| 口蹄疫病毒非结构蛋白VPg1-2、VPg2-3、VPg3基因的克隆、表达及VPg1-2抗体持续时间的研究 |
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| 引用本文： | 孙明,王宏伟,田克恭,王传彬,陈西钊,遇秀玲,金萍,曹振,许燕辉,赵心力.口蹄疫病毒非结构蛋白VPg1-2、VPg2-3、VPg3基因的克隆、表达及VPg1-2抗体持续时间的研究[J].中国农业科学,2007,40(3):601-607. |
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| 作者姓名： | 孙明 王宏伟 田克恭 王传彬 陈西钊 遇秀玲 金萍 曹振 许燕辉 赵心力 |
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| 作者单位： | 1. 农业部兽医诊断中心,北京,100094
2. 内蒙古自治区兽医工作站,呼和浩特,010010 |
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| 基金项目： | 国家科技部口蹄疫防治科技攻关资助项目 |
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| 摘 要： | 【目的】表达口蹄疫病毒（Foot-and-Mouth Disease Virus，FMDV）非结构蛋白VPg1-2、VPg2-3、VPg3，用于FMDV感染与灭活疫苗动物鉴别诊断和其蛋白功能研究。【方法】对O 型口蹄疫病毒太保毒株3ABC 基因片段中的 VPg1-2 (3B1-2)、VPg2-3 (3B2-3)、VPg3 (3B3)基因进行扩增和亚克隆，并将它们分别插入pGEX-4T-1 表达载体构建了重组表达质粒，经IPTG诱导后，进行Western blotting 分析。以纯化的VPg1-2表达蛋白为抗原建立间接ELISA，对背景清楚的实验牛血清进行检测。【结果】表达的VPg1-2、VPg2-3蛋白可与FMDV阳性血清发生特异性反应，表达的VPg3蛋白没有反应。VPg1-2表达蛋白与对照组（C组）和灭活疫苗反复免疫组（CI组）牛血清均不发生反应，与未免疫直接攻毒组（D组）血清均发生反应，与部分免疫后攻毒组（I组）（4/7）血清发生反应，与部分I组（3/7）血清不发生反应；D组和I组VPg1-2表达蛋白抗体持续时间最长可达90 d以上，最早检出相应抗体的时间为攻毒后第10天。【结论】表达的VPg1-2抗原用于健康牛群、免疫后未感染牛群以及未免疫感染牛群的鉴别诊断的特异性、敏感性达到100%，对免疫后感染牛群进行检测时可能有一定的漏检现象。
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| 关 键 词： | 口蹄疫病毒 非结构蛋白 抗体消长 |
| 收稿时间： | 2005-8-29 |
| 修稿时间： | 2005-08-29 |
Research on Cloning and Expression of VPg1-2, VPg2-3, VPg3 Gene Fragments of Foot-and-Mouth Disease Virus and the Antibody Dynamics of Expressed VPg1-2 Protein |
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| SUN Ming,WANG Hong-wei,TIAN Ke-gong,WANG Chuan-bin,CHEN Xi-zhao,YU Xiu-ling,JIN Ping,CAO Zhen,XU Yan-hui,ZHAO Xin-li.Research on Cloning and Expression of VPg1-2, VPg2-3, VPg3 Gene Fragments of Foot-and-Mouth Disease Virus and the Antibody Dynamics of Expressed VPg1-2 Protein[J].Scientia Agricultura Sinica,2007,40(3):601-607. |
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| Authors: | SUN Ming WANG Hong-wei TIAN Ke-gong WANG Chuan-bin CHEN Xi-zhao YU Xiu-ling JIN Ping CAO Zhen XU Yan-hui ZHAO Xin-li |
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| Institution: | 1.National Veterinary Diagnostic Center, Beijing 100094; 2.Veterinary Service oflnnermongolia, Huhehaote 010010 |
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| Abstract: | Abstract VPg1-2, VPg2-3, VPg3 gene fragments were successfully subcloned from 3ABC gene of the foot-and-mouth disease virus ( FMDV ) Taibao strain and their expressing plasmids were constructed by inserting the target gene fragments into pGEX-4T-1 vector. The expressed proteins were analysed by Western blotting. The results showed that the expressed VPg1-2, VPg2-3 proteins reacted with FMDV positive serum while the reaction of the VPg3 protein was negative. An indirect ELISA was set up using purified VPg1-2 protein as antigen, and 4 groups of background-known cattle sera were tested. Negative results were obtained for the non-vaccinated control group and the vaccinated group and a part of the infected post-vaccination group(3/7). On the contrary, positive results were achieved for the infected group and a part of the infected post-vaccination group(4/7). In the infected group and infected post-vaccination group, the longest duration of the antibody against VPg1-2 lasted for at least 90 days. The earliest appearing time of VPg1-2 antibody was10 days after infection. |
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| Keywords: | Foot-and-mouth disease virus Non-structural protein Antibody dynamics |
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