Difference between revisions of "Part:BBa K5115085"

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__TOC__
 
__TOC__
 
===Introduction===
 
===Introduction===
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 B/B Homodimerizer, which is a small molecule inducers. The addition of B/B Homodimerizer to live cells expressing a DmrB-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>
  
 
===Usage and Biology===
 
===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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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 [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.
 
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 11:32, 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 DmrB-tagged fusion protein induces self-association of the fusion protein by promoting the interaction of the dimerization domains.[1]

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.

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.