Team:Hong Kong HKUST/characterization/cmv
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Revision as of 16:23, 27 September 2013
![](https://static.igem.org/mediawiki/igem.org/c/c7/BANNER1_%281%29.png)
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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
![](https://static.igem.org/mediawiki/parts/thumb/c/c6/Final_CMV_annotated_no_ABC.jpg/600px-Final_CMV_annotated_no_ABC.jpg)
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