Gerlitz Offer, Nellen Denise, Ottiger Michael, Basler Konrad
Institut für Molekularbiologie, Universität Zürich, Switzerland.
Int J Dev Biol. 2002 Jan;46(1):173-6.
The development of Drosophila imaginal discs serves as a model system to understand how genes determine the shape and size of an organ. The identification of genes involved in this process is an important step towards this goal. Here we describe a P-element based enhancer trap screen for genes expressed in the larval imaginal discs. Our aim was to establish a large collection of enhancer trap lines each showing expression of Gal4 in imaginal discs. To this end, we improved the well established P-element vector pGawB in order to obtain higher in vivo transposition frequencies. In addition we chose an F1-screening approach using UAS-GFP as a reporter gene. This system permits the efficient screening of larval and pupal stages of living animals and the detection of imaginal gene expression patterns through the transparent cuticle. The procedure has been optimized for high-throughput. 2'000 P-element insertions have been established which exhibit expression in imaginal discs.
果蝇成虫盘的发育是理解基因如何决定器官形状和大小的一个模型系统。鉴定参与这一过程的基因是朝着这个目标迈出的重要一步。在这里，我们描述了一种基于P因子的增强子陷阱筛选方法，用于筛选在幼虫成虫盘中表达的基因。我们的目标是建立大量的增强子陷阱品系，每个品系都在成虫盘中显示Gal4的表达。为此，我们改进了已得到广泛应用的P因子载体pGawB，以获得更高的体内转座频率。此外，我们选择了一种使用UAS-GFP作为报告基因的F1筛选方法。这个系统允许对活体动物的幼虫和蛹阶段进行高效筛选，并通过透明角质层检测成虫基因的表达模式。该程序已针对高通量进行了优化。现已建立了2000个在成虫盘中表现出表达的P因子插入系。
