Part:BBa_K5048046

Neae

Codes for the surface presentation system containing an export tag, a LysM sequence, a β-barrel, and a Spacer sequence.

Design

We take the N-terminal domain of intimin of enterohemorrhagic Escherichia coli as our surface display system in the adhesion module. This system has been proven to be effective and safe by papers.

Source

N-terminal domain of intimin of enterohemorrhagic Escherichia coli.

Usage and Biology

A surface presentation system that enables proteins to be displayed on the outer membrane of the cell.

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 Flag tag at the C-terminal of the protein forming Neae-FLAG(BBa_K5048047) and inserted this improved sequence into pET-28a backbone to create the plasmid Neae-FLAG.

Applications

Neae can be used as a surface display system in E. coli., conjugating with different proteins and presenting them on the cell's outer membrane. In our project, Neae is used to present adhesion protein LAP and HSP60 on the surface of engineered E. coli. to enable the engineered bacteria to adhere to the intestine, and also realize cell-cell adhesion simultaneously.

References

Glass, D. S., & Riedel-Kruse, I. H. (2018). A Synthetic Bacterial Cell-Cell Adhesion Toolbox for Programming Multicellular Morphologies and Patterns. Cell, 174(3), 649-658.e616. doi:https://doi.org/10.1016/j.cell.2018.06.041

Sequence and Features

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1244