Team:Valencia Biocampus/TheSeat

From 2014.igem.org

The Seat of the ST²OOL

By checking how standard, stable, orthogonal or patentable Biobricks are we have performed an analysis of four engineering pillars. But we wanted to combine these pillars, these four legs of our ST2OOL, with a truly synthetic and constructive approach.

In The Seat of the ST2OOL we are developing a functional metagenomic strategy in which we aim at isolating promoter sequences from natural environments. We will then characterize these sequences in order to select those with the highest stability, standard and orthogonal behaviors. We are working with soil and compost samples, from which we will isolate total metagenomic DNA, which will then be digested, ligated into an expression vector and used to built a library in E. coli. Transformants –clones with a promoter-like natural sequence- will be subjected to a range of stresses and subtransformation steps to find out their stability and standard/orthogonal behavior. To the best of our knowledge, this is the first Synthetic Biology functional metagenomic approach aiming at selecting biological parts not because of their “strength” but because of their compatibility with other parts and suitability for engineering purposes.

Finally, one technical –yet important- detail: we are not using neither commercial kits nor manual protocols for isolating metagenomic DNA. We are building our own robot…

Robot

Our robot design