炭疽芽孢杆菌EA1蛋白的融合表达和纯化

目的：原核表达重组炭疽芽孢杆菌EA1蛋白。方法：用PCR方法从炭疽芽孢杆菌A16R疫苗株染色体中扩增编码EA1蛋白的eag基因序列,经过纯化、酶切后克隆到含有GST标签的原核表达载体pGEX-6P-2中,构建重组载体pGEX-EA1;将空载体（作为对照）、重组载体转化大肠杆菌BL21（DE3）菌株获得表达工程菌株,对其表达和纯化条件进行优化;利用Western印迹检测融合蛋白的表达。结果：构建了EA1蛋白的融合表达载体,并在大肠杆菌中获得高效表达;经Glutathione Sepharose 4B纯化获得了EA1蛋白;Western印迹表明,此蛋白可与GST标签抗体反应。结论：在原核表达系统中表达并纯化得到EA1融合蛋白,为进一步对其进行功能研究奠定了基础。

Prokaryotic Fusion Expression and Purification of EA1 Protein of Bacillus anthracis

Objective： To express and purify EA1 protein of Bacillus anthracis. Methods： The eag gene coding EA1 protein was amplified by PCR from B.anthracis A16R and inserted into plasmid pGEX-6P-2 to construct a recombinant vector pGEX-EA1. Both control vector and the recombinant vector were transformed into E.coil BL21（DE3）. The conditions of expression and purification were optimized. The fusion protein was characterized by Western blot. Results： The pGEX- EA1 vector was successfully constructed and the recombinant EA1 was purified with Glutathione Sepharose 4B affinity chromatography. Western blot analysis showed that the fusion protein interacted with GST-tagged antibody. Conclusion： The recombinant EA1 protein was successfully expressed in E.coli and purified. This will be helpful to study the function of EA1 protein.