Surface display of recombinant protein on the cell surface of Bacillus subtilis by the CotB anchor protein |
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Authors: | Han Meimei Enomoto Keiichi |
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Institution: | (1) Department of Environmental Systems Engineering, Kochi University of Technology, 185 Miyanokuchi, Tosayamada, Kami, Kochi 782-8502, Japan;(2) Institutes of Biomedical Sciences, Fudan University, 138 Yixueyuan Road, 200032 Shanghai, China |
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Abstract: | We developed a novel surface display system based on the CotB anchoring motif in order to express foreign protein on the surface
of vegetative Bacillus subtilis cells. CotB is a protein in the B. subtilis spore coat. In this system, three repeats of the immunodominant ovalbumin T-cell epitope (OVA323–339) were linked with the cholera toxin B subunit (CTB) to construct a fusion protein, CTB-OVA epi, which was then fused to the
C-terminal of the CotB protein so that CTB-OVA epi was expressed in vegetatively-growing B. subtilis. The expression and localization of the CTB-OVA epi protein was confirmed by western blotting, immunofluorescence microscopy,
and flow cytometry. The results indicated that a CotB-based surface display system was successfully used to express the CTB-OVA
epi protein on the surface of vegetative B. subtilis cells. |
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