猪囊尾蚴头节单链抗体-PE40重组蛋白的表达及抗虫活性分析

Expression of fusion protein of single-chain antibody against Cysticercus Cellulosae Scolex antigen and PE40

将含有抗猪囊尾蚴头节单链抗体基因(ScFv)和绿脓杆菌外毒素基因(PE40)的重组质粒pET28-ScFv-PE40转化到大肠杆菌BL21中,经IPTG诱导,表达产物通过SDS-PAGE和western blot鉴定,其表达蛋白的相对分子量约为68ku,表达量约占菌体总蛋白的13.8%。将融合蛋白通过金属Ni2+亲和层析柱纯化后,目的蛋白纯度在95%以上,可溶性蛋白的回收率约为50%,回收量为5mg/L。本研究所进行的纯化重组蛋白的体外杀伤六钩蚴试验表明,重组蛋白对六钩蚴具有较强的杀灭作用,而且与其剂量呈正相关性。

In this study, recombinant plamid pET28a containing single-chain antibody gene （ScFv） and Pseudomonas exotoxin gene （PE40） was transformed to E.coli BL21 and induced for protein expression. The fusion protein was detected by SDS-PAGE and western blot. The expressed protein had a molecular weight of 68 ku and accounted for 13.8 % of total protein. The protein was purified by Ni2＋ affinity column and tested in a kill and wound experiment in vitro. Strong killing action against oncosphere was detected for recombinant toxinurn, which acted in a dose-dependent manner.