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鹅坦布苏病毒非结构蛋白NS1的原核表达及纯化

DOI: 10.3969/j.issn.1000-7091.2012.06.003, PP. 11-14

Keywords: 鹅坦布苏病毒,NS1蛋白,原核表达

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Abstract:

根据鹅坦布苏病毒JS804株非结构蛋白NS1基因序列,设计1对特异性引物,利用PCR方法扩增得到完整的NS1基因,并将其克隆至原核表达载体pET28a和pET32a上,构建出重组表达质粒pET28a-NS1和pET32a-NS1,转化至BL21(DE3)中,经IPTG诱导得到NS1融合蛋白(His-NS1),其分子质量约分别为44,58kDa,均在诱导后6h达到表达量高峰。分析显示,2种融合蛋白均以包涵体形式存在,包涵体经过变性和复性后均可获得单一、高表达量的目的蛋白,为进一步开展关于鹅坦布苏病毒NS1蛋白的研究奠定了基础。

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