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. 2003 Mar;23(6):1968–1982. doi: 10.1128/MCB.23.6.1968-1982.2003

FIG. 8.

FIG. 8.

K88R mutant protein has decreased DNA binding to the bicoid consensus site. (A) Schematic indicating the lysine-to-arginine change in PITX2a K88R. (B) Increasing amounts (100, 200, and 300 ng) of WT PITX2a and K88R proteins were incubated with the Bcd 1x probe (containing the bicoid consensus sequence TAATCC) in an EMSA. The bound complex is indicated. The lower band seen with the probe alone is most likely a multimer of the free probe that has been run off this gel. (C) Silver-stained gel of the PITX2a and K88R protein preparations following removal of the GST moiety, with WT and K88R proteins (arrow) and molecular weight markers indicated. (D) SAAB results for GST-PITX2a or GST-K88R proteins with a probe containing a degenerate (TAATNN) binding sequence are summarized. The overall frequency of each nucleotide occurring at either position 5 or position 6 is shown. For WT PITX2a, a total of 44 sequences were analyzed. For K88R, a total of 43 sequences were analyzed.