| PRRSV变异株ORF6基因与IL-18共表达真核质粒的构建及其表达 |
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| 引用本文： | 王晓莉,李志杰,丁壮,张泉鹏,宣华,王贵平.PRRSV变异株ORF6基因与IL-18共表达真核质粒的构建及其表达[J].中国兽医学报,2010,30(10). |
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| 作者姓名： | 王晓莉 李志杰 丁壮 张泉鹏 宣华 王贵平 |
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| 作者单位： | 1. 吉林大学,畜牧兽医学院,吉林,长春,130062
2. 广东省农科院,兽医研究所,广东,广州,510000 |
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| 基金项目： | 广东省科技攻关重大项目 |
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| 摘 要： | 针对白细胞介素18(IL-18)基因和本实验室分离的PRRSVJL/07/SW毒株全基因序列,分别设计了2对特异性引物,扩增出IL-18基因和ORF6基因。将2个目的基因克隆于真核表达载体pEGFP-N1中,成功构建了重组真核质粒pEGFP-ORF6和pEGFP-IL18-ORF6,阳性质粒转染Marc-145细胞进行筛选。通过表达载体在转染细胞中的高效转染,增强型绿色荧光蛋白(EGFP)快速、准确的反映出阳性质粒在细胞质中的正确表达,再通过SDS-PAGE和Western blot鉴定构建的真核表达质粒pEGFP-IL18-ORF6的正确性。结果表明,ORF6基因和IL-18基因在Macr-145细胞中能有效表达。结论,所构建的真核质粒pEGFP-IL18-ORF6结构正确,能够在Marc-145细胞中高效表达,而且表达产物具有特异免疫学反应。
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| 关 键 词： | PRRSV 真核质粒pEGFP-IL18-ORF6 表达 |
Construction and identification of eukaryotic expression plasmid for IL-18 and ORF6 of porcine reproductive and respiratory syndrome virus |
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| WANG Xiao-li,LI Zhi-jie,DING Zhuang,ZHANG Quan-peng,XUAN Hua,WANG Gui-ping.Construction and identification of eukaryotic expression plasmid for IL-18 and ORF6 of porcine reproductive and respiratory syndrome virus[J].Chinese Journal of Veterinary Science,2010,30(10). |
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| Authors: | WANG Xiao-li LI Zhi-jie DING Zhuang ZHANG Quan-peng XUAN Hua WANG Gui-ping |
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| Institution: | WANG Xiao-li1,LI Zhi-jie1,DING Zhuang1,ZHANG Quan-peng1,XUAN Hua2,WANG Gui-ping2(1.College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China,2.Guangdong Veterinary Research Institute,Guangzhou 510000,China) |
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| Abstract: | The genes of IL-18 and ORF6 were amplified using two pairs of primers based on the gene of interleukin 18(IL-18) and the sequence of mutant PRRSV JL/07/SW strain which was isolated in our laboratory.The products were cloned into the eukaryotic expression vector pEGFP-N1 to construct the recombinant eukaryotic expression plasmids pEGFP-ORF6 and pEGFP-IL18-ORF6,which were then used to transfer to Marc-145 cells for screening.The enhanced green fluorescent protein(EGFP) could quickly and exactly reflect the ex... |
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| Keywords: | PRRSV eukaryotic plasmid pEGFP-IL18-ORF6 expression |
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