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研究生:黃育誠
研究生(外文):Yu-Cheng Huang
論文名稱:果蠅CG9669在細胞中的位置和功能
論文名稱(外文):Localization and function of the Drosophila CG9669
指導教授:黃敏郎黃敏郎引用關係
指導教授(外文):Min-Lang Huang
口試委員:陳永恩沈國愉
口試委員(外文):Michael ChanKuo-Yu Shen
口試日期:2011-07-21
學位類別:碩士
校院名稱:國立中正大學
系所名稱:分子生物研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:英文
論文頁數:34
外文關鍵詞:CG9669
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果蠅的CG9669是一個沒人研究過的基因,在不同物種中普遍可見到與CG9669相類似的基因。CG9669有兩個未經證實的transmembrane domain,並且目前已知道CG9669會和一個核膜蛋白Klarsicht (Klar)結合,因此CG9669可能是一個核膜蛋白。在我們的實驗裡,我們利用過度表達CG9669於不同的組織裡來研究CG9669在細胞中的位置。我們發現CG9669的位置會和核膜蛋白Lamin Dm0 (Lamin) 及Klar重疊,因此推論CG9669應該位於在細胞核膜上。另外我們也發現CG9669出現在細胞質的膜狀結構內,總結上面的結果,推測出CG9669應該可以出現在細胞核膜和細胞質中一些膜狀的結構上面。此外我們發現過度或是降低表達CG9669時會增加細胞核膜的周長,而當降低表達CG9669時則是使改變細胞核的結構。最終我們推測CG9669是一個細胞核膜和細胞質裡都有的膜蛋白,在細胞膜上的CG9669可能有調節細胞核大小的功能。
The Drosophila CG9669 is a novel gene conserved in different species. CG9669 has two putative transmembrane domains. It is known that CG9669 interacts to the Klarsicht (Klar), which locates in outer nuclear envelope and required for nuclear positioning of photoreceptors. We studied the localization of CG9669 in different tissues by overexpression of CG9669. We found that CG9669 colocalized with the nuclear membrane protein Lamin and Klar, suggesting that CG9669 locates in nuclear envelope. CG9669 was also localized at the membranous structures in the cytoplasm. We speculated that CG9669 could integrate into the nuclear membrane and some membranous structures in the cytoplasm. We found that overexpressing and silencing CG9669 increased the nuclear perimeter. Silencing CG9669 could alter the nuclear structure. We demonstrated that CG9669 located at the nuclear membrane and membranous structure in cytoplasm, and CG9669 on the nuclear membrane could regulate the size of the nuclei.
Introduction............................................................................................1
Result..................................................................................................4
The flies of overexpressing and silencing CG9669 .......................................................4
The localization of CG9669 in the neurons ..............................................................4
The localization of CG9669 in the muscles ..............................................................6
The phenotype of overexpressing or silencing CG9669 ....................................................7
Discussion..............................................................................................9
The localization of CG9669 .............................................................................9
The localization of CG9669 and Lamin with treating digitonin ...........................................9
The Rab4 changed the localization of CG9669 ...........................................................10
The phenotype of silencing CG9669......................................................................12
Overexpressing or silencing CG9669 induced the increase of nuclear size................................12
The stunned flies of overexpression of CG9669..........................................................13
Figures................................................................................................14
Figure. 1 The transmembrane domain of CG9669...........................................................14
Figure. 2 The localization of CG9669 in eye imaginal discs.............................................15
Figure. 3 The localization of CG9669 in PNS neurons....................................................16
Figure. 4 The localization of CG9669 and endosome in PNS neurons.......................................17
Figure. 5 The localization of CG9669 in the muscles....................................................18
Figure. 6 The localization of CG9669 and Lamin after treating digitonin in the muscles.................19
Figure. 7 The localization of CG9669, Rab4 and Rab5 in the muscles.....................................20
Figure. 8 The appearance of mislocated Lamin in overexpressing CG9669 and Rab4.........................21
Figure. 9 The phenotype of CG9669-Ri...................................................................22
Figure. 10 The nuclear size of WT, 24B>CG9669-CFP, 24B>CG9669-Ri and 24B-Gal4..........................23
Figure. 11 The spots on the surface of nuclear membrane of WT, 24B>CG9669-CFP and 24B>CG9669-Ri........24
Figure. 12 The lifespan test of overexpressing CG9669 and control......................................25
Materials and Methods..................................................................................26
Fly stocks.............................................................................................26
The transgenic flies for overexpressing or silencing CG9669............................................26
Immunostaining of eye imaginal disc ...................................................................27
Immunostaining of larval PNS...........................................................................28
Immunostaining of Muscle...............................................................................28
Preabsorbtion of the antibody .........................................................................29
Treatment of digitonin.................................................................................29
Lifespan Test..........................................................................................29
Calculation of the nuclear size in larva muscle........................................................30
References.............................................................................................31

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