Difference between revisions of "Part:BBa K2510112"
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__NOTOC__ | __NOTOC__ | ||
− | <partinfo> | + | <partinfo>BBa_K2510108 short</partinfo> |
− | //////////////// | + | <p> This part encodes for TetR in wtdronpa </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"> | ||
+ | |||
+ | <b>Figure 1 : </b> the experiment conducted with the repressors caged with Dronpa | ||
+ | |||
+ | </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 | ||
+ | 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"> | ||
+ | <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 | ||
+ | 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 | ||
+ | version(BBa_K2510113).<b>Bottom</b>: Testing with HKcI caged with either wt- | ||
+ | Dronpa (BBa_K2510110) or a mutated version(BBa_K2510111) </span> | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2017/5/5f/Wtvs_mut_repressors.png" > | ||
+ | <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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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
− | <partinfo> | + | <partinfo>BBa_K2510108 SequenceAndFeatures</partinfo> |
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
− | <partinfo> | + | <partinfo>BBa_K2510108 parameters</partinfo> |
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Latest revision as of 03:29, 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="">
Figure 1 : the experiment conducted with the repressors caged with Dronpa </span>
<img src=" ">
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="">
<img src="">
<img src="">
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="" > 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:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]