Difference between revisions of "Part:BBa K3885126"
(→Characterization) |
|||
Line 33: | Line 33: | ||
} | } | ||
</style> | </style> | ||
− | <div | + | <div style="margin: 0 auto; width: 100%;"> |
− | <div | + | <div style="border: 1px solid #000;width: 80%; background-color: #f9f9f9;"> |
+ | <div class="flex" style="width:100%;"> | ||
<img src="https://static.igem.org/mediawiki/parts/d/d0/337-2.png" width=350 style=" margin: 10px auto;"> | <img src="https://static.igem.org/mediawiki/parts/d/d0/337-2.png" width=350 style=" margin: 10px auto;"> | ||
<img src="https://static.igem.org/mediawiki/parts/d/da/337-3.png" width=288 style=" margin: 10px auto;"/> | <img src="https://static.igem.org/mediawiki/parts/d/da/337-3.png" width=288 style=" margin: 10px auto;"/> | ||
+ | </div> | ||
+ | </div> | ||
<p style="text-align: center:">Figure 2. Schematic of gene circuits and kinetics of tetR inhibition in the Cell-Free system.<br> | <p style="text-align: center:">Figure 2. Schematic of gene circuits and kinetics of tetR inhibition in the Cell-Free system.<br> | ||
(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. <br> | (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. <br> | ||
(B) The gene circuits contains an additional part P70-ClpXP that makes the tetR repressor degrade, and degfp expresses.<br> | (B) The gene circuits contains an additional part P70-ClpXP that makes the tetR repressor degrade, and degfp expresses.<br> | ||
(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.</p> | (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.</p> | ||
− | |||
</div> | </div> | ||
TetR with ssrA can be degraded by ClpXP, so that the flourescence intensity increased as a result of the inhibition released from tet Repressor. <br> | TetR with ssrA can be degraded by ClpXP, so that the flourescence intensity increased as a result of the inhibition released from tet Repressor. <br> |
Revision as of 12:51, 20 October 2021
tetR-ssrA
SsrA is a degradation tag, added at the 3 end of tetR that degrades the repressor protein tetR
Usage and Biology
![](https://static.igem.org/mediawiki/parts/2/28/337-1.png)
Figure 1. Schematic of hydrolysis by ClpXP protein.
As the improvement of the 10_Heidelberg registered part (BBa_K337028: tet Repressor) , the degradation tag ssrA is added to the 3' end of the tetR (Figure 1) , hence the newly registered partBBa_K3885126.
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
Assembly Compatibility:
- 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]