| 猪瘟病毒E~(rns)和E2基因在昆虫细胞中的表达研究 |
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| 引用本文： | 韩建强,信爱国.猪瘟病毒E~(rns)和E2基因在昆虫细胞中的表达研究[J].西南农业大学学报,2011(2). |
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| 作者姓名： | 韩建强 信爱国 |
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| 作者单位： | 玉溪市农业职业技术学院;云南省畜牧兽医科学院; |
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| 基金项目： | 云南省农业科技创新工程资助项目(2008LA019); 国家农业公益性行业科研专项基金资助项目(200803026) |
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| 摘 要： | 利用昆虫细胞/杆状病毒系统表达猪瘟病毒(classical swine fever virus,CSFV)Erns和E2基因,用于CSFV新型疫苗以及建立相关的血清学诊断方法等研究.从猪瘟病毒石门株血液样品中提取总RNA,对Erns及E2基因进行扩增,将产物回收后分别连接于T载体,酶切及测序分析后,将其亚克隆至真核表达转移载体pFastBac HT经重组筛选后获得杆状病毒重组质粒,重组质粒转染昆虫细胞sf9后连续传3~4代,分别收获细胞上清液及沉淀,用于SDS-PAGE及Western-blotting试验,检测重组Erns,E2基因表达蛋白.用抗组氨酸(His)单克隆抗体在表达细胞中检测到Erns重组的His标签蛋白;应用抗E2蛋白单克隆抗体在表达细胞及培养上清液中检测到E2基因表达蛋白.结果表明在杆状病毒系统中成功表达了CSFV Erns和E2蛋白.
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| 关 键 词： | 猪瘟病毒 Erns基因 E2基因 杆状病毒表达系统 |
Expression of E~(rns) and E2 Genes of Classical Swine Fever Virus in Insect Cells |
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| HAN Jian-qiang,XU Wen-hua.Yuxi Agricultural Vocational Technique College,Yuxi Yunnan ,China,.Yunnan Animal Science , Veterinary Institute,Kunming.Expression of E~(rns) and E2 Genes of Classical Swine Fever Virus in Insect Cells[J].Journal of Southwest Agricultural University,2011(2). |
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| Authors: | HAN Jian-qiang XU Wen-huaYuxi Agricultural Vocational Technique College Yuxi Yunnan China Yunnan Animal Science Veterinary Institute Kunming |
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| Institution: | HAN Jian-qiang1,XU Wen-hua21.Yuxi Agricultural Vocational Technique College,Yuxi Yunnan 653100,China,2.Yunnan Animal Science and Veterinary Institute,Kunming 650224 |
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| Abstract: | The Erns and E2 genes of classical swine fever virus(CSFV) were expressed in insect cells/Baculovirus expression system to establish CSFV serological diagnostic method and develop CSFV subunit vaccine.The Erns gene and E2 gene of CSFV shimen strain were amplified from total the RNA of the blood sample,and purified products were cloned into T-easy vector.After identification with enzyme digestion and sequence analysis,two genes were subcloned into the transfer vector pFastBacHT.Recombinant plasmids were obta... |
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| Keywords: | classical swine fever virus Erns gene E2 gene baculovirus expression system |
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