FIGURE 5.

Characterization of novel GST from A. nidulans. A, SDS-PAGE of purified enzyme. Purified enzymes (1 μg) were resolved by SDS-PAGE on 10% polyacrylamide gels and stained with Coomassie Brilliant Blue. Lane 1, markers (Novex Sharp Protein Standard kit, Invitrogen); lane 2, rGstB. B, Quantitative PCR analysis of gstB gene transcript. A. nidulans A26 was grown at 30 °C for 20 h, the indicated compounds were added or temperatures were shifted, and then incubation proceeded for further 3 h. Bars indicate values reported as relative expression rates obtained by RT-PCR and normalized to 18 S rRNA. C, phylogenetic relationships among GstB-like GSTs from various organisms. Multiple alignment and neighbor joining were performed using Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0. Accession numbers are from Broad Institute. GstA involved in cluster 2 GST from A. nidulans served as outgroup.