摘要
将合成的CRM197基因片段克隆到表达载体pBAD-DEST49中,转化到大肠杆菌菌株(ATCCMC1061).经阿拉伯糖诱导,CRM197获得高效表达.目的蛋白主要以包涵体形式存在,变性、复性后经DEAE阴离子交换后获得高于95%纯度的CRM197样品.用该样品做Western blotting鉴定后,能得到单一的特异性条带.本实验的表达策略,为CRM197进一步生产及应用奠定基础.
The gene of CRM197 （cross reacting material 197, CRM197） is synthesized and then expressed in Escherichia coli （ATCCMC1061） driven by pBAD-DEST49. CRM197 is efficiently expressed by Arabinose induced. The recombinant protein is expressed mainly as inclusion body. After denaturation, renaturation and purification by DEAE anion-exchange column, CRM197 with the purity higher than 95% is obtained. For Western blotting, the protein can get single specific bands. Given these characteristics, the CRM197 expressed in Escherichia coli is suitable for further research.
出处
《天津师范大学学报（自然科学版）》
CAS
2012年第3期78-80,共3页
Journal of Tianjin Normal University：Natural Science Edition
