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

PABLO RAVASI AND HUGO MENZELLA

Estambul

Congreso; 15 European Congress of Biotechnology; 2012

European Federation of Biotechnology

Design and testing of a multi-species synthetic vector for tunable gene expression Pablo Ravasi and Hugo Menzella Genetic Engineering & Fermentation Technology. CONICET. Facultad de Ciencias Bioquimicas y Farmacéuticas. Universidad Nacional de Rosario. Suipacha 531 Rosario 2000. Republica Argentina. Phone: 54-9341-3304478. Abstract Synthetic biology is an emerging discipline that aims to crate novel organisms containing designed genetic circuits. These circuits are built from standard biological parts, known as biobricks, that in most of the cases are provided by nature. So far, the vast majority of synthetic biology tools have been developed for Escherichia coli. Here we aim to extend the principles by designing approaches for rapid engineering of other species for which available molecular tools are scarce. For this purpose, we have developed strategies to: (i) rapidly identify and characterize parts involved in gene expression regulation from microorganisms of biotechnological interest and (ii) create tools for the facile assembly of genetic circuits to manipulate pathways in such microorganisms. Our approach use a plasmid, where all parts are flanked by a standard set of restriction sites enabling the rapid testing regulatory sequences. The approach was validated for Corynebacterium glutamicum, a microorganism with great biotechnological interest. Only a few characterized vectors are available for this biotechnology work horse. The design of standard tools for exploring the potential of novel parts to regulate gene expression and for the assembly of genetic circuits for metabolic engineering may allow for the improvement of the capacity of C. glutamicum as a producer of valuable compounds with a consequent economic impact. Additionally, we demonstrate that our approach can be extended to multiple bacterial species by using a shuttle vector with an E. coli origin of replication, and the second origin of replication contained within an exchangeable cassette.