Composite

Part:BBa_K4998036

Designed by: Asimenia Ioannidou   Group: iGEM23_thessaloniki   (2023-10-09)


Composite SilA gene

Usage and Biology

This composite part consists of the PgroES promoter (BBa_K4998020), the signal peptide sequence LipA (BBa_K4998019), the SilA gene (BBa_K4998018) that expresses the laccase protein lac7, and a terminator (BBa_K780000). The part was expressed in Bacillus subtilis.

The PgroES promoter is a strong, constant promoter that we used to express our protein [1]. LipA is a signal sequence that is part of the twin-arginine pathway system (Tat) and is recognized by the cell and results in the translocation of folded proteins extracellularly [2]. Laccases are multicopper enzymes that oxidize a broad spectrum of substrates, like polyphenolic compounds, lignin/lignin-like structures, humus, etc. SilA gene originates from Streptomyces ipomonae and expresses a 335 amino acid protein [3].




Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 480
    Illegal XhoI site found at 353
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 284
    Illegal NgoMIV site found at 946
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 230


Characterization

The results for this part are shown in BBa_K4998018

References

[1] Song, Y., Nikoloff, J. M., Fu, G., Chen, J., Li, Q., Xie, N., Zheng, P., Sun, J., & Zhang, D. (2016). Promoter screening from bacillus subtilis in various conditions hunting for synthetic biology and Industrial Applications. PLOS ONE, 11(7). https://doi.org/10.1371/journal.pone.0158447

[2]Zhang, W., Yang, M., Yang, Y., Zhan, J., Zhou, Y., & Zhao, X. (2016). Optimal secretion of alkali-tolerant xylanase in bacillus subtilis by signal peptide screening. Applied Microbiology and Biotechnology, 100(20), 8745–8756. https://doi.org/10.1007/s00253-016-7615-4

[3] Janusz, G., Pawlik, A., Świderska-Burek, U., Polak, J., Sulej, J., Jarosz-Wilkołazka, A., & Paszczyński, A. (2020). Laccase properties, physiological functions, and evolution. International Journal of Molecular Sciences, 21(3), 966. https://doi.org/10.3390/ijms21030966

[edit]
Categories
Parameters
None