congresos y reuniones científicas
Transient expression of subunit vaccine candidates against Canine Parvovirus and Canine Distemper Virus in Nicotiana tabaco.
LAGUIA BECHER MELINA; GALLO CALDERÓN MARINA; ROMANUTTI CARINA; ALVAREZ MARIA ALEJANDRA; MARCONI PATRICIA L
Congreso; 11th Congress of International Plant Molecular Biology; 2015
The use of plants for the production of recombinant antigens is gaining great interest as an alternative for the development of effective, safe and economical subunit vaccines. In particular, high yields of recombinant proteins can be obtained quickly with transient expression systems. In this work, we propose to transiently express vaccine candidates against Canine parvovirus (CPV) and Canine distemper virus (CDV) in Nicotiana tabacum. CPV and CDV are two common pathogens of domestic and wild carnivores that have a worldwide distribution. CPV is a non-enveloped single-stranded DNA virus with a capsid formed by three structural viral proteins (VP). VP2 is the most abundant protein of the capsid and exhibits good immunogenicity. On the other hand, CDV is an enveloped negative-strand RNA virus. Due to its immunogenic properties the nucleoprotein (NP) is considered a potential vaccine candidate. Full length VP2 and NP genes of local and vaccine strains were amplified using primers that include the Kozak sequence, the signal peptide 2S2 from Arabidopsis thaliana which delivers the antigen via the secretory pathway and the C-terminal KDEL sequence for retrieving the antigen to the endoplasmic reticulum. The amplified products were digested, purified and cloned into pENTR4 vector. Finally, these plasmids and the pK7WG2D or p35SGATFH binary vectors were recombined using the LR clonase enzyme. The obtained plant expression vectors were introduced into Agrobacterium tumefaciens bacteria and the T-DNA carrying the VP2 or NP gene was agroinfiltrated into tobacco leaves. The transient expression of the recombinant antigens will be analyze by western blot.