Difference between revisions of "Team:CMUQ/Parts"

 
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                 <a class="dropdown-toggle" data-toggle="dropdown" href="#">PROJECT <span class="caret"> </span></a>
 
                 <a class="dropdown-toggle" data-toggle="dropdown" href="#">PROJECT <span class="caret"> </span></a>
 
                 <ul class="dropdown-menu">
 
                 <ul class="dropdown-menu">
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                            <li><a href="https://2017.igem.org/Team:CMUQ/Description">Description</a></li>
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  <li><a href="https://2017.igem.org/Team:CMUQ/Description">Description</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Design">Design</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Design">Design</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Safety">Safety</a></li>
 
                             <li><a href="https://2017.igem.org/Team:CMUQ/Safety">Safety</a></li>
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<div class="centered"> <h1></h1> </div>
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<div class="centered"> <h1>Parts</h1> </div>
  
 
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        <p style="font-family: 'Montserrat', sans-serif; margin-right: 120px; margin-left:60px;font-size:25px;margin-top:100px;">We collaborated with the <a href="https://2017.igem.org/Team:Pittsburgh" >Pitt iGEM team </a> to characterize osmolarity sensitive promoters. Sulfate Reducing Bacteria (SRB) require high salt and low oxygen conditions to grow. Therefore genetically engineering a strain of bacteria that can report osmolarity in environments such as oil pipelines and then activate a remediation circuit would be beneficial.
 
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<p style="font-family: 'Montserrat', sans-serif;  margin-right: 120px; margin-left:60px;font-size:25px;margin-top:30px;">Firstly the promoter was tested by making a sensor with a fluorescent protein reporter. The sensor will induce expression of RFP when osmolarity is high. The sensor can be utilized to measure salt concentrations to determine if SRB can survive. The promoter can then be used to produce Dispersin B enzyme to degrade the biofilm where SRB  can accumulate and ultimately, corrode the pipes. With the deadlines approaching, we shared the sequences for the osmolarity sensors with Pitt iGEM.
 
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<p style="font-family:'Montserrat', sans-serif; margin-right: 120px; margin-left:60px;font-size:25px;margin-top:30px;">The two versions of the sensor, which were designed, include the native wild-type promoter sequence of Escherichia coli proU operon (REF) and the optimized Consensus promoter sequence (Lucht and Bremer, 1990), which is a synthetic sequence with mutations previously discovered to increase expression of the operon that codes for a binding protein-dependent transport system.  
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<p style="font-family: 'Montserrat', sans-serif;margin-right: 120px; margin-left:120px;font-size:25px;margin-top:30px;">The following table includes details of all Basic and Composite parts in our project.
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<p style="font-family:'Montserrat', sans-serif; margin-right: 120px; margin-left:60px;font-size:25px;margin-top:30px;">In this collaboration, Pitt iGEM team cloned the constructs and transformed them into MACH1-T1 cells with the help from Dr. Telmer , who is our PI. Then they proceeded to test growth of the cells in LB media with varying concentrations of sodium chloride. Finally, RFP fluorescence levels of these different cultures were measured to determine osmolarity regulated RFP expression in cells with WT and the Consensus promoters compared to the negative control.
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<table style="width:50%; margin-left: 250px;">
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 +
  <tr>
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 +
    <th>Part Number</th>
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    <th>Description </th>
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    <th>Comments</th>
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 +
   
 +
 
 +
  </tr>
 +
  <tr>
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    <td>BBa_K2467000</td>
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    <td>pRSET-DsbADspB-emGFP</td>
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    <td>ntermediate to then clone DsbA DspB emGFP
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into pSB1C3</td>
 +
   
 +
 
 +
 
 +
  </tr>
 +
  <tr>
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    <td>BBa_K2467001</td>
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    <td>WT proU promoter</td>
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    <td>native wild type promoter sequence and a new
 +
part</td>
 +
 
 +
 
 +
  </tr>
 +
 
 +
  <tr>
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    <td>BBa_K2467002</td>
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    <td>Consensus proU-RBS-RFP</td>
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    <td>optimized synthetic proU, strong RBS and mRFP1
 +
as a reporter making the osmolarity sensor</td>
 +
   
 +
 
 +
  </tr>
 +
 
 +
  <tr>
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    <td>BBa_K2467003</td>
 +
    <td>WT proU-RBS-RFP</td>
 +
    <td>wild type native proU, strong RBS and mRFP1
 +
as a reporter making the osmolarity sensor</td>
 +
   
 +
 
 +
  </tr>
 +
 
 +
    <tr>
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    <td>BBa_K2467004</td>
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    <td>Consensus proU promoter</td>
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    <td>optimized synthetic proU promoter sequence</td>
 +
   
 +
 
 +
  </tr>
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  <tr>
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  <td>BBa_K2467005</td>
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  <td>emGFP</td>
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  <td>emerald GFP a variant that is more photostable
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  em/ex 489/507</td>
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</tr>
 +
 +
<tr>
 +
  <td>BBa_K1659200</td>
 +
  <td>DspB coding</td>
 +
  <td></td>
 +
  </tr>
 +
 
 +
  <tr>
 +
    <tr>
 +
    <td>BBa_K1195000</td>
 +
    <td>DspB coding</td>
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    <td></td>
 +
 
 +
 
 +
  </tr>
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 +
    <tr>
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    <td>BBa_K1659201</td>
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    <td>DsbA DspB</td>
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    <td></td>
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 +
  </tr>
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    <tr>
 +
    <td>BBa_K1659211</td>
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    <td>DsbA DspB</td>
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    <td></td>
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 +
 
 +
  </tr>
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 +
 
 +
</table>
  
 
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Latest revision as of 03:16, 2 November 2017

Parts




The following table includes details of all Basic and Composite parts in our project.



Part Number Description Comments
BBa_K2467000 pRSET-DsbADspB-emGFP ntermediate to then clone DsbA DspB emGFP into pSB1C3
BBa_K2467001 WT proU promoter native wild type promoter sequence and a new part
BBa_K2467002 Consensus proU-RBS-RFP optimized synthetic proU, strong RBS and mRFP1 as a reporter making the osmolarity sensor
BBa_K2467003 WT proU-RBS-RFP wild type native proU, strong RBS and mRFP1 as a reporter making the osmolarity sensor
BBa_K2467004 Consensus proU promoter optimized synthetic proU promoter sequence
BBa_K2467005 emGFP emerald GFP a variant that is more photostable em/ex 489/507
BBa_K1659200 DspB coding
BBa_K1195000 DspB coding
BBa_K1659201 DsbA DspB
BBa_K1659211 DsbA DspB