Surface display of recombinant proteins on Bacillus subtilis spores
Surface display of recombinant proteins on Bacillus subtilis spores
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DOI：
10.1128/jb.183.21.6294-6301.2001
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发表时间：
2001-11-01
影响因子：
3.2
通讯作者：
Ricca, E
中科院分区：
文献类型：
--
作者：
Isticato, R;Cangiano, G;Ricca, E
我们开发了一种新的表面展示系统的基础上使用的细菌孢子。枯草芽孢杆菌孢子外壳的蛋白质CotB被发现位于孢子表面，并被用作融合伴侣来表达破伤风毒素（TTFC）的459个氨基酸的C-末端片段。蛋白质，斑点印迹和荧光激活细胞分选分析用于监测TTFC表面表达纯化的孢子。我们估计，超过1.5 × 10（3）TTFC分子暴露在每个孢子的表面上，并被TTFC特异性抗体识别。异源蛋白质的有效表面呈递，连同简单的纯化程序和B的高稳定性和安全性记录。枯草芽孢杆菌孢子，使这种基于孢子的展示系统的生物活性分子的表面表达的潜在的强大的方法。
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.