Team:Glasgow/PathwayTools/Intro

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<html><h1>Tools for Pathway Engineering</h1>
<html><h1>Tools for Pathway Engineering</h1>
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<h6><a href="https://2011.igem.org/Team:Glasgow/Results">Back to Results</a></h6>
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It is difficult to tune protein expression levels within pathway. <br/>
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The modular nature of synthetic biology makes it ideal for pathway engineering for the purpose of compound production.  This means that engineered microorganisms can be used to manufacture a variety of useful chemicals, including pharmaceuticals and drug intermediates (e.g. Opiates), fine chemicals and biofuels. <br/><Br/>
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<center><img src=https://static.igem.org/mediawiki/2011/f/fe/Glasgowpathway.png></center><br/>
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There are many possible reasons why protein expression level can vary along a pathway.  For example, if enzyme 2 were rate limiting, then the intermediate product B would build up within the cell. This is undesirable and a waste of resources.<br/>
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In order to optimise this, the carbon flux needs to be rapid.  To do this, you would need to be able to control the levels of each enzyme in the system.<Br/><br/>
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This is why we have created a library of promoters with an RBS. This allows expression levels of enzyme to be readily controlled at each step in the pathway. <Br/>
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We have also created a Multiple Cloning site biobrick, which is ideal for use with our library of promoters.<Br/><Br/>
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These biobricks represent an invaluable set of tools for anyone working on expression levels in pathway engineering.<br/><br/>
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When engineering a pathway, there are many considerations such as correctly inserting the pathway genes into your chosen host organism. However one of the most important considerations is the precise control of protein expression levels in order to control the carbon flux in the system.<Br/><br/>
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If the protein is over expressing, it can have detrimental effects on the levels of cellular resources (such as levels of cofactors)and may even be cytotoxic.  Overall yield is also affected by variance in levels of gene expression.  This is why it is essential to ensure that gene expression over the whole pathway is optimal for the system. <br/><br/>
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One way of controlling the flux through pathways is to engineer the strength of promoters and ribosome binding sites (RBSs) to regulate gene expression. These can be used in a modular combinatorial approach to create diverse variations of the pathway of interest.
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To this end we have created a library of combined promoters and RBS of different strengths in biobrick format which can be used to create and test different expression levels of individual metabolic pathway genes for optimum product formation.<br/><Br/>
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We have also created a Multiple Cloning site biobrick. It contains a double terminator and five restriction sites, so is ideal for use with our library of promoters.  The aim of this biobrick is to significantly simplify the testing of genes by reducing the number of overall ligations that must be performed.<br/><br/>
To read more about the creation of the Promoter + RBS library, click <a href=https://2011.igem.org/Team:Glasgow/Results/PromoterLibrary>here.</a><br/>
To read more about the creation of the Promoter + RBS library, click <a href=https://2011.igem.org/Team:Glasgow/Results/PromoterLibrary>here.</a><br/>
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To read more about the creation of the Multiple Cloning Site, click <a href=https://2011.igem.org/Team:Glasgow/Results/MCS>here.</a>
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To read more about the creation of the Multiple Cloning Site, click <a href=https://2011.igem.org/Team:Glasgow/Results/MCS>here.</a><Br/>
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To read more about potential modular pathways, click <a href=https://2011.igem.org/Team:Glasgow/PathwayTools/Pathways>here.</a><br/>

Latest revision as of 04:53, 22 September 2011

Tools for Pathway Engineering

Back to Results
The modular nature of synthetic biology makes it ideal for pathway engineering for the purpose of compound production. This means that engineered microorganisms can be used to manufacture a variety of useful chemicals, including pharmaceuticals and drug intermediates (e.g. Opiates), fine chemicals and biofuels.

When engineering a pathway, there are many considerations such as correctly inserting the pathway genes into your chosen host organism. However one of the most important considerations is the precise control of protein expression levels in order to control the carbon flux in the system.

If the protein is over expressing, it can have detrimental effects on the levels of cellular resources (such as levels of cofactors)and may even be cytotoxic. Overall yield is also affected by variance in levels of gene expression. This is why it is essential to ensure that gene expression over the whole pathway is optimal for the system.

One way of controlling the flux through pathways is to engineer the strength of promoters and ribosome binding sites (RBSs) to regulate gene expression. These can be used in a modular combinatorial approach to create diverse variations of the pathway of interest. To this end we have created a library of combined promoters and RBS of different strengths in biobrick format which can be used to create and test different expression levels of individual metabolic pathway genes for optimum product formation.

We have also created a Multiple Cloning site biobrick. It contains a double terminator and five restriction sites, so is ideal for use with our library of promoters. The aim of this biobrick is to significantly simplify the testing of genes by reducing the number of overall ligations that must be performed.

To read more about the creation of the Promoter + RBS library, click here.
To read more about the creation of the Multiple Cloning Site, click here.
To read more about potential modular pathways, click here.

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