An efficient promoter trap for detection of patterned gene expression and subsequent functional analysis in Drosophila
An efficient promoter trap for detection of patterned gene expression and subsequent functional analysis in Drosophila
Transposable elements have been used in Drosophila to detect gene expression, inactivate gene function, and induce ectopic expression or overexpression. We have combined all of these features in a single construct. A promoterless GAL4 cDNA is expressed when the construct inserts within a transcriptional unit, and GAL4 activates a GFP-encoding gene present in the same transposon. In a primary screen, patterned gene expression is detected as GFP fluorescence in the live progeny of dysgenic males. Many animals expressing GFP in distinct patterns can be recovered with relatively little effort. As expected, many insertions cause loss of function. After insertion at a genomic location, specific parts of the transposon can be excised by FLP recombinase, thus allowing it to induce conditional misexpression of the tagged gene. Therefore, both gain- and loss-of-function studies can be carried out with a single insertion in a gene identified by virtue of its expression pattern. Using this promoter trap approach, we have identified a group of cells that innervate the calyx of the mushroom body and could thus define a previously unrecognized memory circuit.
- King's College London United Kingdom
- Kings College London, University of London United Kingdom
- UCLA Clinical and Translational Science Institute United States
- University of California, Los Angeles United States
Male, 570, Transcription, Genetic, Recombinant Fusion Proteins, Gene Expression Regulation, Developmental, Promoter Regions, Mutagenesis, Insertional, Drosophila melanogaster, Gene Expression Regulation, Genetic, Mutagenesis, Insertional, Larva, DNA Transposable Elements, Animals, Developmental, Female, Promoter Regions, Genetic, Transcription, Transcription Factors
Male, 570, Transcription, Genetic, Recombinant Fusion Proteins, Gene Expression Regulation, Developmental, Promoter Regions, Mutagenesis, Insertional, Drosophila melanogaster, Gene Expression Regulation, Genetic, Mutagenesis, Insertional, Larva, DNA Transposable Elements, Animals, Developmental, Female, Promoter Regions, Genetic, Transcription, Transcription Factors
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