Difference between revisions of "Part:BBa K5115085"
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| − | | <html><img style="width:400px" src="https://static.igem.wiki/teams/5115/registry/f1v | + | | <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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| '''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. | | '''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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| + | | <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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| + | | '''Figure 2. While using our iDimerize kit, the explanation of AP20187 in the kit's User Manual. | ||
| + | ''' | ||
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| + | |} | ||
===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]. | 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]. | ||
| − | For convenient connections between the Fiv and NixA, we delete the stop codon of F1v. | + | 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. |
| − | + | ||
| + | ===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
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]
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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.
|
|
| 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
- ↑ 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.