Part:BBa_K4765108
Twister P1 + T7_RBS + INPNC-Nb3 fusion + stem-loop
introduction
INPNC-Ag3 fusion is composed of a surface display system (INPNC+linker) and the coding sequence of a nanobody. INPNC exhibits compatibility with the translocation and surface display of proteins containing multiple cofactors and disulfide bond-containing passengers[1].Ag3 is a corresponding antigen of Nb3[2]. The interaction between Ag-Nb can mediate specific adhesion of Escherichia coli. A flexible protein domain linker of 10 aa was introduced between INPNC and Ag3 to ensure independent functionality of Ag3 and INPNC with minimal mutual disruption. We’ve constructed this fusion protein into our ribozyme-assisted polycistronic co-expression system：pRAP.
Usage and Biology
The surface-displayed antigen can specifically interact with the nanobody produced by ribozyme+strong RBS+INPNC-Ag3 fusion+stem-loop.
Charaterization
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 542
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 391
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 133
Illegal NgoMIV site found at 466
Illegal AgeI site found at 490 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1199
Reference
- ↑ van Bloois, E., Winter, R. T., Kolmar, H., & Fraaije, M. W. (2011). Decorating microbes: Surface display of proteins on Escherichia coli. Trends in Biotechnology, 29(2), 79–86. https://doi.org/10.1016/j.tibtech.2010.11.003
- ↑ 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.e16. https://doi.org/10.1016/j.cell.2018.06.041
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