Delivery of the C-terminal region of the rotavirus VP6 protein by Junín virus Z VLPS

BERGIER, J.A.; BORIO, C.S.; BENTANCOR, L.V.; BILEN, M.F.; LOZANO, M.E.

Mar del Plata

Congreso; LI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2015

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

The ability to boost the viral particle budding of viral matrix proteinshas been studied in several RNA viruses. For arenaviruses, the only expressionof the Z protein in mammalian cells allows the generation of virus-likeparticles (VLPs) containing this protein inside. In previous studies, we haveshown that the Junín Z protein, belonging to the Arenaviridae family, is ableto maintain its activity even when fused to a heterologous sequence at itsC-terminal region. We have also demonstrated the in vivo immunogenicityof these chimeric virus-like particles using GFP and pther viral antigens. Theaim of this study is to evaluate this antigen presentation system using theC-terminal region of the Rotavirus VP6 protein. We have cloned the C-terminalregion of the VP6 protein (332-397) in an eukaryotic expression vector fused tothe Z protein, pZ-CT-VP6. We have also made a reporter construction using GFP,which allow cellular localization studies in mammalian cells (pZ-CT-VP6-GFP).In all cases the expression of the cloned proteins was confirmed, and theformation of VLPs was analyzed. Before testing the immunogenicity of the CT-VP6VLPs, performed an immunization schedule using the Z-EGFP chimera. And fromthese results, we will select the best condition for the stimulation of theimmune response against CT-VP6.