title: Autodisplay of streptavidin.
abstract: Streptavidin was expressed on the outer membrane of E. coli as a recombinant fusion protein with an autotransporter domain called AIDA-I (adhesin involved in diffuse adherence) using autodisplay technology. The autodisplay of streptavidin was confirmed by SDS-PAGE of the outer membrane proteins, and the number of autodisplayed streptavidin molecules on a single E. coli cell was evaluated with densitometric analysis. The biotin-binding activity of the autodisplayed streptavidin was estimated after treatment with fluorescently labeled biotin by fluorescence microscopy and flow cytometry. The biotin-binding activity of the E. coli with autodisplayed streptavidin was compared with the activity of streptavidin immobilized on magnetic beads. Finally, the outer membrane presenting autodisplayed streptavidin was isolated and layered on a 96-well microplate for an immunoassay.
authors: School of Materials and Sciences, College of Engineering, Yonsei University, 134 Shin-chon-dong, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea.
authors: Min
authors: M
authors: Park
authors: Joachim
authors: J
authors: Jose
authors: Sarah
authors: S
authors: Thömmes
authors: Jo-Il
authors: JI
authors: Kim
authors: Min-Jung
authors: MJ
authors: Kang
authors: Jae-Chul
authors: JC
authors: Pyun
mesh_terms: D018830:Adhesins, Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001709:Biotechnology / Q000379:methods; D001710:Biotin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry; D007118:Immunoassay; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D019809:Streptavidin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C075559:AIDA-I protein, E coli; D018830:Adhesins, Escherichia coli; D011993:Recombinant Fusion Proteins; D001710:Biotin; D019809:Streptavidin