Integrated platform for the production and purification of recombinant proteins in plant systems

SMITH E; MARTÍNEZ CA; GIULIETTI AM; RODRÍGUEZ TALOU J

Mendoza

Congreso; XLVIII Reunion Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2012

Plant systems have emerged as safe, effective and inexpensive platforms for the production of recombinant proteins with biotechnological application. Low protein accumulation levels and the lack of efficient purification methods are major problems to be solved. Hydrophobins (HFB) are fungal proteins that alter the hydrophobicity of their fusions partners enabling efficient purification by surfactant-based aqueous two-phase system. Furthermore, they enhanced the accumulation of their fusion proteins. Dengue virus (DV) envelope protein (E) is the major structural component and the most immunogenic protein of the virus and it is involved in the induction of a protective immunity. The aim of this work is to produce E protein as a fusion with HFB in plant systems to be used for the potential production of a vaccine and for the generation of a diagnostic reagent for rapid detection of DV. A gene encoding DV serotype 2 E protein fused with HFB was successfully cloned in a binary vector and expressed in Nicotiana Benthamiana plants and cell suspensions via Agrobacterium tumefaciens-mediated transformation. GFP fused with HFB was used as a control. The results obtained indicate that fusion proteins are able to be expressed transiently in plants and stably in cell suspensions cultures of N. benthamiana. Currently, we are evaluating the integrity and the expression levels of these proteins.