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研究生:陳玫束
研究生(外文):Mei-Shu Chen
論文名稱:果蠅CG17064為HURP的同源基因會影響眼睛發育時的細胞增生及凋亡
論文名稱(外文):Drosophila CG17064, a putative homologue of HURP, affects cell proliferation and apoptosis in eye development
指導教授:范聖興
指導教授(外文):Seng-Sheen fan
學位類別:碩士
校院名稱:東海大學
系所名稱:生物學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:58
中文關鍵詞:果蠅眼睛發育肝癌基因細胞增生細胞凋亡
外文關鍵詞:Drosophilaeye developmentHURPcell proliferationapoptosis
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果蠅CG17064基因為一嶄新的基因,此基因位於第二號染色體50B1-2的位置。全長的CG17064 cDNA共含有3053個核苷酸,轉譯後的蛋白質由921個胺基酸所組成。經由BLAST搜尋比對的結果顯示,CG17064胺基酸序列的486-694含有一個NES motif,且此胺基酸序列與人類HURP有較高度的相似性。我們使用GMR-Gal4誘發基因轉殖果蠅表現過量的CG17064於形態發生溝之後,結果會增加分裂中的細胞數目及細胞凋亡的現象。過量表現CG17064基因對早期眼睛發育所造成的影響,最後會導致果蠅成蟲的眼睛變得小而粗糙。

Drosophila CG17064 is a novel gene that was identified in the Drosophila genome project. This gene was mapped at 50B1-2 on the second chromosome. The full-length cDNA contains 3053 nucleotides and translates to protein with 921 amino acids. BLAST result showed the central region (486-694) of CG17064 peptides contains a NES motif, which is highly conserved in human HURP. Using GMR-Gal4 line to drive transgenic flies to overexpress CG17064 in the eye disc increase the mitotic cells and apoptotic cells behind the furrow. The consequence of the early defect in eye disc impacts the normal eye development and resulted in a small and rough eye in GMR>CG17064 adult fly.

目錄……………………………………………………………………..1
中文摘要………………………………………………………………..3
英文摘要………………………………………………………………..4
前言……………………………………………………………………..5
材料與方法…………………………………………………………….11
果蠅品系………………………………………………………….11
CG17064基因之定序……………………………………………11
建構pUAST-CG17064轉殖基因………………………………..12
基因轉殖果蠅之選殖…………………………………….………13
組織免疫染色…………………………………………………….14
1.果蠅三齡幼蟲之免疫染色………………………………..14
2.果蠅蛹期眼睛免疫染色…………………………………..15
Propidum iodide染色…………………………………………….16
Acridine orange染色……………………………………………..16
原位雜合………………………………………………………….17
1.RNA探針之製備…………………………………………17
2.胚胎之原位雜合…………………………………………..17
3.眼盤之原位雜合…………………………………………..19
掃瞄式電子顯微鏡觀察………………………………………….21
結果…………………………………………………………………….22
果蠅CG17064基因………………………………………………22
CG17064在胚胎發育早期的表現……………………………….23
建構UAS-CG17064基因轉殖果蠅……………………………...24
GMR>CG17064導致果蠅複眼外觀形態的改變……………….25
GMR>CG17064會破壞早期果蠅眼睛的發育………………….26
GMR>CG17064導致分裂中的細胞數目增加………………….27
GMR>CG17064會促使細胞凋亡……………………………….28
GMR>CG17064對小眼發育後期的影響……………………….28
討論…………………………………………………………………….30
CG17064影響細胞週期的可能機制…………………………….30
CG17064導致細胞凋亡的可能機制….…………………………32
CG17064與細胞分裂之關係…………………...………………..34
參考文獻………………………………………………………………..38
圖表目錄………………………………………………………………..43
圖表說明………………………………………………………………..44

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