Part:BBa_K2387018
Cpx system reporter + Affinity molecule-CpxP inhibitor
The Cpx envelope stress reporter consists of the subunits CpxA, a membrane receptor, CpxR, the response regulator, and CpxP, an auxiliary periplasmic inhibitor of CpxA. If E. coli cells undergo stress on their envelope, the Cpx system will be activated due to the CpxA sensing stress related signals. In turn it will activate CpxR which acts as a transcription factor for genes involved in the alleviation of this stress and reinstatement of homeostasis. In non-stress conditions CpxP will inhibit activation of CpxA by direct physical interaction.
This part consists of the CpxP gene fused N-terminally to an affinity molecule specific for IgG (Aff-CpxP), under control of tac promoter, which was used to suppress the Cpx system. Furthermore it includes reporter mRFP1 under control of a CpxR promoter, leading to fluorophore production in case the Cpx system is activated. In our project this was used as a fundamental test to verify whether expression of Aff-CpxP fusion could inhibit the system to the same extend as the native CpxP can.
Usage and Biology
In the Wageningen iGEM 2017 project Mantis, this biobrick was used to measure the Cpx inhibitory capacity of CpxP and affinity body fusions in E. coli ΔCpxP strain, together with parts BBa_K2387014, BBa_K2387017. Specifically, this biobrick was tested for it's capacity to inhibit the Cpx system under non-stress conditions. Furthermore, BBa_K339007 was used as a positive control to show fluorescence resulting from Cpx system activation.
Figure 1 shows that the fusions of CpxP with the affinity bodies were able to inhibit the Cpx system equally as well as the native CpxP gene.
For more information about this experiment see the Cpx Signal Transduction page of the 2017 Wageningen iGEM team.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1373
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 636
Illegal AgeI site found at 748 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1092
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