GENERATION OF A P10-BASED BACULOVIRUS EXPRESSION VECTOR IN YEAST WITHINFECTIVITY FOR INSECT LARVAE AND INSECT CELLS

A new, versatile baculovirus Vector was developed for the generation o f recombinants in the yeast Saccharomyces cerevisiae and for the expre ssion of foreign proteins in both insect larvae and in insect cells. T his vector is based on Autographa californica multiple nucleocapsid nu cleopolyhedrovirus (AcMNPV) and exploits the 10-kDa protein promoter ( p10) for the expression of the foreign gene. The p10 locus was used fo r the insertion of a yeast-selectable marker system (ARS-URA-URA3) and of a gene for screening and titration of recombinants in insect cells (beta-galactosidase). The polyhedron-positive phenotype of this vecto r is maintained allowing its use in insect larvae, by feeding polyhedr a, and in insect cells, by infecting with budded virus. The generation of this baculovirus vector requires a single recombination step in ye ast prior to infection of insect cells, but has the advantage over the vector designed previously (Patel et al., A new method for the isolat ion of recombinant baculovirus, Nucleic Acids Research 20 (1992) 97-10 4) that these vectors can also be used in insects. (C) 1997 Elsevier S cience B.V.