Part:BBa_K3955101:Design
DB3DB3 peptide fused with slpmod signal peptide
- 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
In order for DB3DB3 to be successfully secreted extracellularly, it needed to be distinguishable from SLPmod by being larger than it was. We therefore repeated the sequence 3 times and introduced cleavege sites for pancreatic elastase in between the repeats, by adding an alanine. Pancreatic elastase was chosen as a peptidase since it naturally occurs in the colon, which would be the target organ of our potential future implementation.
Codons were chosen manually to reduce the sequence repeats in the nucleotide sequence but still mainly use the most common codons for L. reuteri, our intended chassi.
Source
Peptide sequence: Klein A, Ziehm T, Tusche M, Buitenhuis J, Bartnik D, Boeddrich A et al. Optimization of the All-D Peptide D3 for Aβ Oligomer Elimination. PLOS ONE. 2016;11(4):e0153035.
SLPmod: https://parts.igem.org/Part:BBa_K3183008
References
1. Klein A, Ziehm T, Tusche M, Buitenhuis J, Bartnik D, Boeddrich A et al. Optimization of the All-D Peptide D3 for Aβ Oligomer Elimination. PLOS ONE. 2016;11(4):e0153035.
2. Perov S, Lidor O, Salinas N, Golan N, Tayeb- Fligelman E, Deshmukh M et al. Structural Insights into Curli CsgA Cross-β Fibril Architecture Inspire Repurposing of Anti-amyloid Compounds as Anti-biofilm Agents. PLOS Pathogens. 2019;15(8):e1007978.
3. Fernandez A, Horn N, Wegmann U, Nicoletti C, Gasson M, Narbad A. Enhanced Secretion of Biologically Active Murine Interleukin-12 by Lactococcus lactis. Applied and Environmental Microbiology. 2009;75(3):869-871.