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. 2016 Mar 16;6(5):1327–1343. doi: 10.1534/g3.116.028506

Figure 5.

Figure 5

Recombinant SEB-1 binds specifically to the N. crassa STRE motif in vitro. (A) Schematic representation of the gsn gene and the STRE positions in the promoter region. The sequences of the STRE1 and STRE2 DNA elements are represented. The STRE1 DNA oligonucleotide containing the motif sequence (underlined) and the STRE1 probe with mutations in the STRE core sequence (underlined) are also represented. T: transcription start site (Freitas and Bertolini 2004). (B) DNA probes containing both STRE motifs present at the 5′-flanking region of the gene gsn (STRE1 and STRE2) were assayed for DNA-binding activity using the GST-tagged recombinant SEB-1 as a protein source. The specificity of the DNA shifts was demonstrated by the prior addition of specific competitors: the cold probes STRE1 and STRE2, the dsDNA oligonucleotide STRE1, and the dsDNA fragment containing the mutated STRE1 motif. SC: specific competitor (unlabeled probes STRE1 and STRE2); SC oligo STRE1: specific competitor corresponding to a short dsDNA oligonucleotide (18 bp) containing the STRE1 motif; mSRE1: probe STRE1 containing the mutated STRE1 motif. dsDNA, double-stranded; GFP, green fluorescent protein; STRE, stress response element.