Difference between revisions of "Part:BBa K4765108"

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===introduction===
 
===introduction===
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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<ref>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</ref>.Ag3 is a corresponding antigen of [https://parts.igem.org/Part:BBa_K4765007 Nb3]<ref>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</ref>. 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.
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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<ref>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</ref>.Ag3 is a corresponding antigen of [https://parts.igem.org/Part:BBa_K4765007 Nb3]<ref>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</ref>. 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.
 
We’ve constructed this fusion protein into our ribozyme-assisted polycistronic co-expression system：pRAP.
  

Revision as of 15:03, 1 October 2023


Twister P1 + T7_RBS + INPNC-Nb3 fusion + stem-loop

contributed by Fudan iGEM 2023

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 542
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 391
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 133
    Illegal NgoMIV site found at 466
    Illegal AgeI site found at 490
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1199


Reference

  1. ↑ 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
  2. ↑ 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