INVESTIGADORES
RODRIGUEZ TALOU Julian
congresos y reuniones científicas
Título:
Efficient production of a kdel-tagged Dengue virus protein in plant cell suspension cultures.
Autor/es:
MARTÍNEZ, CA; GIULIETTI, AM; RODRIGUEZ TALOU, J
Lugar:
Puerto Madryn, Argentina
Reunión:
Congreso; XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010
Institución organizadora:
SAIB
Resumen:
Dengue virus envelope glycoprotein (DV-E) is the antigen associated with immunity induction and it is an effective candidate for the development of a subunit vaccine and a promising antigen for diagnostic kits. As a part of a project to develop a plant-made dengue virus vaccine, we explored the ability of plant cells to produce DV serotype 2 (DV-2) E protein in Nicotiana tabacum and Morinda citrifolia cell suspension cultures. DV-E cDNA was cloned with a signal peptide at its 5’ end and with and without the addition of KDEL endoplasmic retention sequence at its 3’ end to analyze its influence in recombinant protein accumulation levels. The expression cassette was sub-cloned into pCAMBIA 1305.2 binary vector and the cell suspension culture transformation was carried out using A. tumefaciens LBA4404. The maximum accumulation levels (0,71 ± 0,06 mg DV-E/L) were obtained by tobacco cells at 5 days of culture when KDEL tetrapeptide was fused. It represents 0,3% of the total soluble protein. Its integrity was confirmed by western blot. The recombinant protein was reactive with anti-E monoclonal and polyclonal antibodies. Our results demonstrate for the first time that plant cell in vitro cultures represents a low cost expression system suitable for the production of recombinant DV-E protein in which biosafety conditions are guaranteed.