Coding

Part:BBa_K3955101:Design

Designed by: Camilla Lane   Group: iGEM21_Lund   (2021-09-30)


DB3DB3 peptide fused with slpmod signal peptide


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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.