A modular misexpression screen in Drosophila detecting tissue-specific phenotypes
A modular misexpression screen in Drosophila detecting tissue-specific phenotypes
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DOI：
10.1073/pnas.93.22.12418
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发表时间：
1996-10-29
影响因子：
11.1
通讯作者：
Rorth, P
中科院分区：
文献类型：
--
作者：
Rorth, P
在果蝇中进行的基因筛选已经导致发现了许多对不同生物体的模式形成和信号转导非常重要的基因。传统上，人们分析的是功能缺失突变的表型效应。作为一种将基因与功能联系起来的替代方法，我在果蝇中开发了一种通用的异位表达筛选方法，这是在高等真核生物中的首个此类筛选。该筛选可鉴定出那些当以一种感兴趣的模式过度表达或异位表达时，会产生一种特定表型或调节一种现有突变表型的基因。它基于一个可移动的增强子和启动子的Gal4反式激活作用，该增强子和启动子“靶向”随机的内源基因以使其表达。这种筛选的模块化设计允许以任何时间或空间模式进行定向表达。当在发育中的眼睛中被激活时，4%的靶标插入产生了显性表型。其中一个插入位于编码Ras GTP酶激活蛋白的基因中；其过度表达表型被Ras1的一个突变强烈增强。因此，利用这种方法可以鉴定出具有生物学相关性的表型和遗传相互作用。这种筛选是发育遗传学的一种强大的新工具；类似的方法也可应用于其他生物体。
Genetic screens in Drosophila have Lead to the discovery of many genes important for patterning and signal transduction in diverse organisms. Traditionally, the phenotypic effects of loss-of-function mutations are analyzed. As an alternative way to link genes and function, I have developed a versatile misexpression screen in Drosophila, the first such screen in higher eukaryotes. The screen identifies genes that, when over- or misexpressed in a pattern of interest, give a specific phenotype or modulate an existing mutant phenotype. It is based on Gal4 transactivation of a mobile enhancer and promoter that ''targets'' random endogenous genes for expression. The modular design of the screen allows directed expression in any temporal or spatial pattern. When activated in the developing eye, 4% of target inserts gave dominant phenotypes. One insertion was in the gene encoding Ras GTPase-activating protein; its overexpression phenotype was strongly enhanced by a mutation in Ras1. Thus, biologically relevant phenotypes and genetic interactions are identified using this method. The screen is a powerful new tool for developmental genetics; similar approaches can also be developed for other organisms.