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

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<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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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</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>
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<h1>Composite Parts</h1>
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A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
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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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<p>New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).</p>
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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>
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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 width=40%><b>Source</b>: <i>Ralstonia eutropha H16</i></th>
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<th><b>Biobrick</b>: <a href="http://parts.igem.org/Part:BBa_K2260001"> BBa_K2260001</a></th>
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                <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>Part type</b>: Composite</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>: <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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<h3>Best Composite Part Special Prize</h3>
 
  
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<p>New BioBrick devices can be made by combining existing BioBrick Parts. For example, Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on. To be eligible for this award, this part must adhere to <a href="http://parts.igem.org/DNA_Submission">Registry sample submission guidelines</a> and have been sent to the Registry of Standard Biological Parts. If you have a part you wish to nominate your team for this <a href="https://2017.igem.org/Judging/Awards">special prize</a>, make sure you add your part number to your <a href="https://2017.igem.org/Judging/Judging_Form">judging form</a> and delete the box at the top of this page.
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<h2>Secretion</h2>
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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 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><b>Part type</b>: Composite</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>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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|REFERENCES=
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<!-- If you want to included references, please include a heading (h2) titles "Works Cited" followed by all your references in separate paragraph tags -->
  
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<b>Please note:</b> Judges will only look at the first part number you list, so please only enter ONE (1) part number in the judging form for this prize. </p>
 
 
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<h4>Note</h4>
 
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<p>This page should list all the composite parts your team has made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page. Remember judges will only look at the first part in the list for the Best Composite Part award, so put your best part first!</p>
 
  
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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