风疹病毒JR23株包膜糖蛋白E2重组体的构建表达与抗原性分析

Expression Analysis of Envelope Glycoprotein E2 Protein of Rubella virus JR23 Strain

风疹病毒(Rubella virus, RV)是人类最重要、最常见的致畸病毒之一,但其致畸机理至今尚未完全明了[1].RV的包膜上镶嵌着两个糖蛋白E2和E1,E2/E1以异二聚体的形式在病毒体表面形成刺突结构[2].E2的糖基化程度很高,存在O-联及N-联糖基化位点,糖类在成熟的E2蛋白中所占比重大于三分之一,E2蛋白的糖基化程度影响蛋白功能,糖基化位点的改变可能与RV减毒有关[3].

To construct the recombinant plasmid containing envelope glycoprotein E2 gene of RV JR23 strain, to express E2 protein in BHK21 cells, and to analyze the antigenic sites of E2 protein of RV JR23 strain, E2 gene of RV JR23 strain was amplified by RT-PCR. The PCR product was cloned into the expression vector pBluescript Ⅱ SK^+ and transformed into E. coli JM109, and the recombinant plasmid was transfected into BHK21 cells with recombinant vaccinia virus VTF-7. The expression was detected by immunohistochemistry and indirect immunofluorescence. The recombinant plasmid contained glycoprotein E2 gene, and expressed well in BHK21 cells. The expressed E2 protein mainly accumulated in plasma near the nuclei. There are immunological epitopes of E2 protein on RV JR23 virions. The results showed that RV JR23 E2 transient expression system was successfully constructed. This study offers the basis for investigation of the relationship between the structure and function of RV proteins, and interaction of virus and host cells.