title: Co-autodisplay of Z-domains and bovine caseins on the outer membrane of E. coli.
abstract: In this work, two proteins, Z-domains and bovine casein, were auto-displayed on the outer membrane of the same Escherichia coli cells by co-transformation of two different auto-display vectors. On the basis of SDS-PAGE densitometry, Z-domains and bovine casein were expressed at 3.12 × 10⁵ and 1.55 × 10⁵ proteins/E. coli cell, respectively. The co-auto-displayed Z-domains had antibody-binding activity and the bovine casein had adhesive properties. E. coli with co-auto-displayed proteins were analyzed by fluorescence assisted cell sorting (FACS). E. coli with co-auto-displayed Z-domains and bovine casein aggregated due to hydrophobic interaction. For application to immunoassays, the Z-domain activity was estimated after (1) immobilizing the E. coli and (2) forming an OM layer. E. coli with co-auto-displayed two proteins that were immobilized on a polystyrene microplate had the same antibody-binding activity as did E. coli with auto-displayed Z-domains only. The OM layer from the co-transformed E. coli had Z-domains and bovine casein expressed at a 1:2 ratio from antibody-binding activity measurements.
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-ro, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea.
authors.forename: Gu
authors.initials: G
authors.lastname: Yoo
authors.affiliation: Institute of Pharmaceutical and Medical Chemistry, University of Muenster, Muenster, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Saenger
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-ro, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea.
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Bong
authors.affiliation: Institute of Pharmaceutical and Medical Chemistry, University of Muenster, Muenster, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Jose
authors.affiliation: Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea.
authors.forename: Min-Jung
authors.initials: MJ
authors.lastname: Kang
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-ro, Seo-dae-mun-gu, Seoul 120-749, Republic of Korea. Electronic address: jcpyun@yonsei.ac.kr.
authors.forename: Jae-Chul
authors.initials: JC
authors.lastname: Pyun
mesh_terms: D000818:Animals; D002364:Caseins / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005434:Flow Cytometry; D007118:Immunoassay
chemical_list: D002364:Caseins
keywords: Autodisplay; Casein; FACS; Immunoassay; Z-domain