Characterization of a nucleopolyhedrovirus with a deletion of the baculovirus core gene Bm67

Ge, Jun-Qing; Yang, Zhenzhen; Tang, Xudong; Xu, Haijun; Hong, Jian; Chen, Jianguo; Zhang, Chuan-Xi

doi:10.1099/vir.0.83398-0

Cited by 19 publications

(11 citation statements)

References 38 publications

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“…However, this phenotype was rescued by the ectopic expression of the Bm67 gene, suggesting that this gene is critical for the formation of infectious viral particles. These findings are consistent with those published by Ge et al (2008) who demonstrated that the deletion of Bm67 reduces viral nucleocapsid assembly, and hinders the formation of BV in enveloped viruses. Our transmission electron microscopy results also confirmed that Bm67 deletion affects the capacity for virus assembly, leading to a failure to form mature viral particles.…”

Section: Discussionsupporting

confidence: 93%

“…All the sequenced baculovirus genomes include Bm67, one of the core baculovirus genes; its homologous gene in AcMNPV is ORF81 (Ge et al, 2008). The role of this protein in viral replication is as yet unclear, the understanding the biological function of Bm67, however, would clarify the mechanism of transcriptional regulation in the insect nuclear polyhedrosis viruses.…”

Section: Introductionmentioning

confidence: 99%

“…Bm68 encodes a structural protein of the budded virus (BV), and its deletion reduces the viral budding rate, leading to delayed viral DNA replication (Iwanaga et al, 2002). Although Bm67 is essential to early viral DNA replication and Bm67-deleted viruses do not form infectious virus particles (Ge et al, 2008), the role of Bm67 in viral gene transcription has not yet been determined.…”

Section: Introductionmentioning

confidence: 99%

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The effect of BM67 gene deletion on Bombyx mori nuclear polyhedrosis virus replication

Shi¹,

Zhang²,

Xie³

et al. 2015

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Summary. -Homologs of Bombyx mori nuclear polyhedrosis virus (BmNPV) Bm67 gene ORF67 have been found in the genome of all lepidopteran nuclear polyhedrosis viruses, but their function is still not very clear. In order to analyze it we employed a bacmid harboring the complete BmNPV genome including the Bm67 gene and expressing infectious virus (wtBacmid) for the construction of its Bm67-deficient variant (Bm67-KO-Bacmid) using the Red recombination system and the Bm67-repaired variant (Bm67-Re-Bacmid) using the Bac-to-Bac system. By transfecting BmN cells with these bacmids we demonstrated that the Bm67-deficient virus did not generate infectious virus, while the repaired virus restored its infectivity, indicating that the Bm67 gene is essential for the formation of infectious budding virus (BV). Electron microscopy of BmN cells transfected with the abovementioned bacmids showed many mature rodshaped virus particles in both wtBacmid-and Bm67-Re-Bacmid-transfected cells but none in Bm67-KO-Bacmid-transfected ones. Moreover, the real-time RT-PCR showed that the deletion of Bm67 from wtBacmid significantly reduced the levels of viral genomic DNA and transcripts of viral early genes dnapol, ie-1 and lef-3 but not those of transcripts of late gene vp39 and very late gene p10. The finding that the Bm67-deficient virus generated reduced levels of infectious virus and transcripts of early dnapol gene but not those of late genes indicates that the Bm67 gene is essential for BmNPV replication.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The effect of BM67 gene deletion on Bombyx mori nuclear polyhedrosis virus replication

Shi¹,

Zhang²,

Xie³

et al. 2015

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“…Construction of Bm54-deleted, repair and wild-type bacmids A Bm54-null bacmid was constructed using k Red homologous recombination in E. coli as described previously [6,33]. The reconstructed Bm54 locus and ph locus are shown in Fig.…”

Section: Cellular Localization Of Bm54mentioning

confidence: 99%

Bombyx mori nucleopolyhedrovirus ORF54, a viral desmoplakin gene, is associated with the infectivity of budded virions

et al. 2012

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Bombyx mori nucleopolyhedrovirus (BmNPV) ORF54 (Bm54), a member of the viral desmoplakin N-terminus superfamily, is homologous to Autographa californica nucleopolyhedrovirus (AcMNPV) ORF66, which is required for the efficient egress of nucleocapsids from the nucleus and occlusion body formation. In this paper, we generated a bacmid with the Bm54 gene deleted via homologous recombination in Escherichia coli and characterized the mutant virus using a transfection-infection assay and transmission electron microscopy analysis. Our results demonstrated that the cells transfected with viral DNA lacking Bm54 produced non-infectious budded viruses (BVs). Electron microscopy showed that although the deletion of Bm54 did not affect assembly and release of nucleocapsids, it severely affected polyhedron formation. In conclusion, deletion of Bm54 resulted in non-infectious BV and defective polyhedra. Although the sequences of Bm54 and Ac66 are very similar, the two genes function quite differently in the regulation of viral life cycle.

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“…Because the genome of Bombyx mori nucleopolyhedrovirus (BmNPV) (T3 strain) was completely sequenced [8], numerous genes from BmNPV have been characterized, including Bm79 [9], Bm67 [10,11], Bm122 [12], Bm60 [13], DBP [14], and Bm68 [15]. However, the function of many genes in BmNPV genome remains unknown.…”

Section: Introductionmentioning

confidence: 99%

Bombyx mori nucleopolyhedrovirus ORF51 encodes a budded virus envelope associated protein

Tian

Tang

et al. 2008

Virus Genes

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Bombyx mori nucleopolyhedrovirus (BmNPV) ORF 51 (Bm51) is a gene present in many lepidopteran NPVs, but its function is unknown. In this study, Bm51 was characterized. Transcripts of Bm51 were detected from 4.5 through 72 hour post infection (h p.i.) by RT-PCR. The corresponding protein was detected from 6 to 72 h p.i. in BmNPV-infected BmN cells by western blot analysis using a polyclonal antibody against Bm51. Western blot assay of occlusion-derived virus and budded virus (BV) preparations revealed that Bm51 encodes a 23-kDa structural protein that is associated with BV and is located in the envelope fraction of budded virions. The protein was temporarily called BV-E23. In addition immunofluorescence microscopy demonstrated that the protein was present within the cytoplasm and nuclei in virus-infected cells. In conclusion, the available data suggest that Bm51 is a functional ORF of BmNPV and encodes a protein expressed in the early stage of the infection cycle that is associated with the BV envelope.

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Characterization of a nucleopolyhedrovirus with a deletion of the baculovirus core gene Bm67

Cited by 19 publications

References 38 publications

The effect of BM67 gene deletion on Bombyx mori nuclear polyhedrosis virus replication

The effect of BM67 gene deletion on Bombyx mori nuclear polyhedrosis virus replication

Bombyx mori nucleopolyhedrovirus ORF54, a viral desmoplakin gene, is associated with the infectivity of budded virions

Bombyx mori nucleopolyhedrovirus ORF51 encodes a budded virus envelope associated protein

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