Stowers R S, Schwarz T L
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California 94305, USA.
Genetics. 1999 Aug;152(4):1631-9. doi: 10.1093/genetics/152.4.1631.
The genetic analysis of a gene at a late developmental stage can be impeded if the gene is required at an earlier developmental stage. The construction of mosaic animals, particularly in Drosophila, has been a means to overcome this obstacle. However, the phenotypic analysis of mitotic clones is often complicated because standard methods for generating mitotic clones render mosaic tissues that are a composite of both mutant and phenotypically normal cells. We describe here a genetic method (called EGUF/hid) that uses both the GAL4/UAS and FLP/FRT systems to overcome this limitation for the Drosophila eye by producing genetically mosaic flies that are otherwise heterozygous but in which the eye is composed exclusively of cells homozygous for one of the five major chromosome arms. These eyes are nearly wild type in size, morphology, and physiology. Applications of this genetic method include phenotypic analysis of existing mutations and F(1) genetic screens to identify as yet unknown genes involved in the biology of the fly eye. We illustrate the utility of the method by applying it to lethal mutations in the synaptic transmission genes synaptotagmin and syntaxin.
如果一个基因在发育早期是必需的,那么在发育后期对该基因进行遗传分析可能会受到阻碍。构建镶嵌动物,尤其是在果蝇中,一直是克服这一障碍的一种方法。然而,有丝分裂克隆的表型分析往往很复杂,因为产生有丝分裂克隆的标准方法会产生镶嵌组织,这些组织是突变细胞和表型正常细胞的复合体。我们在此描述一种遗传方法(称为EGUF/hid),该方法利用GAL4/UAS和FLP/FRT系统克服果蝇眼睛的这一局限性,通过产生基因镶嵌果蝇,这些果蝇在其他方面是杂合的,但眼睛完全由五个主要染色体臂之一的纯合细胞组成。这些眼睛在大小、形态和生理上几乎是野生型的。这种遗传方法的应用包括对现有突变的表型分析以及F(1)遗传筛选,以鉴定果蝇眼睛生物学中尚未知晓的基因。我们通过将该方法应用于突触传递基因突触结合蛋白和 syntaxin 中的致死突变来说明该方法的实用性。