1887

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Volume 83, Issue 4

Phenotypic and genotypic analysis of nucleopolyhedrovirus serially passaged in cell culture Free

Abstract

Rapid accumulation of few polyhedra (FP) mutants was detected during serial passaging of nucleopolyhedrovirus (HaSNPV) in cell culture. 100% FP infected cells were observed by passage 6. The specific yield decreased from 178 polyhedra per cell at passage 2 to two polyhedra per cell at passage 6. The polyhedra at passage 6 were not biologically active, with a 28-fold reduction in potency compared to passage 3. Electron microscopy studies revealed that very few polyhedra were produced in an FP infected cell (<10 polyhedra per section) and in most cases these polyhedra contained no virions. A specific failure in the intranuclear nucleocapsid envelopment process in the FP infected cells, leading to the accumulation of naked nucleocapsids, was observed. Genomic restriction endonuclease digestion profiles of budded virus DNA from all passages did not indicate any large DNA insertions or deletions that are often associated with such FP phenotypes for the extensively studied nucleopolyhedrovirus and nucleopolyhedrovirus. Within an HaSNPV gene homologue, a single base-pair insertion (an adenine residue) within a region of repetitive sequences (seven adenine residues) was identified in one plaque-purified HaSNPV FP mutant. Furthermore, the sequences obtained from individual clones of the gene PCR products of a late passage revealed point mutations or single base-pair insertions occurring throughout the gene. The mechanism of FP mutation in HaSNPV is likely similar to that seen for nucleopolyhedrovirus, involving point mutations or small insertions/deletions of the gene.

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© Microbiology Society
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2002-04-01
2024-05-05
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Phenotypic and genotypic analysis of Helicoverpa armigera nucleopolyhedrovirus serially passaged in cell culture
J. Gen. Virol. 83, 945 (2002); https://doi.org/10.1099/0022-1317-83-4-945
/content/journal/jgv/10.1099/0022-1317-83-4-945
/content/journal/jgv/10.1099/0022-1317-83-4-945
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