Characterization of baculovirus constructs lacking either the Ac 101, Ac 142, or the Ac 144 open reading frame

Adam L. Vanarsdall, Margot N. Pearson, George F. Rohrmann

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39 Scopus citations

Abstract

To investigate the role of the gene products encoded from the open reading frames 101, 142, and 144 of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), a set of bacmid knockout and repair constructs were generated. The repair genes were engineered to contain an HA epitope tag at their C-termini. The results of transfection-infection assays and growth curve analyses showed that the Ac 101, 142, and 144 genes were required for infectious virus production. To better characterize the role of these genes in the baculovirus replication cycle, quantitative DNA replication assays were performed and demonstrated that in cells transfected with the Ac 101, 142, or 144 knockouts, DNA replicated with similar kinetics as a control virus. Western blot analyses of budded virus from cells infected with the repair viruses showed that these proteins are associated with the viral nucleocapsid. Furthermore, immunoelectron microscopy of cells transfected with the knockout bacmids revealed defects in nucleocapsid production for all three constructs. From these results we concluded that the gene products encoded from these open reading frames are essential for virus production and may be involved in DNA processing, packaging, or nucleocapsid morphogenesis.

Original languageEnglish (US)
Pages (from-to)187-195
Number of pages9
JournalVirology
Volume367
Issue number1
DOIs
StatePublished - Oct 10 2007

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Keywords

  • AcMNPV
  • Baculovirus
  • DNA replication
  • Insect virus
  • Nucleoployhedrovirus
  • Virus structure

ASJC Scopus subject areas

  • Virology

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