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J. Biochem, 2004, Vol. 135, No. 3 285-288
© 2004 The Japanese Biochemical Society

BIOTECHNOLOGY

DNA Display of Biologically Active Proteins for In Vitro Protein Selection

Masato Yonezawa1,2, Nobuhide Doi1, Toru Higashinakagawa2 and Hiroshi Yanagawa*,1

1 Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522; 2 Department of Biology, School of Education, Waseda University, 1-6-1 Nishi-waseda, Shinjuku-ku, Tokyo 169-8050

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.

* To whom correspondence should be addressed. Tel: +81-45-566-1775, Fax: +81-45-566-1440, E-mail: hyana{at}bio.keio.ac.jp


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