• 研究论文 •

### 灵活操控果蝇翅芽中wingless基因表达水平的策略

1. (山西大学应用生物学研究所, 太原 030006)
• 出版日期:2019-09-20 发布日期:2019-09-03

### A strategy for flexibly modulating wingless expression levels in Drosophila wing disc

ZHANG Xu-Bo, WU Wen-Jun, LI Kai-Xia, MA En-Bo, WU Hai-Hua, ZHANG Jian-Zhen, DONG Wei*

1.  (Institute of Applied Biology, Shanxi University, Taiyuan 030006, China)
• Online:2019-09-20 Published:2019-09-03

【目的】灵活操控靶基因的表达水平对于研究基因的功能十分重要。Gal4/UAS系统已被广泛应用于调控基因表达，可研究果蝇Drosophila等模式生物复杂的生物学问题。受采用载体的特性及插入位点的影响，Gal4或UAS转基因品系在构建好之后，其调控靶基因的能力基本是确定的。本研究旨在在现有Gal4/UAS系统的基础上，开发一种新的策略，实现在果蝇翅芽中灵活操控wingless(wg)基因的表达水平。【方法】用遗传学手段将黑腹果蝇Drosophila melanogaster品系的UAS-wg和UAS-wg-RNAi转基因重组到同一黑腹果蝇品系中。将该重组黑腹果蝇品系与dpp-Gal4黑腹果蝇品系杂交，同时驱动UAS-wg和UAS-wg-RNAi在果蝇幼虫翅芽中共表达。杂交子代幼虫分别放置在不同的温度(18, 25和30℃)下培养。将幼虫翅芽解剖并进行免疫组化染色，测量染色的荧光强度，分析翅芽中wg的表达水平。【结果】在低温(18℃)下，UAS-wg在基因表达调控中起主要作用，wg表现为超表达，但其超表达的效率可被UAS-wg-RNAi有效地削弱。相反，在高温(30℃)下，UAS-wg-RNAi起主导作用，wg的表达受到抑制。并且通过转换温度，可实现wg在翅芽发育的不同阶段在超表达和抑制之间相互转化，从而灵活地操控wg基因在翅芽中的表达水平。【结论】该方法可以灵活操控果蝇翅芽中wg基因的表达水平，对于调控转基因的表达有重要的意义。

Abstract:  【Aim】 Flexibly controlling the expression levels of target genes is critical for the study of gene function. Gal4/UAS system has been intensively used to manipulate gene expression and to study complex biological problems of model organisms such as Drosophila. Affected by the properties of transposon vector constructs and the insertion site in the transgenic stock, the ability of Gal4 or UAS transgenic lines to regulate target genes is roughly predefined after construction. This study aims to develop a new strategy based on the conventional GAL4/UAS system to flexibly modulate the expression level of wingless (wg) gene in Drosophila wing discs. 【Methods】 UAS-wg and UAS-wg-RNAi transgenes of Drosophila melanogaster were recombined into the same D. melanogaster line by genetic approach. And this line was crossed with dpp-Gal4 line to co-express UAS-wg and UAS-wg-RNAi in larval wing discs. The progeny larvae were cultured at different temperatures (18, 25 and 30℃). The larval wing discs were dissected and stained by immunohistochemistry. The fluorescence intensity was measured to analyze the expression level of wg in the wing discs. 【Results】 UAS-wg played a dominant role in the regulation of gene expression at low temperature (18℃). In this case, the efficiency of overexpression of wg was attenuated effectively by UAS-wg-RNAi. On the contrary, UAS-wg-RNAi predominated in the regulation of gene expression, and the expression of wg was inhibited at high temperature (30℃). Besides, by switching temperature, the wg expression could be transformed between overexpression and repression in wing disc at different developmental stages, and the expression of wg could be modulated flexibly in wing disc. 【Conclusion】 By this approach, we can flexibly modulate the expression of wg in wing disc of Drosophila. It has great significance in target gene regulation.