pEF1α-IRES-ZsGreen1 is a bicistronic mammalian expression vector that allows the simultaneous, constitutive expression of a protein of interest and the green fluorescent protein ZsGreen1. Constitutive expression of the bicistronic transcript is driven by the human elongation factor 1 alpha (EF1α) promoter.
Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter
Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).
Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter
Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter. IRES: internal ribosome entry sequence; FP1: fluorescent protein (AcGFP1, DsRed-Monomer, or mCherry); FP2: fluorescent protein (mCherry or ZsGreen1); MCS: multiple cloning site.
pEF1α-IRES-ZsGreen1 is a bicistronic mammalian expression vector that allows the simultaneous, constitutive expression of a protein of interest and the green fluorescent protein ZsGreen1. Constitutive expression of the bicistronic transcript is driven by the human elongation factor 1 alpha (EF1α) promoter.
Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter
Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).
Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter
Map schematic of the plasmid choices that are available carrying the EF1-alpha promoter. IRES: internal ribosome entry sequence; FP1: fluorescent protein (AcGFP1, DsRed-Monomer, or mCherry); FP2: fluorescent protein (mCherry or ZsGreen1); MCS: multiple cloning site.