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。
In this study, the eukaryotic expression recombinant plasmid of pVAXI-Vp4 was transfected into MA104 cells with Lipofectamine^TM2000 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.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2006年第3期306-311,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
国家863计划(No.2003AA241121002)
国家自然科学基金项目(No.30200199)
吉林省科技厅项目(No.20020220)
吉林省杰出青年基金(No.20030118)
中国博士后科学基金项目(No.2002031156)
吉林农业大学青年教师启动基金(No.2002-QQN-002)
