汉滩病毒核蛋白部分片段与囊膜糖蛋白G1在昆虫细胞中的融合表达

Fused expression of hantaan virus partial NP and glycoprotein G1 in insect cells

目的 应用Bac -to -Bac杆状病毒表达系统融合表达汉滩病毒囊膜糖蛋白G1与核蛋白部分片段。方法 构建含有汉滩病毒G1S0 7嵌合基因的杆状病毒表达载体 pFBD -G1S0 7,转化DH10Bac致敏菌 ,利用其含有的细菌Tn7转座系统将嵌合基因重组至穿梭质粒Bacmid上 ,快速筛选出含有G1S0 7嵌合基因的重组杆状病毒 ,在昆虫细胞中表达该融合蛋白 ,利用间接免疫荧光、ELISA和免疫印迹对表达产物进行检测。结果 构建的含G1S0 7嵌合基因之重组杆状病毒可在昆虫细胞中表达出融合蛋白 ,该蛋白可被抗汉滩病毒核蛋白及糖蛋白G1特异性单抗所识别 ,表达产物主要集中在细胞内。结论 在昆虫细胞中表达出具有生物学活性的G1S0 7融合蛋白 。

Aim To obtain the fused expression of hantaan virus glycoprotein G1 and partial NP by using the Bac to Bac baculovirus expression system Methods A recombinant baculovirus expression vector pFBD G1S0 7 carrying chimeric gene G1S0 7 of hantaan virus was constructed The gene was inserted into shuttle plasmid(Bacmid) in E coli DH10Bac with the help of Tn7 transposition system The recombinant baculovirus was selected and the fusion protein was expressed in insect cells The expression was identified by ELISA,immunofluorescence and Western blot Results The recombinant baculovirus containing the chimeric gene G1S0 7 was constructed successfully and could express the fusion protein in insect cells This protein could be recognized by the hantaan virus nucleoprotein specific mAb and glycoprotein G1 specific mAb Most production was within cells Conclusion The biologically active fusion protein G1S0 7 has been expressed in insect cells It could be used for further research on its immunological characteristic