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)

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。

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.