Coding
Part:BBa_K4722020
Designed by: Anqi Chen Group: iGEM23_BJEA-China (2023-10-09)
SP-Histag-NicX-BrkA
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 971
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 971
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 971
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 971
Illegal NgoMIV site found at 1130
Illegal NgoMIV site found at 1823
Illegal NgoMIV site found at 2252
Illegal NgoMIV site found at 2888 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2856
Usage and Biology
SP[1][2], Histag, NicX[3][4], BrkA[5] were fused together
Design Consideration
The construction includes: NicX with SP, BrkA and Histag NicX was genetically connected with SP and BrkA 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-Histag-NicX-BrkA. The part contains Histag and does not require Histag on the plasmid.
References
- ↑ 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
- ↑ 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
- ↑ 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
- ↑ 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
- ↑ Sun, F., Pang, X., Xie, T., Zhai, Y., Wang, G., & Sun, F. (2015). BrkAutoDisplay: functional display of multiple exogenous proteins on the surface of Escherichia coli by using BrkA autotransporter. Microbial Cell Factories, 14, 1-12. https://doi.org/10.1186/s12934-015-0316-3
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Categories
Parameters
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