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目的在大肠埃希菌中高效表达重组人干扰素λ2(rhIFNλ2),并对其抗病毒活性进行初步研究。方法根据大肠埃希菌的偏爱密码子人工设计合成人干扰素λ2(hIFNλ2)的高表达基因,将其克隆到原核表达载体pBV220,在大肠埃希菌中进行表达,表达产物经变性、复性、阳离子交换层析及分子筛层析纯化,测定其抗病毒活性、种属特异性及抗HBV活性。结果在大肠埃希菌表达的目的蛋白占细胞总蛋白量的15%左右,纯化后的纯度达到90%以上,在WISH细胞上抗VSV活性约为1.5×106IU/mg。与rhIFNα2b比较,表达产物在非灵长类细胞上的抗病毒活性较在灵长类细胞上弱,目的蛋白在HepG2.2.15细胞上表现有与rhIFNα2b相似的抗HBV活性。结论hIFNλ2可以在大肠埃希菌内实现高效表达,表达产物具有抗病毒活性,有与rhIFNα2b相似的抗HBV活性,本型IFN有较严格的种属特异性。
Objective To express recombinant human interferon λ2 (rhIFNλ2) efficiently in Escherichia coli and to study its antiviral activity. Methods According to the preferred codons of Escherichia coli, human interferon λ2 (hIFNλ2) highly expressed gene was cloned into prokaryotic expression vector pBV220 and expressed in Escherichia coli. The expressed product was denatured and renatured , Cation exchange chromatography and molecular sieve chromatography to determine its antiviral activity, species-specific and anti-HBV activity. Results The expressed protein in Escherichia coli accounted for about 15% of the total cellular protein and the purified purity was over 90%. The anti-VSV activity on WISH cells was about 1.5 × 106 IU / mg. Compared with rhIFNα2b, the antiviral activity of the expressed product on non-primate cells was weaker than that on primate cells, and the target protein displayed anti-HBV activity similar to that of rhIFNα2b on HepG2.2.15 cells. Conclusions hIFNλ2 can be highly expressed in Escherichia coli. The expressed product possesses antiviral activity and anti-HBV activity similar to that of rhIFNα2b. This IFN has more strict genus-specificities.