Dengue protein production in plant using Agrobacterium-mediated and biolistic transformation

MARTÍNEZ, CA; PERASSOLO, M; CARDILLO, AB; BUSTO, VD; QUEVEDO, CV; GIULIETTI, AM; RODRÍGUEZ TALOU, JULIÁN

Carlos Paz, Córdoba, Argentina

Congreso; XLIV Reunión Anual de la SAIB; 2008

Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB)

Production of recombinant proteins in plant systems has emerged as a new alternative platform. Plant cells can be maintained in simple and economic media, without risk of contamination with bacterial toxins, virus or prions. Moreover, the plant cells can glycosilate and do the post-transcriptional arrangements need for complex glycoproteins. The envelope protein (E) is the major structural component and the most immunogenic of dengue virus proteins and it is involved in the induction of a protective immunity. E protein needs to be directed to the secretory pathway through the N-terminus signal peptide (SP), to be N-glycosilated. The presence of an ER retention signal in the C-terminus of some proteins increases their stability and the yield of recombinant protein in plant systems.                           The aim of this work is to produce E protein in a plant system to serve as a diagnostic reagent in the rapid detection of dengue virus. A gene encoding dengue type 2 E protein was successfully cloned in a binary vector and expressed in N. tabacum plant using Biolistic and Agrobacterium-mediated expression systems. The result indicates that expression systems used can produce the dengue virus antigen in plant. We are currently evaluating the integrity and the expressions level of this protein. The effect of different genetic sequences on the expression levels of the E protein will be assessed.