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Fig. 1 | Hereditas

Fig. 1

From: Overexpression of the Drosophila ATR homologous checkpoint kinase Mei-41 induces a G2/M checkpoint in Drosophila imaginal tissue

Fig. 1

Generation of a UAS-mei-41 overexpression line. (a) Cloning scheme of the pUAST-mei-41 construct. Coloured line corresponds to the mei-41 gene; intron positions are depicted as pale dashes. Segments derived from plasmid vectors are depicted in black and are not to scale. Genomic mei-41 DNA was PCR-amplified in fragments I-IV, which were eventually merged and shuttled into pUAST transformation vector in the correct 5′-3′ orientation. (b) UAS-mei-41 flies were crossed to en-Gal4-GFP, and induction of mei-41 expression was visualized in the posterior compartment of a wing imaginal disc by in situ hybridization (arrow). Size bar represents 100 μm. (c) Overexpression of mei-41 was quantified by qRT-PCR. To this end, UAS-mei-41 was ubiquitously induced with da-Gal4, and mRNA isolated from third instar larvae. Compared to control (da::lacZ), mei-41 is about 500-fold overexpressed. Data were assembled from three biological and two technical replicates. Mini-max depicts 95% confidence, median corresponds to expression ratio. Gapdh and beta-Tubulin56D served as reference genes. Efficiencies for mei-41 (0.92), for gapdh (0.94) and for beta-Tubulin56D (0.95) were accounted for determining relative quantities [44]

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