| 水貂肠炎病毒VP2基因在昆虫杆状病毒表达系统中的表达 |
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| 引用本文： | 倪佳,张蕾,柴秀丽,高晗,张海玲,赵建军,白雪,邵西群,闫喜军.水貂肠炎病毒VP2基因在昆虫杆状病毒表达系统中的表达[J].中国畜牧兽医,2011,38(9):97-100. |
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| 作者姓名： | 倪佳 张蕾 柴秀丽 高晗 张海玲 赵建军 白雪 邵西群 闫喜军 |
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| 作者单位： | 中国农业科学院特产研究所,预防兽医实验室,吉林长春 130122 |
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| 基金项目： | 科技部科研院所社会公益研究专项“野生动物重要传染病生态学发生,监测和控制技术”(2005DIB4J048) |
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| 摘 要： | 利用昆虫杆状病毒表达系统表达水貂肠炎病毒(mink enteritis virus,MEV) VP2 基因,表达的蛋白能够形成病毒样粒子,具有反应原性,为研究MEV新型疫苗奠定基础。采用PCR方法扩增MEV VP2 基因,将PCR产物连接到pMD18-T载体,目的基因定向克隆到pFast-BacⅠ载体中,构建重组转座载体后转化DH10 Bac感受态细胞,获得重组Bacmid 质粒后转染Sf9昆虫细胞,传毒3代,对表达蛋白进行 Western blotting鉴定。结果成功克隆MEV VP2基因。Western blotting结果证实,表达蛋白能够被MEV单克隆抗体识别。在 Bac-To-Bac 杆状病毒表达系统中成功的表达了MEV VP2蛋白,目的蛋白具有较好的反应原性。
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| 关 键 词： | 水貂肠炎病毒 VP2基因 病毒样粒子 杆状病毒表达系统 |
| 收稿时间： | 2011-01-25 |
Expression of VP2 Gene of Mink Enteritis Virus in Baculovrius Expression System |
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| NI Jia,ZHANG Lei,CHAI Xiu-li,GAO Han,ZHANG Hai-ling,ZHAO Jian-jun,BAI Xue,SHAO Xi-qun,YAN Xi-jun.Expression of VP2 Gene of Mink Enteritis Virus in Baculovrius Expression System[J].China Animal Husbandry & Veterinary Medicine,2011,38(9):97-100. |
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| Authors: | NI Jia ZHANG Lei CHAI Xiu-li GAO Han ZHANG Hai-ling ZHAO Jian-jun BAI Xue SHAO Xi-qun YAN Xi-jun |
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| Institution: | Laboratory of Preventive Veterinary Medicine, Institute of Special Wild Economic Animal and Plant Science, CAAS, Changchun 130122, China |
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| Abstract: | To obtain expression of VP2 gene of MEV in Bac-To-Bac expression system and develop MEV subunit vaccine. MEV VP2 gene was amplified by PCR method and then cloned into pMD18-T vetcor. The VP2 gene was inserted into pFast-BacⅠ vector,then transferred the recombinant plasmid pFast-BacⅠ-VP2 into DH10 Bac to get recombinant Bacmid. Subsequently,the recombinant bacmid was transfected into Sf9 cells. The expressed recombinant protein was identified by Western blotting after three cell passages.VP2 gene of MEV was successfully cloned. The obtained recombinant VP2 protein reacted with MEV monoclonal antibody was identified by Western blotting. The VP2 gene of MEV is successfully expressed in Bac-To-Bac expression system and shows well reactivity with MEV monoclonal antibody. |
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| Keywords: | mink enteritis virus VP2 gene virus-like particles baculovirus expression system |
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