PCV2b Cap蛋白重组杆状病毒的构建及鉴定

猪圆环病毒(PCV)能够引起多种综合征和疾病,给养猪业造成了较大的经济损失。PCV2毒株在2003年后以PCV2b取代PCV2a成为临床感染的猪群中最为流行的基因型。PCV基因组为单股负链环状DNA,其中ORF2编码病毒核衣壳蛋白Cap蛋白,Cap蛋白是主要的免疫原性蛋白。本研究利用Bac-to-Bac杆状病毒表达系统将PCV2b的Cap蛋白基因克隆到杆状病毒载体中,通过制备杆粒、转染sf9细胞,获得表达PCV2b Cap蛋白的重组杆状病毒毒株,从IFA、SDS-PAGE及Western-blotting分析结果可以看出,该重组杆状病毒毒株成功表达了PCV2b Cap蛋白,为制备PCV2b Cap蛋白亚单位疫苗打下基础。

Construction and identification of a recombinant baculovirus expressing swine PCV2b Cap protein

Porcine circovirus(PCV)can cause a variety of syndromes and diseases,which poses a significant threat to the economics of global swine industry.After 2003,PCV2b strain instead of PCV2a has becoming a prevalent genotype in clinical infection pig population.The PCV genome is a single circular negative-stranded DNA and ORF2 encodes the virus nucleocapsid Cap protein,which is the main immunogenic protein.In this study,the Bac-to-Bac baculovirus expression system was used to clone the Cap protein gene.The recombinant baculovirus strain expressing PCV2b Cap protein was obtained by the methods of preparing bacmids and transfecting Sf9 cells et al.The results of IFA,SDS-PAGE and Western-blotting analysis showed that the recombinant baculovirus strain successfully expressed PCV2b Cap protein which lays a foundation for the study of the PCV2b subunit vaccine.