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Rice
Fig. 3 | Rice

Fig. 3

From: Genome-wide identification of grain filling genes regulated by the OsSMF1 transcription factor in rice

Fig. 3

Identification of OsSMF1 binding motifs in vitro. a, c, e EMSA-based competition analysis of OsSMF1. Sequences of oligonucleotides used as probes and competitors are depicted. Wild type, 21-bp sequences containing the GCN4 (a), ACGT (c), and ATGA motifs (e); and M1 to M6, 21-bp sequences with three nucleotide mutations. The GCN4, ACGT, and ATGA motifs are underlined. The OsSMF1-DsRed fusion protein was used for EMSA with the 21-bp sequences. Competitors were added in 100-fold molar excess. Lane 1, no protein; Lane 2, no competitor; and lanes 3 to 9, with competitors. b, d, f Accurate determination of the dissociation constant K d by gel shift assays using SYBR Gold. The DNA-binding band intensities determined using Gel-Pro Analyzer program were plotted against the various DNA substrate concentrations. The measured K d values were 0.6458 μM for the binding of OsSMF1 to the GCN4 motif (b), 0.3353 μM for ACGT (d), and 1.117 μM for ATGA (f)

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