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

作　　者：郑君[1] 赵权[1] 王石[1] 种法政[1] 徐辞[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118

出　　处：《中国预防兽医学报》2008年第12期969-972,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30571396)

摘　　要：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.

关 键 词：猪囊尾蚴头节 ScFv-PE40 重组蛋白 表达 纯化 活性分析 

分 类 号：S852.7[农业科学—基础兽医学]