Team:NTU-Taida/Project/Target

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Project Target Molecules
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==Project Target Molecules==
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Quorum sensing signals can be transmitted through several types of molecules. In our project, we use Rhl, Las and PQS quorum sensing signals as our target. Both Rhl and Las systems use acyl homoserine lactones (AHLs) as quorum sensing targets, while PQS uses 2-heptyl-3-hydroxy-4-quinolone as its QS target.  
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Quorum sensing (QS) signals can be transmitted through several types of molecules. In our project, we used Rhl, Las and PQS quorum sensing signals as our target. Both Rhl and Las systems use acyl homoserine lactones (AHLs) as quorum sensing targets, while PQS uses 2-heptyl-3-hydroxy-4-quinolone as its QS target.  
[[File:NTU-Taida-target-1.jpg|600px|thumb|center|Figure.1]]
[[File:NTU-Taida-target-1.jpg|600px|thumb|center|Figure.1]]
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==Acyl-homoserine lactone==
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==Acyl-homoserine lactone and quorum sensing transcriptional regulator==
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Environmentally, when bacterial concentration is low, quorum sensing related genes are expressed at minimum level. As bacterial density increases, AHL concentration also increases and therefore binds with their target receptors which act as transcriptional regulators to enhance target gene expression. In gram negative bacteria, we use AHLsIn our project, we constitutively express these AHL receptors(such as RhlR and LasR), and add the promoter sequences of these receptors in front of fluorescent reporters.
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For gram negative bacteria, Acyl-homoserine lactones (AHLs) are used as quorum sensing signals. AHL consists of a five-member homoserine lactone ring and an acyl side chain. The length and the substitution group of the side chain determine the specificity of AHL-receptor binding. On the other hand, the transcriptional regulator consists of an N-terminal ligand-binding domain (to bind AHL) and a C-terminal DNA binding domain.
[[File:NTU-Taida-target-2.jpg|700px|thumb|center|Figure.2]]
[[File:NTU-Taida-target-2.jpg|700px|thumb|center|Figure.2]]
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==AHL binding and quorum sensing transcriptional regulator==
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==Long Term Project: Quorum Sensing array==
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Rhl transcriptional regulator senses C-12 acyl homoserine lactone. The structure of the transcriptional regulator consists of an N-terminal ligand-binding domain and a C-terminal DNA binding domain. The length of the AHL acyl group plays a critical role in quorum sensing specificity. Las transcriptional regulator has similar characteristics.  
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Since the structure of the ligand-binding domain enables it to have certain interaction with AHLs that have different side chain length, and the interaction is still obvious when the length difference is in 2 carbons, a multi-QS system with their AHL of 4, 8, 12-C side chain could detect and identify AHL from 2~14 carbons; applying the idea of linear combination.
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What we have used, Rhl, Las and PQS are all original from ''Pseudomonas aeruginosa''. According to the fact that strains with different drug-resistance also have different combination of QS expression level, our system may not only distinguish bacteria between species but also within species.  
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Due to the fact that quorum sensing genes are highly related to bacterial resistant factors, we want to sense the expression of AHL of resistant bacteria and create novel data for further bacterial identification. Our final goal is to collect enough data to create a quorum sensing array that provides instant identification of bacterial species.
 
[[File:NTU-Taida-target-3.jpg|700px|thumb|center|Figure.3]]
[[File:NTU-Taida-target-3.jpg|700px|thumb|center|Figure.3]]
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==jjj==
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==Reference==
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{{:Team:NTU-Taida/Templates/Reference|Author=Ling EA, ''et al''|Essay=A novel plasmid for detection of N-acyl homoserine lactones|Journal=Elsevier}}
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==jjjp==
 
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Latest revision as of 03:28, 28 September 2013

Target



Contents

Project Target Molecules

Quorum sensing (QS) signals can be transmitted through several types of molecules. In our project, we used Rhl, Las and PQS quorum sensing signals as our target. Both Rhl and Las systems use acyl homoserine lactones (AHLs) as quorum sensing targets, while PQS uses 2-heptyl-3-hydroxy-4-quinolone as its QS target.

Figure.1

Acyl-homoserine lactone and quorum sensing transcriptional regulator

For gram negative bacteria, Acyl-homoserine lactones (AHLs) are used as quorum sensing signals. AHL consists of a five-member homoserine lactone ring and an acyl side chain. The length and the substitution group of the side chain determine the specificity of AHL-receptor binding. On the other hand, the transcriptional regulator consists of an N-terminal ligand-binding domain (to bind AHL) and a C-terminal DNA binding domain.

Figure.2

Long Term Project: Quorum Sensing array

Since the structure of the ligand-binding domain enables it to have certain interaction with AHLs that have different side chain length, and the interaction is still obvious when the length difference is in 2 carbons, a multi-QS system with their AHL of 4, 8, 12-C side chain could detect and identify AHL from 2~14 carbons; applying the idea of linear combination.

What we have used, Rhl, Las and PQS are all original from Pseudomonas aeruginosa. According to the fact that strains with different drug-resistance also have different combination of QS expression level, our system may not only distinguish bacteria between species but also within species.


Figure.3

Reference

Ling EA, et al. A novel plasmid for detection of N-acyl homoserine lactones, Elsevier.