New vector for recombinant protein expression in eukaryotic cells

RODRIGUEZ ARIEL; VER LORENA; BILEN MARCOS; GHIRINGHELLI DANIEL

Villa Carlos Paz

Congreso; XXXVIII Reunión Anual de SAIB; 2002

Several members of the family Baculoviridae are widely used as vectors for recombinant protein expression. One of the most used is the multicapsid nucleopolyhedrovirus of Autographa californica. Based on it genome, a big collection of different expression vectors were developed in the last decade. In order to increase the availability of alternative baculovirus vectors, we developed new plasmids based on the genome of Anticarsia gemmatalis multiple nuclear polyhedrovirus (AgMNPV). We designed and constructed several cassettes that allow us to clone heterologous genes under the control of different promoters: ie-1 (immediate early promoter), gp64 (early and late promoter) and polyhedrin (very late promoter). These variants will permit to optimize the heterologous protein production through the analysis of expression patterns at different times in the viral cycle. On the other hand, the vectors could be used as recombinant plasmids alone, in transient expression assays, or as transfer vectors to generate recombinant viruses, using the polyhedrin locus as target for homologous recombination. In addition to the general strategy, we present the cloning and expression of heterologous proteins, using reporter genes.