OmpA fusion proteins for presentation of foreign antigens on the bacterial outer membrane.
The ompA genes of Escherichia coli and Shigella dysenteriae have been used to construct a group of enterobacterial surface expression vectors for foreign genes. Linker oligonucleotides were inserted into the sequence corresponding to the third or fourth outer domain to allow in-frame sandwich fusion of foreign genes or epitopes into ompA. Influenza haemagglutinin was inserted without its leader peptide and anchor sequences and shown to be transferred as an ompA fusion protein to the bacterial surface in large amounts. The stability of this system depends on the stem structure (i.e. the bottom part) of the haemagglutinin unit which apparently initiates the folding process that extends into the ompA segment. This fusion construct can be used as a vector system and has been used to transfer to the bacterial surface several other proteins inserted into it, including beta-galactosidase, foot-and-mouth disease virus (FMDV) and malaria antigens. All are exported from the cytoplasm across both the inner and outer membranes to become exposed on the bacterial surface. Very hydrophobic segments or inserts with distinct secondary structures, such as the capsid protein, VP1 of FMDV, will, however, block this process.
Hobom
G
G
Institut für Mikrobiologie und Molekularbiologie, Universität Giessen, Germany.
Arnold
N
N
Ruppert
A
A
D000818:Animals; D017951:Antigen Presentation / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000276:immunology; D001428:Bacterial Vaccines / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012762:Shigella dysenteriae / Q000235:genetics / Q000276:immunology; D014613:Vaccines, Attenuated / Q000235:genetics; D014614:Vaccines, Synthetic / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D006389:Hemagglutinins, Viral; D011993:Recombinant Fusion Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic