炭疽芽胞杆菌A16R株FtsE蛋白的可溶性表达和纯化  

Soluble Expression and Purification of FtsE Protein of Bacillus anthracis

作　　者：李强[1] 刘先凯[2] 冯尔玲[2] 朱力[2] 王沛[1] 王红[1] 杨新华[1] 赵美玲[1] 姜永强[3] 王恒樑[2] 

机构地区：[1]白求恩医务士官学校,河北石家庄050081 [2]军事医学科学院生物工程研究所,北京100071 [3]军事医学科学院微生物流行病研究所,北京100071

出　　处：《生物技术通讯》2014年第1期45-48,共4页Letters in Biotechnology

基　　金：国家自然科学基金(81271785)

摘　　要：目的:构建炭疽芽胞杆菌FtsE蛋白的原核表达载体,实现其在原核表达系统中的可溶性表达,并纯化融合蛋白。方法:用PCR方法从炭疽芽胞杆菌A16R株扩增得到ftsE基因片段,酶切后连接到pET28a原核表达载体,构建重组表达质粒pET28a-ftsE,转化大肠杆菌BL21(DE3)菌株,筛选可溶性诱导表达与纯化融合蛋白的条件,以获得高纯度融合蛋白。结果:构建了FtsE蛋白的融合表达载体,并在大肠杆菌中获得高效表达;在20℃下,经0.1 mmol/L IPTG诱导3 h表达的产物主要是可溶性蛋白,经Ni-NTA亲和层析纯化获得了高纯度的FtsE融合蛋白,经Western印迹检测,目的蛋白表达正确。结论:实现了炭疽芽胞杆菌FtsE蛋白原核表达系统的可溶性表达并获得了高纯度融合蛋白,为后续研究奠定了基础。Objective： To construct risE recombinant expression plasmid and optimize the expression conditions, and to purify the fusion protein. Methods： The amplified rise gene fragment by PCR was inserted into pET28a to construct fusion protein expression plasmid. Then the recombinant plasmid was transformed into E.coli BL21（DE3）, and the conditions of induction and purification of FtsE fusion protein were optimized. Results： The pET28a-ftsE recombinant plasmid was constructed successfully. Soluble fusion protein was obtained by induction of 0.1 mmol/L IPTG at 20℃ lasted 3 hours. And the FtsE fusion protein was purified by Ni-NTA affinity chromatography and verified by Western blot. Conclusion： Soluble FtsE fusion protein was achieved, and high purity of the FtsE fu sion protein was obtained. This work may lay a foundation for further study of the functions of FtsE protein.

关 键 词：炭疽芽胞杆菌 FtsE蛋白 融合表达 

分 类 号：Q78[生物学—分子生物学]