Part:BBa_K3955050:Design

Repeat of CsgF peptide blocking CsgG mutant

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 155
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 160
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

As the csgF peptide is only 17 amino acid residues long, we expected to have difficulties with secretion. Therefore the part is designed as 5 repeats of the peptide, with alanine added in between each repeat. The alanine makes a cleavage site for pancreatic elastase, which is found in the gut. The gut was the target organ for team Lund 2021 which designed this part.

The part is also manually codon optimized for Limosilactobacillus reuteri.

Source

The sequence was made by combining the findings from the article https://www.nature.com/articles/s41467-019-14145-7 with our idea to produce several CsgF mutant peptides at once.

References

1. Zhaofeng Yan, Meng Yin, Jianan Chen, Xueming Li. Assembly and substrate recognition of curli biogenesis system. Nature Communications [Internet]. 2020 Jan 1 [cited 2021 Oct 9];11(1):1–10. Available from: https://search.ebscohost.com/login.aspx?direct=true&db=edsdoj&AN=edsdoj.19f841225ae740398e4682f0391a5534&site=eds-live&scope=site