| 非洲马瘟病毒VP7基因拼接、表达及重组ELISA方法的建立与初步应用 |
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| 引用本文： | 高志强,张鹤晓,赖平安,谷强,蒲静,汪琳,乔彩霞,吴丹,柏亚铎,张伟. 非洲马瘟病毒VP7基因拼接、表达及重组ELISA方法的建立与初步应用[J]. 畜牧兽医学报, 2008, 39(11) |
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| 作者姓名： | 高志强 张鹤晓 赖平安 谷强 蒲静 汪琳 乔彩霞 吴丹 柏亚铎 张伟 |
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| 作者单位： | 北京出入境检验检疫局动物检疫中心,北京,101113 |
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| 基金项目： | 奥运科技行动计划专项基金 |
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| 摘 要： | 采用人工拼接的方式拼接了非洲马瘟病毒（AHSV）含有绝大多数线性抗原表位的VP7编码基因片段，克隆于pET-30a构建重组质粒pET-30a-VP7，将pET-30a—VP7转化BL21（DE3），经1．0mmol／LIPTG诱导，外源基因以包涵体的形式获得高效表达。通过Dot-EuSA以及ELISA试验证明表达产物具有良好的反应原性。以纯化后表达产物作为诊断抗原包被酶标板建立了检测AHSV抗体的间接ELISA方法。结果表明，抗原的最佳包被浓度为0．25μg／mL，血清的最佳稀释度为1：40，待检血清阳性临界值初步定为0．25。用此方法和商品化ELISA试剂盒检测了184份血清样品，结果完全符合。
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| 关 键 词： | 非洲马瘟 基因拼接 重组ELISA |
Development and Application of the Recombinant ELISA Based on the Assembling and Expression of VP7 Gene of African Sickness Virus |
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| GAO Zhi-qiang,ZHANG He-xiao,LAI Ping-an,GU Qiang,PU Jing,WANG Lin,QIAO Cai-xia,WU Dan,BAI Ya-duo,ZHANG Wei. Development and Application of the Recombinant ELISA Based on the Assembling and Expression of VP7 Gene of African Sickness Virus[J]. Chinese Journal of Animal and Veterinary Sciences, 2008, 39(11) |
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| Authors: | GAO Zhi-qiang ZHANG He-xiao LAI Ping-an GU Qiang PU Jing WANG Lin QIAO Cai-xia WU Dan BAI Ya-duo ZHANG Wei |
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| Affiliation: | GAO Zhi-qiang,ZHANG He-xiao,LAI Ping-an,GU Qiang,PU Jing,WANG Lin,QIAO Cai-xia,WU Dan,BAI Ya-duo,ZHANG Wei(Animal Quarantine Center of Beijing Entry-Exit Inspection , Quarantine Bureau,Beijing 101113,China) |
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| Abstract: | A DNA fragment,which encodes the most linear epitopes of VP7 of African sickness virus(AHSV),was assembled artificially,then cloned into pET-30a.Target protein expressed at very high level as inclusion body after the recombinant pET-30a-VP7 plasmid was transformed into BL21(DE3),and induced with 1.0 mmol/L IPTG.The indirect ELISA for detecting AHSV VP7 protein antibody was established after the reaction activity of the recombinant protein was determined by Dot-ELISA and ELISA.The results obtained indicated ... |
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| Keywords: | african horse sickness gene assembling recombinant ELISA |
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