Study on expression,purification and biological characteristics of recombinant human interferon-epsilon in E.coli / 中华微生物学和免疫学杂志

ABSTRACT

Objective To construct a novel recombinant human IFN-ε,and to analyze its physi-cal,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 BL2l(DE3).The form of ex-pression product was inclusion bodies.After purification and renaturation.high purity active protein IFN-εwas achieved.The final product was tested for its physical.chemical properties and biological characteristics including anti-viral and anti-proliferative acfivities.Results IFN-ε was expressed in inclusion body in E.coli.After the protein renaturation and purification,the purity was more than 95%.The rhIFN-ε protein had a specific anti-viral activity of about 1.2×103 IU/mg.Its anti-poliferative activity is obvious and can in-duce cells to produce anti-viral protein MxA.Conclusion Human IFN-ε protein was expressed successful-ly.and this protein has anti-virus and anti-prolireration activity.