A new strategy for efficient in vivo screening of mutagenized Drosophila embryos.
The analysis of mutants is an indispensable approach towards characterizing gene function. Combining several tools of Drosophila genetics, we designed a new strategy for a mutagenesis screen which is fast, easy-to-apply, and cheap. The combination of a cell-specific Gal4 line with an upstream activating sequence-green fluorescent protein (UAS-GFP) allows the in vivo detection of the cells or tissues of interest without the need for fixation and staining. To further simplify and accelerate the screening procedure, we generated recombinant flies that carry the Gal80 transgene in balancer chromosomes. Gal80 inactivates Gal4; and thus prevents GFP-expression during embryonic and postembryonic development in all individuals carrying the balancer chromosomes. This allows for an easy distinction in vivo between heterozygous and homozygous mutants, the latter being the only ones expressing GFP. Since most of the fly strains and balancer chromosomes can be substituted, this method is suitable for nearly any mutagenesis screen that does not have major restrictions.
Vef
Olaf
O
Institute of Genetics, University of Mainz, Germany.
Cleppien
Diana
D
Löffler
Thomas
T
Altenhein
Benjamin
B
Technau
Gerhard M
GM
D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D002460:Cell Line; D002875:Chromosomes / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D004268:DNA-Binding Proteins; D004330:Drosophila / Q000196:embryology / Q000235:genetics* / Q000254:growth & development; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology / Q000737:chemistry; D004742:Enhancer Elements, Genetic / Q000235:genetics; D017344:Genes, Insect / Q000235:genetics*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D016296:Mutagenesis; D012097:Repressor Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; C055002:GAL80 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins