融合干扰素-BLA(IFN-BLA)的构建、表达、纯化及抗病毒活性测定

摘要

A fusion protein, Interferon-BLA (IFN-BLA), was constructed with IFN-beta-1 b 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.%干扰素-BLA(IFN-BLA)由干扰素-beta-1b(IFN-beta-1b)和干扰素-alpha-2b(IFN-alpha-2b)通过连接肽-GGGS-融合而成.优化了其在大肠杆菌BL21 CodonPlus(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lFN-BLA的复性及纯化方法,纯化后的蛋白产量约为45 mg/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.