Coding

Part:BBa_K4722015

Designed by: Anqi Chen   Group: iGEM23_BJEA-China   (2023-10-09)

SP-NicX-Histag-AIDAI

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 945
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 945
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2790
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 945
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 945
    Illegal NgoMIV site found at 1104
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

SP[1][2], enzymes NicX[3][4], Histag, and E. coli autotransporter AIDAI were fused together

Design Consideration

The construction includes: NicX with SP, Histag and AIDAI NicX was genetically connected with SP and AIDAI to enable its direct translation onto the surface of BL21. This innovation eliminated the need for protein purification steps, allowing for the direct utilization of E. coli as a host for enzymes in various applications. Enzymatic cleavage was performed at the NcoI and XhoI restriction sites, allowing for the precise integration of SP-NicX-Histag-AIDAI. The part contains Histag and does not require Histag on the plasmid.

References

  1. Wang, S. N., Liu, Z., Tang, H. Z., Meng, J., & Xu, P. (2007). Characterization of environmentally friendly nicotine degradation by Pseudomonas putida biotype A strain S16. Microbiology, 153(5), 1556-1565. https://doi.org/10.1099/mic.0.2006/005223-0
  2. Wang, W., Xu, P., & Tang, H. (2015). Sustainable production of valuable compound 3-succinoyl-pyridine by genetically engineering Pseudomonas putida using the tobacco waste. Scientific Reports, 5(1), 16411. https://doi.org/10.1038/srep16411
  3. Chen, B., Sun, L., Zeng, G., Shen, Z., Wang, K., Yin, L., ... & Jiang, C. (2022). Gut bacteria alleviate smoking-related NASH by degrading gut nicotine. Nature, 610(7932), 562-568. https://doi.org/10.1038/s41586-022-05299-4
  4. Jiménez, J. I., Canales, Á., Jiménez-Barbero, J., Ginalski, K., Rychlewski, L., García, J. L., & Díaz, E. (2008). Deciphering the genetic determinants for aerobic nicotinic acid degradation: the nic cluster from Pseudomonas putida KT2440. Proceedings of the National Academy of Sciences, 105(32), 11329-11334.https://doi.org/10.1073/pnas.080227310
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