重组人干扰素λ1的表达、纯化及生物活性研究  

Expression,Purification and Biological Activities of Recombinant Human Interferon Lambda 1

作　　者：张钫[1] 郑丽舒[1] 袁武梅[1] 王翔[1] 薛鹏浩[1] 麻粉莲[1] 张骞[1] 

机构地区：[1]中国疾病预防控制中心病毒病防控所病毒基因工程国家重点实验室,北京100052

出　　处：《生物技术通讯》2010年第3期310-314,共5页Letters in Biotechnology

摘　　要：目的:利用大肠杆菌系统表达重组人Ⅲ型干扰素λ1(rhIFN-λ1),并进行纯化,以比较其与重组人干扰素-α2b(rhIFN-α2b)生物学活性的差异。方法:密码子优化的rhIFN-λ1基因与pET-44a载体连接构建原核表达质粒,在大肠杆菌BL21(DE3)中表达,表达产物经系列层析纯化;采用细胞病变抑制法比较纯化的rhIFN-λ1与rhIFN-α2b、市售rhIFN-λ1的抗病毒活性;采用MTS法比较这些干扰素的抗肿瘤细胞增殖活性。结果:基因优化后的rhIFN-λ1在大肠杆菌中得以高效表达,用所建立的纯化工艺可制备得到纯度达95.7%的rhIFN-λ1;rhIFN-λ1的抗病毒比活性为3.2×105 U/mg,比市售rhIFN-λ1的比活性(1.1×105 U/mg)略高,抗肿瘤细胞增殖活性为rhIFN-α2b的1.7倍。结论:获得的rhIFN-λ1具有剂量依赖的抗病毒活性和抗增殖活性,其抗病毒活性比市售rhIFN-λ1略高,抗细胞增殖活性高于rhIFN-α2b,提示rhIFN-λ1有较大的应用前景。Objective：To prepare recombinant human interferon-lambda 1（rhIFN-λ1） expressed in Escherichia coli,and compare its biological activity with rhIFN-α2b determine by different assays.Methods：The optimized rhIFN-λ1 gene was inserted into pET-44a vector and transformed into E.coli BL21（DE3） strain.The colony was induced with IPTG,and the expressed protein was purified with chromatographic methods.The antiviral activity of the purified rhIFN-λ1 was evaluated by cytopathic effect inhibition and anti-proliferation activity by MTS assay.Results：The rhFN-λ1 was highly expressed in E.coli and its purity reached 95.7%.The recombinant rhIFN-λ1 showed comparable anti-VSV activity in WISH cells with IFN-α2b,and its anti-proliferation activity was 1.7 fold stronger than that of commercial rhIFN-λ1.Conclusion：The prepared rhIFN-λ1 demonstrates both dose-dependent antiviral activity and anti-proliferation activity in vitro.It shows more potent anti-VSV activity than commercial rhIFN-λ1,and robust anti-proliferation activity than rhIFN-α2b.These results imply its potential clinical applica-tions.

关 键 词：重组人干扰素λ1 表达 纯化 抗病毒活性 抗增殖活性 

分 类 号：Q78[生物学—分子生物学]