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Journal of Bacteriology, August 2002, p. 4197-4204, Vol. 184, No. 15
0021-9193/02/$04.00+0 DOI: 10.1128/JB.184.15.4197-4204.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Antigen 43-Mediated Autotransporter Display, a Versatile Bacterial Cell Surface Presentation System
Kristian Kjærgaard, Henrik Hasman, Mark A. Schembri, and Per Klemm*
Microbial Adhesion Group, Section of Molecular Microbiology, BioCentrum-DTU, Technical University of Denmark, DK-2800 Lyngby, Denmark
Received 25 February 2002/
Accepted 7 May 2002
Antigen 43 (Ag43), a self-recognizing outer membrane protein of Escherichia coli, has been converted into an efficient and versatile tool for surface display of foreign protein segments. Ag43 is an autotransporter protein characterized by the feature that all information required for transport to the outer membrane and secretion through the cell envelope is contained within the protein itself. Ag43 consists of two subunits (
and ß), where the ß-subunit forms an integral outer membrane translocator to which the
-subunit is noncovalently attached. The simplicity of the Ag43 system makes it ideally suited as a surface display scaffold. Here we demonstrate that the Ag43
-module can accommodate and display correctly folded inserts and has the ability to display entire functional protein domains, exemplified by the FimH lectin domain. The presence of heterologous cysteine bridges does not interfere with surface display, and Ag43 chimeras are correctly processed into
- and ß-modules, offering optional and easy release of the chimeric
-subunits. Furthermore, Ag43 can be displayed in many gram-negative bacteria. This feature is exploited for display of our chimeras in an attenuated Salmonella strain.
* Corresponding author. Mailing address: Section of Molecular Microbiology, BioCentrum-DTU, Bldg. 301, Technical University of Denmark, DK-2800 Lyngby, Denmark. Phone: 45 45 25 25 06. Fax: 45 45 93 28 09. E-mail:
per.klemm{at}biocentrum.dtu.dk.
Journal of Bacteriology, August 2002, p. 4197-4204, Vol. 184, No. 15
0021-9193/02/$04.00+0 DOI: 10.1128/JB.184.15.4197-4204.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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