Cell surface display of recombinant proteins on Staphylococcus carnosus.
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.
Samuelson
P
P
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Hansson
M
M
Ahlborg
N
N
Andréoni
C
C
Götz
F
F
Bächi
T
T
Nguyen
T N
TN
Binz
H
H
Uhlén
M
M
Ståhl
S
S
D000953:Antigens, Protozoan / Q000096:biosynthesis* / Q000235:genetics; D000954:Antigens, Surface / Q000096:biosynthesis / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors / Q000235:genetics; D008049:Lipase / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D016253:Microscopy, Immunoelectron; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013205:Staphylococcal Protein A / Q000096:biosynthesis / Q000235:genetics; D013210:Staphylococcus / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D013499:Surface Properties
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D001426:Bacterial Proteins; D002352:Carrier Proteins; C034284:IgG Fc-binding protein, Streptococcus; D008565:Membrane Proteins; D010446:Peptide Fragments; D011498:Protein Precursors; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; C050256:ring-infected erythrocyte surface antigen (RESA), Plasmodium falciparum; D008049:Lipase