猪圆环病毒2型Cap蛋白在杆状病毒表面的展示

Baculovirus surface display Cap protein of PCV2

本试验将猪圆环病毒2型(PCV2)去核定位区ORF2基因克隆到杆状病毒表面展示转座载体pBacSC中,转化DH10Bac大肠杆菌感受态,经抗性及蓝白斑筛选得到含ORF2基因的重组杆状病毒DNA,以脂质体介导法将此重组DNA转染Sf9昆虫细胞,获得重组杆状病毒。对重组病毒进行Western-blot分析和免疫金电子显微镜检测。结果表明,PCV2 Cap蛋白在重组杆状病毒中获得表达;免疫金电子显微镜观察表明,重组蛋白展示在杆状病毒囊膜上。本试验成功构建表面展示PCV2 Cap蛋白的重组杆状病毒,为下一步应用重组杆状病毒作为亚单位疫苗奠定基础。

In this study,the ORF2 gene of Porcine circovirus typeⅡ（PCV2） deleting N-terminal nuclear localization region was cloned into vector pBacSC to form the recombinant plasmid.The recombinant plasmid was transformed into E.coli DH10Bac.The recombinant baculovirus DNA was screened and identified by blue-white plaque assay and antibotic resisitant selection.Recombinant baculovirus were obtained from Sf9 insect cells in which recombinant baculovirus DNA was transfected by the liposome.Cap protein was expressed and displayed on the viral surface,as revealed by western blot and immunogold electron microscopy.The successful construction of recombinant baculovirus BacSC-Cap would be useful for production of gene vaccination in the future.