Published January 20, 2023 | Version v1

Comparison of tetracycline and temperature sensitive GAL80 transgenes with UAS-lacZ

  • 1. Queen's University

Description

ß-galactosidase specific activity was measured during development and aging (Experimental workflow) as described in STAR Protocols 3, 101843, 2022. Two different muscle-specific GAL4 drivers, Mef2-Gal4 (BDSC:27390) and DJ694 (BDSC: 8176), were each crossed with a second chromosome insertion of UAS-lacZ (Bg2) (BDSC: 1776), Bg2 recombined with third chromosome insertions of TetOFF-GAL80 (3.3+1077, PeerJ 5:e4167), and Bg2 recombined with third chromosome insertions of GAL80ts (BDSC:7017). Flies were raised at 20˚C and 29℃. Two independent sets of parents were used for each cross (biological replicates 1 and 2). After 2-3 days, parents were flipped into a new bottles (technical replicates 1 and 2 reported as replicate 3 and 4 in the dataset). CPRG and Bradford assays were performed on whole animal for the third instar larvae (L3), early pupae (EP) and late pupae (LP) stages, and on dissected thoraces for adult stages. Five extracts (1 individual in each) were measured per experimental replicate. Raw microplate readings are available upon request.

Notes

A small subset of this dataset (29˚C L3, EP and adults) has been previously used in the publication of the protocol to measure ß-galactosidase in Drosophila extracts (STAR Protocols 3, 101843, 2022)(https://data.mendeley.com/datasets/k2m2m3jgmn).

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CPRG GAL80ts repression-induction final DATASET.pdf

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Additional details

References

  • Barwell T, DeVeale B, Poirier L, Zheng J, Seroude F, Seroude L. 2017. Regulating the UAS/GAL4 system in adult Drosophila with Tet-off GAL80 transgenes. PeerJ 5:e4167 https://doi.org/10.7717/peerj.4167
  • Barwell T and Seroude L. 2022. Protocol to measure ß-galactosidase in Drosophila extracts using a CPRG assay. STAR Protocols 3:101843 https://doi.org/10.1016/ j.xpro.2022.101843