Regulatory

Part:BBa_K4115004:Design

Designed by: Kaijun Wang   Group: iGEM22_ShanghaiTech_China   (2022-09-21)


PcstA Mutant1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This promoter is found to be activated by CRP under a carbon source starvation. It can be used as a sensor for the carbon source condition in the culture medium. After having mutated, this promoter is much more sensitive and strong compared with its original version(BBa_K118011). So it can also be used in protein production.


Source

The origin PcstA was amplified from E.coli DH5-alpha genome through PCR. After inserting this promoter into a plasmid, we use PCR to do the mutant at its CRP binding site.

References

[1] Soberon-Chavez, G., Alcaraz, L.D., Morales, E., Ponce-Soto, G.Y., and Servin-Gonzalez, L. (2017). The Transcriptional Regulators of the CRP Family Regulate Different Essential Bacterial Functions and Can Be Inherited Vertically and Horizontally. Front Microbiol 8. ARTN 959 10.3389/fmicb.2017.00959.
[2] Shimada, T., Fujita, N., Yamamoto, K., and Ishihama, A. (2011). Novel roles of cAMP receptor protein (CRP) in regulation of transport and metabolism of carbon sources. PLoS One 6, e20081. 10.1371/journal.pone.0020081.
[3] Gasperotti, A., Going, S., Fajardo-Ruiz, E., Forne, I., and Jung, K. (2020). Function and Regulation of the Pyruvate Transporter CstA in Escherichia coli. Int J Mol Sci 21. 10.3390/ijms21239068.