| C型口蹄疫病毒型特异性抗原的表达与鉴定 |
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| 引用本文： | 高闪电,独军政,常惠芸,丛国正,邵军军,林彤,宋帅,谢庆阁.C型口蹄疫病毒型特异性抗原的表达与鉴定[J].农业科学与技术,2009,10(5):79-81,95. |
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| 作者姓名： | 高闪电 独军政 常惠芸 丛国正 邵军军 林彤 宋帅 谢庆阁 |
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| 作者单位： | 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州730046 |
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| 基金项目： | 国家"十一五"重大科技支撑计划 |
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| 摘 要： | 目的]为C型FMDV型特异性多克隆抗体、单克隆抗体的制备和FMDV定型提供理论依据。方法]以含有C型口蹄疫病毒（FMDV）结构蛋白基因VP1的重组质粒pGEM—CP1为模板，设计特异性表达引物，扩增VP1及其c端编码区。对C型口蹄疫病毒VP1及其c端进行原核表达，并测定反应原性。利用纯化的C型VP1及其C端融合蛋白建立间接EHSA，分别对0、A、C、Asia1四型豚鼠阳性血清进行检测，确定C型VP1及其C端与其他3型FMDV抗体的型间交叉反应性。结果]构建了pPR0-CVP1、pPR0-CVP1c重组原核表达质粒，实现了C型口蹄疫病毒VP1及其C端的高效表达，目的蛋白的分子量大小分别为33kD和20kD。Westernblot显示，VP1及其C端融合蛋白均可与对应血清型的豚鼠阳性血清反应。C型VP1及其C端与其他血清型的FMDV阳性血清均未发生交叉反应，且以VP1C端的型特异性最好。结论]获得了C型FMDV特异性抗原。
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| 关 键 词： | C型口蹄疫病毒 血清学 交叉反应 血清分型 |
Expression and Identification of the Type-specific Antigen of FMDV of Serotype C |
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| Institution: | GAO Shan-dian, DU Jun-zheng, CHANG Hui-yun, CONG Guo-zheng, SHAO Jun-jun, LIN Tong, SONG Shuai, XlE Qing-ge (Chinese Academy of Agricultural Sciences, Lanzhou Veterinary Research Institute, State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou 730046) |
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| Abstract: | Objective]The aim was to provide a theoretical basis for preparation of polyclonal antibodies and monoclonal antibody that of type specificity, as well as Foot-and-mouth Disease Virus （FMDV） typing.Method] The recombinant plasmid pGEM-CP1 that contained VPI gene of FMDV of serotype C was used as template for the VP1 and its C terminus coding fragments of FMDV of serotypes C amplification. The coding fragments of VP1 and its C terminus were respectively cloned into prokaryotic expression vector for prokaryotic expression and the reactionogenicity was detected. The purified fusion protein of FMDV VP1 and its C terminus of serotype C were used to construct the indirect ELISA method to detect positive sera of four serotypes A, O, C and Asia 1 of guinea pig and determine the cross reactivity of FMDV antibody of VP1 and its C terminus of serotype C with other three serotypes. Result] The recombinant prokaryotic expression plasmids of PPRO-CVP1 and pPRO-CVPlc were constructed, FMDV VP1 and its C terminus of serotype C were expressed in high level, and the molecular weight of target proteins was 33 kD and 20 kD respectively. Western blot result showed that the fusion protein of VP1 and its C terminus could react with the positive sera of guinea pig of the same serotype. ELISA results revealed that VP1 and its C terminus of serotype C are type-specific and no cross-reactivities were shown between guinea pig positive sera of FMDV of serotype C with the other three serotypes, and the C terminus showed better type-specificity. Conclusion] FMDV specific antigen of serotype C was obtained. |
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| Keywords: | Foot-and-mouth disease virus of serotype C Serology Cross reactivity Serotyping |
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