R-BphP

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(The BphP / PpsR system)
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== The BphP / PpsR system ==
 
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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.
 
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PpsR2 is a transcriptional repressor, regulated by BphP.
 
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PpsR1 and PpsR2 bind to TGTN_12ACA possibly arranged in tandem with a 7 base spacer. The affinity for both PpsRs are around 100nM.
 
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BphP (or BrBphP for the Bradyrhizobium variant) is sensitive to far-red light (~770nm) and controls PpsR1.
 
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==To find==
 
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- Define genetic circuit <b><font size=3><span style="color:green"> Tú </span></font size></b>
 
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<br> - Look for exact wavelength for the corresponding light receptor<font size=3><span style="color:Blue"> Basile</span></font size>
 
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<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>
 
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==Implementation==
 
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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).
 
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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. 
 
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==To do==
 
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* contact Eric Giraud (giraud@mpl.ird.fr) and ask if he could send us the following material:
 
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** Bradyrhizobium ORS278
 
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** The following plasmids:
 
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*** pBAD::ppsR1
 
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*** pBAD::ppsR2
 
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*** pGEM-T::ppsR2/BrbphP
 
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** the PpsR1-C429S construct
 
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==Found==
 
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- Bacterium: Bradyrhizobium  can be ordered on  [http://www.lgcstandards-atcc.org/LGCAdvancedCatalogueSearch/ProductDescription/tabid/1068/Default.aspx?ATCCNum=10244&Template=bacteria ATCC]
 
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
 
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!align="center"|[[Team:EPF-Lausanne|Home]]
 
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!align="center"|[[Team:EPF-Lausanne/Team|The Team]]
 
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!align="center"|[[Team:EPF-Lausanne/Project|The Project]]
 
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!align="center"|[[Team:EPF-Lausanne/Parts|Parts Submitted to the Registry]]
 
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!align="center"|[[Team:EPF-Lausanne/Modeling|Modeling]]
 
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!align="center"|[[Team:EPF-Lausanne/Notebook|Notebook]]
 
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!align="center"|[[Team:EPF-Lausanne/Lectures|Lectures]]
 
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!align="center"|[[Team:EPF-Lausanne/Team Management|Team Management]]
 
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!align="center"|[[Team:EPF-Lausanne/References|References]]
 
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|}
 

Latest revision as of 20:21, 10 July 2009

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