Exploration of the variance in exogenous gene expression driven by the different promoters in leukemia cells
Yan Zhou, Qiuhua Huang
Abstract
Yan Zhou, Qiuhua Huang
Abstract
To investigate the variance in exogenous gene expression driven by the different promoters in leukemia cells, four GFP reporter lentivirus vectors carrying different promoters, including EF1α, PGK, Ubiquitin and CMV, were selected. Leukemia cell lines NB4, THP1, HL60 and Kasumi were infected with lentivirus produced from these reporter vectors, respectively. Then, fluorescence microscope, flow cytometry and fluorescence quantitative PCR were used to detect the GFP expression strength. The results of this study clearly showed that the expression levels of the reporter genes with four different promoters were significantly different. Among them, EF1α drove the highest level of GFP expression, while CMV promoter induced the lowest level. Our results suggested that promoters should be carefully chosen in order to get the appropriate exogenous expression level in leukemia cells according to the study need.
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To investigate the variance in exogenous gene expression driven by the different promoters in leukemia cells, four GFP reporter lentivirus vectors carrying different promoters, including EF1α, PGK, Ubiquitin and CMV, were selected. Leukemia cell lines NB4, THP1, HL60 and Kasumi were infected with lentivirus produced from these reporter vectors, respectively. Then, fluorescence microscope, flow cytometry and fluorescence quantitative PCR were used to detect the GFP expression strength. The results of this study clearly showed that the expression levels of the reporter genes with four different promoters were significantly different. Among them, EF1α drove the highest level of GFP expression, while CMV promoter induced the lowest level. Our results suggested that promoters should be carefully chosen in order to get the appropriate exogenous expression level in leukemia cells according to the study need.
Key concepts: Promoter, Reporter gene, Green fluorescent protein, Flow cytometry, Molecular biology, Biology, Leukemia, Fluorescence microscope