Background Camplylobacter jejuni is a leading cause of bacterial gastroenteritis in the United States, with estimated loss of productivity due to illness being eight billion dollars annually. Previous studies have indicated that the CadF protein of C. jejuni is important in adherence to host epithelial cells. Further research indicates that the FRLS domain of the CadF protein possesses Fibronectin (Fn) binding ability, necessary for adhesion. A novel system for expressing foreign peptides in the S-layer of Caulobacter crescentus has been developed, which allows for rapid creation and purification of peptides.
Study Design and Methods PCR sequencing was performed on various isolates of C. jejuni to see if the cadF gene is conserved across different isolates. Monoclonal antibodies were created and screened by ELISA to determine their binding efficiency to the FRLS domain of the CadF protein. Recombinant DNA methods were used to create a recombinant plasmid that upon electroporation into C. crescentus expressed a portion of the CadF protein, including the FRLS domain.
Results The cadF gene is well conserved across the 23 isolates tested. A monoclonal antibody that binds near the FLRS domain with good specificity was identified. A C. crescentus strain expressing the FRLS domain of the CadF gene from C. jejuni was created and isolated.
Conclusion The cadF gene is conserved across 22 isolates at the nucleotide level. A system for rapid expression, purification, and testing for the FRLS domain of the CadF gene has been created.
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