Translational_Unit
Part:BBa_K2387078
Designed by: Bart Scholten Group: iGEM17_Wageningen_UR (2017-10-27)
Revision as of 12:58, 1 November 2017 by Bart scholten (Talk | contribs)
TEV cleavable CpxA-eYFP[1-154]-nZIP and CpxA-eYFP[155-238]-cZIP+ araC/pBAD promoter
In order to visualize Cpx pathway activation, transmembrane signal transducing protein CpxA is fused to respectively eYFP(1-154) and N-Leucine Zipper [1] ,as well as eYFP(155-238) and C-Leucine Zipper [2]. CpxA and eYFP[1-154] /eYFP[155-238] are fused with a TEV cleavable peptide. When cotransformed with CpxR-TEV protease fusion activation of the Cpx pathway is visualized in combination with CpxA-eYFP[1-154]-N-Leucine Zipper fusion due to eYFP reassembly.
Usage and Biology
This BioBrick was used following this protocol.
Figure 1: CpxA-CpxR protein interaction visualized with different L-arabinose concentrations over time. Upon Cpx activation, eYFP-termini are cleaved off of CpxA by TEV protease and reassemble in the cytoplasm using leucine zipper's natural affinity for each other.
More information can be found here
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
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Categories
Parameters
| None |