Difference between revisions of "Part:BBa K5115085"
 
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__TOC__
 
__TOC__
 
===Introduction===
 
===Introduction===
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This part is FKBP with F36V mutation, which is a signal transducer. Binding with F1v, the target protein can get dimerized with B/B Homodimerizer in the culture solution.
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This part is FKBP with F36V mutation, which is a signal transducer. It's used to bind with the target protein. The working procedure of F1v need the use of AP20187, which is a small molecule inducers. The addition of AP20187 to live cells expressing a F1v-tagged fusion protein induces self-association of the fusion protein by promoting the interaction of the dimerization domains.<ref>Clackson, T., Yang, W., Rozamus, L. W., Hatada, M., Amara, J. F., Rollins, C. T., Stevenson, L. F., Magari, S. R., Wood, S. A., Courage, N. L., Lu, X., Cerasoli, F., Gilman, M., & Holt, D. A. (1998). Redesigning an FKBP–ligand interface to generate chemical dimerizers with novel specificity. Proceedings of the National Academy of Sciences of the United States of America, 95(18), 10437–10442.</ref>
  
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===Usage and Biology===
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{|
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In our experiment, we adopted the iDimerize kit, to learn fully about this kit, please check [https://www.takarabio.com/documents/User%20Manual/iDimerize%20Inducible%20Homodimer%20System%20User%20Manual%20%28PT5178/iDimerize%20Inducible%20Homodimer%20System%20User%20Manual%20%28PT5178-1%29_111115.pdf?srsltid=AfmBOoqcK3gD-UsdZMD_JLop5oY-5L8W9a82H6GI4TmsKvN2gEBf37iH iDimerize™ Inducible Homodimer System User Manual]
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| <html><img style="width:400px" src="https://static.igem.wiki/teams/5115/registry/f1v.jpg" alt="contributed by Fudan iGEM 2024"></html>
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|-
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| '''Figure 1. The working procedure of AP20187 and F1v according to the iDimerize™ Inducible Homodimer System User Manual. The F1v is combined with the target protein, which is NixA in our experiment, and the AP20187 can induce self-association of the NixA.
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'''
  
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In [https://parts.igem.org/Part:BBa_K5115086 BBa_K5115086(NixA-F1v)] and [https://parts.igem.org/Part:BBa_K5115087 BBa_K5115087(F1v-NixA)], we link this part with [https://parts.igem.org/Part:BBa_K5115071 BBa_K5115071(NixA)] in two different ways to find the more efficient way of dimerizing the NixA protein.
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|}
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{|
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| <html><img style="width:400px" src="https://static.igem.wiki/teams/5115/registry/ap20187-in-the-manual.png" alt="contributed by Fudan iGEM 2024"></html>
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|-
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| '''Figure 2. While using our iDimerize kit, the explanation of AP20187 in the kit's User Manual.
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'''
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|}
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===Usage and Biology===
 +
In our experiment, we adopted the iDimerize kit. To learn fully about this kit, please check [https://www.takarabio.com/documents/User%20Manual/iDimerize%20Inducible%20Homodimer%20System%20User%20Manual%20%28PT5178/iDimerize%20Inducible%20Homodimer%20System%20User%20Manual%20%28PT5178-1%29_111115.pdf?srsltid=AfmBOoqcK3gD-UsdZMD_JLop5oY-5L8W9a82H6GI4TmsKvN2gEBf37iH iDimerize™ Inducible Homodimer System User Manual].
  
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For convenient connections between the Fiv and NixA, we delete the stop codon of F1v. We link this part with [https://parts.igem.org/Part:BBa_K5115071 BBa_K5115071(NixA)] in two different ways to find the more efficient way of dimerizing the NixA protein. It turns out that linking F1v with the C end of NixA is a better choice, please check [https://parts.igem.org/Part:BBa_K5115086 BBa_K5115086(NixA-F1v)]for more details about our experiment.
  
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===Sequence and Features===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 12:04, 2 October 2024


F1v (FKBP with F36V mutation) without stop codon

contributed by Fudan iGEM 2023

Introduction

This part is FKBP with F36V mutation, which is a signal transducer. It's used to bind with the target protein. The working procedure of F1v need the use of AP20187, which is a small molecule inducers. The addition of AP20187 to live cells expressing a F1v-tagged fusion protein induces self-association of the fusion protein by promoting the interaction of the dimerization domains.[1]

contributed by Fudan iGEM 2024
Figure 1. The working procedure of AP20187 and F1v according to the iDimerize™ Inducible Homodimer System User Manual. The F1v is combined with the target protein, which is NixA in our experiment, and the AP20187 can induce self-association of the NixA.


contributed by Fudan iGEM 2024
Figure 2. While using our iDimerize kit, the explanation of AP20187 in the kit's User Manual.

Usage and Biology

In our experiment, we adopted the iDimerize kit. To learn fully about this kit, please check iDimerize™ Inducible Homodimer System User Manual.

For convenient connections between the Fiv and NixA, we delete the stop codon of F1v. We link this part with BBa_K5115071(NixA) in two different ways to find the more efficient way of dimerizing the NixA protein. It turns out that linking F1v with the C end of NixA is a better choice, please check BBa_K5115086(NixA-F1v)for more details about our experiment.

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ Clackson, T., Yang, W., Rozamus, L. W., Hatada, M., Amara, J. F., Rollins, C. T., Stevenson, L. F., Magari, S. R., Wood, S. A., Courage, N. L., Lu, X., Cerasoli, F., Gilman, M., & Holt, D. A. (1998). Redesigning an FKBP–ligand interface to generate chemical dimerizers with novel specificity. Proceedings of the National Academy of Sciences of the United States of America, 95(18), 10437–10442.