2013
DOI: 10.1099/jmm.0.057372-0
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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 … Show more

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Cited by 43 publications
(50 citation statements)
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References 30 publications
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“…Recombinant spores expressing bacterial antigens elicited specific immune responses and provided protection following mucosal immunization of mice (Mauriello et al, 2004;Negri et al, 2013;Zhou et al, 2008a). Here we reported the first application, we believe, of spores expressing the H. pylori urease B antigen as a vaccine candidate.…”
Section: Discussionmentioning
confidence: 99%
“…Recombinant spores expressing bacterial antigens elicited specific immune responses and provided protection following mucosal immunization of mice (Mauriello et al, 2004;Negri et al, 2013;Zhou et al, 2008a). Here we reported the first application, we believe, of spores expressing the H. pylori urease B antigen as a vaccine candidate.…”
Section: Discussionmentioning
confidence: 99%
“…Each pair consists of one version of vector in which recombinant protein is linked directly to Cot protein and the other with short peptide linker (-GGGGS-). The vectors with cotZ and cgeA genes are designed in a way enabling for obtaining fusions with heterologous protein attached to the C-terminus of the coat protein directly or via alpha-helical linker (-GGGEAAAKGGG-) [21]. Such set of vectors gives a flexibility in construction of strains producing recombinant spores.…”
Section: Resultsmentioning
confidence: 99%
“…The resulting plasmid, pKH143, was verified by restriction analysis and nucleotide sequencing. The protein was purified and used for antibody production following method described previously [21]. …”
Section: Methodsmentioning
confidence: 99%
“…Samples were fixed directly in the medium as described by Negri A, et al [12]. Briefly, intact spores were incubated overnight at 4°C with GA2B anti-influenza A matrix protein 1 mAb (Pierce), followed by incubation with anti-mouse Cy3 (Jackson Immuno Research) overnight at 4°C.…”
Section: Immunofluorescence Microscopymentioning
confidence: 99%