RRC ID 44832
著者 Tsuji S, Yamashita M, Kageyama T, Ohtsu T, Suzuki K, Kato S, Akitomi J, Furuichi M, Waga I.
タイトル Hishot display--a new combinatorial display for obtaining target-recognizing peptides.
ジャーナル PLoS One
Abstract 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-α). 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.
巻・号 8(12)
ページ e83108
公開日 2013-1-1
DOI 10.1371/journal.pone.0083108
PII PONE-D-13-33067
PMID 24386149
PMC PMC3873924
MeSH Amino Acid Sequence Base Sequence Cell Surface Display Techniques / methods* Enzyme-Linked Immunosorbent Assay / methods* Escherichia coli / genetics* Escherichia coli / metabolism Genetic Vectors Green Fluorescent Proteins / analysis Molecular Sequence Data Peptide Library
IF 2.74
引用数 1
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 RK13(RCB0183)