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https://hdl.handle.net/2440/28157
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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 1574-695X |
Statement of Responsibility: | 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 T-Lymphocytes Fimbriae, Bacterial Animals Mice, Inbred BALB C Mice Escherichia coli Alkaline Phosphatase Cyclin-Dependent Kinases Escherichia coli Proteins Protein Subunits Recombinant Fusion Proteins Bacterial Toxins Antigens, Surface Epitopes Immunodominant Epitopes Epitope Mapping Protein Folding Female Male |
Description: | The definitive version is available at www.blackwell-synergy.com |
DOI: | 10.1111/j.1574-695X.2002.tb00599.x |
Published version: | http://www.blackwell-synergy.com/doi/abs/10.1111/j.1574-695X.2002.tb00599.x |
Appears in Collections: | Aurora harvest 6 Molecular and Biomedical Science publications |
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