Difference between revisions of "Part:BBa K2510113"
 
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<p> This part encodes for TetR in wtdronpa </p>
 
<p> This part encodes for TetR in wtdronpa </p>
  
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<p> In our project we developed a logic gate at the promoter level by creating dually repressed promoters using different combinations of the operators for TetR, P22 c2 and HK CI, the logic gate was tested with Dronpa-repressors fusions, the fluorescence levels increased over 1000-fold when violet light is shone    </p>
 
  
 +
<p> In our project we developed a logic gate at the promoter level by creating dually repressed promoters using different combinations of the operators for TetR, P22 c2 and HK CI, the logic gate was tested with Dronpa-repressors fusions</p>
 +
<p> By illuminating our system with violet light, caging increase the repression strength while uncaging with cyan light decreases the repression strength. Our results also show that the mutDronpa can better control the activity of TetR than the wtDronpa. However, HKC1 and C2P22 are better controlled by the wtDronpa. </p>
  
  
  
 +
                      <img src="https://static.igem.org/mediawiki/2017/b/b1/96_conditions.png">
  
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                            <b>Figure 1 : </b> the experiment conducted with the repressors caged with Dronpa
 +
                                       
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                                    </span>
 +
 +
                                      <img src="https://static.igem.org/mediawiki/2017/7/7d/Repression_strength_of_closed_state.png ">
 +
 +
                                      <b>Figure 2 : </b> An over view of the results of the cell-free experiment. Each
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                                        promoter was tested with its cognate repressors. Our results show that in 90% of our
 +
                                        constructs caging the repressors with Dronpa has increased the repression strength.
 +
                                        We have also obtained a wide linear range of repression strength-indicated by the red
 +
                                        slope-.
 +
                                    </span>
 +
                                    <img src="https://static.igem.org/mediawiki/2017/2/22/TetR_dronpa.png">
 +
                                    <img src="https://static.igem.org/mediawiki/2017/e/ed/P22c2_dronpa.png">
 +
                                    <img src="https://static.igem.org/mediawiki/2017/5/5e/HKCI_caged_dronpa.png">
 +
                                    <span  class="image-span text-center">
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                                            <b>Figure 3:</b> detailed Results of the cell-free experiment. Each promoter was
 +
                                            tested with its cognate repressors. <b>Top</b>: Testing with TetR caged with
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                                            either wt-Dronpa (BBa_K2510108) or a mutated version(BBa_K2510109)<b>Middle</b>:
 +
                                            Testing with P22c2 caged with either wt-Dronpa (BBa_K2510112) or a mutated
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                                            version(BBa_K2510113).<b>Bottom</b>: Testing with HKcI caged with either wt-
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                                            Dronpa (BBa_K2510110) or a mutated version(BBa_K2510111)          </span>
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                                  <img src="https://static.igem.org/mediawiki/2017/5/5f/Wtvs_mut_repressors.png" >
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                                    <b>Figure 4:</b> the wtDronpa has a better control of HKC1 and C2P22 repressors while the mutDronpa has a better control of TetR repressors </span>
  
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<img src="https://static.igem.org/mediawiki/2017/e/e2/Logic_tetR_PB.png">
 
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<img src="https://static.igem.org/mediawiki/2017/8/8e/Logic_P22c2_PB.png">
 
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<img src="https://static.igem.org/mediawiki/2017/1/19/Logic_HKcIfigure_PB.png">
 
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                                            <span  class="image-span text-center">
 
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                                            <b>Figure 1:Results of the cell-free experiment. Each promoter was tested with its cognate repressors. Top: Testing with TetR caged with either wt-Dronpa (BBa_K2510108) or a mutated version(BBa_K2510109)Middle: Testing with P22c2 caged with either wt-Dronpa (BBa_K2510112) or a mutated version(BBa_K2510113).Bottom: Testing with HKcI caged with either wt-Dronpa (BBa_K2510110) or a mutated version(BBa_K2510111)
 
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 03:30, 16 December 2017


pdDronpa wt TetR

This part encodes for TetR in wtdronpa


In our project we developed a logic gate at the promoter level by creating dually repressed promoters using different combinations of the operators for TetR, P22 c2 and HK CI, the logic gate was tested with Dronpa-repressors fusions

By illuminating our system with violet light, caging increase the repression strength while uncaging with cyan light decreases the repression strength. Our results also show that the mutDronpa can better control the activity of TetR than the wtDronpa. However, HKC1 and C2P22 are better controlled by the wtDronpa. 


                      <img src="96_conditions.png">

                            Figure 1 :  the experiment conducted with the repressors caged with Dronpa
                                        
                                   </span>

                                     <img src="Repression_strength_of_closed_state.png ">

                                      Figure 2 :  An over view of the results of the cell-free experiment. Each 
                                       promoter was tested with its cognate repressors. Our results show that in 90% of our 
                                       constructs caging the repressors with Dronpa has increased the repression strength. 
                                       We have also obtained a wide linear range of repression strength-indicated by the red 
                                       slope-.
                                   </span>
                                   <img src="TetR_dronpa.png">
                                   <img src="P22c2_dronpa.png">
                                    <img src="HKCI_caged_dronpa.png"> 
                                   
                                           Figure 3: detailed Results of the cell-free experiment. Each promoter was 
                                           tested with its cognate repressors. Top: Testing with TetR caged with 
                                           either wt-Dronpa (BBa_K2510108) or a mutated version(BBa_K2510109)Middle: 
                                           Testing with P22c2 caged with either wt-Dronpa (BBa_K2510112) or a mutated 
                                            version(BBa_K2510113).Bottom: Testing with HKcI caged with either wt-
                                           Dronpa (BBa_K2510110) or a mutated version(BBa_K2510111)           

                                  <img src="Wtvs_mut_repressors.png" >
                                   Figure 4: the wtDronpa has a better control of HKC1 and C2P22 repressors while the mutDronpa has a better control of TetR repressors </span>

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]