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小鼠白细胞介素6的cDNA克隆及其在昆虫细胞中的表达

Cloning and Expression in Spodoptera frugiperda 9(Sf9)of murine IL-6 cDNA
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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
关键词 RT-PCR 杆状病毒 CDNA 白细胞介素6 IL-6 IL-6 cDNA RT-PCR baculovirus
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