Part:BBa_K2557033
EF1α promoter
The elongation factor-1 promoter is a high expression synthetic promoter in mammalian cells.
Characterization
Fig. 1 Fluorescence microscope observation of HEK 293T only transfect with plasmids containing promoters with tetO sequence.
Fig. 1 shows that the strength of the three promoters in HEK 293T cells is from strong to weak: Ubc > EF1 α > miniCMV.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Contribution:BM-AMU 2019
CAG&EF1a
Summary:BM-AMU 2019 made contributions to the quantitative characterization data of pEF1a. We measured the strength of EF1a promoter (pEF1a) and compare it with other three promoters.
Our team built four kinds of plasmids which are built based on pGuide, with the pEF1a, pCAG, pCMV and pUbiquitin activating the expression of GFP. The ex-pression of GFP can reflect the promoter strength after building complete four plas-mids. Transfect the plasmids respectively into 293FT cells by liposome transfec-tion.The resistance of plasmids was used to screen the cells, then we collected samples after 24/48 hours. Finally, compare GFP expression of pEF1a, pCAG, pCMV and Ubiquitin in 293FT cells by flow cytometry. The experimental procedures used in this assay involved measuring fluorescence using Median Fluorescence Intensity . Our experiment, however, does reveal the relative strength of the EF1a promoter device as compared to three promoters. we obtained the result of the diagram below:
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