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) |