生命科学研究2017，Vol.21Issue(2)：125-129,5.DOI:10.16605/j.cnki.1007-7847.2017.02.006
过表达KLF4重组质粒的构建及其在白血病K562细胞中的表达
Construction of Recombinant Plasmid for Over-expressing KLF4 and Its Expression in K562 Cells
摘要
Abstract
To construct the recombinant plasmid for over-expressing Krüppel-likefactor 4 (KLF4) and to study its expression in K562 cells,human KLF4 gene was amplified by RT-PCR,and then cloned into pEGFP-C1 vector to construct pEGFP-KLF4.After confirmed by restriction analysis and sequencing,the recombinant plasmid was transfected into K562 cells by electroporation.The K562 cells with the recombinant plasmid were obtained as an experimental group (K562/pEGFP-KLF4),and the cells with the vector (K562/pEGFPC1) and the untreated cells were both used as control.The expression of pEGFP-KLF4 fusion protein in K562 cells was observed by fluorescence microscopy,and the G418-resistant clones of K562 cells were screened to establish stably transfected K562 cell line.RT-PCR was performed to detect the level of KLF4 mRNA in each group of cells.The results showed that the pEGFP-KLF4 plasmid was constructed successfully.Compared with K562/pEGFP-C1 group and K562 group,the experimental group had a significantly increased level of KLF4 mRNA,with over-expression ratio up to 65.71％ (P＜0.05).The recombinant plasmid proved to be able to over-express KLF4 and the expression level could be high in K562 cells,which lays a foundation for further study on the function of KLF4 over-expression in leukemia.关键词
锌指转录因子Krüppel样因子4基因(KLF4)/过表达质粒/电穿孔转染/白血病/K562细胞Key words
Krüppel-like factor 4 (KLF4)/over-expressing plasmid/electroporation/leukemia/K562分类
医药卫生引用本文复制引用
李波,胡海艳,谭明贤..过表达KLF4重组质粒的构建及其在白血病K562细胞中的表达[J].生命科学研究,2017,21(2):125-129,5.基金项目
重庆市万州区市政府计划项目(201303015) （201303015）
