Team:Hong Kong HKUST/characterization

From 2013.igem.org

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<br><br><br><div id="slide"><center><h3 class="title">Characterizations</h3></center><br>  
<br><br><br><div id="slide"><center><h3 class="title">Characterizations</h3></center><br>  
<h3>Mitochondrial Leader Sequence</h3>
<h3>Mitochondrial Leader Sequence</h3>
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<p id="yo">For one of our human practice projects, our team investigated the current development of synthetic biology in some Asian countries. While doing research, we found out that synthetic biology is really a new field in science and not well-known in Asia. In fact, when some of our members tried to research information about synthetic biology in Cantonese, we could not find any webpages dedicated to share information about synthetic biology for the public in Hong Kong. To address this observation, we have decided to use an easily accessible platform called Google Blogger. This blog will be used as means to introduce synthetic biology, share our experience and our project, and report current events related to synthetic biology. It will be written in Chinese and English so most people in Hong Kong can read it easily and pay more attention to this new field in science. By providing holistic and informational blog posts, we aim at raising public awareness to synthetic biology.</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>
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<p id="yo">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).
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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.
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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).
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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>
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<div class="hr"><hr /></div><br>
<h3>CMV Promoter</h3><br><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>
<h3>CMV Promoter</h3><br><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>

Revision as of 11:20, 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







EF1-alpha Promoter






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