| EV71型vP1蛋白表达、纯化及免疫原性的初步评价 |
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| 引用本文： | 李晓楠,赵忠鹏,段跃强,赵晓燕,王希良,李培锋.EV71型vP1蛋白表达、纯化及免疫原性的初步评价[J].免疫学杂志,2010(3). |
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| 作者姓名： | 李晓楠 赵忠鹏 段跃强 赵晓燕 王希良 李培锋 |
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| 作者单位： | 内蒙古农业大学动物科学与医学学院;军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室; |
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| 摘 要： | 目的运用Bac-to-Bac杆状病毒表达系统表达新肠道病毒EV71 vP1蛋白(EV71vP1),并对EV71 vP1蛋白的免疫原性进行初步评价。方法利用RT-PCR技术获得EV71 vP1基因,将基因序列克隆到载体pFastBac HTA,通过转座反应,与Bacmid DNA重组,重组后转染Sf9昆虫细胞。采用透射电镜观察法、SDS-PAGE、Western-blot方法对vP1蛋白进行验证,利用ELISA、微量中和抗体方法对蛋白的免疫原性进行初步评价。结果 EV71 vP1蛋白相对分子量约为33KD,表达量约10mg/L;ELISA抗体效价为1∶900;中和抗体效价1∶800。结论利用Bac-to-Bac杆状病毒表达系统成功的表达了EV71 vP1蛋白,并对其免疫原性做了初步评价,为今后EV71 vP1亚单位疫苗的研究提供参考。
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| 关 键 词： | EV71 vP1蛋白 Bac-to-Bac表达系统 中和抗体实验 免疫原性 |
Expression of enterovirus 71 vP1 in baculovirus expression system and evaluation of its immunogenicity |
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| LI Xiaonan,ZHAO Zhongpeng,DUAN Yueqiang,ZHAO Xiaoyan,WANG Xiliang,LI Peifeng College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot ,China.Expression of enterovirus 71 vP1 in baculovirus expression system and evaluation of its immunogenicity[J].Immunological Journal,2010(3). |
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| Authors: | LI Xiaonan ZHAO Zhongpeng DUAN Yueqiang ZHAO Xiaoyan WANG Xiliang LI Peifeng College of Veterinary Medicine Inner Mongolia Agricultural University Hohhot China |
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| Institution: | LI Xiaonan,ZHAO Zhongpeng,DUAN Yueqiang,ZHAO Xiaoyan,WANG Xiliang,LI Peifeng College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China |
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| Abstract: | Enterovirus 71 is one of viruses causing hand-foot-mouth disease.In this study,we expressed enterovirus 71 vP1 by Bac-to-Bac(r) Baculovirus Expression System,and evaluated its immunogenicity.Firstly,EV71 vP1 was amplified by RT-PCR and cloned into pFast HTA plasmid,and then the plasmid was recombined with Bacmid DNA by transposition reaction and transfected into Sf9 cell.The expressed protein was analyzed by ElectroMicrocopy examinations,SDS-PAGE,and Western blotting,while its immunogenicity was measured by... |
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| Keywords: | Enterovirus 71 vP1 Bac-to-Bac(r) Baculovirus Expression System Microneutralization Immunogenicity |
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