Affinity Selection of DNA-Binding Proteins from Yeast Genomic DNA Libraries by Improved λ Phage Display Vector

Abstract

Phage display is a useful means of identifying and selecting proteins of interest that bind specific targets. In order to examine the potential of phage display for the genomewide screening of DNA-binding proteins, we constructed yeast genomic libraries using λfoo-based vectors devised in this work. After affinity selection using GAL4 UAS_G as a probe, phages expressing GAL4 were enriched approximately 5×10⁵-fold from the library. Approximately 90% of polypeptides encoded in correct translation reading frames by the selected phages were known or putative polynucleotide-binding proteins. This result clearly indicates that the modified lambda phage display vector in combination with our enrichment technique has great potential for the enrichment of DNA-binding proteins in a sequence-specific manner.