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O型FMDV VP1基因的真核表达及其
产物的生物学活性
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李刚， 朱丽杰， 曲哲会， 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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根据已克隆的O型口蹄疫病毒VP1基因序列，设计了1对带有SacⅠ和HindⅢ酶切位点的引物,用其将pMD18-T-VP1质粒中的VP1基因亚克隆到真核表达载体pBlueBacHis2A中,成功地构建了重组表达质粒pBlueBacHis2A-VP1(633bp)。将pBlueBacHis2A-VP1(633bp)质粒与Bac-N-BlueTMDNA共转染sf9昆虫细胞，蚀斑筛选重组病毒后，将纯化的重组病毒感染sf9细胞，获得了32.6ku的目的蛋白条带。Dot-ELISA分析结果表明，该表达产物具有反应活性,可用于建立间接ELISA方法，进行口蹄疫病毒抗体检测。 |
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关键词：
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口蹄疫病毒； 昆虫细胞； 真核表达； 转染； 酶联免疫吸附试验 |
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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)03-0183-06 |
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Eukaryotic expression of the VP1 gene of foot-and-mouth disease virus
type O and characterization of expressed fusion protein |
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LI Gang, ZHU Li-jie， QU Zhe-hui, WANG Jun-wei |
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(College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China) |
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Abstract: According to the published VP1 gene sequence, one pair of primers with SacⅠ and HindⅢ endonuclease sites was designed. A VP1 gene in the plasmid
pMD18-T-VP1 was subcloned into an eukaryotic vector pBlueBacHis2A, and the recombinant plasmid
pBlueBacHis2A-VP1(633bp) was constructed successfully. The plasmid
pBlueBacHis2A-VP1(633bp) and Bac-N-BlueTM DNA were co-transfected into sf9 cells. After screening the plaques, sf9 cells were infected with the recombinant virus. The target protein of 32.6ku in size was obtained.
Dot-ELISA analysis showed that the protein had good antigenicity. |
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Key words: foot-and-mouth disease virus; insect cell; eukaryotic expression; transfection; ELISA |
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