Difference between revisions of "Team:ShanghaitechChina/Proof"

 
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  As figure illustrated, His-CsgA-SpyCatcher-Histag mutant incubated with mCherry-SpyTag show a clear biofilm-associated mcherry fluorescence signal, which indicating the accurate conformation and function of the SpyTag and SpyCatcher linkage system. The distinct localization highlight of red fluorescence on E.coli, which to a large extent prove the specificity of our desired linkage between SpyTag and SpyCatcher system.  
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  As figure illustrated, His-CsgA-SpyCatcher-Histag mutant incubated with mCherry-SpyTag show a clear biofilm-associated mcherry fluorescence signal, which indicating the accurate conformation and function of the SpyTag and SpyCatcher linkage system. The distinct localization highlight of red fluorescence on <i>E.coli</i>, which to a large extent prove the specificity of our desired linkage between SpyTag and SpyCatcher system.  
  
 
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<h3>So far above, we proved that two engineered biofilm devices function properly. Later, we tested different inducer concentration gradient to find out the best induction condition.</h3>
 
<h3>So far above, we proved that two engineered biofilm devices function properly. Later, we tested different inducer concentration gradient to find out the best induction condition.</h3>
 
<h3><b>Inducer concentration optimization</b></h3>
 
<h3><b>Inducer concentration optimization</b></h3>
We cultured all E.coli mutants in multi-wells with increasing inducer gradient. The result demonstrated in accordance that 0.25 μg ml-1 of aTc will induce the best expression performance of biofilm, which is exactly the inducer concentration we applied in the project.  
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We cultured all <i>E. coli</i> mutants in multi-wells with increasing inducer gradient. The result demonstrated in accordance that 0.25 μg ml-1 of aTc will induce the best expression performance of biofilm, which is exactly the inducer concentration we applied in the project.  
 
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<h4><b>Hydrogenese gene clusters</b></h4><p></p>
 
<h4><b>Hydrogenese gene clusters</b></h4><p></p>
  <p>High-activity hydrogenase is necessary for our system. To achieve efficient enzymatic activities, we codon-optimized and constructed the whole hydrogenase gene clusters (from Clostridium Acetobutylicum) by leveraging the multi-expression Acembl System.  Please refer to <b><a href="https://2016.igem.org/Team:ShanghaitechChina/Hydrogen">Hydrogenase Session</a></b> for more details. </p>   
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  <p>High-activity hydrogenase is necessary for our system. To achieve efficient enzymatic activities, we codon-optimized and constructed the whole hydrogenase gene clusters (from <i>Clostridium acetobutylicum</i>) by leveraging the multi-expression Acembl System.  Please refer to <b><a href="https://2016.igem.org/Team:ShanghaitechChina/Hydrogen">Hydrogenase Session</a></b> for more details. </p>   
 
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Latest revision as of 22:53, 19 October 2016

igem2016:ShanghaiTech