Expression of foreign epitopes in P-fimbriae of Escherichia coli.
Hypervariable regions (HRs) of the major subunit of F11 fimbriae were exploited for insertion of foreign epitopes. Two insertion vectors were created that contain a unique cloning site in HR1 or HR4 respectively. Several oligonucleotides, coding for antigenic determinants derived from different pathogens, were cloned in both insertion vectors. Hybrid fimbrial subunits were generally shown to be assembled in fimbriae when the length of the inserted peptide did not exceed 14 amino acids. The inserted peptides appeared to be exposed in the fimbrial filament. One hybrid fimbrial protein induced detectable levels of antibodies against the inserted epitope if injected into mice.
van Die
I
I
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
van Oosterhout
J
J
van Megen
I
I
Bergmans
H
H
Hoekstra
W
W
Enger-Valk
B
B
Barteling
S
S
Mooi
F
F
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004279:DNA, Viral; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D010861:Fimbriae, Bacterial / Q000276:immunology*; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation; D055503:Protein Multimerization; D015183:Restriction Mapping; D020798:Two-Hybrid System Techniques
D000914:Antibodies, Viral; D000942:Antigens, Bacterial; D000956:Antigens, Viral; D004269:DNA, Bacterial; D004279:DNA, Viral; D000939:Epitopes