【摘要】 本研究采用LipofectamineTM2000将真核重组表达质粒pVAXI-Vp4转染至MA-104细胞中,得到了表达。以荧光抗体检测法和RT-PCR法双重检测了真核重组表达质粒pVAXI-Vp4在MA-104细胞中的表达情况。并将pVAXI-Vp4免疫BALB/c鼠,检测其血清抗体和脾脏中的淋巴细胞增殖情况及CD4+,CD8+T细胞的数量变化情况。结果表明,猪A组轮状病毒Vp4全基因不但在哺乳动物细胞中获得了表达,而且将真核重组表达质粒pVAX-I-Vp4给动物免疫后,获得了免疫效果。这为pVAXI-Vp4DNA核酸疫苗的进一步推广应用提供了科学依据。更多还原

【Abstract】 In this study,the eukaryotic expression recombinant plasmid of pVAXI-Vp4 was transfected into MA104 cells with LipofectamineTM2000 and the Vp4 gene was expressed and detected with immunofluorescence and RT-PCR,which indicated that the Vp4 gene was expressed in MA-104 cells.To explore the Vp4 protein protective efficacy,the BALB/c mice was immunied with the recombinant plasmid of pVAXI-Vp4.The protective efficacy of recombinant plasmid of pVAXI-Vp4 was detected by the serum antibody titer,lymphocyte cells content and CD4+,CD8+ T cells determination.The result showed that the protective efficacy of mice vaccined with recombinant plasmid of pVAXI-Vp4 was better than the control mice vaccined with pVAXI plasmid and saline.This laid a solid foundation for the development of pVAXI-Vp4 DNA vaccine.更多还原