"

From 2011.igem.org

Tools for Pathway Engineering

The modular nature of synthetic biology makes it ideal for pathway engineering and chemical manufacture. However one of the biggest concerns with this is fine tuning protein expression level.

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. 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. 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.
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.

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.
We have also created a Multiple Cloning site biobrick, which is ideal for use with our library of promoters.

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.

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.