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.
Larsen
Camilla
C
University of California, Life Sciences Building 4214, Los Angeles, CA 90025, USA.
Franch-Marro
Xavier
X
Hartenstein
Volker
V
Alexandre
Cyrille
C
Vincent
Jean-Paul
JP
D000818:Animals; D004251:DNA Transposable Elements; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000196:embryology / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D007814:Larva / Q000033:anatomy & histology / Q000502:physiology; D008297:Male; D016254:Mutagenesis, Insertional; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D004251:DNA Transposable Elements; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors