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Team:Hong Kong HKUST/characterization/cmv
From 2013.igem.org
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<h3>Introduction</h3> | <h3>Introduction</h3> | ||
| + | <p>CMV (Cytomegalovirus) promoter is a constitutive mammalian promoter. </p> | ||
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<p>In our characterization, CMV promoter was assembled with GFP reporter (BBa_K648013) and hGH polyA terminator (BBa_K404108). | <p>In our characterization, CMV promoter was assembled with GFP reporter (BBa_K648013) and hGH polyA terminator (BBa_K404108). | ||
The pCMV-GFP was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope. | The pCMV-GFP was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope. | ||
Revision as of 15:39, 27 September 2013
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Mitochondrial Leader Sequence
- Introduction
- Result
- Conclusion
- Reference
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Characterization
- Mitochondrial Leader Sequence
- CMV Promoter
- EF1-alpha Promoter
CMV Promoter
Introduction
CMV (Cytomegalovirus) promoter is a constitutive mammalian promoter.
In our characterization, CMV promoter was assembled with GFP reporter (BBa_K648013) and hGH polyA terminator (BBa_K404108). The pCMV-GFP was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope. The positive control was pEGFP-N1 (Clontech) that contains CMV promoter and EGFP reporter. A negative control was made by GFP generator (BBa_K648013) that does not contain the CMV promoter. The detailed protocol of our characterization can be found in HKUST iGEM 2013 Wiki.
Result
Figure 1. CMV promoter drives expression of GFP. HEK cells transfected with pCMV-GFP gave GFP signals. HEK cells transfected with the commercial pEGFP-N1 showed similar results, while the same construct without any promoter did not give any GFP signals. Scale bar = 10 microns
