| 新城疫病毒F基因在大肠埃希菌中的表达及其抗原性分析 |
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| 引用本文： | 王青,徐彦召,胡建和,王选年,杭柏林.新城疫病毒F基因在大肠埃希菌中的表达及其抗原性分析[J].动物医学进展,2009,30(11):9-12. |
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| 作者姓名： | 王青 徐彦召 胡建和 王选年 杭柏林 |
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| 作者单位： | 1. 河南科技学院,河南新乡,453003
2. 中国农业科学院上海兽医研究所,上海,200241 |
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| 摘 要： | 构建新城疫病毒融合蛋白F基因的原核表达载体,并将其在宿主菌BL21(DE3)感受细胞中表达.以含有融合蛋白F基因的重组质粒pMD19T-F为模板,设计特异性引物,应用PCR技术扩增获得F基因的F1片段,定向插入原核表达载体pET-28a,构建重组质粒pET-F1,将构建成功的重组质粒pET-F1转化宿主菌BL21(DE3)感受态细胞经IPTG诱导,将其在宿主菌细胞中表达,表达产物用SDS-PAGE和Western blot方法检测.结果显示,成功克隆出了新城疫病毒F1基因片段序列852 bp.构建的pET-F1载体经PCR、双酶切、测序鉴定均无误,转化表达宿主菌后经SDS-PAGE和Western blot检测结果显示,目的基因被成功表达,并具有良好的反应原性.成功构建了新城疫病毒的融合蛋白F基因原核表达载体,转化宿主细胞后成功表达了融合蛋白F1重组蛋白片段.
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| 关 键 词： | 新城疫病毒 F基因 原核表达 抗原性 |
Prokaryotic Expression of F Gene of NDV and Antigenicity of Expressed Product |
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| WANG Qing,XU Yan-zhao,HU Jian-he,WANG Xuan-nian,HANG Bo-lin.Prokaryotic Expression of F Gene of NDV and Antigenicity of Expressed Product[J].Progress In Veterinary Medicine,2009,30(11):9-12. |
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| Authors: | WANG Qing XU Yan-zhao HU Jian-he WANG Xuan-nian HANG Bo-lin |
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| Institution: | WANG Qing1,XU Yan-zhao2,HU Jian-he1,WANG Xuan-nian1,HANG Bo-lin1(1.Henan Institute of Science and Technology,Xinxiang,Henan,453003,China,2.Shanghai Veterinary Institute,Chinese Academy of Agricultural Sciences,Shanghai,200241,China) |
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| Abstract: | Newcastle disease virus fusion protein F gene was subcloned into a prokaryotic expression vetor,and then using E.coli BL21(DE3) to express this protein.F gene was amplified from plasmid pMD19T-E2 by PCR,then the gene was subcloned into pET-28a;PCR,digestion and sequencing were used to identify positive plasmid,which named pET-F1;the identified recombined plasmid was expressed by E.coli BL21(DE3) and induced by IPTG.The expressed products were identified by SDS-PAGE and Western blot.The sequence of F1 gene f... |
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| Keywords: | Newcastle disease virus F gene prokaryotic expression antigenicity |
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