title: Combinatorial parallel display of polypeptides using bacteriophage T7 for development of fluorescent nano-bioprobes.
abstract: Application of nano-particles to diagnostic fields has attracted much attention. Biotechnology can contribute to produce useful nano-materials by engineering bacteriophage nano-particles, which are easily prepared by infecting phages to bacterial host cells. In this study, establishment of nano-bioprobes was demonstrated, based on the T7 phage display system, by constructing phage particles displaying a ligand polypeptide S-tag and a green fluorescent protein (GFP) at the same time on the surface of phage head. To achieve this purpose, two types of phage particles were tested: One displayed S-tag and GFP as a single polypeptide (tandem display), and another displayed these molecules as two different polypeptides (parallel display). Only the parallelly displayed phage could be detected with ligand blotting using S-protein and with immunoblotting using an anti-GFP antibody. S-protein-coated magnetic beads and nano-particles were successively labeled with fluorescence using the parallelly displayed phage but could not be labeled with the tandemly displayed phage. Thus, the parallel display of a ligand molecule and fluorescent protein on the head surface of bacteriophage T7 could provide a new scheme of producing fluorescent nano-bioprobes for diagnostic applications.
authors: Graduate School of Agricultural Science, Utsunomiya University, 350 Minemachi, Utsunomiya 321-8505, Japan.
authors: Mie
authors: M
authors: Tsuboyama
authors: Isamu
authors: I
authors: Maeda
mesh_terms: D017123:Bacteriophage T7 / Q000737:chemistry / Q000235:genetics*; D036022:Capsid Proteins / Q000235:genetics; D061505:Cell Surface Display Techniques; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D008024:Ligands; D010455:Peptides / Q000032:analysis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D005456:Fluorescent Dyes; D008024:Ligands; D010455:Peptides; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins