Difference between revisions of "Part:BBa K5115085"
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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].
 
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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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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For convenient connections between the Fiv and NixA, we delete the stop codon of F1v. 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.
  
  

Revision as of 12:43, 29 September 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 B/B Homodimerizer, which is a small molecule inducers. The addition of B/B Homodimerizer 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 B/B Homodimerizer 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 B/B Homodimerizer can induce self-association of the NixA.


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. In BBa_K5115086(NixA-F1v) and BBa_K5115087(F1v-NixA), we link this part with BBa_K5115071(NixA) in two different ways to find the more efficient way of dimerizing the NixA protein.


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.