摘要
本文采用RT-PCR技术从LPS刺激后的小鼠腹腔巨噬细胞,扩增小鼠IL-6cDNA,并克隆构建pGEM-3Zf(+)IL-6质粒,继将小鼠所得cDNA克隆到pVL1392载体上,利用杆状病毒AcNPV表达系统在Sf9中进行瞬间表达,用IL-6依赖细胞株MH60·BSF2检测其表达活性,结果表明,感染后48h后就具有明显IL-6表达,由此可见,利用该系统可成功地表达小鼠IL-6基因。
Murine IL-6 cDNA was amplified with RT-PCR technique from LPS-stimulated murine peritoneal macrophages and cloned to pGEM-3Zf(+). Mouse IL-6 gene was ligated into a baculovirus transfer vector pVL-1392. Sf9 cells were transfected transiently with the wild-type baculovirus AcNPV DNA and transfer vector. Activity of IL-6 from the expression system was detected with IL-6 dependent cell line, MH60·BSF2.It was observed that the activity of IL-6 was detected 48h after transfecting significantly. This result suggests that murine IL-6 gene was successfully expressed with the Sf9 expression system.
出处
《上海免疫学杂志》
CSCD
北大核心
1995年第2期69-73,共5页
Shanghai Journal of Immunology