Part:BBa_K4854026

CsgA+Mfp5+GFP

This composite part contains lac promoter(BBa_R0010)^[1], RBS(BBa_B0034)^[2], CsgA(BBa_K4854013)^[3], GS linker(BBa_K4854012)^[4], Mfp5(BBa_K4854001)^[5], a flexible linker(BBa_K105012)^[6] and GFP(BBa_E0040)^[7] gene. It is meant to express a fusion protein of the E. coli membrane protein CsgA, the mussel foot protein Mfp5, and the green fluorescent protein (GFP).
Mussel foot protein Mfp5 is an adhesive protein from the Mediterranean mussel (Mytilus galloprovincialis). Mussels can secrete special proteins to adhere to surfaces under turbulent environments.^[8]
CsgA protein is the major component of biofilms of E. coli. It can self-assemble into a network of amyloid nanofibers outside the cell. Because of this characteristic, CsgA can be a platform to display recombinant protein on the surface of E. coli.^[9]
We designed GFP as the functional protein linked at the C-terminal end of the CsgA+Mfp5 fusion protein.
We performed codon optimization to improve gene expression in E. coli.

Figure 1. The circuit of the GFP+CsgA+Mfp5 protein.

Reference
[1] https://parts.igem.org/Part:BBa_R0010
[2] https://parts.igem.org/Part:BBa_B0034
[3] https://parts.igem.org/Part:BBa_K4854013
[4] https://parts.igem.org/Part:BBa_K4854012
[5] https://parts.igem.org/Part:BBa_K4854001
[6] https://parts.igem.org/Part:BBa_K105012
[7] https://parts.igem.org/Part:BBa_E0040
[8] https://www.uniprot.org/uniprotkb/Q8WTE8/entry
[9] Fei Li, Luona Ye, Longyu Zhang, Xiaoyan Li, Xiaoxiao Liu, Jiarui Zhu, Huanhuan Li, Huimin Pang, Yunjun Yan, Li Xu, Min Yang, Jinyong Yan, Design of a genetically programmed barnacle-curli inspired living-cell bioadhesive, Materials Today Bio, Volume 14, 2022.

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.rc site found at 1603