Composite

Part:BBa_K4345004

Designed by: Luka Van den Berghe   Group: iGEM22_KU_Leuven   (2022-09-27)


NarX mutant fused to mPapaya with a rigid linker

For more information on NarX, go to this part page: BBa_K4345000. The mutant was designed based on a study of Cavicchioli et al., (1995). The histidine on place 399 was replaced by glutamic acid. This allows the NarX mutants to dimerize but blocks it from phosphorylating NarL (the second messenger).

This composite part consists of BBa_K4345008 (NarX H399-E), BBa_K4345002 (Rigid linker) and BBa_K4345003 (mPapaya).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2153
    Illegal PstI site found at 659
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2153
    Illegal PstI site found at 659
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2153
    Illegal XhoI site found at 260
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2153
    Illegal PstI site found at 659
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2153
    Illegal PstI site found at 659
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

This particular narX protein was derived from E. coli.


NarX nitrate Cheung&Hendrickson2009.jpeg

Image obtained from Cheung & Hendrickson, 2009

References

Cavicchioli, R., Schröder, I., Schröder, S., Constanti, M., & Gunsalus, R. P. (1995). The NarX and NarQ Sensor-Transmitter Proteins of Escherichia coli Each Require Two Conserved Histidines for Nitrate-Dependent Signal Transduction to NarL. JOURNAL OF BACTERIOLOGY, 177(9), 2416–2424.

Cheung, J., & Hendrickson, W. A. (2009). Structural Analysis of Ligand Stimulation of the Histidine Kinase NarX. Structure, 17(2), 190–201. https://doi.org/10.1016/J.STR.2008.12.013

narX sensor histidine kinase NarX [ Escherichia coli str. K-12 substr. MG1655 ]. (2022, September 22). National Library of Medicine - National Center for Biotechnology Information. https://www.ncbi.nlm.nih.gov/gene/945788

[edit]
Categories
//cds/enzyme/phosphorylation
//cds/membrane/receptor
Parameters
activation_coeficient
protein
signalling_molecule