Part:BBa_K5322005:Design
NO-inducible Mfp3 Expression System
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 552
Illegal NheI site found at 836
Illegal NotI site found at 800 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
To ensure that the animal-derived mussel foot protein Mfp3 can be successfully expressed in Escherichia coli Nissle 1917, we performed codon optimization on the Mfp3 sequence.
Source
The plasmid pET29a-J23119-SoxR-T-pSoxS-RBS-Mfp3-T7 utilizes the pET29a vector, which is commonly used for high-level gene expression in Escherichia coli. The components used in this design, including the J23119 promoter, Mfp3, SoxR protein, and SoxS promoter, are derived from standard biological parts registered in the iGEM registry (BBa_J23119, BBa_K4854000, BBa_K554003 , BBa_K5322004 ), with the mussel foot protein Mfp3 sourced from natural marine mussels. These parts were selected due to their validated functions and compatibility in synthetic biology applications, facilitating controlled expression within bacterial cells under specific conditions.
References
1. Hidalgo, E.; Demple, B. An iron-sulfur center essential for transcriptional activation by the redox-sensing SoxR protein EMBO J. 1994, 13, 138– 146.
2. Rafa H, Saoula H, Belkhelfa M, Medjeber O, Soufli I, Toumi R, de Launoit Y, Moralès O, Nakmouche M, Delhem N, et al. IL-23/IL-17A axis correlates with the nitric oxide pathway in inflammatory bowel disease: immunomodulatory effect of retinoic acid. J Interferon Cytokine Res. 2013;33:355–368.
3. Rafa H, Amri M, Saoula H, Belkhelfa M, Medjeber O, Boutaleb A, Aftis S, Nakmouche M, Touil-Boukoffa C. Involvement of interferon-γ in bowel disease pathogenesis by nitric oxide pathway: a study in Algerian patients. J Interferon Cytokine Res. 2010;30:691–697.