"
Team:ESBS-Strasbourg/Results/Biobricks
From 2010.igem.org
(Difference between revisions)
| Line 638: | Line 638: | ||
<br> | <br> | ||
<br> | <br> | ||
| + | <p><b>Background:</b></p> | ||
| + | PIF3 is a downstream transcription factor in a well studied signalling pathway of A. thaliana, upon stimulation with red (650 nm) light, it binds directly to PhyB and translocates to the nucleus as a heterodimer where it modulates the transcription of response genes. PIF3 binds only the red-light-exposed form of phytochrome, Pfr, and shows no-measurable binding affinity for the dark- or infrared-exposed Pr state.<br> | ||
| + | In our system target proteins are fused to PIF3 and tagged with the DAS degradation sequence which, through light activation, brings the degradation tag in proximity to ClpX.<br> | ||
| + | <br> | ||
| + | <p><b>Conception:</b></p> | ||
| + | The light-sensitive interaction with PhyB has been mapped to the first 100-residue N-terminal activated phytochrome binding (APB) domain of PIF3 <a href="https://2010.igem.org/Team:ESBS-Strasbourg/Project/Reference">(Lim & Voigt, 2009)</a>.<br> | ||
| + | We chose this sequence, as it has already been successfully used in different synthetic in vitro applications that benefitted from its light-sensitive interactions with PhyB. The original sequence contains an XbaI restriction site. | ||
| + | |||
| + | |||
| + | |||
... | ... | ||
<br> | <br> | ||
Revision as of 17:19, 24 October 2010
{|
|
Biobricks
|
Parts Submitted to Registry
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Phytochrome Interacting Factor-3 (PIF3) - BBa_K365000
Background: PIF3 is a downstream transcription factor in a well studied signalling pathway of A. thaliana, upon stimulation with red (650 nm) light, it binds directly to PhyB and translocates to the nucleus as a heterodimer where it modulates the transcription of response genes. PIF3 binds only the red-light-exposed form of phytochrome, Pfr, and shows no-measurable binding affinity for the dark- or infrared-exposed Pr state.In our system target proteins are fused to PIF3 and tagged with the DAS degradation sequence which, through light activation, brings the degradation tag in proximity to ClpX. Conception: The light-sensitive interaction with PhyB has been mapped to the first 100-residue N-terminal activated phytochrome binding (APB) domain of PIF3 (Lim & Voigt, 2009).We chose this sequence, as it has already been successfully used in different synthetic in vitro applications that benefitted from its light-sensitive interactions with PhyB. The original sequence contains an XbaI restriction site. ... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Phytochrome Interacting Factor-6 (PIF6) - BBa_K365001
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Phytochrome B (aa 1-908) - BBa_K365002
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Phytochrome B (aa 1-642) - BBa_K365003
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
∆N-ClpX (aa 61-425) - BBa_K365004
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Linker (aa 20) - BBa_K365005
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
LAA tag - BBa_K365006
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
DAS tag - BBa_K365007
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Lambda tag - BBa_K365008
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
GFP (super fold) - BBa_K365009
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
PhyB642-(linker-∆NClpX)3 - BBa_K365010
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
PhyB908-(linker-∆NClpX)3 - BBa_K365011
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Full-length ClpX - BBa_K365012
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
∆NClpX-linker-∆NClpX-linker-∆NClpX - BBa_K365013
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
(linker-∆NClpX)3 - BBa_K365014
... |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
16
... |
