Abstract |
The N-terminal region of the gene encoding polyhedrin, the major occlusion protein of the insect baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV), has been fused to DNA encoding Escherichia coli beta-galactosidase. The fused gene was inserted into the AcNPV DNA genome by cotransfection of insect cells with recombinant plasmid DNA and wild-type AcNPV genomic DNA. Recombinant viruses were selected as blue plaques in the presence of a beta-galactosidase indicator, 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside. Studies of one such virus, L1GP-gal3, indicated that the synthesis of beta-galactosidase is temporally controlled beginning late (20 h) in infection after the release of infectious virus particles from the cell. By 48 h postinfection, a remarkably high level of expression is achieved. On the basis of these results, AcNPV should be a useful vector for the stable propagation and expression of passenger genes in a lepidopteran cell background. A generalized transplacement vector that facilitates the construction and selection of recombinant viruses carrying passenger genes under their own promoter control has also been developed.
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Authors | G D Pennock, C Shoemaker, L K Miller |
Journal | Molecular and cellular biology
(Mol Cell Biol)
Vol. 4
Issue 3
Pg. 399-406
(Mar 1984)
ISSN: 0270-7306 [Print] United States |
PMID | 6325875
(Publication Type: Journal Article)
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Chemical References |
- DNA Restriction Enzymes
- Galactosidases
- beta-Galactosidase
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Topics |
- Amino Acid Sequence
- Animals
- Base Sequence
- Cell Line
- DNA Restriction Enzymes
- Escherichia coli
(enzymology, genetics)
- Galactosidases
(genetics)
- Genes
- Genes, Bacterial
- Genetic Vectors
- Insect Viruses
(genetics)
- Lepidoptera
- Plasmids
- beta-Galactosidase
(genetics, metabolism)
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