表达猪圆环病毒2型核衣壳蛋白重组杆状病毒的构建与鉴定

本研究根据GenBank数据库中的猪圆环病毒2型全基因组序列设计并合成了1对能扩增完整Cap序列的引物,采用PCR扩增猪圆环病毒2型YZ株的Cap基因,并将其克隆到杆状病毒转移载体pFastBacHTA中,获得阳性质粒pFastBacHTA-Cap;测序后将该质粒转化到大肠杆菌DH10Bac感受态细胞中,获得含有Cap基因的重组杆状病毒质粒Bacmid-Cap;最后将获得的重组杆状病毒质粒Bacmid-Cap转染sf9昆虫细胞并成功拯救了能稳定表达猪圆环病毒2型Cap蛋白的重组杆状病毒rBacmid-Cap,该重组病毒的成功拯救为研制猪圆环病毒2型的基因工程亚单位疫苗奠定了基础.

Construction and Identification of Recombinant Baculovirus Expressing Cap Protein of Porcine Circovirus Type 2

A pair of primers was designed base on the genome of porcine circovirus type 2 (PCV2) retrieved from GenBank, which could amplify the whole coding sequence of Cap gene. The PCR product was cloned into pFastBacHTA and then sequenced. Subsequently, the positive plasmid was transformed into DH10Bac cells and the recombinant plasmid with Cap gene of PCV2 (Bacmid-Cap) was identified by PCR method. The recombinant plasmid was extracted and transfected into sf9 cells and the recombinant baculovirus expressing Cap protein of PCV2 (rBacmid-Cap) was rescued, which established the basis for the further research on subunit vaccine against PCV2.