%0 Journal Article
%T Hishot Display¡ªA New Combinatorial Display for Obtaining Target-Recognizing Peptides
%A Shoutaro Tsuji
%A Makiko Yamashita
%A Taihei Kageyama
%A Takashi Ohtsu
%A Katsuo Suzuki
%A Shintaro Kato
%A Joe Akitomi
%A Makio Furuichi
%A Iwao Waga
%J PLOS ONE
%D 2013
%I Public Library of Science (PLoS)
%R 10.1371/journal.pone.0083108
%X Display technologies are procedures used for isolating target-recognizing peptides without using immunized animals. In this study, we describe a new display method, named Hishot display, that uses Escherichia coli and an expression plasmid to isolate target-recognizing peptides. This display method is based on the formation, in bacteria, of complexes between a polyhistidine (His)-tagged peptide including random sequences and the peptide-encoding mRNA including an RNA aptamer against the His-tag. When this system was tested using a sequence encoding His-tagged green fluorescent protein that included an RNA aptamer against the His-tag, the collection of mRNA encoding the protein was dependent on the RNA aptamer. Using this display method and a synthetic library of surrogate single-chain variable fragments consisting of VpreB and Ig heavy-chain variable domains, it was possible to isolate clones that could specifically recognize a particular target (intelectin-1 or tumor necrosis factor-¦Á). These clones were obtained as soluble proteins produced by E. coli, and the purified peptide clones recognizing intelectin-1 could be used as detectors for sandwich enzyme-linked immunosorbent assays. The Hishot display will be a useful method to add to the repertoire of display technologies.
%U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0083108