DNA Display of Biologically Active Proteins for In Vitro Protein Selection

Masato Yonezawa, Nobuhide Doi, Toru Higashinakagawa, Hiroshi Yanagawa

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)285-288
Number of pages4
JournalJournal of Biochemistry
Volume135
Issue number3
DOIs
Publication statusPublished - 2004 Mar

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Display devices
Screening
Streptavidin
DNA
Peptides
Proteins
Peptide Library
Biotin
Glutathione Transferase
Emulsions
Glutathione
Labels
Genes
Technology
In Vitro Techniques

Keywords

  • Biotin
  • DNA display
  • In vitro translation
  • Protein engineering
  • Streptavidin

ASJC Scopus subject areas

  • Biochemistry

Cite this

DNA Display of Biologically Active Proteins for In Vitro Protein Selection. / Yonezawa, Masato; Doi, Nobuhide; Higashinakagawa, Toru; Yanagawa, Hiroshi.

In: Journal of Biochemistry, Vol. 135, No. 3, 03.2004, p. 285-288.

Research output: Contribution to journalArticle

Yonezawa, Masato ; Doi, Nobuhide ; Higashinakagawa, Toru ; Yanagawa, Hiroshi. / DNA Display of Biologically Active Proteins for In Vitro Protein Selection. In: Journal of Biochemistry. 2004 ; Vol. 135, No. 3. pp. 285-288.
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