Journal of Virology (2002) 76, 5605-5611

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Gorben P. Pijlman, Jos C.F.M. Dortmans, Angela M.G. Vermeesch, Kai Yang, Dirk E. Martens, Rob W. Goldbach and Just M. Vlak (2002)
Pivotal role of the non-hr origin of DNA replication in the genesis of defective interfering baculoviruses
Journal of Virology 76 (11), 5605-5611
Abstract: The generation of deletion mutants, including defective interfering viruses, upon serial passage of Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV) in insect cell culture has been studied. Sequences containing the non-homologous region origin of DNA replication (non-hr ori) became hypermolar in intracellular viral DNA within 10 passages in Se301 insect cells, concurrent with a dramatic drop in budded virus and polyhedron production. These predominant non-hr ori-containing sequences accumulated in larger concatenated forms and were generated de novo as demonstrated by their appearance and accumulation upon infection with a genetically homogenous bacterial clone of SeMNPV (bacmid). Sequences were identified at the junctions of the non-hr ori units within the concatemers, which may be potentially involved in recombination events. Deletion of the SeMNPV non-hr ori using RecE/RecT-mediated homologous ET recombination in Escherichia coli resulted in a recombinant bacmid with strongly enhanced stability of virus and polyhedron production upon serial passage in insect cells. This suggests that the accumulation of non-hr oris upon passage is due to the replication advantage of these sequences. The non-hr ori deletion mutant SeMNPV bacmid can be exploited as a stable eukaryotic heterologous protein expression vector in insect cells.
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Database assignments for author(s): Just M. Vlak

Research topic(s) for pests/diseases/weeds:
biocontrol - natural enemies
Research topic(s) for beneficials or antagonists:
molecular biology - genes

Pest and/or beneficial records:

Beneficial Pest/Disease/Weed Crop/Product Country Quarant.

Spodoptera exigua multiple NPV (entomopathogen)