- 學位論文
果蠅傷口癒合過程中puckered表現之研究
Analysis of puckered expression during wound healing of Drosophila
摘要
傷口癒合機制的研究是很重要的，JNK signaling pathway中的puckered會表現在傷口周圍的表皮細胞，而pucE69是puckered基因的enhancer trap line，可以在傷口周圍觀察到lacZ的表現，是一般用來研究傷口癒合的marker。我們利用puckered上游的非基因DNA片段表現GFP，希望使傷口癒合的過程能在活體的狀態下觀察。我們建構的pucF43-pGP包含puckered上游1.9kb的DNA片段可以在果蠅成蟲及幼蟲的傷口周圍表現GFP，而且與Ddc-GFP相較之下，在傷口產生10小時以後，pucF43-pGP有比較大的GFP表現範圍。然而在胚胎dorsal closure過程中的leading edge cells上，即使是含有puckered上游3.7kb DNA片段的pucF63-pGP也無法表現GFP。或許代表著在dorsal closure及傷口癒合過程中puckered受到啟動的方式不一樣。pucF43-pGP在所包含的片段當中，可預測到數個與Ddc同時擁有的轉錄因子的結合位置，或許Ddc與puckered可受到一個共同的轉錄因子啟動。
並列摘要
It is important for revealing the mechanisms of wound healing. It was known that JNK signaling pathway component, puckered, expresses in the epithelium cells around the wound. In genetic research of wound healing process of Drosophila, pucE69 was usually used as a lacZ marker. To observe the dynamic motion of wound healing process of Drosophila, we constructed the GFP reporters by the upstream non-coding DNA sequence of puckered. The GFP reporter containing 1.9kb non-coding DNA sequence upstream of puckered (pucF43-pGP) was sufficient to express around the wound of adults and larva. By the measurement of GFP expression area, pucF43-pGP has larger area than Ddc-GFP after 10 hours. However, even the 3.7kb non-coding DNA sequence could not drive the GFP reporter (pucF63-pGP) to express in the leading edge cells during dorsal closure. It suggests that there are different mechanisms driving puckered expression during wound healing and dorsal closure. Additionally, we predicted some transcriptional factor binding sites in pucF43 sequence, which also exist in upstream of Ddc. It is probably that there was a factor driving both puckered and Ddc during wound healing process.