Team:Groningen:Project:Detection/Pseudonomas

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  Detection of <i>Staphylococcus aureus</i>
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Detection of Pseudomonas aeruginosa
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  Quorum sensing pathway in Staphylococcus aureus
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Quorum sensing pathway in Pseudomonas aeruginosa
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  The quorum sensing system in <i>Staphylococcus aureus</i> consist of four genes (Figure 1). These genes are controlled by leaky inducible promoter (P2), and encode for two membrane proteins (AgrB and AgrC), one regulator protein (AgrA) and a precursor peptide (AgrD). The membrane protein AgrB cleaves the precursor peptide AgrD so the mature signaling peptide is formed. The mature signal peptide is called autoinducing peptide (AIP). After the AIP is secreted from the cell it is able to bind to the second membrane protein AgrC. After the binding the AgrC phosphorylates AgrA, AgrA is able to induce the P2 promoter which leads to increased production of the AIPs.  
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The Quorum sensing system of pseudomonas aeruginosa is a multiple hierarchically ordered complex. In one layer of this complex system, two proteins are synthesized, LasR and LasI. The LasI produces PAIs which can bind with the LasR. The LasR-PAI couple induces expression of virulence factors, LasA protease, alkaline protease and exotoxin A. Moreover, the couple acts as autoinducer on the expression of LasI by influencing the promoter pLasI, increasing the synthesis of PAIs.<sup>3</sup>. PAIs can move from cell to cell and thus influence the QS of other cells in their surrounding.
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</html>{{:Team:Groningen/Template/MODULE/newfigure|Figure 1|2/27/AGRBDCA_SYSTEM_in_staphylococcus_aureus.art.png|Quorum sensing system of <i>Staphylococcus aureus</i> The precursor peptide, AgrD (1), is modified to AIP by the membrane protein AgrB (2) and translocated over the membrane. The AIP binds to AgrC (3) which in turn phosphorylates AgrA (4) the phosphorylated ArgA induces the P2 promoter.}}<html>
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  How LactoAid detects <i>S. aureus</i>
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How LactoAid detects Pseudonomas aeruginosa
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  In our LactoAid want introduced the DNA encoding for the membrane protein AgrC and the regulator protein ArgA. When <i>Staphylococcus aureus</i> starts to produce the AIPs, the LactoAid should be able to sense the molecules via the same system the <i>S. aureus</i> uses. In figure 1 the designed biobrick for the detection of <i>S. aureus</i> is shown. Unfortunately we were not able to assemble the promoter in this BioBrick. We therefore did not send the part to the registry. After the AgrA is phosphorilated it will induce to the P2 promoter which is part of our secretion system. More about the secretion system can be found at the <a href="https://2014.igem.org/Team:Groningen:Project:Secretion">sectretion page</a> 
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Because the AHLs defuse from cell to cell we designed a biobrick which produces LasR. This will bind to the PAI and induce the pLasI promoter. Behind the pLasI promoter we placed our secretion system to produce the infection preventing molecules. (Figure 4).  
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The protein LasR is present in our LactoAid and thus, if the PAI-1 signal molecule is present it will bind to the LasR and it will induce the pLasI whereby the secretion against <i>Pseudonomas aeruginosa</i> is activated.
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</html>{{:Team:Groningen/Template/MODULE/newfigure|Figure 1|b/b2/Groningen_2014_THOMASStpahylococcusaureus4.png|<i>Staphylococcus aureus</i>         picture adopted from center for disease control and prevention}}<html>
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<i>Pseudonomas aeruginosa</i> picture adopted from center for disease control and prevention.
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Latest revision as of 02:27, 18 October 2014

Detection of Pseudomonas aeruginosa
Quorum sensing pathway in Pseudomonas aeruginosa
The Quorum sensing system of pseudomonas aeruginosa is a multiple hierarchically ordered complex. In one layer of this complex system, two proteins are synthesized, LasR and LasI. The LasI produces PAIs which can bind with the LasR. The LasR-PAI couple induces expression of virulence factors, LasA protease, alkaline protease and exotoxin A. Moreover, the couple acts as autoinducer on the expression of LasI by influencing the promoter pLasI, increasing the synthesis of PAIs.3. PAIs can move from cell to cell and thus influence the QS of other cells in their surrounding.
How LactoAid detects Pseudonomas aeruginosa
Because the AHLs defuse from cell to cell we designed a biobrick which produces LasR. This will bind to the PAI and induce the pLasI promoter. Behind the pLasI promoter we placed our secretion system to produce the infection preventing molecules. (Figure 4).
Figure 4
 
Figure 4: The protein LasR is present in our LactoAid and thus, if the PAI-1 signal molecule is present it will bind to the LasR and it will induce the pLasI whereby the secretion against Pseudonomas aeruginosa is activated.
 
 
Figure 5
 
Figure 5: Pseudonomas aeruginosa picture adopted from center for disease control and prevention.
 
 

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