Difference between revisions of "Part:BBa K1404006:Design"

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Lin, X., Li, Z., Li, Y., & Lu, Y. (2021). A robust escherichia coli cell-free expression toolbox driven by sigma factors. Biochemical Engineering Journal, 171, 108031. https://doi.org/10.1016/j.bej.2021.108031
 
Lin, X., Li, Z., Li, Y., & Lu, Y. (2021). A robust escherichia coli cell-free expression toolbox driven by sigma factors. Biochemical Engineering Journal, 171, 108031. https://doi.org/10.1016/j.bej.2021.108031
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Thaller, M. C., Schippa, S., Bonci, A., Cresti, S., & Rossolini, G. M. (2006). Identification of the gene (aphA) encoding the class B acid phosphatase/phosphotransferase of escherichia coli MG1655 and characterization of its product. FEMS Microbiology Letters, 146(2), 191-198. https://doi.org/10.1111/j.1574-6968.1997.tb10192.x
 

Latest revision as of 15:32, 13 October 2022


p70-CsgA, curli generator


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

If you intend to use this part, be aware that CsgA might be toxic to E. coli cells in large quantities. We were unable to clone csgA in front of the strong promoter J23119 or the weaker promoter J23114

Source

A strong RBS desgined by team Paris Bettencourt 2009 and the coding region for the csgA gene as found in the genome of the E. coli strain MC4100 were synthesized by Genecust. An illegal restriction site was removed by introducing a silent mutation. This direct synthesis was then cloned downstream of the K342000 promoter by standard assembly.

References

References added by REC-CHENNAI

Blanco, L. P., Evans, M. L., Smith, D. R., Badtke, M. P., & Chapman, M. R. (2012). Diversity, biogenesis and function of microbial amyloids. Trends in Microbiology, 20(2), 66-73. https://doi.org/10.1016/j.tim.2011.11.005

Lin, X., Li, Z., Li, Y., & Lu, Y. (2021). A robust escherichia coli cell-free expression toolbox driven by sigma factors. Biochemical Engineering Journal, 171, 108031. https://doi.org/10.1016/j.bej.2021.108031