Difference between revisions of "Part:BBa K1378003"
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<html><img style="float:left;width:64px;margin-right:2em" src="https://static.igem.wiki/teams/4765/wiki/2023-b-home.png" alt="contributed by Fudan iGEM 2023"></html>
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===Improved by Fudan iGEM 2023 ===
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Intimin which includes a short N-terminal signal peptide to direct its trafficking to the periplasm, a LysM domain for peptidoglycan binding, and a beta-barrel for transmembrane insertion<ref>Piñero-Lambea, C., Bodelón, G., Fernández-Periáñez, R., Cuesta, A. M., Álvarez-Vallina, L., & Fernández, L. Á. (2015). Programming controlled adhesion of E. coli to target surfaces, cells, and tumors with synthetic adhesins. ACS Synthetic Biology, 4(4), 463–473. https://doi.org/10.1021/sb500252a</ref> , possesses the outer membrane anchoring of MVN.Instead of using INPNC-MVN fusion, we introduce intimin as a surface display system to present MVN, thus facilitating adhesion between ''E. coli'' and ''Microcystis aeruginosa'' PCC7806.
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====Improved part====
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Our improved part is [https://parts.igem.org/Part:BBa_K4765109 BBa_K4765109(Twister P1 + T7_RBS + intimin-MVN fusion + stem-loop)]
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. We introduce intimin as a surface display system to present MVN and construct this part into our ribozyme-assisted polycistronic co-expression system：pRAP.
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=='''Introduction'''==
 
=='''Introduction'''==
  
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<html><img style="float:left;width:64px;margin-right:2em" src="https://static.igem.wiki/teams/4765/wiki/2023-b-home.png" alt="contributed by Fudan iGEM 2023"></html>
 
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=== Fudan iGEM 2023 ===
 
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Intimin which includes a short N-terminal signal peptide to direct its trafficking to the periplasm, a LysM domain for peptidoglycan binding, and a beta-barrel for transmembrane insertion<ref>Piñero-Lambea, C., Bodelón, G., Fernández-Periáñez, R., Cuesta, A. M., Álvarez-Vallina, L., & Fernández, L. Á. (2015). Programming controlled adhesion of E. coli to target surfaces, cells, and tumors with synthetic adhesins. ACS Synthetic Biology, 4(4), 463–473. https://doi.org/10.1021/sb500252a</ref> , possesses the outer membrane anchoring of MVN.Instead of using INPNC-MVN fusion, we introduce intimin as a surface display system to present MVN, thus facilitating adhesion between ''E. coli'' and ''Microcystis aeruginosa'' PCC7806.
 
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====Improved part by Team Fudan 2023====
 
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Our improved part is [https://parts.igem.org/Part:BBa_K4765109 BBa_K4765109(Twister P1 + T7_RBS + intimin-MVN fusion + stem-loop)]
 
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. We introduce intimin as a surface display system to present MVN and construct this part into our ribozyme-assisted polycistronic co-expression system：pRAP.
 
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====Characterization of Twister P1 + T7_RBS + intimin-MVN fusion + stem-loop====
 
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Get details in [https://parts.igem.org/Part:BBa_K4765109 BBa_K4765109(Twister P1 + T7_RBS + intimin-MVN fusion + stem-loop)]
 
  
 
=='''Reference'''==
 
=='''Reference'''==

Revision as of 09:01, 10 October 2023

Microvirin (MVN)---a lectin from Microcystis aeruginosa


contributed by Fudan iGEM 2023

Improved by Fudan iGEM 2023

Intimin which includes a short N-terminal signal peptide to direct its trafficking to the periplasm, a LysM domain for peptidoglycan binding, and a beta-barrel for transmembrane insertion[1] , possesses the outer membrane anchoring of MVN.Instead of using INPNC-MVN fusion, we introduce intimin as a surface display system to present MVN, thus facilitating adhesion between E. coli and Microcystis aeruginosa PCC7806.

Improved part

Our improved part is BBa_K4765109(Twister P1 + T7_RBS + intimin-MVN fusion + stem-loop) . We introduce intimin as a surface display system to present MVN and construct this part into our ribozyme-assisted polycistronic co-expression system：pRAP.

Introduction

This plasmid is a basic part of the mannan binding lectin Microvirin (MVN). Microvirin can be expressed in E. coli and it can bind to Microcystis aeruginosa PCC7806 specifically.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 23

Usage and Biology

MVN is a lectin isolated from the cyanobacteria Microcystis aeruginosa PCC7806. It is an 108 aa protein that consists of 2 domains. According to a carbohydrate microarray carried out previously, MVN binds to carbohydrate and the highest signals are observed with structures that contain α(1→2) linked mannose residues [1]. A binding partner of MVN was identified in the lipopolysaccharide (LPS) fraction of M. aeruginosa PCC7806 and it possibly represents the O-antigen of a LPS. MVN has been previously expressed in E. coli and its binding effect to cyanobacteria cells is species-specific [1].

Figure 1. Solution structure of MVN. MVN consists of 2 domains, domain A and domain B, which are colored yellow and blue respectively. Domain A is formed by residues 38-93 while domain B is formed by residues 1-37 and 94-108. The mannan binding site is only found in domain A. This figure is taken from Protein Data Bank (PDB ID: 2Y1S).


Reference

1.Kehr, J. C., Zilliges, Y., Springer, A., Disney, M. D., Ratner, D. D., Bouchier, C., ... & Dittmann, E. (2006). A mannan binding lectin is involved in cell–cell attachment in a toxic strain of Microcystis aeruginosa. Molecular microbiology, 59(3), 893-906.

  1. ↑ Piñero-Lambea, C., Bodelón, G., Fernández-Periáñez, R., Cuesta, A. M., Álvarez-Vallina, L., & Fernández, L. Á. (2015). Programming controlled adhesion of E. coli to target surfaces, cells, and tumors with synthetic adhesins. ACS Synthetic Biology, 4(4), 463–473. https://doi.org/10.1021/sb500252a