Part:BBa_K5322007:Design
NO-inducible Mfp53 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 1113
Illegal NotI site found at 1077 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 763
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
To ensure that the animal-derived mussel foot protein Mfp53 can be successfully expressed in Escherichia coli Nissle 1917, we performed codon optimization on the Mfp53 sequence.
Source
The plasmid pET29a-J23119-SoxR-T-pSoxS-RBS-Mfp53-T7 utilizes the pET29a vector, which is typically used for high-level expression of genes in Escherichia coli. The components used in this design, including the J23119 promoter, Mfp3, Mfp5, the protein linker (GGGGS), SoxR protein, and SoxS promoter, are all sourced from the iGEM registry of standard biological parts （BBa_J23119，BBa_K4854000，BBa_K1583002,BBa_K5322001,BBa_K554003，BBa_K5322004）. Among these, the mussel foot protein Mfp5 and Mfp3 are derived from natural marine mussels. These components were selected for their validated functions and compatibility in synthetic biology applications, facilitating controlled expression of genes within bacterial cells under specific conditions.
References
1. Yu LC. Microbiota dysbiosis and barrier dysfunction in inflammatory bowel disease and colorectal cancers: exploring a common ground hypothesis. J. Biomed. Sci. 2018;25:79. doi: 10.1186/s12929-018-0483-8
2. Khanna Reena, Levesque Barrett G, Sandborn William J. IBD: Measuring what counts--endoscopic assessment in IBD[J]. Nature Reviews Gastroenterology & Hepatology. 2014, 11(1): 9-10.
3. Lee BP, Messersmith PB, Israelachvili JN, et al. Musselinspired adhesives and coatings[J]. Annu Rev Mater Res. 2011, 41(1): 99-132.