Part:BBa_K4765107
Twister P1 + T7_RBS + INPNC-Ag3 fusion + stem-loop
Contents
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 BBa_K4765007(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-Nb3 fusion+stem-loop.
Characterization
Sequencing map
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| Figure 1. Sequencing map of INPNC-Nb3 fusion
Sequencing is performed using the primer：Kan-F: 5-ATTCTCACCGGATTCAGT-3. |
Selection through Aggregation Assay
Get details in BBa_K4765108.
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 - 1000COMPATIBLE WITH RFC[1000]
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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