Figure 10.
Binding of the N termini of OsWRKY76 and OsWRKY62 to the W box element in the promoter of OsWRKY76. A, Nucleotide sequence of 81P1 used in DNA-binding experiments, with the core W box sequences underlined. B, OW76.1N, OW76.2N, OW62.1N, and OW62.2N represent the GST-tagged recombinant proteins of OsWRKY76.1N (amino acids 1–230, C-terminal 97 amino acids deleted), OsWRKY76.2N (amino acids 1–158, C-terminal 97 amino acids deleted), OsWRKYW62.1N (amino acids 1–205, C-terminal 113 amino acids deleted), and OsWRKY62.2N (amino acids 1–166, C-terminal 113 amino acids deleted), respectively. 81P1-B, Biotin-labeled 81P1; 50×, addition of 50-fold unlabeled 81P1; +, presence; −, absence. The reaction mixture was separated on a native PAGE gel and blotted on a nylon membrane. The 81P1-B probe was detected by anti-biotin antibody. C, Loading control indicating the amounts of proteins used in the DNA-binding assay, stained by Coomassie Brilliant Blue. M, Protein molecular weight markers.