蓝舌病毒L3基因的克隆与表达

Cloning and expression of bluetongue virus VP3 protein in insect cells

目的 通过高效表达，研究蓝舌病毒（ＢＴＶ） ＶＰ３ 的功能，为后续ＢＴＶ病毒样颗粒的装配作准备。方法 克隆出完整的ＢＴＶ１３ Ｌ３ 基因，将其插入杆状病毒表达载体进行表达。结果 获得了含有全长Ｌ３ 基因的克隆，ＶＰ３ 在昆虫细胞中得到了高效表达，表达蛋白占细胞总蛋白的１０％ ～１５％ ，ＶＰ３ 与ＶＰ７ 共表达可装配出ＢＴＶ 核心样颗粒。结论 在昆虫细胞中表达ＢＴＶＶＰ３ 蛋白具有生物学活性。

Objective To elucidate the biological activities of VP3 protein of bluetongue virus (BTV) expressed in insect cells and to assemble, in future, the BTV core like particles.Methods First, full length of BTV13 L3 cDNA was constructed from 2 sequenced clones. The silver staining of the splicing region proved that the construction was correct. Then, a recombinant baculovirus expression vector pFB1BL3 was constructed to express the L3 gene in insect cells. Results BTV13 VP3 could be highly expressed in Sf 9 cells, the production could occupy up to 10%～15% of the total cell protein. Core like particles could be observed after co expression of the VP3 and VP7 in insect cells.Conclusion VP3 of BTV expressed in insect cells possesses biological activity and can be used to assemble BTV core like particles.