易產生或是促進細胞分裂。我們目前的結果顯示是無關於促進細胞分裂的作用。另外也在clone中表達各種信號傳遞鏈上的相關分子，觀察是否有類似Atg5 RNAi的結果。而這些訊號傳遞鏈包含了EGFR、 DPP、JaK-STAT、JNK、Hippo、Toll、cell polarity以及 Myc。
Flip-Out system is a genetic tool commonly used for gene overexpression and lineage tracing. After heat shock treatment of larvae, FLP recombinase is expressed to induce the excision of marker gene flanked by FRTs that in turn causes the constitutive expression of gal4 to activate UAS-GFP reporter or other gene of interest in the flip-out clones after cell division in the wing disc. We found that the area covered by flip-out clones overexpressing UAS-Atg5 RNAi is much larger compared to UAS-LacZ overexpression. It’s possible that Atg5 RNAi overexpression in the flip-out clone can either cause the increase of clone number or promote cell division. Our data suggest that the increase of area is independent of cell division. In addition, we also test other signal pathway components (EGFR, DPP, JaK-STAT, JNK, Hippo, Toll, cell polarity, and Myc) to see whether any of them, when overexpressed, can have similar effect as Atg5 RNAi on the generation of flip-out clone.
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