Difference between revisions of "Team:HokkaidoU Japan/Proof"

 
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<div id="Project_Description"><img src="https://static.igem.org/mediawiki/2016/5/58/T--HokkaidoU_Japan--proof_of_concept.png" width="300px" height="90px" alt="Proof of concept"></div>
  
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<h3>★  ALERT! </h3>
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We made a functional unit to regulate gene expression. <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> codes constitutive promoter (<a href="http://parts.igem.org/Part:BBa_J23101">BBa_J23101</a>), RBS (<a href="http://parts.igem.org/Part:BBa_B0032">BBa_B0032</a>), LacI (<a href="http://parts.igem.org/Part:BBa_C0012">BBa_C0012</a>), dT (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>), PLac (<a href="http://parts.igem.org/Part:BBa_R0011">BBa_R0011</a>) and RBS (<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>). To insert mRFP at the downstream we identified this construct could regulate the expression (Fig. 1).
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<p>This page is used by the judges to evaluate your team for the <a href="https://2016.igem.org/Judging/Medals">gold medal criterion for proof of concept</a>. </p>
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<p> Delete this box in order to be evaluated for this medal. See more information at <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions"> Instructions for Pages for awards</a>.</p>
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<td style="border-style:none; float:center"><img src="https://static.igem.org/mediawiki/2016/4/43/T--HokkaidoU_Japan--K2015012-RFP.png" alt="result1" height="400px" width="auto"></td>  
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<td style="border-style: none"; align="center"><span class="small">Fig. 1. Change of expression by IPTG induction<br>left: not induced, right: induced</span></td>
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<br>And also we induced <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> + <a href="http://parts.igem.org/Part:BBa_K2015008">BBa_K2015008</a> / <a href="http://parts.igem.org/Part:BBa_K2015012">BBa_K2015012</a> + <a href="http://parts.igem.org/Part:BBa_K2015009">BBa_K2015009</a> on pSB1C3 of <span style="font-style: italic">E. coli</span> (DH5&alpha;).           
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<a href="http://parts.igem.org/Part:BBa_K2015008">BBa_K2015008</a> consists Self Assembling Regions (SAR, RADA16-I) and GFP. And <a href="http://parts.igem.org/Part:BBa_K2015009">BBa_K2015009</a> this part consists Self Assembling Region (SAR, P<span class="sitatuki">11</span>-4) and GFP.
  
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iGEM teams are great at making things work! We value teams not only doing an incredible job with theoretical models and experiments, but also in taking the first steps to make their project real.
 
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<h4> What should we do for our proof of concept? </h4>
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The results are shown the below pictures. The fluorescence of GFPs was not detected any condition.
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You can assemble a device from BioBricks and show it works. You could build some equipment if you're competing for the hardware award. You can create a working model of your software for the software award. Please note that this not an exhaustive list of activities you can do to fulfill the gold medal criterion. As always, your aim is to impress the judges!
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<td style="border-style:none; float:center"><img src="https://static.igem.org/mediawiki/2016/d/da/T--HokkaidoU_Japan--multimerization_BBa_K2015012-BBa_K2015008.png" alt="result1" height="300px" width="auto"></td>
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<td style="border-style: none"; align="center"><span class="small">Fig. 2. BBa_K2015012-BBa_K2015008<br>BBa_K2015012-BBa_K2015008 on pSB1C3 vector were induced by IPTG and incubated at 37&deg;C. (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG</span></td>
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<img src="https://static.igem.org/mediawiki/2016/b/be/T--HokkaidoU_Japan--multimerization_BBa_K2015012-BBa_K2015009_25.png" alt="result2" height="300px" width="auto">
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<img src="https://static.igem.org/mediawiki/2016/f/fa/T--HokkaidoU_Japan--multimerization_BBa_K2015012-BBa_K2015009.png" alt="result3" height="300px" width="auto">
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<span class="small">Fig. 3. BBa_K2015012-BBa_K2015009<br>BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25&deg;C for 24 h (left) <br>and at 37&deg;C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG</span>
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<a href="mailto:igemhokkaidou@gmail.com">
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Latest revision as of 03:10, 20 October 2016

Team:HokkaidoU Japan - 2016.igem.org

 

Team:HokkaidoU Japan

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Proof of concept

We made a functional unit to regulate gene expression. BBa_K2015012 codes constitutive promoter (BBa_J23101), RBS (BBa_B0032), LacI (BBa_C0012), dT (BBa_B0015), PLac (BBa_R0011) and RBS (BBa_B0034). To insert mRFP at the downstream we identified this construct could regulate the expression (Fig. 1).
result1
Fig. 1. Change of expression by IPTG induction
left: not induced, right: induced


And also we induced BBa_K2015012 + BBa_K2015008 / BBa_K2015012 + BBa_K2015009 on pSB1C3 of E. coli (DH5α). BBa_K2015008 consists Self Assembling Regions (SAR, RADA16-I) and GFP. And BBa_K2015009 this part consists Self Assembling Region (SAR, P11-4) and GFP.


The results are shown the below pictures. The fluorescence of GFPs was not detected any condition.
result1
Fig. 2. BBa_K2015012-BBa_K2015008
BBa_K2015012-BBa_K2015008 on pSB1C3 vector were induced by IPTG and incubated at 37°C. (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG

result2 result3
Fig. 3. BBa_K2015012-BBa_K2015009
BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25°C for 24 h (left)
and at 37°C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG