Design and testing of a multi-species synthetic vector for tunable gene expression

PABLO RAVASI; SALVADOR PEIRU; HUGO G MENZELLA

Estambul

Congreso; 15th EUROPEAN CONGRESS on BIOTECHNOLOGY ?bio-crossroads?; 2012

European Federation of Biotechnology

Synthetic biology approaches can make a significant contribution to the advance of metabolic engineering by reducing the development time of recombinant organisms. However, the vast majority of synthetic biology tools for genetic engineering have been developed for Escherichia coli. We aim to extend the principles by designing approaches for rapid engineering of other species. As a proof of concept, we have undertaken an endeavor to create tools for the facile assembly of genetic circuits to manipulate pathways in Corynebacterium glutamicum, a microorganism widely used in biotechnology. Here, we have designed and created a synthetic plasmid, where all parts are flanked by a standard set of unique restriction sites enabling the rapid testing of regulatory sequences. We are currently using this tool for exploring the potential of novel parts to regulate gene expression, and for the assembly of genetic circuits for metabolic engineering. Using this strategy, we anticipate to rapidly populate a collection of standard parts for C. glutamicum including: promoters, ribosome binding sites and transcriptional terminators.