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      <h1><strong>Oxygen Control System: PrrA/PrrB  two component system </strong></h1>
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      <p>This regulatory  system can sense oxygen concentration and senda response, under high oxygen tension, the system remains  inactive, when oxygen concentration decreases PrrB (His sensor kinase) turns active  trough an autophosphorylation with help of PrrC, which transmit the signal from  electron transport chain with help. Then PrrB transmit the phosphate group to  PrrA response regulator that directly binds promoter and recruits RNA  polymerase to start GFP transcription.</p>
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      <p><strong>How does it work?</strong></p>
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      <p>This genetic system  consists in two modules, the first one is formed by <strong><em>prra, prrb </em></strong><em>and<strong> prrc</strong></em>coupled to <strong>B0030</strong> RBS  units, <strong>J23104</strong> Strong constitutive  promoter and <strong>B0014</strong> Double terminator  to form the transcription unit. We will evaluate the functioning of J23104,  B0030 and B0014 in a new chassis. The second module consists in <strong>J54103</strong> GFP generator (B0030 RBS + E1010  GFP + B0014 Double terminator) coupled to PpsR repressible promoter.</p>
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      <p>&nbsp;</p>
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      <p><img src="https://static.igem.org/mediawiki/2012/9/93/Oxi.JPG" width="636" height="231" /></p>
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<h2>&nbsp;</h2>
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<p center><stronge>Inspired in…</Stronge>
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This system is inspired in PrrBCA two component system from Rhodobacter sphaeroides, which is a master regulator that is involved in expression of approximately 850 genes, >20% of the genome (Kaplan & Eraso 2005) This system coordinately controls genes involved in the complex switch between aerobic and anaerobic lifestyles and the optimum use of reducing power. It also regulates expression of genes involved in photosynthesis, carbon dioxide fixation, nitrogen fixation, hydrogen uptake, aerotaxis, denitrification, electron transport, aerobic and anaerobic respiration, and heme biosynthesis, among others, thus emphasizing its global role (Elsen et.al 2004, Kaplan & Eraso 2005, Zeilstra-Ryalls & Kaplan 2004).
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    Kaplan S, Eraso J, Roh JH. (2005). Interacting regulatory networks in the facultative
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photosynthetic bacterium, Rhodobacter sphaeroides 2.4.1. Biochem. Soc. Trans. 33:51–55
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    Zeilstra-Ryalls JH, Kaplan S. (2004). Oxygen intervention in the regulation of gene xpression: the photosynthetic bacterial paradigm. Cell. Mol. Life Sci. 61:417–36
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    Eraso JM, Kaplan S (2009) Regulation of gene expression by PrrA in Rhodobacter sphaeroides 2.4.1: role of polyamines and DNA topology. J Bacteriol 2009, 191(13):4341-4352.
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<h1><em>Oxygen Control System: PrrA/PrrB two <br /><br />component regulation system! </em></h1>
 +
<p>This regulatory system is able to sense oxygen concentration and send a response, under high oxygen tension, the system remains inactive, when oxygen concentration decreases PrrB (Histidine sensor kinase) turns active through an autophosphorylation with help of PrrC, which receive the signal from electron transport chain. Then PrrB transfer a phosphate group to PrrA that directly binds promoter and recruits RNA polymerase to start GFP transcription.
 +
 
 +
 
 +
(See the next video for a visual explanation).</p>
 +
<div align="center"><iframe width="480" height="360" src="http://www.youtube.com/embed/pgp6DzpNyA0" frameborder="0" allowfullscreen></iframe></div><br>
 +
<p id="text2">Our biobricks </p>
 +
<p>The first biobrick consists in the complete oxygen dependent system, PrrA, PrrB and PrrC, each one with a Ribosome Binding Site, under a
 +
Medium strength promoter (J23104), this first biobrick also it has the PrrA dependent promoter and GFP as a reporter gene</p>
 +
     <img src="https://static.igem.org/mediawiki/2012/4/45/Rodo04.jpg" alt="rodo04" width="560" height="116"><br>
 +
<p><br>
 +
  The second biobrick is just the PrrA dependent promoter and GFP as a reporter gene.</p>
 +
<p align="center"><img src="https://static.igem.org/mediawiki/2012/1/16/Rodo05.jpg" alt="rodo05" width="450" height="249"></p>
 +
<p id="text2">We were inspired in:</p>
 +
<p>This system is inspired in PrrBCA two component system from <em>R. sphaeroides</em>, which is a master regulator involved in expression of
 +
approximately 850 genes, >20% of the genome (Kaplan & Eraso 2005) This system coordinately controls genes involved in the complex switch
 +
between aerobic and anaerobic conditions and the optimum use of reducing power. It also regulates gene expression involved in
 +
photosynthesis, carbon dioxide fixation, nitrogen fixation, hydrogen uptake, aerotaxis, denitrification, electron transport, aerobic and anaerobic
 +
respiration, and heme biosynthesis, and others. Thus emphasizing its global role (Elsen et.al 2004, Kaplan & Eraso 2005, Zeilstra-Ryalls &
 +
Kaplan 2004).</p>
 +
<hr>
 +
<p id="refe">References<br>
 +
1.
 +
Kaplan S, Eraso J, Roh JH. (2005). <strong>Interacting regulatory networks in the facultative
 +
photosynthetic bacterium, <em>Rhodobacter sphaeroides</em> 2.4.1.</strong> Biochem. Soc. Trans. 33:51–55<br>
 +
 
 +
2.
 +
Zeilstra-Ryalls JH, Kaplan S. (2004). <strong>Oxygen intervention in the regulation of gene xpression: the photosynthetic
 +
bacterial paradigm.</strong> Cell. Mol. Life Sci. 61:417–36<br>
 +
 
 +
3.
 +
Eraso JM, Kaplan S (2009) <strong>Regulation of gene expression by PrrA in <em>Rhodobacter sphaeroides</em> 2.4.1: role of
 +
polyamines and DNA topology.</strong> J Bacteriol 2009, 191(13):4341-4352.
 +
</p>
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Latest revision as of 00:22, 27 October 2012

Rho

Oxygen Control System: PrrA/PrrB two

component regulation system!

This regulatory system is able to sense oxygen concentration and send a response, under high oxygen tension, the system remains inactive, when oxygen concentration decreases PrrB (Histidine sensor kinase) turns active through an autophosphorylation with help of PrrC, which receive the signal from electron transport chain. Then PrrB transfer a phosphate group to PrrA that directly binds promoter and recruits RNA polymerase to start GFP transcription. (See the next video for a visual explanation).


Our biobricks

The first biobrick consists in the complete oxygen dependent system, PrrA, PrrB and PrrC, each one with a Ribosome Binding Site, under a Medium strength promoter (J23104), this first biobrick also it has the PrrA dependent promoter and GFP as a reporter gene

rodo04


The second biobrick is just the PrrA dependent promoter and GFP as a reporter gene.

rodo05

We were inspired in:

This system is inspired in PrrBCA two component system from R. sphaeroides, which is a master regulator involved in expression of approximately 850 genes, >20% of the genome (Kaplan & Eraso 2005) This system coordinately controls genes involved in the complex switch between aerobic and anaerobic conditions and the optimum use of reducing power. It also regulates gene expression involved in photosynthesis, carbon dioxide fixation, nitrogen fixation, hydrogen uptake, aerotaxis, denitrification, electron transport, aerobic and anaerobic respiration, and heme biosynthesis, and others. Thus emphasizing its global role (Elsen et.al 2004, Kaplan & Eraso 2005, Zeilstra-Ryalls & Kaplan 2004).

References
1. Kaplan S, Eraso J, Roh JH. (2005). Interacting regulatory networks in the facultative photosynthetic bacterium, Rhodobacter sphaeroides 2.4.1. Biochem. Soc. Trans. 33:51–55
2. Zeilstra-Ryalls JH, Kaplan S. (2004). Oxygen intervention in the regulation of gene xpression: the photosynthetic bacterial paradigm. Cell. Mol. Life Sci. 61:417–36
3. Eraso JM, Kaplan S (2009) Regulation of gene expression by PrrA in Rhodobacter sphaeroides 2.4.1: role of polyamines and DNA topology. J Bacteriol 2009, 191(13):4341-4352.

 

Rhodofactory 2012

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