Surface display of recombinant proteins on Bacillus subtilis spores.
We developed a novel surface display system based on the use of bacterial spores. A protein of the Bacillus subtilis spore coat, CotB, was found to be located on the spore surface and used as fusion partner to express the 459-amino-acid C-terminal fragment of the tetanus toxin (TTFC). Western, dot blot and fluorescent-activated cell sorting analyses were used to monitor TTFC surface expression on purified spores. We estimated that more than 1.5 x 10(3) TTFC molecules were exposed on the surface of each spore and recognized by TTFC-specific antibodies. The efficient surface presentation of the heterologous protein, together with the simple purification procedure and the high stability and safety record of B. subtilis spores, makes this spore-based display system a potentially powerful approach for surface expression of bioactive molecules.
Isticato
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Dipartimento di Fisiologia Generale ed Ambientale, Sezione di Microbiologia, Università Federico II, Naples, Italy.
Cangiano
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G
Tran
H T
HT
Ciabattini
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A
Medaglini
D
D
Oggioni
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MR
De Felice
M
M
Pozzi
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G
Ricca
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D007074:Immunoglobulin G / Q000096:biosynthesis; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013171:Spores, Bacterial / Q000378:metabolism*; D013745:Tetanus Toxoid / Q000235:genetics / Q000276:immunology
D001426:Bacterial Proteins; C439051:CotB protein, Bacillus subtilis; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D013745:Tetanus Toxoid