EXPLORING DIFFERENT STRATEGIES TO EXPRESS DENGUE VIRUS ENVELOPE PROTEIN IN A PLANT SYSTEM

MARTÍNEZ, CA; GIULIETTI, AM; RODRIGUEZ TALOU, J; MASON, H

San Miguel de Tucuman

Congreso; XLV Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2009

SAIB

The utilization of plants as bioreactors presents several advantages related with its simplicity, biosafety and low cost. It retains the advantages of eukaryotic expression systems, as post-translational modifications, and prokaryotic expression systems, as scalability and economical production. However, long time-scale and low yield of recombinant proteins have been the main bottlenecks in plant-made biopharmaceuticals. A recently developed technique known as Magnifection can offset these disadvantages. This transient expression technology is based on replication of viral vectors delivered to the plant by Agrobacterium and allows very fast production, high recombinant protein expression levels. Dengue virus (DV) serotype 2 envelope glycoprotein (E) is the main protein associated with viral entry and the induction of immunity. In order to produce a candidate for subunit vaccines and to provide an antigen for diagnostic kits, E protein was expressed using deconstructed viral modules. An E truncated version was designed to be expressed alone and co-expressed with DV structural proteins. As well, the critical Domain III of E protein was fused to Hepatitis B core antigen. The recombinant proteins were produced successfully in Nicotiana benthamiana plants and were reactive with anti-E polyclonal antibody and the fusion was reactive with anti-E polyclonal and anti-HBcore antibodies.