| 鸡传染性法氏囊病病毒BC6/85株VP2基因的原核表达、纯化及鉴定 |
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| 引用本文： | 刘丹.鸡传染性法氏囊病病毒BC6/85株VP2基因的原核表达、纯化及鉴定[J].中国兽药杂志,2015,49(5):17-21. |
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| 作者姓名： | 刘丹 |
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| 作者单位： | 中国兽医药品监察所 |
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| 基金项目： | 中国兽医药品监察所所级课题（201421） |
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| 摘 要： | 为获得可用于鸡传染性法氏囊病病毒(IBDV)抗体检测的重组抗原VP2蛋白,根据GenBank中发表的IBDV VP2序列设计一对特异性引物,应用RT-PCR技术克隆IBDV经典标准攻毒株(BC6/85株)的VP2基因,插入质粒pET-32a中构建重组表达质粒pET-32a-VP2,经IPTG诱导后获得了以包涵体形式表达的重组蛋白。重组蛋白纯化后,Western-blot检测表明具有良好的反应原性。本研究为下步建立IBDV抗体的间接ELISA方法及新型疫苗的研制奠定了基础。
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| 关 键 词： | 鸡传染性法氏囊病病毒 VP2基因 克隆 原核表达 |
| 收稿时间： | 2014/11/25 0:00:00 |
| 修稿时间： | 2015/2/10 0:00:00 |
Prokaryotic Expression, Purification and Identification of VP2 Gene of Infectious Bursal Disease Virus BC6/85 Strain |
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| Institution: | China Institute of Veterinary Drug Control |
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| Abstract: | In order to obtain recombinant VP2 antigen for chicken infectious bursal disease virus (IBDV) antibody detection, a pair of specific primers were designed according to the published sequence of IBDV VP2 gene. The VP2 gene was amplified by RT-PCR from IBDV BC6/85 strain and cloned into pET-32a vector, the recombinant plasmid pET-32a-VP2 was induced by IPTG to get the recombinant protein expressed in inclusion body forms. After the recombinant protein was purified, Western blot detection showed that the expressed protein had good antigenicity and could be used for IBDV antibody detection. The research provides a foundation for IBDV antibody detection research and development of new vaccines. |
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| Keywords: | infectious bursal disease virus BC6/85Strain VP2 gene prokaryotic expression |
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