Cloning and expression of Junin virus, Candid#1 strain, proteins and reverse genetic system design.

BORIO, C.S.; ROMANO CHERÑAC, F.; ISERTE, J.A.; STEPHAN, B.I.; PILLOFF, M.G.; GHIRINGHELLI, P.D.; GOÑI, S.E.; LOZANO, M.E.

Mar del Plata, Buenos Aires

Congreso; XLIII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2007

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Arenaviridae comprises 22 recognized virus species with a bipartite ssRNA genome and an ambisense coding strategy. The virions are enveloped and include non-equimolar amounts of each genomic RNA species, designated L (ca. 7200 nt) and S (ca. 3500 nt), coding for four ORFs (N, GPC, L and Z). After expression, GPC protein is processed into three mature peptides (G1. G2 and a signal peptide). Junín virus is the ethiological agent of Argentine hemorrhagic fever (AHF), an acute disease with high mortality rate. In the ´80s, the Candid#1 strain od Junín virus (CD1) was developed as a live attenuated vaccine for AHF. Molecular characterization of CD1, and of its more virulent direct ancestors, XJ-13 and XJ#44, permits a systematic approach to study the basis of Junín virus virulence. We sequenced both genomic RNAs of CD1 and XJ#44 and compared them to XJ-13. Furthermore we analyzed the secondary structure predicted for the complete set of proteins coded by the viral genome. Our results revealed 15 amino acid changes among CD1 proteins and the wild-type ones. We are developing a reverse genetic system in order to test the attenuation hypothesis. We cloned and expressed the four Junín virus ORFs in different systems (bacterial and eukaryote), and generate a set of minigenomes containing Junin virus non-coding sequences and marker genes.