In:
Journal of Virology, American Society for Microbiology, Vol. 76, No. 11 ( 2002-06), p. 5605-5611
Abstract:
The generation of deletion mutants, including defective interfering viruses, upon serial passage of Spodoptera exigua multicapsid nucleopolyhedrovirus (Se M NPV) 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 Se M NPV (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 Se M NPV 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 ori s upon passage is due to the replication advantage of these sequences. The non- hr ori deletion mutant Se M NPV bacmid can be exploited as a stable eukaryotic heterologous protein expression vector in insect cells.
Type of Medium:
Online Resource
ISSN:
0022-538X
,
1098-5514
DOI:
10.1128/JVI.76.11.5605-5611.2002
Language:
English
Publisher:
American Society for Microbiology
Publication Date:
2002
detail.hit.zdb_id:
1495529-5
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