Team:EPF-Lausanne/R-BphP

From 2009.igem.org

(Difference between revisions)
(New page: {{EPF-Lausanne09}} <div CLASS="epfltrick">__TOC__ </div><div CLASS="epfl09"> == The BphP / PpsR system == PpsR1 is a redox sensitive activator. It binds DNA under anaerobic conditions, a...)
 
(19 intermediate revisions not shown)
Line 1: Line 1:
{{EPF-Lausanne09}}
{{EPF-Lausanne09}}
 +
<div CLASS="epfltrick">__TOC__
<div CLASS="epfltrick">__TOC__
</div><div CLASS="epfl09">
</div><div CLASS="epfl09">
-
== 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.
+
<html><center>
 +
<font size="6" color="#007CBC"><i>R-BphP project </i></font>
 +
</center></html>
 +
<br>
 +
----
 +
<br>
 +
<br>
-
PpsR2 is a transcriptional repressor, regulated by BphP.
+
This part of the project was cancelled, due to lack of time. You can read what our idea was on [https://2009.igem.org/Team:EPF-Lausanne/Brainstorming the brainstorming session]
-
 
+
-
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 Bradyrhizobium variant) is sensitive to far-red light (~770nm) and controls PpsR1.
+
-
 
+
-
For more info, see [http://www.jbc.org/cgi/content/full/279/43/44407].
+
-
 
+
-
Similar constituents could possibly be derived from R. Palustris as well with the following model derived from [http://www3.interscience.wiley.com/cgi-bin/fulltext/118512333/HTMLSTART]:
+
-
[[Image:R._palustris_circuit.gif]]
+
-
 
+
-
==To find==
+
-
- Define genetic circuit <big>'''Tú'''</big>
+
-
<br> - Look for exact wavelength for the corresponding light receptor<big><span style="color:Blue"> Basile</span></big>
+
-
<br> - Sequences of the genes involved in the pathway (minimal genes to  => [http://www.ncbi.nlm.nih.gov/ Pubmed])
+
-
<br>[[Media:‎ ORS278_BrBphP_genomic_seq.txt|ORS278 BrBphP genomic sequence]]
+
-
<br>[[Media:‎ ORS278_PpsR1_genomic_seq.txt|ORS278 PpsR1 genomic sequence]]
+
-
<br>[[Media:‎ ORS278_PpsR2_genomic_seq.txt|ORS278 PpsR2 genomic sequence]]
+
-
<br>[[Media:‎ ORS278_PpsR1-C429S_protein_seq.txt|ORS278 PpsR2-C429S protein sequence]]
+
-
 
+
-
==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
+
-
*** pGEM-T::ppsR2/BrbphP
+
-
** 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 ATCC]
+
-
----
+
-
{| 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]]
+
-
|}
+
</div><div CLASS="epfl09bouchon"></div>
</div><div CLASS="epfl09bouchon"></div>

Latest revision as of 08:07, 11 October 2009




R-BphP project



This part of the project was cancelled, due to lack of time. You can read what our idea was on the brainstorming session

Retrieved from "http://2009.igem.org/Team:EPF-Lausanne/R-BphP"