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目的表达纯化一种新型重组人干扰素Epsilon(rhIFNε155ser),并对它的生物学特性进行初步研究。方法人工合成rhIFNε155ser的全基因序列,并按照大肠埃希菌密码子嗜性作适当改造,然后构建到原核表达载体pBV220,在大肠埃希菌DH5α中表达。将表达的包涵体蛋白纯化复性后,对最终产物的抗病毒活性、细胞生长抑制活性和NK细胞刺激活性进行鉴定。同时,利用芯片技术,从基因的水平对rhIFNε155ser的生物机制进行初步的了解。结果rhIFNε155ser以包涵体的形式表达,经纯化蛋白纯度可达95%。复性后rhIFNε155ser在WISH抗VSV系统中的抗病毒活性可达6×105IU/mg。在100～1000pg/ml下rhIFNε155ser具有剂量依赖性的抗增殖活性,而NK细胞刺激活性无剂量依赖性。基因芯片扫描发现,22278个位点中,有283个基因显著上调,另外1894个显著下降。结论成功地表达了rhIFNε155ser,发现此IFN具有一定的抗病毒、抗增殖和NK细胞刺激活性。芯片技术可进一步拓展对IFN的功能研究。
Objective To express and purify a novel recombinant human interferon Epsilon (rhIFNε155ser) and study its biological characteristics. Methods The whole gene sequence of rhIFNε155ser was synthesized and transformed into E.coli DH5α according to the codonophilicity of Escherichia coli. The recombinant plasmid was then constructed into prokaryotic expression vector pBV220 and expressed in Escherichia coli DH5α. After purified and renatured the expressed inclusion body protein, the antiviral activity, cytostatic activity and NK cell stimulating activity of the final product were identified. At the same time, using the chip technology, preliminary understanding of the biological mechanism of rhIFNε155ser from the gene level. Results rhIFNε155ser was expressed in the form of inclusion bodies. The purity of the purified protein was up to 95%. After refolding, the antiviral activity of rhIFNε155ser in WISH anti-VSV system was up to 6 × 105IU / mg. RhIFNε155ser exhibited dose-dependent antiproliferative activity at 100-1000 pg / ml, whereas NK cell-stimulating activity was dose-independent. Gene chip scan found that out of 22278 loci, 283 genes were significantly up-regulated, while 1894 were significantly decreased. Conclusion The rhIFNε155ser was successfully expressed and found to possess anti-viral, antiproliferative and NK cell stimulating activity. Chip technology can further expand the function of IFN research.