| 猪繁殖与呼吸综合征病毒HB-3株GP5-M-N蛋白核酸疫苗的构建及免疫原性 |
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| 引用本文： | 杜冬华,周静,王爱华,陈赛娟,刘涛.猪繁殖与呼吸综合征病毒HB-3株GP5-M-N蛋白核酸疫苗的构建及免疫原性[J].兽医大学学报,2014(2):177-181. |
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| 作者姓名： | 杜冬华 周静 王爱华 陈赛娟 刘涛 |
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| 作者单位： | [1]河北北方学院动物科技学院,河北张家口075131 [2]河北农业大学,河北保定071000 [3]保定瑞普生物药业有限公司,河北保定071000 |
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| 基金项目： | 河北省重大技术创新基金资助项目（07221008Z） |
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| 摘 要： | 为探索猪繁殖与呼吸综合征病毒（PRRSV）核酸疫苗用于免疫预防的可行性，试验用PCR方法扩增出PRRSVHB-3株GP5、M和N基因，通过I，inker序列将GP5和M串联为GP5-M，双酶切后和N基因-起插入真核表达载体构建重组质粒pcDNA-GP5-M-N，经酶切鉴定表明GP5、M和N基因和载体连接正确。然后将重组质粒转染至Marc-145细胞，经间接免疫荧光及Western blot分析证实重组蛋白能在Marc-145细胞中表达。然后用重组质粒pcDNA-GP5-MN免疫Balb／c小鼠，中和抗体检测结果表明，首免后2周即有小鼠产生可检测到的病毒中和抗体（1：4），随后抗体水平快速升高，第8周抗体效价达到最高（1：32）。说明本试验构建的重组质粒pcDNA-GP5-M-N能诱发免疫小鼠产生较高水平的中和抗体，为PRRSV核酸疫苗的研究奠定了基础。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 GP5 M N基因 核酸疫苗 免疫原性 |
Construction and immunogenicity of recombinant plasmid co-expressing GP5, M and N proteins of porcine reproductive and respiratory syndrome virus |
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| Authors: | DU Dong-hua ZHOU Jing WANG Ai-hua CHEN Sai-juan LIU Tao |
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| Institution: | a ( 1. College of Animal Science , Hebei North University, Zhangjiakou , Hebei 075131, China; 2. Hebei Agricultural Uni- versity, Baoding, Hebei 071000, China ; 3. Ringpu ( Baoding ) Biological Pharmaceutical Co. Ltd. ,Baoding, Hebei 071000 ,China) |
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| Abstract: | This expieriment was conducted to study the DNA vaccine for preventing PRRSV infection. The GP5,M and N genes of porcine reproductive and respiratory syndrome virus （PRRSV） were amplified by PCR, and then were subcloned into eukaryotic expression vectors pcDNA3. 1 （＋）. The recombinant named as pcDNA-GP5-M-N was transfected into Marc-145 cells,and its ex pression was detected by indirect immune fluorescence detection. The results showed that GP5 M- N genes of PRRSV HB-3 strain were expressed in Marc-145 cells. Using pcDNA-GP5-M-N to im munize Balb/c mice,anti-PRRSV neutralizing antibodies were detected at two weeks after primary vaccination and reached a peak of titer （1 = 32） at eight weeks. These results showed that recombi- nant plasmid pcDNA-GP5-M-N has been constructed successfully,and with good immunity. These data indicated that pcDNA-GP5-M-N could be used for further development of DNA vaccines for PRRSV. |
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| Keywords: | PRRSV GP5/M/N gene nucleic acid vaccine immunogenicity |
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