Difference between revisions of "Part:BBa K3885126"
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<partinfo>BBa_K3885126 short</partinfo> | <partinfo>BBa_K3885126 short</partinfo> | ||
− | SsrA is a degradation tag, added at the 3 end of tetR that degrades the repressor protein tetR | + | This is an improved version of tetracycline resistance transcriptional repressor (tetR), which is registered by 10_Heidelberg iGEM team (<a href="https://parts.igem.org/Part:BBa_K337028">BBa_K337028</a>: tet Repressor). |
+ | SsrA is a degradation tag, added at the 3' end of tetR that degrades the repressor protein tetR to become tetR-ssrA, thus we endow the original part a new function as improvement. | ||
+ | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
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To ensure the normal operation of the Cell-Free system, an adjustable protein degradation mechanism is crucial. It is degraded by ClpXPAAA+ protease in Escherichia coli . The protein must contain a degradation tag in order to be recognized by our cell-free system .<br> | To ensure the normal operation of the Cell-Free system, an adjustable protein degradation mechanism is crucial. It is degraded by ClpXPAAA+ protease in Escherichia coli . The protein must contain a degradation tag in order to be recognized by our cell-free system .<br> | ||
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Revision as of 13:52, 20 October 2021
tetR-ssrA
This is an improved version of tetracycline resistance transcriptional repressor (tetR), which is registered by 10_Heidelberg iGEM team (<a href="https://parts.igem.org/Part:BBa_K337028">BBa_K337028</a>: tet Repressor). SsrA is a degradation tag, added at the 3' end of tetR that degrades the repressor protein tetR to become tetR-ssrA, thus we endow the original part a new function as improvement.
Usage and Biology
![](https://static.igem.org/mediawiki/parts/2/28/337-1.png)
Figure 1. Schematic of hydrolysis by ClpXP protein.
Characterization
![](https://static.igem.org/mediawiki/parts/d/d0/337-2.png)
![](https://static.igem.org/mediawiki/parts/d/da/337-3.png)
Figure 2. Schematic of gene circuits and kinetics of tetR inhibition in the Cell-Free system.
(A) The gene circuits contains three parts, sigma 28 activates promoter P28 to express tet repressor with ssrA tag, which inhibits the expression of deGFP.
(B) The gene circuits contains an additional part P70-ClpXP that makes the tetR repressor degrade, and degfp expresses.
(C) Group 1 carried tetR without ssrA and ClpXP; Group 2 carried tetR with ssrA; Group 3 didn't carry tetR as positive control; Group 4 carried tetR with ssrA and ClpXP in comparison with Group 1.
Therefore, we gave the tetR a new function and improved this part. Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 10
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 652
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 10
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 10
- 1000COMPATIBLE WITH RFC[1000]