摘要
目的研究重组干扰素-tau的纯化和复性。方法将构建好的重组质粒pBV220转化E.coli BL21,发酵培养后迅速升温至42℃诱导干扰素-tau以包涵体的形式高效表达。裂菌后利用8mol/L尿素溶解目的蛋白质,经凝胶过滤色谱层析纯化后,再透析复性。结果干扰素-tau的表达量占菌体总蛋白质量的20%以上,纯化后纯度可达9 6%以上,活性得到有效恢复。结论建立了干扰素-tau纯化及复性的方法,获得了高表达、高纯度、有活性的干扰素-tau。
Objective To study the purification and renaturation of recombinant interferon-tau（IFN-tau）. Methods Recombinant plasmid pBV220 carrying interferon-tau gene was transformed into E. coli BL21. IFN-tau was highly expressed as the form of inclusion bodies in E. coil BL21 which was cultured by fermentation with quickly raising temperature to 42 ℃. Carbamide （Smol/L） was used to dissolve interest protein after the bacterium broke down, then, IFN-tau was purified by gel filtration chromatography and renatured by dialysis. Results The expression of IFN-tau was above 20% of total bacterial protein. The purity of IFN-tau was about 98% and its activity was well recovered. Conclusion The process of purification and renaturation of IFN-tau is established, and the IFN-tau with high expression, high purity and high activity can be obtained.
出处
《食品与药品》
CAS
2006年第11A期35-38,共4页
Food and Drug
基金
山东省科技厅资助项目(No.0310501010)
