1887

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Volume 69, Issue 4

Functional Studies on the p10 Gene of Nuclear Polyhedrosis Virus Using a Recombinant Expressing a p10-β-Galactosidase Fusion Gene Free

Abstract

Summary

The β-galactosidase gene () of was inserted in phase with the coding sequence of the nuclear polyhedrosis virus (AcMNPV) late-expressed 10000 (p10) gene. The fusion gene was inserted into the AcMNPV genome by cotransfection of a recombinant plasmid pAcR159Z, consisting of the RI P fragment-containing pBR325-derived plasmid pAcR159 and the insert in the p10 gene, and wild-type AcMNPV DNA. Infection of cells by the resulting recombinant AcMNPV/p10Z-2 showed high level expression of a p10- fusion protein, but no synthesis of p10. Therefore, the p10 gene is dispensable for virus replication and the p10 promoter is effective in driving the expression of foreign genes. Cells infected with AcMNPV/p10Z recombinants resembled those infected with wild-type AcMNPV in the amounts of polyhedrin synthesized and polyhedra formed, although p10 was absent. The nucleus and cytoplasm of AcMNPV/p10Z-2-infected cells lacked the fibrous structures that are associated with p10 in wild-type AcMNPV-infected cells. Instead, large granular structures were observed that were found by immunogold labelling to contain the gene product. The electron-dense ‘spacers’, thought to be precursors of the polyhedron membrane, were absent from cells infected by the recombinant virus and the polyhedra did not have a membrane. The recombinant AcMNPV/p10Z-2 was at least twice as virulent for second instar larvae than was wild-type AcMNPV. The increased virulence of the recombinant is an important property for the control of insects.

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Keyword(s): AcMNPV , baculovirus genetic engineering and immunogold labelling
© Society for General Microbiology 1988
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1988-04-01
2024-04-23
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Functional Studies on the p10 Gene of Autographa californica Nuclear Polyhedrosis Virus Using a Recombinant Expressing a p10-β-Galactosidase Fusion Gene
J. Gen. Virol. 69, 765 (1988); https://doi.org/10.1099/0022-1317-69-4-765
/content/journal/jgv/10.1099/0022-1317-69-4-765
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