Affinity selection of DNA-binding proteins from yeast genomic DNA libraries by improved lambda phage display vector.
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.
Hagiwara
Hiroko
H
Research Center for Glycoscience, Advanced Institute of Industrial Science and Technology, Central Higashi, Tsukuba, Ibaraki 305-8566, Japan.
Kunihiro
Sumiko
S
Nakajima
Keiichi
K
Sano
Motoaki
M
Masaki
Haruhiko
H
Yamamoto
Midori
M
Pak
Jeong Won
JW
Zhang
Yan
Y
Takase
Kumiko
K
Kuwabara
Ichiro
I
Maruyama
Ichiro N
IN
Machida
Masayuki
M
D010582:Bacteriophage lambda / Q000235:genetics* / Q000378:metabolism*; D004271:DNA, Fungal / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D016681:Genome, Fungal; D019151:Peptide Library; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D019151:Peptide Library; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors