Difference between revisions of "Team:Calgary/Composite Part"

 
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<h1>Parts</h1>
 
<h1>Parts</h1>
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<p>We submitted three parts to the Registry this year: <b>two parts</b> involved in PHB synthesis and <b>one part</b> involved in PHB secretion.</p>
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<p>To meet our medal requirements, we submitted <a href="http://parts.igem.org/Part:BBa_K2260001">BBa_K2260001</a> as a new part, which uniquely contains the <i>phaJ4</i> gene native to <i>Pseudomonas putida</i>, along with the <i>phaC1</i> gene from <i>Pseudomonas aeruginosa</i>.</p>
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<p>We also improved two parts. The first, <a href="http://parts.igem.org/Part:BBa_K2260000">BBa_K2260000</a>, was an improvement on <a href="http://parts.igem.org/Part:BBa_K1149052">Imperial College's phaCAB operon</a> submitted in 2013. The gene order was changed to improve PHB yield (Hiroe <i>et al.</i>, 2012), codons were optimized for expression in <i>Escherichia coli</i>, and restriction sites were removed to allow part compatibility in all iGEM RFC assembly standards. The second, <a href="http://parts.igem.org/Part:BBa_K2260002">BBa_K2260002</a>, was an improvement on <a href="http://parts.igem.org/Part:BBa_K2018024">SDU Denmark's phasin-HlyA</a> part submitted in 2016. Codons were optimized for expression in <i>E. coli</i>.</p>
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<h2>Synthesis</h2>
 
<h2>Synthesis</h2>
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<th><b>Biobrick</b>: <a src="http://parts.igem.org/Part:BBa_K2260000"> BBa_K2260000</a></th>
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<th><b>Biobrick</b>: <a href="http://parts.igem.org/Part:BBa_K2260000"> BBa_K2260000</a></th>
 
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<img id="medium-image" src="https://static.igem.org/mediawiki/2017/0/0c/CBA_construct.png"></td> </tr>
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<img id="medium-image" src="https://static.igem.org/mediawiki/2017/b/b4/Calgary2017_GlycolysisConstruct.png"></td> </tr>
 
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<th><b>Part type</b>: Composite</th>
 
<th><b>Part type</b>: Composite</th>
 
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<th><b>Description</b>: PhaCBA operon w/ RBS</th>
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<th><b>Description</b>: <i>phaCBA</i> operon with <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a> RBS</th>
 
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<th><b>Biobrick</b>: <a src="http://parts.igem.org/Part:BBa_K2260001"> BBa_K2260010</a></th>
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<th><b>Biobrick</b>: <a href="http://parts.igem.org/Part:BBa_K2260001"> BBa_K2260001</a></th>
 
                 <td rowspan=4>      <img id="medium-image" src="https://static.igem.org/mediawiki/2017/7/75/Calgary2017_BetaOxidationConstruct.png"></td>
 
                 <td rowspan=4>      <img id="medium-image" src="https://static.igem.org/mediawiki/2017/7/75/Calgary2017_BetaOxidationConstruct.png"></td>
 
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<th><b>Description</b>: PhaC1-J4</th>
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<th><b>Description</b>: <i>phaC1-J4</i> operon with <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a> RBS</th>
 
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<th width=40%><b>Source</b>: PhaC1 gene was taken from <i>Pseudomonas aeruginosa</i> <source> and PhaJ4 was from <i>Pseudomonas putida</i>.</th>
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<th width=40%><b>Source</b>: <i>phaC1</i> was taken from <i>P. aeruginosa</i> <source> and <i>phaJ4</i> from <i>P. putida</i>.</th>
 
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<h2>Secretion</h2>
 
<h2>Secretion</h2>
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<th><b>Biobrick</b>:<a src="http://parts.igem.org/Part:BBa_K2260002"> BBa_K2260002</a></th>
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<th><b>Biobrick</b>:<a href="http://parts.igem.org/Part:BBa_K2260002"> BBa_K2260002</a></th>
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               <td rowspan=4><img id="medium-image" src="https://static.igem.org/mediawiki/2017/3/32/Calgary2017_PhasinConstruct.png" alt="Secretion Construct" style="width:100%"></td>
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               <td rowspan=4 style="vertical-align: center;"><img id="medium-image" src="https://static.igem.org/mediawiki/2017/3/32/Calgary2017_PhasinConstruct.png" alt="Secretion Construct" style="width:100%"></td>
 
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<th width=40%><b>Description</b>: Phasin-HlyA fusion protein with T7 promoter</th>
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<th width=40%><b>Description</b>: Phasin-HlyA fusion protein with T7 promoter and <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a> RBS</th>
 
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<th width=40%><b>Source</b>: Phasin (PhaP) is from <i>Ralstonia eutropha</i>, HlyA Tag is from <i>E.coli</i>, and the T7 promoter is from T7 bacteriophage. </th>
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<th width=40%><b>Source</b>: Phasin (PhaP) is from <i>R. eutropha</i>, HlyA tag is from <i>E. coli</i>, while the T7 promoter is from the T7 bacteriophage. </th>
 
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<h2>Works Cited</h2>
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<p>Hiroe, A., Tsuge, K., Nomura, C.T., Itaya, M. & Tsuge, T. (2012). Rearrangement of Gene Order in the phaCABOperon Leads to Effective Production of Ultrahigh-Molecular-Weight Poly[(R)-3-Hydroxybutyrate] in Genetically Engineered Escherichia coli. Appl. Environ. Microbiol., 78(9): 3177-3184</p>
  
 
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Latest revision as of 02:47, 1 November 2017

Header

Parts

We submitted three parts to the Registry this year: two parts involved in PHB synthesis and one part involved in PHB secretion.


To meet our medal requirements, we submitted BBa_K2260001 as a new part, which uniquely contains the phaJ4 gene native to Pseudomonas putida, along with the phaC1 gene from Pseudomonas aeruginosa.


We also improved two parts. The first, BBa_K2260000, was an improvement on Imperial College's phaCAB operon submitted in 2013. The gene order was changed to improve PHB yield (Hiroe et al., 2012), codons were optimized for expression in Escherichia coli, and restriction sites were removed to allow part compatibility in all iGEM RFC assembly standards. The second, BBa_K2260002, was an improvement on SDU Denmark's phasin-HlyA part submitted in 2016. Codons were optimized for expression in E. coli.


Synthesis

Biobrick: BBa_K2260000
Part type: Composite
Description: phaCBA operon with BBa_B0034 RBS
Source: Ralstonia eutropha H16
Biobrick: BBa_K2260001
Part type: Composite
Description: phaC1-J4 operon with BBa_B0034 RBS
Source: phaC1 was taken from P. aeruginosa and phaJ4 from P. putida.

Secretion

Biobrick: BBa_K2260002 Secretion Construct
Part type: Composite
Description: Phasin-HlyA fusion protein with T7 promoter and BBa_B0034 RBS
Source: Phasin (PhaP) is from R. eutropha, HlyA tag is from E. coli, while the T7 promoter is from the T7 bacteriophage.


Works Cited

Hiroe, A., Tsuge, K., Nomura, C.T., Itaya, M. & Tsuge, T. (2012). Rearrangement of Gene Order in the phaCABOperon Leads to Effective Production of Ultrahigh-Molecular-Weight Poly[(R)-3-Hydroxybutyrate] in Genetically Engineered Escherichia coli. Appl. Environ. Microbiol., 78(9): 3177-3184