Part:BBa_K4247019:Design
Mfp151_second-half
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
MFP151 is a long protein and as such, its sequence might be difficult to synthesise. Hence, at the UCopenhagen team, we have split the sequence into two halves such that each half of the sequence would be produced in 2 expression plasmids each. These two halves can be put together by Golden Gate Cloning to obtain the sequence coding for the full MFP151 protein.
Both halves of the sequence coding for MFP151 are contained in an pET24 (+) expression vector having a T7 promoter, terminator, KAN resistance gene and a 6x His-tag in the C-terminus of the protein.
Further, this sequence was codon optimised as per E.coli's codon bias.
Source
The sequence of this part was taken from the following 2 articles:
Kitamura, M., Kawakami, K., Nakamura, N., Tsumoto, K., Uchiyama, H., Ueda, Y., Kumagai, I. and Nakaya, T. (1999), Expression of a model peptide of a marine mussel adhesive protein in Escherichia coli and characterization of its structural and functional properties. J. Polym. Sci. A Polym. Chem., 37: 729-736. https://doi.org/10.1002/(SICI)1099-0518(19990315)37:6<729::AID-POLA8>3.0.CO;2-3
Hwang, D. S., Yoo, H. J., Jun, J. H., Moon, W. K., & Cha, H. J. (2004). Expression of functional recombinant mussel adhesive protein Mgfp-5 in Escherichia coli. Applied and environmental microbiology, 70(6), 3352–3359. https://doi.org/10.1128/AEM.70.6.3352-3359.2004
References
Hwang, D. S., Gim, Y., Yoo, H. J., & Cha, H. J. (2007). Practical recombinant hybrid mussel bioadhesive fp-151. Biomaterials, 28(24), 3560–3568. https://doi.org/10.1016/j.biomaterials.2007.04.039
Lin, Q., Gourdon, D., Sun, C., Holten-Andersen, N., Anderson, T. H., Waite, J. H., & Israelachvili, J. N. (2007). Adhesion mechanisms of the mussel foot proteins mfp-1 and mfp-3. Proceedings of the National Academy of Sciences of the United States of America, 104(10), 3782–3786. https://doi.org/10.1073/pnas.0607852104
Kim, E., Dai, B., Qiao, J. B., Li, W., Fortner, J. D., & Zhang, F. (2018). Microbially Synthesized Repeats of Mussel Foot Protein Display Enhanced Underwater Adhesion. ACS applied materials & interfaces, 10(49), 43003–43012. https://doi.org/10.1021/acsami.8b14890
Mengkui Cui, Susu Ren, Shicao Wei, Chengjun Sun, and Chao Zhong,"Natural and bio-inspired underwater adhesives: Current progress and new perspectives", APL Materials 5, 116102 (2017) https://doi.org/10.1063/1.4985756