整合不同之果蠅雙元基因表現系統用以精確調控基因於特定時間及空間下表現

Through the two binary gene expression systems for spatiotemporal gene expression in Drosophila

塗喬惠(Chiao-Hui Tu)

指導教授 ： 傅在峰

國立暨南國際大學/科技學院/應用化學系/碩士(2011年)

https://doi.org/10.6837/NCNU.2011.00137

摘要

在以果蠅為模式動物的研究中，GAL4/UAS 雙元基因表現系統(binary expression system)是廣泛被運用在限制目標基因於特定空間表現的技術，操作上若與GAL80ts 結合使用(即TARGET system)，經由溫度的變化，則更能將基因限制表達在特定的時間與空間。遺憾的，在一些以果蠅為模式動物的研究證據中顯示，溫度的變化會對某些實驗結果造成影響。因此，在本研究中，我們建立一個新的基因調控策略，利用已被廣泛應用於果蠅的GAL4/UAS 系統，表現一個必須經由RU486(或17β-estradiol)活化之轉錄因子(transactivator)- LexPR(或XVE)，在特定時間下經由藥物活化後，即可藉此為橋樑連結至另一雙元基因表現系統，轉錄LexAop 下游之目標基因。藉此方法，將可以避免因使用GAL80ts 調控基因表現時，因溫度改變對研究結果所產生的不良影響。本論文透過定性及定量分析，證明此方法的確可運用於果蠅模式動物，此方法的建立因保留GAL4 driver之豐富性，並減少因溫度改變而造成實驗分析的影響，達到限制基因於特定空間與時間之表現，可以預見對後續以果蠅為模式動物之基因研究有著重要的影響。

關鍵字

果蠅 ； 雙元基因表現系統 ； LexPR

並列摘要

The GAL4/UAS gene expression system is applied frequently in Drosophila. Combining the conventional GAL4/UAS system and temperature-sensitive Gal80(GAL80ts) is able to regulate gene expression through temperature alteration. However, the target gene expression can never be completely restricted neither with temperature changing nor insertion at low basal activity loci. In this study, I generate a UAS-LexA-progesterone receptor chimera (known as LexPR) which under control by GAL4 and transactivate another binary LexA-LexAop gene-expression system when hormones mifepristone (RU486) is added. This new technique provides guidelines for
acutely control gene expression through drug treatment, rather than altering temperature. Beside, the abundant GAL4 library can be applied to the new system. In brief, I provide a new strategy to acutely control and regulate the gene expression without affecting their physiological homeostasis in Drosophila.

並列關鍵字

Drosophila ； Binary gene expression system ； LexPR

參考文獻

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