FTL-EGFP融合基因真核表达载体的构建及其表达
发布时间：2018-08-10 19:58
【摘要】：[目的]构建小鼠铁蛋白轻链(ferritin light chain,FTL)的增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)真核表达载体并检测其表达。[方法]利用PCR扩增技术得到小鼠560 bp的FTL基因编码序列,将此序列插入到含有增强型绿色荧光基因EGFP的真核表达载体PEGFP/N1多克隆位点区域中的限制内切酶HindⅢ和Bam HⅠ之间,得到真核表达载体m FTL-PEGFP/N1。经酶切和测序鉴定后,将构建成功的m FTL-PEGFP/N1及其对照空载体PEGFP/N1分别转染到小鼠腹腔巨噬细胞RAW264.7细胞中,提取蛋白后利用免疫印迹技术检测细胞中带有PEGFP标签的FTL蛋白的表达。[结果]新构建的m FTL-PEGFP/N1重组质粒经酶切鉴定得到预期片段,进一步的测序结果显示所构建的重组质粒中插入的小鼠FTL基因的c DNA序列正确。[结论]利用分子克隆技术获得了带有绿色荧光蛋白标签的小鼠铁蛋白轻链真核表达载体m FTL-PEGFP/N1,并在细胞中表达良好。
[Abstract]:[objective] to construct the eukaryotic expression vector of enhanced green fluorescent protein (enhanced green fluorescent) and detect its expression in mouse ferritin light chain (ferritin light). [methods] the 560bp FTL gene encoding sequence of mice was obtained by PCR amplification and inserted into the region of restriction endonuclease Hind 鈪，
本文编号：2176040
[Abstract]:[objective] to construct the eukaryotic expression vector of enhanced green fluorescent protein (enhanced green fluorescent) and detect its expression in mouse ferritin light chain (ferritin light). [methods] the 560bp FTL gene encoding sequence of mice was obtained by PCR amplification and inserted into the region of restriction endonuclease Hind 鈪，
本文编号：2176040
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