Part:BBa_K5048048

LAP

Codes for the membrane protein of Lister innocua(L. innocua), which can bind to HSP60 on the membrane of small intestinal epithelial cells, and another type of E. coli that we have engineered.

Design Notes

Adhesion protein from non-pathogenic Listeria.

Source

Listeria innocua strain CFSAN044836 chromosome, complete genome,Sequence ID: CP045743.1,Range 1: 55424 to 58024

Experiments

We handed this original sequence to the biology company Tsingke and implemented condon optimization. To facilitate the detection of protein products, we added a 6xHis tag at the C-terminal of the protein forming LAP-6xHis(BBa_K5048058) and inserted this improved sequence into pET-28a backbone to create the plasmid LAP-6xHis.

Applications

When presented on the outer membrane of non-pathogenic Listeria, it acts as a beneficial protein that binds with HSP60 in the small intestine, occupying the binding site of pathogenic Listeria and avoiding the causing intestinal infection.

References

Drolia, R., Amalaradjou, M. A. R., Ryan, V., Tenguria, S., Liu, D., Bai, X., Bhunia, A. K. (2020). Receptor-targeted engineered probiotics mitigate lethal Listeria infection. Nature Communications, 11(1), 6344. doi:10.1038/s41467-020-20200-5

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 859
  Illegal EcoRI site found at 2206
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 859
  Illegal EcoRI site found at 2206
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 859
  Illegal EcoRI site found at 2206
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 859
  Illegal EcoRI site found at 2206
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 859
  Illegal EcoRI site found at 2206
  
• 1000
  COMPATIBLE WITH RFC[1000]