Reproducibility of ß-galactosidase specific activity measurements by CPRG and Bradford assays

ß-galactosidase specific activity was measured from two Drosophila genotypes. 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), generating Bg2/+;Mef2/+ and DJ694/Bg2 genotypes. Flies were raised at 29℃. Crosses were performed at 29℃ in bottles in duplicate, after 2 days bottles were flipped into a new set of bottles, effectively producing 4 experimental replicates. Progeny was maintained at at 29℃. Measurements were performed on whole third instar larvae (L3) and early pupae (EP), and 15 day old adult male and female thoraces (Adult 15). Five extracts (1 individual in each) were measured per experimental replicate. Raw microplate readings are available upon request.

本研究测定了两种果蝇（Drosophila）基因型的β-半乳糖苷酶（β-galactosidase）比活性。选取两种肌肉特异性GAL4驱动株（GAL4 driver）：Mef2-Gal4（BDSC:27390）与DJ694（BDSC:8176），分别与第二条染色体插入的UAS-lacZ（Bg2）（BDSC:1776）进行杂交，获得Bg2/+;Mef2/+与DJ694/Bg2两种基因型子代。所有果蝇均在29℃条件下饲养。杂交实验于29℃下在培养瓶中以双重复开展，培养2天后将亲本瓶转移至新的一批培养瓶，最终得到4个实验重复样本。子代果蝇均维持在29℃条件下培养。检测样本涵盖完整三龄晚期幼虫（L3）、早期蛹（EP）以及15日龄成虫的雌雄胸肌组织（Adult 15）。每个实验重复样本均取5只个体（每份提取物对应1只果蝇）进行比活性测定。原始酶标板读数可应要求提供。