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High level expression of nucleocapsid protein from Canine Distemper Virus in tobacco transplastomic plants
LAGUIA BECHER MELINA; GALLO CALDERÓN MARINA; ROMANUTTI CARINA; TRENTINI A ; ALVAREZ MARIA ALEJANDRA; VERAMENDI J; MARCONI PATRICIA L
Congreso; International Congress on Plant Science and Molecular Biology; 2018
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Canine Distemper Virus (CDV) produces a highly contagious disease in domestic dogs and other wild animals. Although immunization with attenuated CDV vaccines has been shown to be effective, in recent years there has been an increase in the incidence of CDV infection, including vaccinated dogs. Therefore, a more effective vaccine formulation containing updated immunogens is necessary. The use of plants as biofactories for the expression of antigens is an attractive alternative to produce cost-effective subunits vaccines. In particular, chloroplast transplastomic technology offers stable high-level of recombinant protein expression and transgene containment. In this study, we evaluated the production of the nucleocapsid protein (NP) from an Argentinian CDV strain in transplastomic tobacco plants. Protein expression analysis by Western blot showed the presence of two specific bands in total protein extracts. The expected band of 62 kDa for NP represented approximately 60% of the signal in mature leaves. Furthermore, full-length NP was expressed mainly as an insoluble protein. The concentration of NP in young, mature and old leaves was estimated to be 6.2, 4.4 and 6.7 % of total protein, equivalent to 1.2, 0.7 and 0.4 mg/g fresh weight, respectively. We also evaluated the functionality of the plant-derived NP in a murine model. Oral and subcutaneous immunization with transplastomic extracts elicited an increase of NP-specific serum antibodies compared with control mice. Our results show that NP can be efficiently expressed in transplastomic tobacco plants and maintains its immunogenic properties.