小反刍兽疫病毒F和H蛋白在CHO细胞中的融合表达及其免疫原性的研究

以小反刍兽疫病毒(peste des petits ruminants virus,PPRV)疫苗株Nigeria75/1基因序列为基础,构建了融合表达PPRV囊膜蛋白F和H胞外区的真核重组质粒pcDNA3.1-F-H,在悬浮CHO细胞中进行分泌表达。离心收集转染后的细胞培养上清进行亲和层析纯化,纯化样品经Western-blot鉴定,融合蛋白与PPRV阳性血清和His标签抗体都能发生免疫反应。使用纯化后的融合蛋白免疫BALB/c小鼠,可诱导小鼠体内产生针对PPRV的特异性抗体。上述研究结果为小反刍兽疫新型亚单位疫苗的研制奠定了基础。

Fusion expression and immunogenicity of peste des petits ruminants virus F and H proteins in CHO cells

Based on the genomic sequence of Nigeria75/1 of PPRV vaccine strain,an eukaryotic recombinant plasmid designated pc DNA3.1-F-H,expressing the fused extracellular domain of PPRV F and H proteins,was constructed and transfected into CHO cells.The fusion protein was secreted into the culture supernatant and purified by Ni-NTA affinity chromatography column.After identification by Western-blot,the purified fusion protein presented good antigenic reaction with both His-Tag monoclonal antibodies and PPRV positive serum.The BALB/c mice immunized with purified fusion protein developed high-level anti-PPRV antibodies detected by ELISA.The above-mentioned result lays an important foundation for the development of PPR subunit vaccine.