Part:BBa_K678004
PGK, mammalian promoter
Promoter for expression of genes in mammalian cells
Contribution: NUDT_CHINA 2019
Summary：in this part, we made contributions to the quantitative characterization data of mammalian PGK Promoter. We measured the strength of this promoter by the expression of firefly luciferase reporter gene in HepG2 cells.
Methods
To characterize the PGK promoter, firefly luciferase was used as the reporter gene to quantify the transcriptional strength of promoter. To be specific, plasmids carrying PGK promoter-firefly luciferase cassette was transfected into liver carcinoma cell line HepG2 by lipo3000 under standard proytocol. Cells were cultures under standard DMEM High glucose medium under 5% CO2 and 37 ℃ with 10% FBS. Cells were harvested 0, 6, 12 and 24h after transfection, and firefly luciferase activity was measured by Beyotime™ Dual Luciferase Reporter Gene Assay Kit.
Results
The result showed a significant increase of firefly luciferase within 24 hours of transfection.
Figure 1. PGK-drived firefly luciferase activity, Error bar represents SD of at least 3 biological replicates.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 143
Illegal PstI site found at 400 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 143
Illegal PstI site found at 400 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 143
Illegal PstI site found at 400 - 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 143
Illegal PstI site found at 400 - 1000COMPATIBLE WITH RFC[1000]
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//promoter
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