Cloning and Expression of TrV Viral Proteins

SANCHEZ- EUGENIA, R; SANCHEZ, L; AGUIRRE, J; GOIKOLEA, J; MARTI G. A; FERRER-ORTA, C; VERDAGUER, N; VIGUERA A.R; GUERIN, D.M.A.

Workshop; II International Workshop on Chagas Disease, triatomine vectors, Trypanosoma cruzi, and Triatoma virus.; 2012

Triatoma virus (TrV) is a small spherical, non-enveloped, +ssRNA picorna-like virus that infects gut cells of Triatoma infestans (Muscio et al., 1987. J. Invertebr. Pathol. 9:218-220). TrV belongs to Dicistroviridae family and its crystallographic structure (RCSB PDB code 3NAP) shows an icosahedral T=1 (P=3) pseudo T-3 symmetry, which is very similar to the structure of Cricket Paralysis virus. Natural infective TrV particles are composed of 60 copies of the capsid proteins VP1-4, enclosing a bipartite genome of 9010 nt. (Cziebener et al., 2000. J. Gen. Virol. 81:1149- 1154). In this communication we summarize the current advances in the cloning and expression of TrV structural proteins and the non-structural RNA dependent RNA polymerase, RNA helicase, and 3C-like cystein protease. The ultimate goal of this study is to produce recombinant TrV using a Baculovirus as expression system. Acknowledgments R S-E acknowledges a predoctoral grant from the Basque Government (BG). This work received support from Bizkaia:Xede, UPV/EHU (IT-461-07), MV-2012-2-41 from BG, and MICINN (BFU2007- 62062), Spain. G.A.M is a researcher from the CONICET, Argentina. D.M.A.G. thanks the Ibero- American Programme for Science, Technology and Development - CYTED (209RT0364).