Difference between revisions of "Team:British Columbia/Composite Part"

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<h1>Favorite Composite Part: BBa_k2139019</h1>
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<h1>Favorite Composite Part: BBa_k2139005</h1>
 
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<img class="img-center img-responsive" style="display: table; margin: 0 auto" src="https://static.igem.org/mediawiki/2016/b/b7/T--British_columbia--g12_.png">
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<img class="img-center img-responsive" style="display: table; margin: 0 auto" src="https://static.igem.org/mediawiki/2016/e/e8/T--British_columbia--gluc1c.png">
 
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<p align="justify"><a href= "http://parts.igem.org/wiki/index.php?title=Part:BBa_K2139018"> BBa_K2139019</a> is the expression cassette of the basic part <a href= "http://parts.igem.org/wiki/index.php?title=Part:BBa_K2139018"> BBa_K2139018</a> that was attempted to be used to do the surface expression experiments in <i>C. crescentus</i>. Although we were un-able to functionally characterize it during our project due to time constraints (characterized as fusion protein see data here), it has been well documented in literature which can be found below. This is our favorite part because it was the gene that put up the most fight in getting cloned and attempting expression.</p>
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<p align="justify"><a href= "http://parts.igem.org/wiki/index.php?title=Part:BBa_K2139005"> BBa_K2139005</a> is the expression cassette of the basic part <a href= "http://parts.igem.org/wiki/index.php?title=Part:BBa_K2139002"> BBa_K2139002</a> that was used to do the surface expression experiments in <i>C. crescentus</i>. We were able to functionally characterize it during our project and you can see the data <a href= "https://2016.igem.org/Team:British_Columbia/Project/S-Layer/Cellulases"> here</a> , it has been well documented in literature which can be found below. It originates from the microorganism <i>Paenibacillus sp. MTCC 5639 </i> and the part has been codon optimized for expression in <i>C. crescentus</i>. As such, it contains a high GC content and shows significant difficulty in being amplified by PCR.
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<p align="justify">The construct was designed to contain the Ptac promotor, ribosome binding site, coding region, and double terminator. This was to allow for the constitutive expression of the enzyme. The catalytic function of the enzyme is to convert cellulose to cellobiose (below).
 
<p align="justify">The construct was designed to contain the Ptac promotor, ribosome binding site, coding region, and double terminator. This was to allow for the constitutive expression of the enzyme. The catalytic function of the enzyme is to convert cellulose to cellobiose (below).
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<p align="justify"><b>References:</b> Linger, Jeffrey G., William S. Adney, and Al Darzins. "Heterologous Expression and Extracellular Secretion of Cellulolytic Enzymes by Zymomonas Mobilis." Applied and Environmental Microbiology, vol. 76, no. 19, 2010., pp. 6360-6369doi:10.1128/AEM.00230-10.
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<p align="justify"><b>References:</b> Gupta, S., Adlakha, N., & Yazdani, S. S. (2013). Efficient extracellular secretion of an endoglucanase and a β-glucosidase in E. coli. Protein Expression and Purification, 88(1), 20-25. doi:10.1016/j.pep.2012.11.006.
 
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Revision as of 07:13, 19 October 2016

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Composite Parts

Composite Part

Favorite Composite Part: BBa_k2139005

BBa_K2139005 is the expression cassette of the basic part BBa_K2139002 that was used to do the surface expression experiments in C. crescentus. We were able to functionally characterize it during our project and you can see the data here , it has been well documented in literature which can be found below. It originates from the microorganism Paenibacillus sp. MTCC 5639 and the part has been codon optimized for expression in C. crescentus. As such, it contains a high GC content and shows significant difficulty in being amplified by PCR.

The construct was designed to contain the Ptac promotor, ribosome binding site, coding region, and double terminator. This was to allow for the constitutive expression of the enzyme. The catalytic function of the enzyme is to convert cellulose to cellobiose (below).

References: Gupta, S., Adlakha, N., & Yazdani, S. S. (2013). Efficient extracellular secretion of an endoglucanase and a β-glucosidase in E. coli. Protein Expression and Purification, 88(1), 20-25. doi:10.1016/j.pep.2012.11.006.

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