Doxycycline-induced transgene expression during Drosophila development and aging.
The "reverse" tetracycline repressor (rtR) binds a specific DNA element, the tetracycline operator (tetO), only in the presence of tetracycline, or derivatives such as doxycycline (dox). Fusion of rtR to the transcriptional activation domain of herpes virus protein VP16 produces a eukaryotic transactivator protein (rtTA). rtTA has previously been shown to allow dox-dependent transcription of transgenes linked to tetO sequences in mammals. To adapt this system to Drosophila, the Actin5C promoter was used to drive constitutive expression of rtTA in transgenic flies. Three reporter constructs, each encoding E. coli beta-galactosidase (beta-gal), were also introduced into transgenic flies. In one reporter seven tetO sequences were fused to the Adh core promoter. The other two reporter constructs contain seven tetO sequences fused to the hsp70 core promoter. Feeding of transgenic Drosophila containing the rtTA construct and any one of the three reporter constructs with dox caused up to 100-fold induction of beta-gal. Dox induced beta-gal expression in all tissues, in larvae and in young and senescent adults. Induction of beta-gal in adults had no detectable effect on life span. These results suggest the potential usefulness of this system for testing specific genes for effects on Drosophila development and aging.
Bieschke
E T
ET
Department of Biological Sciences, University of Southern California, Los Angeles 90089-1340, USA.
Wheeler
J C
JC
Tower
J
J
D000199:Actins / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004318:Doxycycline / Q000494:pharmacology*; D004330:Drosophila / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D008136:Longevity / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins / Q000235:genetics; D015533:Transcriptional Activation; D019076:Transgenes; D001616:beta-Galactosidase / Q000235:genetics
D000199:Actins; D000900:Anti-Bacterial Agents; D017405:Herpes Simplex Virus Protein Vmw65; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D001616:beta-Galactosidase; D004318:Doxycycline