Difference between revisions of "Part:BBa K337028"

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	The TetR binds to the Tet Operator and by that causes knockdown auf gene expression. (BB-2, RFC12).		The TetR binds to the Tet Operator and by that causes knockdown auf gene expression. (BB-2, RFC12).
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−	<img src="https://static.igem.org/mediawiki/parts/d/d0/337-2.png" width=95% height=70% style="display: block;margin: 10px auto;"/>	+	<img src="https://static.igem.org/mediawiki/parts/d/d0/337-2.png,https://static.igem.org/mediawiki/parts/d/da/337-3.png" width=95% height=70% style="display: block;margin: 10px auto;"/>
	<img src="https://static.igem.org/mediawiki/parts/d/da/337-3.png" width=95% height=70% style="display: block;margin: 10px auto;"/>		<img src="https://static.igem.org/mediawiki/parts/d/da/337-3.png" width=95% height=70% style="display: block;margin: 10px auto;"/>
	<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>
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	(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><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><br>
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	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>
	Therefore, we gave the tetR a new function and improved this part.		Therefore, we gave the tetR a new function and improved this part.

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Revision as of 10:51, 20 October 2021

Tet Repressor

The TetR binds to the Tet Operator and by that causes knockdown auf gene expression. (BB-2, RFC12).

Improvement by iGEM ZJUT-China 2021

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 .
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.

Results

TetR with ssrA can be degraded by ClpXP, so that the flourescence intensity increased as a result of the inhibition released from tet Repressor.
Therefore, we gave the tetR a new function and improved this part.

Sequence and Features