猪圆环病毒2型ORF2截短基因的原核表达与纯化  

Prokaryotic Expression of the ORF2 Truncated Gene of PCV-2 and Purification of Recombinant Protein

作　　者：林彦星[1] 孙彦伟 刘镇明[1] 徐铮[1] 鲁俊鹏[1] 龚善中[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东省动物防疫监督总所,广东广州510230

出　　处：《动物医学进展》2006年第12期66-70,共5页Progress In Veterinary Medicine

摘　　要：对重组原核表达载体pET-ORF2的表达条件进行优化,结果表明,最佳IPTG诱导浓度为1.0mmol/L,最佳诱导时间为5h。在最佳诱导条件下获得猪圆环病毒2型重组Cap蛋白的基础上,利用HisBind蛋白质纯化试剂盒对表达产物进行纯化,再分别用SDS-PAGE电泳和Westernblotting对纯化产物的纯化效果及特异性进行检测,结果显示表达产物纯化良好,并能被PCV-2阳性血清识别,具有良好的抗原性。The expressive conditions of the recombinant prokaryotic expression vector pET-ORF2 was optimized in this experiment. The results demonstrated that the IPTG optimal induction concentration was 1.0 mmol/L and the best induction time was 5 h, Recombinant capsid protein of Porcine circovirus type 2 was obtained under the optimal induced condition. The expressive product was purified by His Bind Purification Kits. The purification effect and the specificity of the purified recombinant eapsid protein were detected respectively by SDS-PAGE and Western blotting assay. The results showed that the recombinant Cap had been well purified and could react with the polyclonal antibody against PCV-2, and possess good antigenicity.

关 键 词：猪圆环病毒2型 重组原核表达载体 重组CAP蛋白 表达 纯化 

分 类 号：S852.659.2[农业科学—基础兽医学] Q785[农业科学—兽医学]