Expression and display of Clostridium difficile protein FliD on the surface of Bacillus subtilis spores.

Abstract

The endospores of Bacillus subtilis can serve as a tool for surface presentation of heterologous proteins. The unique properties of the spore protective layers make them perfect vehicles for orally administered vaccines. In this study, we successfully displayed a fragment of Clostridium difficile FliD protein on the surface of B. subtilis spores using the CotB, CotC, CotG and CotZ spore coat proteins. The presence of the fusion proteins in the spore coat was verified by Western blotting and immunofluorescence microscopy. The amount of recombinant proteins was assessed by a dot-blot technique. C. difficile is one of the most common infectious agents in nosocomial infections and is especially associated with antibiotic therapies. FliD is a flagellar cap protein of C. difficile and is known to be one of the immunogenic surface antigens of this bacterium. Therefore, its use in vaccine formulations gives a good perspective for successful immunization with a FliD-based vaccine. The recombinant spores presented here may be good candidates for C. difficile oral vaccines.

• Authors: Alessandro Negri, Wojciech Potocki, Adam Iwanicki, Michał Obuchowski, Krzysztof Hinc
• Journal: Journal of medical microbiology (J Med Microbiol) Vol. 62 Issue Pt 9 Pg. 1379-1385 (Sep 2013) ISSN: 1473-5644 [Electronic] England
• PMID: 23475909 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Proteins
• Bacterial Vaccines
• Recombinant Fusion Proteins
• FlaD protein, Bacteria

Topics

• Amino Acid Sequence
• Animals
• Bacillus subtilis (genetics, metabolism)
• Bacterial Proteins (genetics, immunology)
• Bacterial Vaccines (genetics, immunology)
• Clostridioides difficile (genetics, immunology)
• Gene Expression Regulation, Bacterial
• Genes, Bacterial
• Immunization
• Mice
• Mice, Inbred C57BL
• Molecular Sequence Data
• Recombinant Fusion Proteins (genetics, immunology)
• Spores, Bacterial (genetics, metabolism)
• Transformation, Genetic