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Team:Hong Kong HKUST/characterization
From 2013.igem.org
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The detailed protocol of our characterization can be found in HKUST iGEM 2013 Wiki.</p><br><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization/mls"><img src="http://w1ggroup.com/devere/img/more_off.png" style="width:13%;height:45px;padding-right:20px;float:right;"></a><br><br><br> | The detailed protocol of our characterization can be found in HKUST iGEM 2013 Wiki.</p><br><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization/mls"><img src="http://w1ggroup.com/devere/img/more_off.png" style="width:13%;height:45px;padding-right:20px;float:right;"></a><br><br><br> | ||
<div class="hr"><hr /></div><br> | <div class="hr"><hr /></div><br> | ||
| - | <h3>CMV Promoter</h3>< | + | |
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| + | <h3>CMV Promoter</h3> | ||
| + | <p id="yo">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.</p><br> | ||
| + | <a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization/cmv"><img src="http://w1ggroup.com/devere/img/more_off.png" style="width:13%;height:45px;padding-right:20px;float:right;"></a><br><br><br> | ||
<div class="hr"><hr /></div><br> | <div class="hr"><hr /></div><br> | ||
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<h3>EF1-alpha Promoter</h3> | <h3>EF1-alpha Promoter</h3> | ||
<br><br><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization/ef1a"><img src="http://w1ggroup.com/devere/img/more_off.png" style="width:13%;height:45px;padding-right:20px;float:right;"></a><br><br><br> | <br><br><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization/ef1a"><img src="http://w1ggroup.com/devere/img/more_off.png" style="width:13%;height:45px;padding-right:20px;float:right;"></a><br><br><br> | ||
Revision as of 13:58, 27 September 2013
Characterizations
Mitochondrial Leader Sequence
In our characterization, the CDS of MLS was assembled in frame with that of GFP reporter using Freiburg’s RFC25 format(BBa_K648013). The translation unit was driven by CMV promoter (BBa_K1119006) and terminated by hGH polyA signal (BBa_K404108). The MLS-GFP generator (BBa_K1119009) was then transfected into HEK293FT cells. Mitochondria were stained after transfection and co-localization was determined by area of signal that overlapped. To provide a positive control, CDS of EGFP from pEGFP-N1 (Clontech) was inserted downstream and in frame with the CDS of the MLS in the commercial plasmid pCMV/myc/mito, (Invitrogen, Carlsbard, CA). A negative control was made by GFP generator that does not contains the CDS of MLS (BBa_K1119008). The detailed protocol of our characterization can be found in HKUST iGEM 2013 Wiki.
CMV 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.
EF1-alpha Promoter
