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微生物学报

泛素基因介导下的猪繁殖与呼吸综合征病毒M基因对小鼠的免疫试验
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国家高科技发展(863)计划”(2006AA10A203);黑龙江省自然科学基金重点项目(ZJN-0602-01)


Effects of fusion gene of ubiquitin and Porcine Reproductive and Respiratory Syndrome virus M gene on the immune response in inoculated mice
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Supported by the National Programs for High Technology Research and Development of China (2006AA10A203) and the Key Project of National Natural Science Foundation of Heilongjiang Province (ZJN-0602-01)

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    摘要:

    摘要:【目的】获得猪繁殖与呼吸综合征病毒(Porcine Reproductive and Respiratory Syndrome virus, PRRSV)M蛋白基因及其与泛素(Ubiquitin, Ub)基因融合的真核表达质粒,并进一步研究Ub对M基因免疫效果的影响。【方法】利用RT-PCR技术以PRRSV CH-1a株和BALB/c小鼠脾脏组织总RNA为模板,分别扩增出PRRSV M蛋白基因与小鼠的Ub基因,利用SOE技术将PRRSV M基因与小鼠Ub基因进行融合,最终构建真核表达质粒pVAX1-M与pVAX1-U-M。以两种真核表达质粒DNA肌肉免疫BALB/c小鼠后,分别检测体液免疫反应与细胞免疫反应,比较M单基因与Ub-M融合基因DNA免疫所诱生的免疫应答强度【结果】IFA试验表明,pVAX1-M与pVAX1-U-M均能BHK-21细胞内成功表达目的基因;两种重组质粒免疫小鼠后均可诱生PRRSV ELISA抗体与细胞免疫反应,但是pVAX1-U-M诱导的细胞免疫反应明显高于pVAX1-M,差异显著(P<0.05);而其诱导的抗体水平明显低于pVAX1-M,二者差异也显著(P<0.05)。【结论】泛素在一定程度上可以促进PRRSV M基因诱导细胞免疫反应,但对体液免疫未见到同样作用。

    Abstract:

    Abstract: [Objective] To evaluate the effects of the fusion gene of ubiquitin (Ub) and Porcine Reproductive and Respiratory Syndrome virus (PRRSV) M gene on the immune response in inoculated mice. [Methods] Mouse Ub gene and PRRSV M gene were amplified by RT-PCR from BALB/c mice spleen cells and PRRSV Ch-1a strain, respectively, and the M and Ub gene (U-M) was fused by SOE PCR. Therefore, pVAX1-U-M and pVAX1-M recombinant plasmid were constructed for eukaryotic expression. [Results] The fusion U-M and M protein expressions were verified in transfected BHK-21 cells by indirect fluorescence assay. Furthermore, both pVAX1-M and pVAX1-U-M induced specific humoral and cellar immune responses against PRRSV in the recombinant plasmid injected mice. However, pVAX1-U-M was able to induce higher level of T cell response then that of pVAX1-M (P< 0.05), but lower level of antibody (P< 0.05). [Conclusion] Expression of U-M fusion gene had ability to enhance specific T cell response against PRRSV, but no effect on stimulation of humoral response in inoculated mice.

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张维军,童铁钢,白宇,王群,徐树兰,孙庆歌,田野,杨涛,童光志,吴东来. 泛素基因介导下的猪繁殖与呼吸综合征病毒M基因对小鼠的免疫试验. 微生物学报, 2009, 49(6): 799-806

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  • 收稿日期:2008-12-09
  • 最后修改日期:2009-03-02
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