Affinity selection of DNA-binding proteins from yeast genomic DNA libraries by improved lambda phage display vector

J Biochem. 2002 Dec;132(6):975-82. doi: 10.1093/oxfordjournals.jbchem.a003312.

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 genome-wide screening of DNA-binding proteins, we constructed yeast genomic libraries using lambda foo-based vectors devised in this work. After affinity selection using GAL4 UAS(G) as a probe, phages expressing GAL4 were enriched approximately 5 x 10(5)-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage lambda / genetics*
  • Bacteriophage lambda / metabolism*
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Gene Library
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Genome, Fungal*
  • Peptide Library*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • DNA, Fungal
  • DNA-Binding Proteins
  • GAL4 protein, S cerevisiae
  • Peptide Library
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors