基于EV71交叉保护抗原P1的病毒样颗粒的构建

Construction of EV71 virus-like particles based on the cross protective antigen P1

目的构建基于EV71交叉保护抗原P1(CP1)的病毒样颗粒。方法利用昆虫杆状病毒表达系统构建CP1和3CD的重组杆状病毒,两者共同感染Sf9细胞,组装形成CP1的EV71病毒样颗粒,并通过Western blot和电镜进行鉴定。结果CP1、3CD重组杆状病毒共感染表达产物的Western blot结果显示,CP1蛋白经3CD蛋白酶酶切形成了大小约为39 k Da、34 k Da、26 k Da的VP1、VP0、VP3蛋白条带,电镜下可观察到形态规则的大小约为30 nm的CP1病毒样颗粒结构。结论CP1病毒样颗粒构建成功,为下一步探讨CP1病毒样颗粒是否对多种亚型的EV71病毒具有交叉保护作用奠定基础。

Objective To construct EV71 virus-like particles based on the cross protective antigen P1(CP1).Methods CP1 and 3 CD recombinant baculovirus was constructed by Bac-to-Bac baculovirus expression system.After co-infection to the Sf9 cells,the CP1 virus-like particles of EV71 was generated.Western blot and TEM were used for identification.Results Western blot showed three specific proteins bands whose molecular masses were corresponded to those of VP1(39 kDa),VP0(34 kDa)and VP3(26 kDa)were observed.TEM examination showed virus-like particles were observed.These particles measured about 30 nm in diameter,resembling the EV71 viral particles in size and appearance.Conclusion CP1 virus-like particles has been successfully constructed.It will lay a foundation for the immunoidentification to figure out whether CP1 virus-like particles could elicit immune responses neutralizing all EV71 genotypes and sub-genotypes virus.