Difference between revisions of "Part:BBa K4765107"

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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 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
 
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 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.
 
</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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We’ve constructed this fusion protein into our ribozyme-assisted polycistronic co-expression system：pRAP.
 
  
 
===Usage and Biology===
 
===Usage and Biology===
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The surface-displayed antigen can specifically interact with the nanobody produced by [https://parts.igem.org/Part:BBa_K4765108 ribozyme+strong RBS+INPNC-Nb3 fusion+stem-loop].
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The surface-displayed antigen can specifically interact with the nanobody produced by [https://parts.igem.org/Part:BBa_K4765108 BBa_K4765108].
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===Characterization===
 
===Characterization===
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====Sequencing map====
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{|
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| <html><img style="width:640px" src="https://static.igem.wiki/teams/4765/wiki/zsl/ag3-inpnc-sequence-map.png" alt="contributed by Fudan iGEM 2023"></html>
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|-
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| '''Figure 1. Sequencing map of INPNC-Ag3 fusion'''
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Sequencing is performed using the primer：Kan-F: 5-ATTCTCACCGGATTCAGT-3.
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|}
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====Selection through Aggregation Assay====
 
Get details in [https://parts.igem.org/Part:BBa_K4765108 BBa_K4765108].
 
Get details in [https://parts.igem.org/Part:BBa_K4765108 BBa_K4765108].
  
 
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<span class='h3bb'>Sequence and Features</span>
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===Sequence and Features===
 
<partinfo>BBa_K4765107 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4765107 SequenceAndFeatures</partinfo>
  
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==Reference==
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===Reference===

Latest revision as of 15:46, 12 October 2023

Twister P1 + T7_RBS + INPNC-Ag3 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 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.

Usage and Biology

The surface-displayed antigen can specifically interact with the nanobody produced by BBa_K4765108.

Characterization

Sequencing map

contributed by Fudan iGEM 2023
Figure 1. Sequencing map of INPNC-Ag3 fusion

Sequencing is performed using the primer：Kan-F: 5-ATTCTCACCGGATTCAGT-3.

Selection through Aggregation Assay

Get details in BBa_K4765108.

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
    COMPATIBLE WITH RFC[1000]


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