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Characterization of Bombyx mori nucleopolyhedrovirus with a knockout of Bm17
Hongxing Shen1, Yang Zhou, Wen Zhang
1School of Medical Science and Laborarory Medicine, Jiangsu University, Zhenjiang, 212013, People's Republic of China.
Abstract:
Open reading frame 17 (Bm17) gene of Bombyx mori nucleopolyhedrovirus is a highly conserved gene in lepidopteran nucleopolyhedroviruses, but its function remains unknown. In this report, transient-expression and superinfection assays indicated that BM17 localized in the nucleus and cytoplasm of infected BmN cells. To determine the role of Bm17 in baculovirus life cycle, we constructed a Bm17 knockout virus and characterized its properties in cells. Analysis of the production and infection of budded virions, the level of viral DNA replication revealed showed that there was no significant difference among the mutant, the control, and the Bm17 repaired virus strains. These results suggest that BM17 is not essential for virus replication in cultured cells.
Insights
The Bombyx mori nucleopolyhedrovirus (BmNPV) Bm17 gene is conserved in lepidopteran viruses but not essential for viral replication in cell cultures. Knockout studies showed no significant difference in viral DNA replication or virion production.
Area of Science:
- Virology
- Molecular Biology
- Insect Pathology
Background:
- The Bombyx mori nucleopolyhedrovirus (BmNPV) harbors numerous genes, including the conserved open reading frame 17 (Bm17).
- The precise function of the Bm17 gene within the baculovirus life cycle has remained largely uncharacterized despite its high conservation among lepidopteran nucleopolyhedroviruses.
Purpose of the Study:
- To elucidate the functional role of the Bm17 gene in the life cycle of Bombyx mori nucleopolyhedrovirus.
- To investigate the necessity of Bm17 for viral replication and virion production in infected insect cells.
Main Methods:
- Transient-expression and superinfection assays were employed to determine the subcellular localization of the BM17 protein.
- A Bm17 knockout virus mutant was constructed and its replication properties were characterized in BmN cells.
- Viral DNA replication and budded virion production were quantitatively analyzed for the mutant, control, and repaired virus strains.
Main Results:
- Transient expression and superinfection assays confirmed that the BM17 protein localizes to both the nucleus and cytoplasm of infected BmN cells.
- Comparative analysis revealed no significant differences in viral DNA replication levels between the Bm17 knockout mutant, control virus, and Bm17-repaired virus.
- Production and infectivity of budded virions were also unaffected in the Bm17 knockout mutant compared to control and repaired strains.
Conclusions:
- The Bm17 gene product is not essential for the replication of Bombyx mori nucleopolyhedrovirus in cultured insect cells.
- These findings indicate that Bm17 may play a non-essential or redundant role in the viral life cycle under these experimental conditions.
