狂犬病病毒NP基因的克隆及真核表达  

Cloning and Eukaryotic Expression of NP Gene of Rabies Virus

作　　者：李敏[1] 袁菊芳[1] 丁璐静[1] 杨丽萍[1] 张广州[1] 邵军石[1] 马兰芝[1] 曾林[1] 

机构地区：[1]军事医学科学院实验动物中心,北京100071

出　　处：《实验动物与比较医学》2009年第4期220-222,227,共4页Laboratory Animal and Comparative Medicine

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PVAXl真核表达载体上：将PVAX1-NP转染Vero细胞，进行SDS．PAGE,TLWestern—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。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 pVAX1 to construct recombinant plasmid pVAX1- NP.The Vero cells were transfected with plasmid pVAX1-NE Expression of the NP protein was detected by SDS-PAGE and Western-blot assay. The mice were immuned with plasmid pVAXI-NP. Result The plasmid PVAX1-NP was obtained and the NP gene was expressed in Veto cells .The expression products had immunogenicity and the antibody level of immuned mice was higher. Conclusion The NP protein of RV had immunogenicity.

关 键 词：狂犬病毒 NP基因 真核表达 

分 类 号：S852.65[农业科学—基础兽医学]