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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
引用本文:马文哲,沈琼,韩威,陆晨轶,杨胜利,龚毅.融合干扰素-BLA(IFN-BLA)的构建、表达、纯化及抗病毒活性测定[J].遗传,2005,27(3):451-456.
作者姓名:马文哲  沈琼  韩威  陆晨轶  杨胜利  龚毅
作者单位:中国科学院上海中科伍佰豪生物工程有限公司, 上海 200233
摘    要: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。优化了其在大肠感菌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IFN-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。

关 键 词:抗病毒活性  纯化  IFN-BLA  表达  
收稿时间:2004-09-20
修稿时间:2004-09-20

Construction, Expression, Purification and Antiviral Activity of Fusing Interferon-BLA (IFN-BLA)
MA Wen-zhe,SHEN Qiong,HAN Wei,LU Chen-Yi,YANG Sheng-Li,GONG Yi.Construction, Expression, Purification and Antiviral Activity of Fusing Interferon-BLA (IFN-BLA)[J].Hereditas,2005,27(3):451-456.
Authors:MA Wen-zhe  SHEN Qiong  HAN Wei  LU Chen-Yi  YANG Sheng-Li  GONG Yi
Institution:Shanghai CASB Biotechnology CO., LTD, Chinese Academy of Sciences, Shanghai 200233, China
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.
Keywords:IGN-BLA  IFN-BLA  expression  purification  antiviral activity
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