|
|
| (30 intermediate revisions not shown) |
| Line 1: |
Line 1: |
| - | == The BphP / PpsR system ==
| |
| | | | |
| - | PpsR1 is a redox sensitive activator. It binds DNA under anaerobic conditions, and forms a tetramer via a disulfide bond. This interaction is ablated in the mutant PpsR1-C429S; meaning that we should be able to mimic an anaerobic system even under aerobic conditions.
| |
| - |
| |
| - | PpsR2 is a transcriptional repressor, regulated by BphP.
| |
| - |
| |
| - | PpsR1 and PpsR2 bind to TGTN_12ACA possibly arranged in tandem with a 7 base spacer. The affinity for both PpsRs are around 100nM.
| |
| - |
| |
| - | BphP (or BrBphP for the Brayrhizobium variant) is sensitive to far-red light (~770nm) and controls PpsR1.
| |
| - |
| |
| - | ==To find==
| |
| - | - Define genetic circuit <b><font size=3><span style="color:green"> Tú </span></font size></b>
| |
| - | <br> - Look for exact wavelength for the corresponding light receptor<font size=3><span style="color:Blue"> Basile</span></font size>
| |
| - | <br> - Sequences of the genes involved in the pathway (minimal genes to => [http://www.ncbi.nlm.nih.gov/ Pubmed])<b><font size=3><span style="color:green"> Tú </span></font size></b>
| |
| - |
| |
| - | ==Implementation==
| |
| - |
| |
| - | There are two modes in which we can use the system. The minimal complement requires PpsR2 and its regulator BphP. We can generate a hybrid system that uses a well known activator or transcription or a constitutively active promoter which will be repressed via PpsR2 upon exposure to far-red light (770nm).
| |
| - |
| |
| - | It is also possible to reconstitute the entire system including PpsR1-C429S, which will serve as the activator and will be de-coupled from the oxidative state of the cell due to the ablation of the disulfide bond formation.
| |
| - |
| |
| - | ==To do==
| |
| - | * contact Eric Giraud (giraud@mpl.ird.fr) and ask if he could send us the following material:
| |
| - | ** Bradyrhizobium ORS278
| |
| - | ** The following plasmids:
| |
| - | *** pBAD::ppsR1
| |
| - | *** pBAD::ppsR2
| |
| - | ** the PpsR1-C429S construct
| |
| - |
| |
| - | ==Found==
| |
| - | - Bacterium: Bradyrhizobium can be ordered on [http://www.lgcstandards-atcc.org/LGCAdvancedCatalogueSearch/ProductDescription/tabid/1068/Default.aspx?ATCCNum=10244&Template=bacteria ATTC]
| |
| - | ----
| |
| - | {| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
| |
| - | !align="center"|[[Team:EPF-Lausanne|Home]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Team|The Team]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Project|The Project]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Parts|Parts Submitted to the Registry]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Modeling|Modeling]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Notebook|Notebook]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Lectures|Lectures]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/Team Management|Team Management]]
| |
| - | !align="center"|[[Team:EPF-Lausanne/References|References]]
| |
| - | |}
| |
"
