EMSA of wild type and mutant Czf1p binding to the <i>CZF1</i> promoter region.

Abstract

<p>(Panel A) SDS PAGE analysis of 500 ng of purified wild type and mutant GST-Czf1 fusion proteins stained with Coomassie Blue. (Panel B) Diagram of the <i>CZF1</i> promoter region (not drawn to scale). The DNA fragment used in this study (fragment E, 565 bp) is represented as a black rectangle and is located −3381 to −2816 from the ATG start of the <i>CZF1</i> ORF. The transcriptional start site is represented as an arrow and is located at −2065 from the ATG <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039624#pone.0039624-Vinces1" target="_blank">[23]</a>. (Panels C–E) Increasing amounts of GST-Czf1p were incubated with ³²P end-labelled fragment E. The samples were analyzed by electrophoretic mobility shift assay and phosphorimaging. Brackets indicate shifted fragments. All EMSA experiments were repeated at least 3 times and a representative experiment is shown.</p