山羊痘病毒P32蛋白的原核表达及其免疫原性研究
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S851.3

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甘肃省科技支撑计划(1104NKCL097)


Prokaryotic Expression and Immunogenicity Research on P32 Protein of Goatpox Virus
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    摘要:

    为表达山羊痘病毒P32蛋白并研究其免疫原性,构建了含有P32基因的重组表达质粒pET-P32,诱导表达和纯化了P32蛋白;分别运用SDS-PAGE和Western blot,对该蛋白进行了分析和生物活性鉴定;用分析鉴定后的P32蛋白免疫BALB/c小鼠,然后采集小鼠血清,用间接ELISA进行抗体检测;用小鼠脾淋巴细胞增殖试验,检测该蛋白对机体的细胞免疫活性。结果显示:表达纯化的含有GST标签的P32重组蛋白,大小约39 kDa;Western blot显示 P32重组蛋白具有较好的特异性;间接ELISA抗体检测和小鼠脾淋巴细胞增殖试验表明P32重组蛋白具有良好的免疫原性。本试验结果为深入研究P32蛋白的生物学功能以及对山羊痘的诊断与防治奠定了基础。

    Abstract:

    To study the immunogenicity and expression of P32 protein from goatpox virus,a recombinant plasmids was constructed and named as pET-P32.The P32 protein was induced expression and purified,and was analyzed by SDA-PAGE,its biological activity was determined by Western blot. BALB/c mice were immunized with the purified and determined the recombinant P32 protein to produce immune serum. Polyclonal antibody from immune serum was detected by enzyme linked immunosorbent assay(ELISA),cellular immune function of P32 protein induced organic immune system was detected by experiment of spleen lymphocyte proliferation of mice. The results indicated that the expressed P32 protein,with about 39 KDa molecular weight,showed a good antibody specificity by Western blot,the P32 protein showed a good immunogenicity by enzyme linked immunosorbent assay(ELISA)and experiment of spleen lymphocyte proliferation of mice. The results laid the solid foundation for the further study on biological function of P32 protein and diagnosis and prevention of Goatpox virus.

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成伟伟,陈伯祥,杨明,李杰.山羊痘病毒P32蛋白的原核表达及其免疫原性研究[J].《中国动物检疫》编辑部,2017,34(11):94-98.

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  • 在线发布日期: 2017-11-01
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