Coding
Part:BBa_K3582026
Designed by: Saillesh Aravindhan C Group: iGEM20_IISER-Pune-India (2020-10-20)
Revision as of 09:37, 24 October 2020 by Sailleshac (Talk | contribs)
Inhibitory sequence 1 of PfEMP1 - ICAM1 interaction grafted into a cyclotide
This biobrick is able to synthesize the grafted cyclotide that could inhibit the 5MZA interaction between the host protein (ICAM-1) and the parasite protein (PfEMP1). The different components of this biobrick are described below:
- 1] BBa_K3582020: C intein, which is the modified version of the biobrick used by the 2014 iGEM Heidelberg team.
- 2] BBa_K2333015: Bsa 1 Reversed recognition site.
- 3] BBA_K3582023(The cyclotide precursor): The gene containing the cyclotide Kalata B1(29 amino acids), which is inserted between the C intein and the N intein so that it will circularize after translation.
- Strep Ⅱ tag (inbuilt): This tag is placed within loop 3 of the cyclotide construct which could reduce the hemolytic activity of the cyclotide. This tag is also used to purify the molecule.
- 4] BBa_K3582024: Inhibitory sequence 1, which is thought to be able to inhibit the interaction between ICAM-1 and PfEMP1, is grafted into loop 6 of the cyclotide. This inhibitor has a higher affinity for interaction as compared to the wild type.
- 5] BBa_K3582021: Bsa 1 recognition site
- 6] BBa_K3582022: N intein, which is the modified version of the biobrick used by the Heidelberg 2014 team for circularization of the protein.
This final product is thought to be able to act like a drug that is capable of preventing the ICAM-1 and PfEMP1 interaction.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 112
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 143
Illegal BsaI.rc site found at 109
References
- 1] Gould, A., Ji, Y., Aboye, T. L., & Camarero, J. A. (2011, December). Cyclotides, a novel ultrastable polypeptide scaffold for drug discovery. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3330703/
- 2] DJ;, P. A. (n.d.). Cyclotides as grafting frameworks for protein engineering and drug design applications. Retrieved from https://pubmed.ncbi.nlm.nih.gov/23893608/
- 3] Camarero, J. A., & Campbell, M. J. (2019, April 19). The Potential of the Cyclotide Scaffold for Drug Development. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631875/
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Categories
Parameters
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