Fig 8.

Zfhep HD and ZHD cannot select a DNA binding site. (A) Zfhep HD did not select a DNA binding site from a pool of random oligonucleotides. Oligonucleotides obtained after eight rounds of binding were radiolabeled with ³²P and used in EMSA with either 5 μl (first lane) or 10 μl (second lane) of GST-HD or MBP-ZFC (positive control) fusion proteins. (B) The Zfhep ZHD did not select a DNA-binding site after five rounds of binding. GST-ZHD or GST-POU (the positive control) were used for five rounds of oligonucleotide selection. The resulting oligonucleotides were labeled and incubated with either 5 μl (first lane) or 10 μl (second lane) of the indicated GST-fusion protein. The binding reactions were separated on a 5% non-denaturing gel, and exposed to film. Labeled oligonucleotides without protein are run on adjacent lanes (DNA). Unbound oligonucleotides migrate to the bottom of the gel (Free DNA).