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Type: Journal article
Title: Epitope analysis of the FanC subunit protein of the K99 (F5) fimbriae of enterotoxigenic Escherichia coli using a recombinant fusion technique
Author: Ogunniyi, A.
Kotlarski, I.
Morona, R.
Manning, P.
Citation: FEMS Immunology and Medical Microbiology, 2002; 34(1):23-31
Publisher: Elsevier Science BV
Issue Date: 2002
ISSN: 0928-8244
Statement of
Abiodun D Ogunniyi, Ieva Kotlarski, Renato Morona, Paul A Manning
Abstract: We have used a recombinant approach to characterise the B- and T-cell epitopes of FanC, the major subunit polypeptide of K99 (F5) fimbriae of enterotoxigenic Escherichia coli strains. This involved the fusion of FanC and its carboxy-terminal truncated derivatives to a reporter, the E. coli alkaline phosphatase (PhoA), generating stable, recombinant fusions. The B-cell epitopes of FanC were characterised by Western blotting of FanC::PhoA fusion proteins with a polyclonal mouse antiserum directed against K99 fimbrial antigen, and with a panel of monoclonal antibodies generated to the K99 antigen. An attempt to characterise the T-cell epitopes of the fimbrial subunit was made by standard in vitro T-cell proliferation assay. Our results suggest that the B-cell epitopes of FanC are likely to be continuous, with a potentially immunodominant epitope at the carboxy-terminus. However, T-cell proliferation assays with the FanC::PhoA fusion proteins did not indicate any immunodominant T-cell epitope(s). We hypothesise that fusion of FanC peptides to PhoA had resulted in altered folding of the peptides for antibody and T-cell recognition, highlighting the potential problems and drawbacks of the recombinant fusion technique in defining the epitopes of certain proteins.
Keywords: B-Lymphocytes
Fimbriae, Bacterial
Mice, Inbred BALB C
Escherichia coli
Alkaline Phosphatase
Cyclin-Dependent Kinases
Escherichia coli Proteins
Protein Subunits
Recombinant Fusion Proteins
Bacterial Toxins
Antigens, Surface
Immunodominant Epitopes
Epitope Mapping
Protein Folding
Description: The definitive version is available at
DOI: 10.1111/j.1574-695X.2002.tb00599.x
Published version:
Appears in Collections:Aurora harvest 6
Molecular and Biomedical Science publications

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