臺灣博碩士論文加值系統

在多細胞生物發育的過程中，有部分的細胞會發生計畫性細胞死亡（細胞凋亡），而這些凋亡的細胞殘骸必須被吞噬清除，以免產生免疫或發炎反應。在線蟲的研究中已發現一系列的基因參與凋亡細胞的吞噬作用，其吞噬作用的發生在於吞噬細胞的表面受器會辨識死細胞表面的磷脂醯絲胺酸（phosphatidylserine），而這些磷脂醯絲胺酸則參與了ced-2、ced-5、ced-12、ced-10以及ced-7、ced-1、ced-6兩組功能部份重疊的基因路徑上，但目前對於ced-7、ced-1、ced-6這條路徑的研究則尚未透徹。在本研究中，我們建構了接有GFP基因的斑馬魚 ced-6 promoter質體，且發現薑黃素（curcumin）（一種促進癌症細胞凋亡的化合物）在加到已經顯微注射pEGFP-1 / zfced-6 promoter的斑馬魚胚胎後，會加強zfced-6 promoter的螢光表現，利用這個發現可以找出curcumin在zfced-6 promoter上的作用區域。除了薑黃素外，我們也將海洋菌的萃取物加到已經顯微注射pEGFP-1 / zfced-6-promoter的斑馬魚胚胎，並觀察這些萃取物對斑馬魚胚胎的影響，同時找出其它可和zfced-6-promoter作用的化合物。最後，我們將建立具有ced-6 promoter的轉基因斑馬魚，當做一個可以篩選與ced-6 promoter作用的化合物的篩選平台。

In the course of multicellular organism development, some cells the planned cell's death will take place (apoptosis). And these cell remains must be engulfed and removed, so as not to induce the immunity or the inflammation. In the research of Caenorhabditis elegans have already found a series of genes been participated the phagocytosis of the death cell. The surface receptor of macrophage can distinguish superficial phosphatidylserine of death cell and induce phagocytosis happened. These phosphatidylserine have participated in ced-2, ced-5, ced-12, ced-10 and ced-7, ced-1, ced-6 two groups of functions part overlap, but the research of the route to ced-7, ced-1, ced-6 at present is not thorough. In this experiment, we have constructed a plasmid which has zebrafish ced-6 promoter and GFP gene. And we found curcumin (one kind of chemical compounds which induce cancer cell death) can enhance the fluoresce intensity in zebrafish embryos, which have injected pEGFP-1 / zfced-6-promoter. Utilizes this result we identified the curcumin activity site on zebrafish ced-6 promoter. Except curcumin, we added marine bacteria extracts in zebrafish embryo which have injected pEGFP-1 / zfced-6-promoter, and measured the effect of these extracts. We attempt to find out another chemical compounds which can induce zfced-6-promoter. Finally, we will establish the transgenic zebrafish with ced-6 promoter, which can be used as a screening platform for selection the chemical compound which effect ced-6 promoter activity.

誌謝 ……………………………………Ⅰ
摘要 ………………………………………Ⅱ
Abstract ……………………………………Ⅲ
目錄 ………………………………………Ⅳ
圖索引 ………………………………Ⅴ
表索引 ………………………………………Ⅵ
壹、前言 ……………………………………………1
貳、材料與方法 ……………………………………12
參、結果 ………………………………………………26
肆、討論 ………………………………………………37
伍、結論 …………………………………………………43
陸、參考文獻 ………………………………………………44
圖 ……………………………………………………………49
表 ………………………………………………………66
柒、附錄 …………………………………………72

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