Internal ribosome entry site (IRES) from Encephalomyocarditis virus (EMCV) as a tool for shuttle expression plasmids

Abstract

In eukaryotes, IRES sequences aid the recruitment of factors needed for translation to occur, enabling protein production independent of 5′ capped mRNA. Many patents and commercially available plasmids exploit their properties for polycistronic expression of recombinant proteins. However, these applications have been restricted to eukaryotic organisms, since it was thought that elements of this origin were essential for their activity. Here, using two tricistronic vectors designed for expression in mammalian hosts, we present evidence of EMCV IRES activity in prokaryotes. This finding enables the development of new and more versatile plasmid vectors for the production of recombinant proteins in multiple hosts from a single construct. Additionally, it provides new hints for the elaboration of alternative models describing the molecular mechanism of EMCV IRES mediated translation, in the absence of eukaryotic elements that were considered indispensable for its function.