重组绿脓杆菌外毒素A受体结合区蛋白动物免疫保护试验

Animal Protection Experiment of the Immunization of Recombinant Receptor Binding Protein of Exotoxin A of P.aeruginosa

将绿脓杆菌外毒素 A( PEA)受体结合区基因重组质粒 p ET-2 8( c) +B1 0 转入大肠杆菌 BL2 1 ( DE3)plys,诱导表达出的重组 PEA受体结合区蛋白包涵体经脲素洗涤后 ,Sephacryl-2 0 0分子筛层析 ,DEAE-Sepharose FF离子交换层析获得纯度为 95.8%重组蛋白包涵体。将该蛋白作免疫原免疫小鼠后 ,用 PEA进行不同时间攻毒 ,观察到最佳保护时间为 1 4 d左右 ;

Expressing plasmid PET 28(c)+B 10 containing receptor binding domain of exotoxin A P.aeruginosa (PEA), was transformed into E.coli BL21 (DE3) plys. By induction of IPTG, a recombinant protein which formed inclusion body and contained receptor binding domain of PEA was expressed in E.coli The inclusion body was washed with urea, then it was purified by sephacryl 200 gel filter and DEAE sepharose FF ion exchange chromatography with the purity of 95.8%. The mice were immuned with the purified inclusion body and attacked with PEA at different times. The results showed that the optimum protective time was about 14 days,and the major pathologic changes occured in the liver and kidney.