Pivotal role of the non-hr origin of DNA replication in the genesis of defective interfering baculoviruses

Pijlman, Gorben P., Dortmans, Jos C. F. M., Vermeesch, Angela M. G., Yang, Kai, Martens, Dirk E., Goldbach, Rob W. and Vlak, Just M. (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. doi:10.1128/JVI.76.11.5605-5611.2002

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Author Pijlman, Gorben P.
Dortmans, Jos C. F. M.
Vermeesch, Angela M. G.
Yang, Kai
Martens, Dirk E.
Goldbach, Rob W.
Vlak, Just M.
Title Pivotal role of the non-hr origin of DNA replication in the genesis of defective interfering baculoviruses
Journal name Journal of Virology   Check publisher's open access policy
ISSN 0022-538X
Publication date 2002-06
Year available 2002
Sub-type Article (original research)
DOI 10.1128/JVI.76.11.5605-5611.2002
Open Access Status File (Publisher version)
Volume 76
Issue 11
Start page 5605
End page 5611
Total pages 7
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Language eng
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 on units within the concatemers, which may be potentially involved in recombination events. Deletion of the SeMNPV non-hr on 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 on deletion mutant SeMNPV bacmid can be exploited as a stable eukaryotic heterologous protein expression vector in insect cells.
Keyword Virology
Nuclear Polyhedrosis-virus
Exigua Multicapsid Nucleopolyhedrovirus
Recombinant Baculoviruses
Deletion Mutants
Putative Origins
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Clinical Medical Virology Centre Publications
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Citation counts: TR Web of Science Citation Count  Cited 59 times in Thomson Reuters Web of Science Article | Citations
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Created: Wed, 19 Sep 2007, 15:19:27 EST