pEF1alpha-AcGFP1-C1 Vector

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10 ug

pEF1α-AcGFP1-C1 is a mammalian expression vector that constitutively expresses a protein of interest fused to the C-terminus of the green fluorescent protein AcGFP1, even after stable integration of the vector into the host cell genome. Stable, constitutive expression of the bicistronic transcript is driven by the human elongation factor 1 alpha (EF1α) promoter, allowing the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. The unmodified vector can be used to express modified AcGFP1 in mammalian cells.

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

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).