DNA display of biologically active proteins for in vitro protein selection.
In vitro display technologies are powerful tools for screening peptides with desired functions. We previously proposed a DNA display system in which streptavidin-fused peptides are linked with their encoding DNAs via biotin labels in emulsion compartments and successfully applied it to the screening of random peptide libraries. Here we describe its application to functional and folded proteins. By introducing peptide linkers between streptavidin and fused proteins, we achieved highly efficient (>95%) formation of DNA-protein conjugates. Furthermore, we successfully enriched a glutathione-S-transferase gene by a factor of 20-30-fold per round on glutathione-coupled beads. Thus, DNA display should be useful for rapidly screening or evolving proteins based on affinity selection.
Yonezawa
Masato
M
Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522.
Doi
Nobuhide
N
Higashinakagawa
Toru
T
Yanagawa
Hiroshi
H
D001710:Biotin / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D019020:Directed Molecular Evolution; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D017510:Protein Folding; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D019151:Peptide Library; D011506:Proteins; D011993:Recombinant Fusion Proteins; D001710:Biotin; D004247:DNA; D019809:Streptavidin; D005982:Glutathione Transferase; D005978:Glutathione