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本实验将基于埃博拉病毒表面糖蛋白抗原GP基因序列,按照哺乳动物密码子使用频率优化,并与人IgG恒定区构建融合蛋白GP-Fc基因序列,重组蛋白序列插入基因疫苗载体pVR中,构建重组蛋白基因疫苗pVRmodGP-Fc。通过基因疫苗导入系统免疫小鼠,用间接ELISA和间接免疫荧光对抗体效价进行评估。实验结果显示,重组蛋白GP-Fc的基因疫苗可以很好的刺激小鼠产生特异性抗体,免疫周期结束后,小鼠血清中结合效价终点达到高剂量组1∶300 000及低剂量组1∶180 000,同时血清中的抗体可以很好地结合表达GP抗原的细胞表面,表现出很强的荧光信号。通过该研究我们验证重组蛋白基因疫苗pVR-modGP-Fc可以很好地诱导BALB/c小鼠产生较高滴度的抗原特异性IgG,具有较好的免疫原性。
Based on the GP gene sequence of Ebola virus surface glycoprotein antigen, we optimized the frequency of mammalian codon usage and constructed the fusion protein GP-Fc gene sequence with human IgG constant region. The recombinant protein sequence was inserted into the gene vaccine vector pVR, Construction of Recombinant Protein Gene Vaccine pVRmodGP-Fc. Mice were immunized by gene vaccine introduction system and antibody titer was evaluated by indirect ELISA and indirect immunofluorescence. The experimental results showed that GP-Fc gene vaccine can stimulate mice to produce specific antibodies. After the end of the immunization period, the final binding titer of mouse serum reached 1: 300 000 in the high-dose group and 1: : 180 000, at the same time, the antibodies in serum can well bind to the cell surface expressing GP antigen and show strong fluorescence signal. Through this study, we verified that the recombinant protein gene vaccine pVR-modGP-Fc can induce BALB / c mice to produce high titer of antigen-specific IgG, and has good immunogenicity.