title: Hishot display--a new combinatorial display for obtaining target-recognizing peptides.
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.
authors.affiliation: Molecular Diagnostic Project, Kanagawa Cancer Center Research Institute, Yokohama, Japan.
authors.forename: Shoutaro
authors.initials: S
authors.lastname: Tsuji
authors.affiliation: Molecular Diagnostic Project, Kanagawa Cancer Center Research Institute, Yokohama, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Yamashita
authors.affiliation: Molecular Diagnostic Project, Kanagawa Cancer Center Research Institute, Yokohama, Japan.
authors.forename: Taihei
authors.initials: T
authors.lastname: Kageyama
authors.affiliation: Anticancer Drug Project, Kanagawa Cancer Center Research Institute, Yokohama, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohtsu
authors.affiliation: Molecular Diagnostic Project, Kanagawa Cancer Center Research Institute, Yokohama, Japan.
authors.forename: Katsuo
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: VALWAY Technology Center, NEC Soft Ltd., Tokyo, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kato
authors.affiliation: VALWAY Technology Center, NEC Soft Ltd., Tokyo, Japan.
authors.forename: Joe
authors.initials: J
authors.lastname: Akitomi
authors.affiliation: VALWAY Technology Center, NEC Soft Ltd., Tokyo, Japan.
authors.forename: Makio
authors.initials: M
authors.lastname: Furuichi
authors.affiliation: VALWAY Technology Center, NEC Soft Ltd., Tokyo, Japan.
authors.forename: Iwao
authors.initials: I
authors.lastname: Waga
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D061505:Cell Surface Display Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D019151:Peptide Library
chemical_list: D019151:Peptide Library; D049452:Green Fluorescent Proteins