Open AccessCell surface display of recombinant proteins on Staphylococcus carnosus
Author(s) -
Patrik Samuelson,
Magnus Hansson,
Niklas Ahlborg,
C. Andréoni,
Friedrich Götz,
Thomas Bächi,
Thien Ngoc Nguyen,
Hans Binz,
Mathias Uhlén,
Stefan Ståhl
Publication year - 1995
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.177.6.1470-1476.1995
Subject(s) - biology , microbiology and biotechnology , recombinant dna , staphylococcus aureus , immunogold labelling , biochemistry , bacteria , antibody , gene , genetics , immunology
A novel expression system for surface display of heterologous proteins on Staphylococcus carnosus cells has been developed. Taking advantage of the promoter and secretion signals, including a propeptide region, from the lipase gene of Staphylococcus hyicus and the cell wall-spanning and membrane-binding region of protein A from Staphylococcus aureus, efficient surface display of an 80-amino-acid peptide from a malaria blood stage antigen could be achieved. A serum albumin binding protein from streptococcal protein G was used both as a general reporter molecule and to increase the accessibility of the surface-displayed proteins. Immunoblotting, immunogold staining, and immunofluorescence on intact recombinant S. carnosus cells verified the presence of the propeptide, the malaria antigen, and the albumin-binding reporter protein on the bacterial surface. For the first time, fluorescence-activated cell sorting was used to analyze the presence of surface-displayed hybrid receptors on gram-positive bacteria.