甲型肝炎病毒VP4-VP2在昆虫细胞膜表面的表达

Surface Display of Hepatitis A Virus VP4-VP2 in Insect Cells

目的:利用杆状病毒表面展示系统将甲型肝炎病毒(Hepatitis A virus,HAV)的衣壳蛋白VP4-VP2展示在昆虫细胞膜表面,为甲型肝炎病毒的免疫检测奠定基础。方法:将HAV VP4-VP2基因片段插入杆状病毒膜蛋白基因gp64信号肽序列和水疱口膜炎病毒G蛋白跨膜区基因片段之间得到重组质粒pBac-sp-VP0-VSVtm。利用Bac-to-Bac系统获得含有该融合基因的重组病毒Ac-VP0-VSV。重组病毒感染昆虫Sf9细胞,免疫荧光分析目的蛋白在昆虫细胞的表达。结果:免疫荧光结果显示HAV VP4-VP2蛋白可在Sf9细胞的膜表面大量表达。结论:昆虫细胞膜表面展示系统构建成功,利用该系统HAV VP4-VP2蛋白可在昆虫细胞内表达并被展示到了细胞膜表面。

Objective： Hepatitis A virus （HAV） VP4 - VP2 was displayed on surface of insect cells using baculovirus surface display sys- tem, which will lay the foundation for the diagnosis of HAV. Method： The gene fragment of HAV VP4 - VP2 was cloned in frame between the sequences of gp64 signal peptide and VSV G transmembrane region to achieve the recombinant plasmid pBac - sp - VP0 - VSVtm. A recombinant virus Ac -VP0 - VSV containing the fusion gene （GP64 - VPd - VP2 - VSV G） was produced using Bac -to - Bac sys- tem. St9 cells were infected with the recombinant virus and analyzed the expression of proteins by immunofluorescence. Result：The results of immunofluorescence showed that HAV VP4 - VP2 could be expressed in Sf9 cells and transported to the cell surface. Conclusion： The insect cell surface display system was successfully constructed and displayed HAV VP4 -VP2 on the cell surface.