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

Abstract

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.

Effect of Promoters and Signal Sequences on the Production of Secreted HIV-1 gp120 Protein in the Baculovirus System

“…Although there have been reports of enhanced protein production from promoters active earlier than the very late polyhedrin and p10 promoters (15)(16)(17), we found that production of secreted protein driven by either the very late polyhedrin promoter or the late-very late hybrid promoter was approximately equivalent. However, production of secreted protein varied significantly according to the signal sequence used.…”

Using cell size kinetics to determine optimal harvest time for Spodoptera frugiperda and Trichoplusia ni BTI-TN-5B1-4 cells infected with a baculovirus expression vector system expressing enhanced green fluorescent protein

“…Suzuki et al 1997;Monsma and Scott 1997), while others have looked at the possibility of using different promoters to replace the commonly used polyhedrin promoter that is activated at the very late phase of a baculovirus infection (e.g. Bonning et al 1994;Hill-Perkins and Possee 1990;Kost et al 1997;Lawrie et al 1995).…”

Analysis of the Complete Genome Sequence of the Hz-1 Virus Suggests that It Is Related to Members of the Baculoviridae

“…JHE inactivates juvenile hormone, which regulates the outcome of an insect molt, and is an essential enzyme for normal insect development. It has been an attractive targets for biorationally designed, environmentally safe pesticides (8,21). The expression of JHE by the Hz-1 virus may influence the developmental characteristics, weight gain, and time of mortality of the insects.…”