髓系细胞触发受体-1真核表达质粒的构建及鉴定
发布时间：2018-11-10 13:02
【摘要】：研究目的：构建髓系细胞触发受体-1(TREM-1)的真核表达质粒。 实验方法：利用RT-PCR从人外周血总RNA中扩增TREM-1基因的全长cDNA,插入克隆载体pUCm-T,经内切酶BamH I和PstⅠ酶切后,将目的片段插入真核表达载体pEGFP-C3,构建含有TREM-1的重组质粒pEGFP-TREM-1,转染293细胞,用荧光显微镜观察荧光、Western-blot鉴定蛋白表达。将pEGFP-TREM-1重组质粒转染THP-1细胞,经100ng/ml LPS刺激24h后,半定量RT-PCR检测TNF-α和IL-1β基因的mRNA水平。 研究结果：经酶切和基因测序证实,克隆的基因片段为TREM-1基因；转染了TREM-1基因表达载体的293细胞在荧光显微镜下可见荧光,并能表达TREM-1蛋白；重组TREM-1蛋白具有生物学活性,能增强细胞因子TNF-α和IL-1β的mRNA表达水平。 结论：成功构建了能够表达TREM-1的真核表达质粒pEGFP-C3-TREM-1,为进一步研究该基因的功能奠定了实验基础。
[Abstract]:Objective: to construct eukaryotic expression plasmid of myeloid cell trigger receptor-1 (TREM-1). Methods: the full-length cDNA, of TREM-1 gene was amplified from human peripheral blood total RNA by RT-PCR and inserted into the cloned vector pUCm-T, by endonuclease BamH I and Pst 鈪，
本文编号：2322547
[Abstract]:Objective: to construct eukaryotic expression plasmid of myeloid cell trigger receptor-1 (TREM-1). Methods: the full-length cDNA, of TREM-1 gene was amplified from human peripheral blood total RNA by RT-PCR and inserted into the cloned vector pUCm-T, by endonuclease BamH I and Pst 鈪，
本文编号：2322547
本文链接：https://www.wllwen.com/xiyixuelunwen/2322547.html
最近更新
教材专著
