人干扰素ω(huIFN-ω)的高效表达、纯化及生物学活性

Expression and Purification of Human Interferon-ω and Bioactivity of the Expressed Product

为了获得人干扰素 ω(huIFN ω)在大肠杆菌中的高效表达,根据大肠杆菌的偏爱密码子,人工设计并合成了huIFN ω高表达基因,并克隆到原核表达载体pBV220,在大肠杆菌DH5α中实现了高效表达,目的蛋白占细胞总蛋白的15%左右,以包涵体形式存在。表达产物经变性、复性及阳离子和分子筛层析纯化,纯度达90%以上,产物的抗病毒活性约为1.0×108IU/mg,并具有体外促NK杀伤活性。此结果为进一步研究和开发重组huIFN ω奠定了基础。

According to preferred codons using in E.coli, the highly-expressed human interferon-ω gene was designed,synthesized and cloned into expression vector pBV220 and transferred to E.coli DH5α.The recombinant plasmid pBV220-huIFN-ω was highly expressed in E.coli DH5α.The expressed product existed in inclusion body and contained about 15% of the total somatic protein.The expressed product was purified by CM FF column and size exclusion chromatography and reached a purity above 90%.The anti-virus activity of the product was about 1.0×10~ 8IU/mg and it could promote NK cytotoxicity in vitro. The obtaining of rhuIFN-ω with bioactivity lays a foundation for its further study and exploitation.