Abstract：Abstract: To construct the recombinant plasmid for over-expressing Krüppel-like factor 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/pEGFP-C1) 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.
引用本文:
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.
LI Bo, HU Hai-Yan, TAN Ming-Xian. Construction of Recombinant Plasmid for Over-expressing KLF4 and Its Expression in K562 Cells. Life Science Research, 2017, 21(2): 125-129.
2017, Vol. 21 
