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. 2008 Jul 23;82(19):9700–9716. doi: 10.1128/JVI.00862-08

FIG. 3.

FIG. 3.

ORF50(1-590) purified from E. coli displays limited DNA-binding activity compared to its counterpart with the KK/EE mutation. (A) Coomassie blue staining and immunoblotting (WB) of purified ORF50(1-590) without or with the KK/EE mutation. (B) Binding of purified ORF50(1-590), ORF50(1-590)(KK/EE), or ORF50(333-691) to the PAN promoter. Increasing amounts (50, 150, 450, and 1,500 ng) of purified protein were used in an EMSA. (C) Binding specificity of purified ORF50(1-590) with the KK/EE mutation to DNA. Wild-type or mutant PAN elements served as a cold competitor (20×, 50×, and 100×) in EMSA. Antibody to ORF50(230-250) was used for supershift analysis.