Gpdh gene duplication and deletion by piggyBac FLP–FRT site-specific recombination and effects on ethanol tolerance. (A) The GPDH activities as ΔOD units for seven piggyBac (pB) alleles recovered in the FRT–FLP recombination crosses and confirmed by diagnostic PCR and direct sequencing. All deletions are semilethal and balanced over CyO. (B) Experiment 5 showing the percentage of survival of three Gpdh genotypes possessing 100% (n = 34), 125% (n = 28), and 150% (n = 24) relative GPDH activity to normal. Among genotypes, differences are statistically significant by ANOVA on arc–sine transformed values (F2,83 = 7.44, ***P < 0.001). Error bars represent ±1 SE.