Part:BBa_K2751015
Human DKK1 promoter
Usage and Biology
The human DKK1 promoter is the promoter regulating the human DKK1 gene. The promoter is located on human chromosome 10, and will be activated upon DKK1 gene activation.
Characterization
We used PCR technique to amplify a segment of human chromosome 10 (from 52311944 to 52314294). The estimated length of our DKK1 promoter segment is around 2356bp.
Functionality Test
To test the functionality of the DKk1 promoter, we transfected HEK 293 cells with a plasmid (BBa_K2751024) containing the DKK1 promoter and a mCherry reporter protein. We added various concentrations of testosterone to stimulate the DKK1 promoter. The expression of mCherry was read by IX83 confocal microscope using the same exposure time, and we selected a field full of cells from each condition group to control the amount of cells observed. Quantification of cells expressing the mCherry protein was determined via Metamorph multi-wavelength cell scoring. Intensity 10 graylevels above local background was determined as real fluorescence emission.
| Picture taken under 10x | Concentration of Testosterone | Number of cells expressing mCherry protein determined by Metamorph |
|---|---|---|
 |
0.05ug/mL | 14 |
 |
0.5ug/mL | 72 |
 |
5ug/mL | 77 |
As the concentration of testosterone rises from 0.05ug/mL to 5ug/mL, the number of cells that express mCherry protein also increases. This shows that testosterone level within a range can stimulate the activation of DKK1 promoter, resulting in increasing expression of mCherry protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1800
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1895
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1247
Illegal SapI site found at 1674
| None |