Difference between revisions of "Part:BBa K4765107"

 
m
 
(16 intermediate revisions by 5 users not shown)
Line 1: Line 1:
 +
__NOTOC__
 +
<partinfo>BBa_K4765107 short</partinfo>
  
 +
<html><img style="float:right;width:128px" src="https://static.igem.wiki/teams/4765/wiki/2023-b-home.png" alt="contributed by Fudan iGEM 2023"></html>
 +
__TOC__
 +
 +
===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<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.
 +
 +
===Usage and Biology===
 +
The surface-displayed antigen can specifically interact with the nanobody produced by [https://parts.igem.org/Part:BBa_K4765108 BBa_K4765108].
 +
 +
===Characterization===
 +
====Sequencing map====
 +
{|
 +
| <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>
 +
|-
 +
| '''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 [https://parts.igem.org/Part:BBa_K4765108 BBa_K4765108].
 +
 +
<!-- -->
 +
===Sequence and Features===
 +
<partinfo>BBa_K4765107 SequenceAndFeatures</partinfo>
 +
 +
<!-- Uncomment this to enable Functional Parameter display
 +
===Functional Parameters===
 +
<partinfo>BBa_K4765107 parameters</partinfo>
 +
<!-- -->
 +
 +
===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