A genetic method for generating Drosophila eyes composed exclusively of mitotic clones of a single genotype.
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.
Stowers
R S
RS
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California 94305, USA.
Schwarz
T L
TL
D000818:Animals; D002135:Calcium-Binding Proteins; D019070:Cell Lineage; D002875:Chromosomes / Q000235:genetics / Q000648:ultrastructure; D002999:Clone Cells; D003433:Crosses, Genetic; D004331:Drosophila melanogaster / Q000235:genetics*; D004596:Electroretinography; D005123:Eye / Q000166:cytology*; D005804:Genes, Lethal; D005821:Genetic Techniques; D005838:Genotype; D019476:Insect Proteins / Q000235:genetics / Q000502:physiology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology; D008855:Microscopy, Electron, Scanning; D009030:Mosaicism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology; D050765:Qa-SNARE Proteins; D011995:Recombination, Genetic; D050857:Synaptotagmins
D002135:Calcium-Binding Proteins; D019476:Insect Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050765:Qa-SNARE Proteins; D050857:Synaptotagmins