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牛病毒性腹泻病毒Erns基因的真核表达及抗原性检测
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吴立君，师东方，王丹娜，何海娟，吴明福，王君伟
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（东北农业大学 动物医学院，黑龙江 哈尔滨150030）
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摘要：
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为研究牛病毒性腹泻病毒(BVDV) Erns基因的生物学功能，将含有牛病毒性腹泻病毒Erns基因的质粒pMD18-T-Erns经BamHⅠ/HindⅢ双酶切，获得了Erns片段，再与杆状病毒转移载体pBlueBacHis2A连接，构建成重组质粒。将重组质粒pBlueBacHis2A-Erns与Bac-N-BlueTMDNA共转染至sf9昆虫细胞中，获得了重组病毒，经噬斑筛选纯化，感染sf9昆虫细胞进行表达。SDS-PAGE分析结果表明，表达的目的蛋白大小约30ku；Western-blotting检测表明，该蛋白具有良好的抗原性。 |
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关键词：
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牛病毒性腹泻病毒；Erns基因；重组杆状病毒 |
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中图分类号：S 852.659.6：Q 786 文献标识码：
A 文章编号：1673-4696(2006)06-0439-05 |
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Eukaryotic expression of Erns gene of bovine viral diarrhea virus
and antigenic detection of the expressed protein |
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WU Li-jun，SHI Dong-fang，WANG Dan-na，HE Hai-juan, WU Ming-fu, WANG Jun-wei |
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(College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China) |
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Abstract: In order to study the biological functions of Erns gene of bovine viral diarrhea virus(BVDV), pMD18-T-Erns was digested with BamHⅠand HindⅢ, and Erns gene fragment was obtained. The Erns gene was subcloned into the baculovirus transfer vector pBlueBacHis2A, and the recombinant plasmid pBlueBacHis2A-Erns was constructed. pBlueBacHis2A-Erns and Bac-N-BlueTM DNA were co-transfected into sf9 cells. After the plaques were screened, sf9 cells were infected with the recombinant virus. SDS-PAGE analysis showed that the target protein of 30ku in size was expressed. The protein was proved to have good antigenicity by Western-blotting analysis. |
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Key words:bovine viral diarrhea virus; Erns gene; recombinant baculovirus |
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