| 猪O型口蹄疫病毒重组结构蛋白的纯化及间接ELISA方法的建立 |
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| 引用本文： | 宋妮,王凤雪,孙娜,师新川,温永俊,武华. 猪O型口蹄疫病毒重组结构蛋白的纯化及间接ELISA方法的建立[J]. 中国畜牧兽医, 2013, 40(12): 20-26 |
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| 作者姓名： | 宋妮 王凤雪 孙娜 师新川 温永俊 武华 |
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| 作者单位： | 1. 中国农业科学院特产研究所, 特种经济动物分子生物学国家重点实验室, 吉林长春 130112;2. 华威特(北京)生物科技有限公司, 北京 100085 |
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| 基金项目： | 国家高技术研究发展计划（863计划）（2011AA10A213）；吉林省特种经济动物疫病防控研究创新团队（20121823）。 |
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| 摘 要： | 将猪O型口蹄疫病毒（foot and mouth disease virus，FMDV）的VP0、VP1、VP3基因，通过SUMO融合表达载体在大肠杆菌BL21（DE3）中成功表达，获得大小为55、48和40 ku的融合蛋白，Western blotting检测结果表明，表达的蛋白质具有良好的生物学活性。采用亲和层析法纯化表达产物，分别以纯化的SUMO-VP0、SUMO-VP1、SUMO-VP3蛋白为抗原建立了FMDV的间接ELISA检测方法。200份田间血清样品的检测结果表明建立的SUMO-VP0-ELISA、SUMO-VP1-ELISA、SUMO-VP3-ELISA检测方法均具有良好的特异性和敏感性。
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| 关 键 词： | 猪O型口蹄疫病毒 重组结构蛋白 SUMO 纯化 间接ELISA |
| 收稿时间： | 2013-05-07 |
Purification of the Recombinant Structural Protein of Foot and Mouth Disease Virus Serotype O in Swine and Establishment of the Indirect ELISA |
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| SONG Ni,WANG Feng-xue,SUN Na,SHI Xin-chuan,WEN Yong-jun,WU Hua. Purification of the Recombinant Structural Protein of Foot and Mouth Disease Virus Serotype O in Swine and Establishment of the Indirect ELISA[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(12): 20-26 |
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| Authors: | SONG Ni WANG Feng-xue SUN Na SHI Xin-chuan WEN Yong-jun WU Hua |
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| Affiliation: | 1. State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;2. Sinovet (Beijing) Biotechnology Co., Ltd., Beijing 100085, China |
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| Abstract: | The complete genes encoding the structural protein of foot and mouth disease virus (FMDV)(VP0,VP1 and VP3) were subcloned into expression vector SUMO,resulting in the fusion expression plasmid SUMO-VP0,SUMO-VP1 and SUMO-VP3,and then transformed into E.coli BL21(DE3) and induced by IPTG,the fusion proteins were expressed at high level.Western blotting was performed to confirm that the expressed fusion proteins could specifically react with antiserum against FMDV.The expressed fusion proteins were purified by affinity chromatography. Based on the fusion protein further purification,novel indirect ELISA (SUMO-VP0-ELISA,SUMO-VP1-ELISA,SUMO-VP3-ELISA) were developed to detect FMDV antibodies in swine by detecting 200 serum samples,indicating that the indirect SUMO-VP0-ELISA,SUMO-VP1-ELISA,SUMO-VP3-ELISA were specific and sensitive. |
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| Keywords: | foot and mouth disease virus serotype O in swine recombinant structural protein SUMO purification indirect ELISA |
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