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Figure 3
Promoter-binding activity of osAca2. (a) Surface electrostatic features of dimeric osAca2. The scale bar ranges from −81.0 kT/e (red) to 81.0 kT/e (blue). The direction of the dimeric structure of osAca2 observed at the start of the rotation is shown by the cartoon figure on the left. (b) The overall shape of the surface of osAca2. (c) Putative osAca2 binding sites in the promoter region of the acr6 gene of O. smirnovii. Two inverted-repeat sequences (P1 and P2) upstream of the acrIF6 gene were identified by sequence analysis. (d) Mobility shift of the synthesized P1–P2 DNA probe containing two inverted sequences (P1 and P2), with increasing concentrations of osAca2, on an agarose gel. The synthesized nucleotide sequences are shown on the abovementioned gel. Concentration dependence is indicated by the black triangle. Lane M contains molecular size markers. The shifted DNA bands are indicated by a black arrow. (e) Sequence-specific interaction of osAca2. N1, N2 and N3 indicate negative controls representing DNA fragments containing no inverted sequences. (f, g) Mobility shift of the synthesized DNA probes P1 (f) and P2 (g) observed on an agarose gel. (h) Mobility shift of synthesized DNA probes observed on an agarose gel with increasing concentrations of A8K mutant osAca2. The DNA probes used in the experiment are indicated below the gel. Concentration dependence is indicated by the black triangle. Lane M contains molecular size markers. The concentrations of osAca2 protein are indicated for each lane in the gel.

Journal logoSTRUCTURAL
BIOLOGY
ISSN: 2059-7983
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