Generation of a p10-based baculovirus expression vector in yeast with infectivity for insect larvae and insect cells.
A new, versatile baculovirus vector was developed for the generation of recombinants in the yeast Saccharomyces cerevisiae and for the expression of foreign proteins in both insect larvae and in insect cells. This vector is based on Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) and exploits the 10-kDa protein promoter (p10) for the expression of the foreign gene. The p10 locus was used for 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 vector is maintained allowing its use in insect larvae, by feeding polyhedra, and in insect cells, by infecting with budded virus. The generation of this baculovirus vector requires a single recombination step in yeast prior to infection of insect cells, but has the advantage over the vector designed previously (Patel et al., A new method for the isolation of recombinant baculovirus, Nucleic Acids Research 20 (1992) 97-104) that these vectors can also be used in insects.
Heldens
J G
JG
Wageningen Agricultural University, Department of Virology, The Netherlands.
Kester
H A
HA
Zuidema
D
D
Vlak
J M
JM
D000818:Animals; D002460:Cell Line; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D007763:Lac Operon; D007814:Larva / Q000235:genetics; D009036:Moths / Q000235:genetics*; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D000077183:Occlusion Body Matrix Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*; D015678:Viral Structural Proteins
D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; C078072:p10 protein, Baculovirus