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融合干扰素-BLA(IFN-BLA)的构建、表达、纯化及抗病毒活性测定(英文)

Construction, Expression, Purification and Antiviral Activity of Fused Interferon-BLA (IFN-BLA)

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【作者】 马文哲沈琼韩威陆晨轶杨胜利龚毅

【Author】 MA Wen-Zhe, SHEN Qiong, HAN Wei, LU Chen-Yi, YANG Sheng-Li, GONG Yi (Shanghai CASB Biotechnology CO., LTD, Chinese Academy of Sciences, Shanghai 200233, China)

【机构】 中国科学院上海中科伍佰豪生物工程有限公司中国科学院上海中科伍佰豪生物工程有限公司 上海200233上海200233上海200233

【摘要】 干扰素BLA(IFN BLA)由干扰素beta1b(IFN beta1b)和干扰素alpha2b(IFN alpha2b)通过连接肽GGGS融合而成。优化了其在大肠杆菌BL21CodonPlus(DE3)RIL中的实验室表达条件,表达的目的蛋白占菌体总蛋白的35%以上并且主要以包涵体的形式存在。对包涵体的复性条件进行了摸索,建立了IFN BLA的复性及纯化方法,纯化后的蛋白产量约为45mg/L,纯度在90%以上。抗病毒活性分析表明这一新的融合蛋白可能具有协同或加成活性。

【Abstract】 A fusion protein, Interferon-BLA (IFN-BLA), was constructed with IFN-beta-1b and IFN-alpha-2b separating by a linker -GGGS-. The laboratory-scale expression conditions in E.coli BL21 CodonPlus (DE3)-RIL were optimized and IFN-BLA was expressed higher than 35% of total protein in the cells mainly as inclusion body. The inclusion body of IFN-BLA was denatured and refolded by dialysis and purified by ion-exchange chromatography. The overall yield of IFN-BLA was about 45 mg/L with purity higher than 90%. Antiviral activity assay suggested that this newly fused (protein) may have synergetic or additive antiviral activities.

【关键词】 IFN-BLA表达纯化抗病毒活性
【Key words】 IFN-BLAexpressionpurificationantiviral activity
  • 【分类号】Q78
  • 【下载频次】114
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