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Team:LMU-Munich/TEV-System/Functional Principle
From 2010.igem.org
(New page: {{:Team:LMU-Munich/Templates/Page Header}} If the plasmid has been incorporated by the cell, both protein complexes should be synthesized. Both protein-complexes are supposed to attach bec...) |
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| - | If the plasmid has been incorporated by the cell, both protein complexes should be synthesized. | + | If one has the Cellline which is stable transformed with TetinduciblePromoter+TEVrecognition+Ndegron+SF3B+bak one can transform these cells with the Construct containing the Protein of interest. |
| - | Both protein-complexes are supposed to attach because p14* and | + | If the plasmid has been incorporated by the cell, both protein complexes should be synthesized when the Tet inducible Promoter is induzed by doxycyclin. |
| - | The TEV-protease is supposed to cut the recognition sites. Now the protein of interest (eGFP) is split apart and the N-terminus of the N-degron is uncovered. This is a signal for the cell to deplete the protein complex. So all protein should be destroyed except the protein of interest. | + | Both protein-complexes are supposed to attach because p14* and SF3B are interacting proteins. |
| + | The TEV-protease is supposed to cut the recognition sites. Now the protein of interest (eGFP) is split apart and the N-terminus of the N-degron is uncovered. This is a signal for the cell to deplete the protein complex including the deadly bak. So all protein should be destroyed except the protein of interest. | ||
If no plasmid has been incorporated, the cell will die because of Bak. | If no plasmid has been incorporated, the cell will die because of Bak. | ||
{{:Team:LMU-Munich/Templates/Page Footer}} | {{:Team:LMU-Munich/Templates/Page Footer}} | ||
Revision as of 18:29, 6 August 2010
If one has the Cellline which is stable transformed with TetinduciblePromoter+TEVrecognition+Ndegron+SF3B+bak one can transform these cells with the Construct containing the Protein of interest.
If the plasmid has been incorporated by the cell, both protein complexes should be synthesized when the Tet inducible Promoter is induzed by doxycyclin.
Both protein-complexes are supposed to attach because p14* and SF3B are interacting proteins.
The TEV-protease is supposed to cut the recognition sites. Now the protein of interest (eGFP) is split apart and the N-terminus of the N-degron is uncovered. This is a signal for the cell to deplete the protein complex including the deadly bak. So all protein should be destroyed except the protein of interest.
If no plasmid has been incorporated, the cell will die because of Bak.
