Analysis of very late gene expression by Autographa californica nuclear polyhedrosis virus and the further development of multiple expression vectors

Weyer, Ulrike; Knight, Stuart W.; Possee, Robert D.

doi:10.1099/0022-1317-71-7-1525

Cited by 92 publications

(43 citation statements)

References 18 publications

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“…That high levels of JHE expression were obtained from the pl0-negative virus AcMP1 .JHE suggests that the absence of pl0 in AcPR1 is not the main factor contributing to decreased yield of JHE relative to AcUW2(B).JHE. It may be dangerous to extrapolate too far from this result because of other minor differences in the viral constructs which may influence expression yields (Vlak et al, 1990;Weyer et al, 1990;Min & Bishop, 1991; Table 1). …”

Section: Discussionmentioning

confidence: 99%

“…The construct of the recombinant transfer vector was verified by restriction enzyme analysis. Cotransfection of the plasmid pAcMP1 .JHE with DNA from the virus AcUWl-PH (Weyer et al, 1990) resulted in a basic protein-positive, polyhedrin-positive, pl0-negative virus, the polyhedrin being under control of the p 10 promoter at the pl0 locus.…”

Section: Methodsmentioning

confidence: 99%

“…Two viruses expressing JHE under control of the pl0 promoter were used: AcPR1 is a polyhedrin-positive, pl0-negative virus (Roelvink et al, 1992) which also expresses fl-galactosidase (Vlak et al, 1990; Table 1) and AcUW2(B).JHE is a polyhedrin-positive, pl0-positive virus . In AcUW2(B).JHE, JHE expression is driven by a duplicated pl0 promoter which is adjacent to the polyhedrin promoter in the AcMNPV genome (Weyer et at., 1990). A virus expressing modified JHE altered by site-directed mutagenesis of the active site serine to produce inactive enzyme, was used as a negative control.…”

Section: Methodsmentioning

confidence: 99%

“…JHE. All transfer vectors used in the construction of these recombinant viruses had previously been optimized for high level expression by analysis of sequences upstream of the viral promoter used (Weyer et al, 1990).…”

Section: Methodsmentioning

confidence: 99%

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Superior expression of juvenile hormone esterase and beta-galactosidase from the basic protein promoter of Autographa californica nuclear polyhedrosis virus compared to the p10 protein and polyhedrin promoters

Bonning

Roelvink

Vlak

et al. 1994

Journal of General Virology

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The expression characteristics of the pl0, polyhedrin and basic protein promoters of Autographa californica nuclear polyhedrosis virus were compared using two reporter enzymes, juvenile hormone esterase (JHE) and fl-galactosidase. In these systems, JHE is exported from the cell and fl-galactosidase is localized to the cytosol. Expression of JHE from the basic, pl0 and polyhedrin promoters was first detected in the medium at 13, 19 and 27 h post-infection respectively. The basic protein promoter yielded the highest expression of the three promoters tested for both enzymes, as determined by protein and enzyme activity assays. In addition, yields of fl-galactosidase and JHE under control of the pl0 promoter are higher relative to expression under control of the polyhedrin promoter. These data highlight the importance of investigation of viral promoters other than the polyhedrin promoter for high yield protein expression in vitro, and for insecticidal use of recombinant baculoviruses requiring high levels of expression. The results support revision of the current concept that very late viral promoters are always optimal for high yield recombinant protein expression.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Superior expression of juvenile hormone esterase and beta-galactosidase from the basic protein promoter of Autographa californica nuclear polyhedrosis virus compared to the p10 protein and polyhedrin promoters

Bonning

Roelvink

Vlak

et al. 1994

Journal of General Virology

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“…A rather different approach was used by Weyer et al (1990), who examined the consequences of locating the polyhedrin coding sequence and promoter at heterologous positions in the AcMNPV genome. Polyhedrin was expressed at normal levels when the gene coding sequence was under the control of the p 10 promoter.…”

Section: Baculoviruses: Molecular Biology and Expression Vectorsmentioning

confidence: 99%

Some Highlights of Virus Research in 1990

Elliott

Crook

Desselberger

et al. 1991

Journal of General Virology

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Purification and reassessment of ligand binding by the recombinant, putative juvenile hormone receptor of the tobacco hornworm,Manduca sexta

Charles

Wojtasek

Lentz

et al. 1996

Arch. Insect Biochem. Physiol.

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The 29 kDa protein from the larval epidermis of the tobacco hornworm, Manduca sexta, that specifically bound photoaffinity analogs of JH I and JH II was produced by a recombinant baculovirus (rJP29). The higher of the two molecular weight forms made corresponded to a protein that could be formed by read‐through of the TGA termination codon to the following TAA. The previously reported, apparent high affinity binding of [methyl‐3H]‐JH I by rJP29 as measured by the dextran‐coated charcoal (DCC) assay [Palli et al., Proc Natl Acad Sci USA 91:6191–6195 (1994)] was found to be artifactual due to endogenous cellular esterases that co‐purified with rJP29 through both DEAE cellulose and MonoQ chromatography. These esterases converted the 10–20 nM labelled JH to JH I acid and [3H]‐methanol during the 1 h incubation at room temperature. Additionally, DEAE fractions containing rJP29 or from wild‐type virus‐infected cells were found to bind nonspecifically high amounts of 12, 13‐3H]‐JH I acid in the DCC assay. Neither rJP29 nor the cellular esterases had JH esterase activity when assayed on a series of thioether surrogate substrates. When separated from these contaminating esterases either by hydroxylapatite or affinity chromatography, rJP29 showed little or no detectable binding of [12,13‐3H]‐JH I. Yet purified rJP29 bound EBDA, the photoaffinity analog of JH I; and this binding was prevented by retinol, JH I, methyl farnesoate, methoprene, and xanthophyll, but not by farnesol and 20‐hydroxyecdysone. Therefore, JP29 is not a high affinity JH receptor. © 1996 Wiley‐Liss, Inc.

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Analysis of very late gene expression by Autographa californica nuclear polyhedrosis virus and the further development of multiple expression vectors

Cited by 92 publications

References 18 publications

Superior expression of juvenile hormone esterase and beta-galactosidase from the basic protein promoter of Autographa californica nuclear polyhedrosis virus compared to the p10 protein and polyhedrin promoters

Superior expression of juvenile hormone esterase and beta-galactosidase from the basic protein promoter of Autographa californica nuclear polyhedrosis virus compared to the p10 protein and polyhedrin promoters

Some Highlights of Virus Research in 1990

Purification and reassessment of ligand binding by the recombinant, putative juvenile hormone receptor of the tobacco hornworm,Manduca sexta

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