人干扰素IFNα-2b在果蝇细胞中的稳定表达

Stable Expression of Human IFNα-2b in Drosophila Cell

以人干扰素IFNα-2b全长基因的质粒为模板,PCR扩增,PCR产物克隆到表达载体获得重组质粒,经转染和筛选,得到稳定表达的果蝇S2细胞株.经CuSO4诱导表达并对表达产物进行检测,结果显示,在果蝇S2细胞中成功地进行了人干扰素IFNα-2b的蛋白表达,表达产物的大小约为26kD.并且,通过体外的抗病毒活性测定,证明所表达蛋白具有抗病毒活性.该研究为进一步利用该系统表达真核细胞蛋白,研究其结构和功能提供了重要的基础.

The entire human IFNα-2b（h IFNα-2b） gene was amplified by PCR and cloned to the Drosophila expression vector pMT/BiP/V5-His A and generated the recombinant plasmid.After transfection and screening, the cell lines Drosophila S2 was obtained,which stably expressed hIFNα-2b.Western blotting analysis showed that the expressed protein was about 26 kD.Furthermore,the in vitro test showed that it has antiviral activity. From these results,important basis were provided for further uses of this system to express other eukaryotic proteins and study their structures and functions.