Difference between revisions of "Team:UiOslo Norway/Collaborations"

 
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<h1  style="text-align:center; font-size:400%;">Collaborations</h1>
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<h1> Experiments </h1>
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<h2> 1. Obtaining the genes </h2>
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<p>The first project part pursues the goal to get or clone all genes that are involved in our project into a plasmid, which allows a rapid and easy amplification of those genes for further experiments. </br>
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<h1  style="text-align:center">iGEM Team Braunschweig 2014</h1>
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For the soluble methane monooxygenase (sMMO) from <i>Methylococcus capsulatus</i> (Bath) we got the genes for the subunits (<i>mmoXYZBCD</i>) cloned into the BioBrick standard vector pSC1B3 from the
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<a href="https://2014.igem.org/Team:Braunschweig/Achievements-content#parts" >  
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<p> The <a href="https://2014.igem.org/Team:Braunschweig">iGEM Team Braunschweig 2014</a>, provided us with their generated BioBricks:
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iGEM 2014 team from Braunschweig, Germany.
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<a href="http://parts.igem.org/Part:BBa_K1390001">BBa_K1390001</a>,  
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</a>  
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<a href="http://parts.igem.org/Part:BBa_K1390002">BBa_K1390002</a>,  
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Those six parts are registered as BioBrick parts under the names; Bba_K1390001 (<i>mmoB</i>), Bba_K1390002 (<i>mmoC</i>), Bba_K1390003 (<i>mmoD</i>), Bba_K1390004 (<i>mmoX</i>), Bba_K1390005 (<i>mmoY</i>), and Bba_K1390006 (<i>mmoZ</i>). </br>
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<a href="http://parts.igem.org/Part:BBa_K1390003">BBa_K1390003</a>,  
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Genes encoding the enzymes for the conversion of methanol into biomass (<i>medh2</i>, <i>hps</i>, and <i>phi</i>) were amplified by PCR from <i>Bacillus methanolicus</i> (MGA3) genomic DNA and TOPO blunt end cloning into the pCR4 vector was performed. Primers were designed in a way that they bind in the 5’ and 3’ untranslated region (UTR) of each gene.</br>
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<a href="http://parts.igem.org/Part:BBa_K1390004">BBa_K1390004</a>,  
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TOPO blunt end cloning of <i>mmoG</i> did not succeed. Instead <i>mmoG</i> was synthesized by IDT as a gBlock gene fragment, codon optimized for protein expression in <i>E. coli</i>.
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<a href="http://parts.igem.org/Part:BBa_K1390005">BBa_K1390005</a>,
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</p>
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<a href="http://parts.igem.org/Part:BBa_K1390006">BBa_K1390006</a>
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, which contain the  subunits of the <i>Methylococcus capsulatus</i> soluble methanemonooxygenase. </p>
  
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<h2> 2. Construction of BioBrick parts</h2>
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<h1  style="text-align:center">iGEM Team Uppsala and iGEM Team Stockholm</h1>
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<p>We joined the Nordic iGEM Conference which took part in the end of July 2015. The Nordic iGEM conference was hosted by the iGEM Teams 
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<a href="https://2015.igem.org/Team:Uppsala">Uppsala</a> and
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<a href="https://2015.igem.org/Team:Stockholm">Stockholm</a>. Teams from
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<a href="https://2015.igem.org/Team:SDU-Denmark">SDU-Denmark</a>,
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<a href="https://2015.igem.org/Team:Chalmers-Gothenburg">Chalmers-Gothenburg</a>,
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<a href="https://2015.igem.org/Team:Linkoping_Sweden">Linköping</a>,
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<a href="https://2015.igem.org/Team:Aalto-Helsinki">Aalto-Helsinik</a>
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joined the conference and it was a great platform to exchange ideas.</p>
  
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<p>The second project part had the intention to create four new basic BioBrick parts. Those basic parts consist of the coding sequences (CDS) of a gene. The codonoptimized <i>mmoG</i> as well as all three genes encoding the enzymes for the methanol to biomass conversion (<i>medh2</i>, <i>hps</i>, and <i>phi</i>) were created as
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<h1  style="text-align:center">iGEM Team Aachen</h1>
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<a href="https://2015.igem.org/Team:UiOslo_Norway/Basic_Part" >  
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<p>We were collaborating with the <a href="https://2015.igem.org/Team:Aachen">iGEM Team Aachen</a>. The iGEM Team Aachen helped us to establish our methane sensor. Further we exchanged thoughts about assays testing the methanol dehydrogenase. </p>
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BioBrick parts.
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</a> </br>
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The CDS of the <i>hps</i> gene, encoding the 3-hexulose-6-phosphate synthase, contains PstI and XbaI restriction sites making it not compatible with the BioBrick system. In two rounds of <i>in vitro</i> mutagenesis both restriction sites were removed and <i>hps</i> was cloned into pSC1B3 and submitted as a
 
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<a href="https://2015.igem.org/Team:UiOslo_Norway/Basic_Part" >
 
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BioBrick part.
 
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</a>
 
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</p>
 
  
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<h2>3. Generation of expression constructs</h2>
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<!---------------sponsors------------------------->
  
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<p>Before assembling the final constructs we wanted to show that each individual protein could be expressed in <i>E. coli</i>. The pET system has been chosen as our preferred system for overexpression of each individual protein. The vector backbones pET-28 and pET-30 were chosen as potential expression vector.</br>
 
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Figure about construct: T7 prom—RBS—GENE Coming soon
 
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With PCR we added restriction enzyme sites at the 5’ and 3’ end of CDS of each gene. Afterwards the gene was cloned either into pET-30 or pET-28 with the use of the listed restriction enzymes (Table 1 coming soon).
 
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<h1> Protocols </h1>
 
  
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<center><h1>iGEM UiOslo 2015 is sponsored by:</h1>
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<hr style="height:2px;border:none;color:black;background-color:black;" />
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<img src="https://static.igem.org/mediawiki/2015/a/ac/UiOslo_Statoil.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/e/e0/UiOslo_uio_logo.png" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/0/08/UiOslo_Enova1.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/e/eb/UiOslo_GATC-Biotech.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/4/4a/UiOslo_IDT.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/d/d9/UiOslo_Norwegian_Biochemical_Society.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/f/f0/UiOslo_SnapGene.jpg" Height="60px" hspace="15">
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</center>
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<hr style="height:2px;border:none;color:black;background-color:black;" />
  
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<!--------end sponsors---------------------------->
  
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Latest revision as of 10:22, 17 September 2015

Collaborations

iGEM Team Braunschweig 2014

The iGEM Team Braunschweig 2014, provided us with their generated BioBricks: BBa_K1390001, BBa_K1390002, BBa_K1390003, BBa_K1390004, BBa_K1390005, BBa_K1390006 , which contain the subunits of the Methylococcus capsulatus soluble methanemonooxygenase.

iGEM Team Uppsala and iGEM Team Stockholm

We joined the Nordic iGEM Conference which took part in the end of July 2015. The Nordic iGEM conference was hosted by the iGEM Teams Uppsala and Stockholm. Teams from SDU-Denmark, Chalmers-Gothenburg, Linköping, Aalto-Helsinik joined the conference and it was a great platform to exchange ideas.

iGEM Team Aachen

We were collaborating with the iGEM Team Aachen. The iGEM Team Aachen helped us to establish our methane sensor. Further we exchanged thoughts about assays testing the methanol dehydrogenase.

iGEM UiOslo 2015 is sponsored by: