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Team:Heidelberg/pages/Circularization Constructs
From 2014.igem.org
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===Introduction=== | ===Introduction=== | ||
| - | - | + | The most promising approaches to [[Team:Heidelberg/Toolbox/Circularization | circularize]] proteins are protein trans-splicing using [[Team:Heidelberg/Project/Background | split inteins]] [[#References[1]]] such as ''Npu'' DnaE [[#References[2]]] and Sortase A-catalyzed cyclization [[#References[3]]]. |
| - | - | + | |
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| + | On DNA level, creating circular proteins equals creating fusion proteins. However, existing protein fusion standards like RFC 23 cause scars. | ||
===NpuDnaE intein RFC [???] circularization constructs=== | ===NpuDnaE intein RFC [???] circularization constructs=== | ||
| + | |||
| + | - Bild: 000, 001 | ||
====Design==== | ====Design==== | ||
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====Cloning==== | ====Cloning==== | ||
| + | |||
| + | mechanismus? | ||
====Results==== | ====Results==== | ||
===Sortase A circularization constructs=== | ===Sortase A circularization constructs=== | ||
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| + | - Bild: 002, 003 | ||
====Design==== | ====Design==== | ||
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====Results?==== | ====Results?==== | ||
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| + | ===References=== | ||
| + | [1] Iwai, H., Lingel, a & Pluckthun, a. Cyclic green fluorescent protein produced in vivo using an artificially split PI-PfuI intein from Pyrococcus furiosus. J. Biol. Chem. 276, 16548–54 (2001). | ||
| + | |||
| + | [2] Zettler, J., Schütz, V. & Mootz, H. D. The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction. FEBS Lett. 583, 909–14 (2009). | ||
| + | |||
| + | [3] Antos, J. M. et al. A straight path to circular proteins. J. Biol. Chem. 284, 16028–36 (2009). | ||
Revision as of 12:16, 15 October 2014
Contents |
Introduction
The most promising approaches to circularize proteins are protein trans-splicing using split inteins [[#References[1]]] such as Npu DnaE [[#References[2]]] and Sortase A-catalyzed cyclization [[#References[3]]].
On DNA level, creating circular proteins equals creating fusion proteins. However, existing protein fusion standards like RFC 23 cause scars.
NpuDnaE intein RFC [???] circularization constructs
- Bild: 000, 001
Design
Cloning
mechanismus?
Results
Sortase A circularization constructs
- Bild: 002, 003
Design
Cloning
Results?
References
[1] Iwai, H., Lingel, a & Pluckthun, a. Cyclic green fluorescent protein produced in vivo using an artificially split PI-PfuI intein from Pyrococcus furiosus. J. Biol. Chem. 276, 16548–54 (2001).
[2] Zettler, J., Schütz, V. & Mootz, H. D. The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction. FEBS Lett. 583, 909–14 (2009).
[3] Antos, J. M. et al. A straight path to circular proteins. J. Biol. Chem. 284, 16028–36 (2009).
