猪圆环病毒2型ORF2截短基因的原核表达与纯化

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。

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

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 capsid 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.