摘要
目的获得用于SIV检测的衣壳蛋白p27重组抗原。方法利用生物信息学软件选择衣壳蛋白p27抗原表位集中的区域,合成SIV p27基因;将该基因与pMAL-p5x载体连接构建pMAL-p5x-p27重组质粒,并转化大肠杆菌BL21中,诱导表达;用Amylose Resin亲和层析柱对表达产物进行纯化。结果 SDS-PAGE分析显示,pMAL-p5x-p27重组质粒可在大肠杆菌中高效表达,表达产物分子量约为70×103;经纯化获得目的蛋白p27纯度可达90%。结论本研究利用原核表达系统成功表达了SIV p27蛋白,为SIV检测方法的建立奠定了基础。
Objective To obtain capsid p27 of SIV for detecting. Methods The p27 gene of SIV was synthesized and inserted into pMAL-p5x vector to construct pMAL-pSx-p27 recombinant plasmid. And then the recombinant plasmid was transformed into E. coli BL21 to express protein and the express product was purified by Amylose Resin maltose-binding protein affinity column. Results SDS-PAGE analysis indicated that pMAL-p5x-p27 can be expressed in E. coli BL21 and the protein molecular weight was about 70 x 103 ; The final p27 protein purification could reach 90%. Conclusion p27 of SIV was successfully expressed in E. coli, which lays a foudation for developing the methods of detecting SIV antibody.
出处
《中国比较医学杂志》
CAS
2013年第1期1-4,共4页
Chinese Journal of Comparative Medicine
基金
十二五重大专项"高致病病原动物模型研究"(2012ZX10004502)
关键词
SIV
P27
表达
纯化
SIV
p27
Prokaryotic expression
Purification
