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Generation of a p10-based baculovirus expression vector in yeast with infectivity for insect larvae and insect cell culture
- Source :
- Journal of Virological Methods 68 (1997), Journal of Virological Methods, 68, 57-63
- Publication Year :
- 1997
-
Abstract
- 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 (β-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.
- Subjects :
- Recombination system
Baculoviridae
p10
viruses
Saccharomyces cerevisiae
Genetic Vectors
Laboratory of Virology
Gene Expression
Moths
Cell Line
Laboratorium voor Virologie
Viral Proteins
Genes, Reporter
Virology
Animals
Baculovirus
Gene
Infectivity
Viral Structural Proteins
biology
fungi
Insect cell culture
Autographa californica
Nuclear Polyhedrosis Virus
biology.organism_classification
PE&RC
Occlusion Body Matrix Proteins
Yeast
Nucleopolyhedroviruses
Lac Operon
Larva
Subjects
Details
- Language :
- English
- ISSN :
- 01660934
- Volume :
- 68
- Database :
- OpenAIRE
- Journal :
- Journal of Virological Methods
- Accession number :
- edsair.doi.dedup.....49840766aba74cc8062d6f12c2f73542