title: Surface display of recombinant proteins on Bacillus subtilis spores.
abstract: 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.
authors.affiliation: Dipartimento di Fisiologia Generale ed Ambientale, Sezione di Microbiologia, Università Federico II, Naples, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Isticato
authors.forename: G
authors.initials: G
authors.lastname: Cangiano
authors.forename: H T
authors.initials: HT
authors.lastname: Tran
authors.forename: A
authors.initials: A
authors.lastname: Ciabattini
authors.forename: D
authors.initials: D
authors.lastname: Medaglini
authors.forename: M R
authors.initials: MR
authors.lastname: Oggioni
authors.forename: M
authors.initials: M
authors.lastname: De Felice
authors.forename: G
authors.initials: G
authors.lastname: Pozzi
authors.forename: E
authors.initials: E
authors.lastname: Ricca
mesh_terms: 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
chemical_list: D001426:Bacterial Proteins; C439051:CotB protein, Bacillus subtilis; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D013745:Tetanus Toxoid