A genetic method for generating Drosophila eyes composed exclusively of mitotic clones of a single genotype.
A genetic method for generating Drosophila eyes composed exclusively of mitotic clones of a single genotype.
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作者：
R. Stowers;T. Schwarz
如果在早期发育阶段需要该基因，则该基因在发育后期的遗传分析可能会受到阻碍。镶嵌动物的构建，特别是果蝇，是克服这一障碍的一种手段。然而，有丝分裂克隆的表型分析通常很复杂，因为产生有丝分裂克隆的标准方法会产生由突变细胞和表型正常细胞组成的嵌合组织。我们在这里描述了一种遗传方法（称为 EGUF/hid），该方法使用 GAL4/UAS 和 FLP/FRT 系统来克服果蝇眼睛的这一限制，方法是产生遗传嵌合果蝇，这些果蝇原本是杂合的，但其中眼睛完全由五个主要染色体臂之一的纯合细胞组成。这些眼睛的大小、形态和生理学都接近野生型。这种遗传方法的应用包括现有突变的表型分析和 F(1) 遗传筛选，以识别与蝇眼生物学有关的未知基因。我们通过将其应用于突触传递基因突触结合蛋白和突触蛋白的致死突变来说明该方法的实用性。
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.