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Journal of Bacteriology, November 2001, p. 6294-6301, Vol. 183, No. 21
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.21.6294-6301.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Surface Display of Recombinant Proteins on Bacillus subtilis Spores

Rachele Isticato,1 Giuseppina Cangiano,1 Hoa T. Tran,1,dagger Annalisa Ciabattini,2 Donata Medaglini,2 Marco R. Oggioni,2 Maurilio De Felice,1 Gianni Pozzi,2 and Ezio Ricca1,*

Dipartimento di Fisiologia Generale ed Ambientale, Sezione di Microbiologia, Università Federico II, Naples,1 and Laboratorio di Microbiologia Molecolare e Biotecnologia, Sezione di Microbiologia, Dipartimento di Biologia Molecolare, Università di Siena, Siena,2 Italy

Received 16 April 2001/Accepted 8 August 2001

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 × 103 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.

* Corresponding author. Mailing address: Dipartimento di Fisiologia Generale ed Ambientale, Università Federico II, via Mezzocannone 16, 80134 Naples, Italy. Phone: 39-081-2534636. Fax: 39-081-5514437. E-mail: ericca{at}unina.it.

dagger Present address: Public Health Research Institute, New York, N.Y.

Journal of Bacteriology, November 2001, p. 6294-6301, Vol. 183, No. 21
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.21.6294-6301.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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