Figure 1.

Identification of a Nuclear Protein That Interacts with the 27-kD γ-Zein Gene Promoter.

(A) EMSA of four probes (P1–P4) from the 27-kD γ-zein promoter with nuclear extract from 15 DAP endosperm. Upper panel shows a schematic representation of the 27-kD γ-zein promoter fragments. Arrowhead points to the nuclear extract-induced shifted bands. P1–P4, Probe1 to Probe 4; N.E., nuclear extract.

(B) EMSA of four probes truncated from P1. Upper panel shows a schematic representation of the P1 fragments. Arrowhead points to the nuclear extract-induced shifted band.

(C) Nuclear proteins from 15-DAP endosperm purified by affinity chromatography using P1-3 (lane 1) and P1-m3 (lane 2) as baits. Proteins were separated by SDS-PAGE and visualized by silver staining. The arrow points to the protein band specifically purified by P1-3. Numbers on the right indicate the positions of the molecular mass standards.

(D) EMSA of ZmbZIP22 with probes containing the second mutation site of P1-m3. DNA probe containing the second mutation site (WT) and a series of probes each with 1-bp point mutations across the 10-bp segment centered around the second mutation site (MP1–MP10) were used. Arrowhead points to the ZmbZIP22-induced shifted bands.