Autodisplay of streptavidin

作者：Park Min; Jose Joachim; Thoemmes Sarah; Kim Jo Il; Kang Min Jung; Pyun Jae Chul^*

来源：Enzyme and Microbial Technology, 2011, 48(4-5): 307-311.

DOI：10.1016/j.enzmictec.2010.12.006

• Autodisplay
• Streptavidin
• E. coli
• Immunoassay

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 SOS-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.

• 出版日期2011-4-7