首页> 外文期刊>Journal of Huazhong University of Science and Technology >Stable Expression of Hantavirus H8205 Strain G1/IL-2 Gene and Immune Protection of the Fusion Gene
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Stable Expression of Hantavirus H8205 Strain G1/IL-2 Gene and Immune Protection of the Fusion Gene

机译:汉坦病毒H8205株G1 / IL-2基因的稳定表达及融合基因的免疫保护

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摘要

To explore the feasibility of stable expression of Hantavirus H8205 strain G1 segment and human IL-2 fusion gene in Vero cells, and to examine the immune protection effects on mice vaccinated with this recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene. With the help of lipofectamine, the Vero cells were transfected with pcDNA3.1/HisB-IL-2-G1 and the positive cells were selected by G418. IFAT and SDS-PAGE electrophoresis were used to determine the stable transfection and expression of recombinant protein. Each mouse was inoculated with plasmids intramuscularly (i.m.) three times, 2 boosts were given at 2-week intervals, serum anti-hantavirus antibodies were detected by ELIS A and neutralizing antibodies (NAb) were detected by Plaque Reduction Neutralization Test. The fusion protein expressed in Vero cells was 78 kD, corresponding to the estimated molecular size. The neutralizing antibody titers of mice with pcDNA3.1/HisB-IL-2-G1 were 1:20-1:80. IL-2/G1 fusion gene could be transferred in Vero cells and stably express the fusion protein. Specific humeral immune responses in mice can be induced with the recombinant eukaryotic expression vector containing the fusion gene, which lays the foundation for further development of therapeutic HTNV vaccine.
机译:探讨汉诺病毒H8205毒株G1片段和人IL-2融合基因在Vero细胞中稳定表达的可行性,并研究接种含汉坦病毒G1和IL-2基因的重组真核表达载体对小鼠的免疫保护作用。在脂质转染胺的帮助下,用pcDNA3.1 / HisB-IL-2-G1转染Vero细胞,并通过G418选择阳性细胞。 IFAT和SDS-PAGE电泳用于确定重组蛋白的稳定转染和表达。每只小鼠经肌内(i.m.)接种质粒三遍,每两周间隔两次加强免疫,通过ELIS A检测血清抗汉坦病毒抗体,通过噬斑减少中和试验检测中和抗体(NAb)。在Vero细胞中表达的融合蛋白为78 kD,与估计的分子大小相对应。具有pcDNA3.1 / HisB-IL-2-G1的小鼠的中和抗体滴度为1:20-1:80。 IL-2 / G1融合基因可在Vero细胞中转移并稳定表达融合蛋白。含有融合基因的重组真核表达载体可诱导小鼠特定的肱骨免疫反应,为进一步开发治疗性HTNV疫苗奠定基础。

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