Expression of a fusion protein, Z (Junin virus)-N (measles), for the generation of recombinant VLPS

TEMPRANA, C.F.; CALVENTE, N.I.; ARGÜELLES, M.H.; ESTEBAN, L.E.; CASTELLO, A.A.; LOZANO, M.E.; GLIKMANN, G.

Ciudad Autónoma de Buenos Aires

Congreso; XLIX Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2013

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

In the last decades, virus like particles (VLPs) have been widely studied for the development of new vaccines, diagnostic kits and nanocarriers. Due to their importance, researchers have focused their study on new proteins, able to induce VLPs formation. It has been previously described that VLPs can be formed when a fusion protein Z (from Junín virus)-GFP is expressed in 293T cells.This work is mainly focused on the study of Junín virus Z protein as a tool to generate quimeric VLPs carrying heterologous viral antigens. In line with this idea a C-terminal fragment of the measles' nucleoprotein (NCT) was cloned in frame with both Z and GFP. The plasmid containing the fusion of the three ORFs (pZ-NCT-GFP) was transfected into Cos-7 cells. Expression was studied through flow cytometry and fluorescence microscopy. Moreover, Western blot analysis was used to comfirm protein identity.Transfection was optimized, and a maximum of 65% efficiency was obtained. The fusion protein was found in these cell lysates as it was detected by anti-Z and anti-GFP antibodies.Furthermore, Z-NCT-GFP was also detected in a clarified transfected cells' supernatant, after concentration by ultracentrifugation through a sucrose cushion. In conclusion, these results suggest that Z-protein induced VLPs can be used as an antigen carrier system