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From 2011.igem.org

Tools for Pathway Engineering

The modular nature of synthetic biology makes it ideal for pathway engineering for the purpose of compound manufacture. However there are a number of factors to consider when synthesizing compounds, chief of which is the precise control of protein expression levels.

Expression of all genes within the pathway at the optimum level is key. If one of the enzymes in this pathway is rate limiting, then a buildup of intermediate products can be extremely undesirable for the cell. Not only is it a waste of resources, but the compound may be cytotoxic.

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.