Surface display of recombinant proteins on Bacillus subtilis spores

被引:193
作者
Isticato, R
Cangiano, G
Tran, HT
Ciabattini, A
Medaglini, D
Oggioni, MR
De Felice, M
Pozzi, G
Ricca, E
机构
[1] Univ Naples Federico II, Dipartimento Fisiol Gen & Ambientale, Sez Microbiol, I-80134 Naples, Italy
[2] Univ Siena, Dipartimento Biol Mol, Sez Microbiol, Lab Microbiol Mol & Biotecnol, I-53100 Siena, Italy
关键词
D O I
10.1128/JB.183.21.6294-6301.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:6294 / 6301
页数:8
相关论文
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