人IFN-ε的表达、纯化及生物学特性研究  

Study on expression, purification and biological characteristics of recombinant human interferon-epsilon in E. coli

作　　者：李莉莉[1] 张振龙[1] 

机构地区：[1]北京生物制品研究所,100024

出　　处：《中华微生物学和免疫学杂志》2008年第8期749-753,共5页Chinese Journal of Microbiology and Immunology

摘　　要：目的表达并纯化人IFN-ε，并对其理化特性和生物学特性进行初步研究。方法利用PCR技术以人基因组DNA为模板，获得目的片段的全基因序列后，克隆到原核表达载体pET-32a（＋）上，构建重组表达载体pET-32a（＋）／IFN-ε，转化到大肠杆菌BL21（DE3）中进行表达，并对表达进行了优化。产物的表达形式为包涵体，经过纯化复性后，获得高纯度的活性蛋白IFN-ε。对IFN-ε的理化性质和抗病毒活性、抑制肿瘤细胞增长活性、刺激细胞产生抗病毒蛋白的活性进行了鉴定。结果IFN-ε以包涵体形式表达，纯化后纯度大于95％，抗病毒活性为1．2×10^5 IU／mg，能够抑制肿瘤细胞的生长，并能诱导细胞产生抗病毒蛋白MxA。结论成功表达了人IFN-ε蛋白，并证明此蛋白质具有抗病毒和抗细胞增殖的活性。Objective To construct a novel recombinant human IFN-ε, and to analyze its physical, chemical properties and biological characteristics. Methods Human genomic DNA was used as the template to synthesize IFN-ε gene by PCR. The sequence was cloned into plasmid vector pET-32a（ ＋ ）, and the recombinant plasmid pET-32a（ ＋ ）/IFN-ε was transformed into E. coli BI21 （DE3）. The form of expression product was inclusion bodies. After purification and renaturation, high purity active protein IFN-ε was achieved. The final product was tested for its physical, chemical properties and biological characteristics including anti-viral and anti-proliferative activities. Results IFN-ε was expressed in inclusion body in E. coll. After the protein renaturation and purification, the purity was more than 95%. The rhIFN-ε protein had a specific anti-viral activity of about 1.2 ×10^5 IU/mg. Its anti-poliferative activity is obvious and can induce cells to produce anti-viral protein MxA. Conclusion Human IFN-ε protein was expressed successfully, and this protein has anti-virus and anti-proliferation activity.

关 键 词：干扰素 包涵体 蛋白复性 生物活性 

分 类 号：R91[医药卫生—药学]