狂犬病病毒NP基因的克隆及真核表达

实验动物与比较医学 ›› 2009, Vol. 29 ›› Issue (4): 220-222.

狂犬病病毒NP基因的克隆及真核表达

军事医学科学院实验动物中心，北京，100071

收稿日期:2009-04-07 出版日期:2009-04-30 发布日期:2009-04-30

Cloning and Eukaryotic Expression of NP Gene of Rabies Virus

Laboratory Animal Center, Academy of Military Medical Science, Beijing 100071, China

Received:2009-04-07 Published:2009-04-30 Online:2009-04-30

摘要/Abstract

摘要： 目的检测狂犬病毒NP蛋白的免疫原性。方法利用RT-PCR扩增狂犬病毒核蛋白（NP）基因，测序后将其克隆到PVAX1真核表达载体上；将PVAX1-NP转染Vero细胞，进行SDS-PAGE及Western-blot分析；以重组质粒PVAX1-NP对小鼠进行免疫试验。结果经酶切分析、鉴定和测序验证，得到重组表达质粒PVAX1-NP，NP蛋白在Vero细胞中获得了表达，且表达产物具有免疫学活性；免疫小鼠抗体水平显著升高。结论狂犬病毒NP蛋白具有免疫原性。

关键词: 狂犬病毒, NP基因, 真核表达

Abstract: Objective To detect the immunogenicity of NP gene of RV. Method NP gene of RV was amplified by RT-PCR and then cloned into plasmid pVAXl to construct recombinant plasmid pVAXl-NP.The Vero cells were transfected with plasmid pVAXl-NP. Expression of the NP protein was detected by SDS-PAGE and Westem-blot assay. The mice were immuned with plasmid pVAXl-NP. Result The plasmid PVAXl-NP was obtained and the NP gene was expressed in Vero cells .The expression products had immunogenic ity and the antibody level of immuned mice was higher. Conclusion The NP protein of RV had immunogenic ity.

Key words: RV, NP gene, Eukaryotic expression

李敏,袁菊芳,丁璐静,等. 狂犬病病毒NP基因的克隆及真核表达[J]. 实验动物与比较医学, 2009, 29(4): 220-222.

LI Min,YUAN Ju-Fang,DING Lu-Jing,et al. Cloning and Eukaryotic Expression of NP Gene of Rabies Virus[J]. Laboratory Animal and Comparative Medicine, 2009, 29(4): 220-222.

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