Skip to main content
. 2011 Mar 31;7(3):e1001328. doi: 10.1371/journal.ppat.1001328

Figure 4. Evidence for specific TgNF7 protein-DNA interactions within the promoter of ENO1.

Figure 4

A) Expression and purification of recombinant TgNF1, TgNF2, TgNF3, TgNF4 and TgNF7 proteins fused to GST. The recombinant factors tested were marked with asterisks. B) Electrophoretic band shift assays using recombinant GST alone (lane 1), free probe (lane 2), rTgNF4 (lane 3), rTgNF1 (lane 4), rTgNF2 (lane 5), rTgNF7 (lane 6), rTgNF3 (lane 7) and the biotinylated probe from ENO1 promoter described in Figure 1. C) Purification of rTgNF7 lacking the GST (lane 1), which has been removed by digestion with the PreScission protease. D) Gel shift binding assay using the 47-bp probe at the ENO1 promoter shown in Figure S1 (red) and pure rTgNF3. Lane 1 corresponds to probe alone; lane 2, DNA-rTgNF7 complex; lane 3, specific unlabelled competitor at 10-fold excess; lane 4, specific unlabelled competitor at 50-fold excess. E) Lane 2, unrelated 27-bp probe containing one G nucleotide with pure rTgNF7; lane 1, probe alone. F) Lane 2, unrelated probe containing 27-bp probe containing one GGGGG motif with pure rTgNF7; lane 1, probe alone. G) unrelated probe containing 27-bp probe containing two GGGGG motifs with pure rTgNF7; lane 1, probe alone.