小鼠白细胞介素6的cDNA克隆及其在昆虫细胞中的表达

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。

Cloning and Expression in Spodoptera frugiperda 9(Sf9)of murine IL-6 cDNA

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.