1. Identification and characterization of vp7 gene in Bombyx mori cytoplasmic polyhedrosis virus
- Author
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Fei Chen, Guangli Cao, Liyuan Zhu, Sulan Kuang, Renyu Xue, Chengliang Gong, Min Zhu, Lei He, Xiaolong Hu, and Zi Liang
- Subjects
0301 basic medicine ,Gene isoform ,Immunoprecipitation ,viruses ,030106 microbiology ,NSP, Non-structural proteins ,NLS, Nuclear localization signal ,Reoviridae ,Virus Replication ,Article ,Virus ,03 medical and health sciences ,Bombyx mori ,RNA interference ,SDS-PAGE, Sodium dodecyl sulfate polyacrylamide gel electrophoresis ,BTV, Bluetongue virus ,Gene expression ,BmCPV ,Genetics ,Viral structural protein ,SD, Standard deviation ,Animals ,VDAC, Voltage-dependent anion-selective channel-like isoform ,Cloning, Molecular ,Antiserum ,BmCPV, Bombyx mori cytoplasmic polyhedrosis virus ,biology ,NTCB, 2-nitro-5-thiocyanobenzoic acid ,fungi ,virus diseases ,CPVs, Cytoplasmic polyhedrosis viruses ,General Medicine ,Nucleocapsid Proteins ,TnCPV-15, Trichoplusiani cypovirus type 15 ,Bombyx ,biology.organism_classification ,Molecular biology ,VP7 ,Intestines ,RNAi, RNA interference ,030104 developmental biology ,FMDV, Foot-and-mouth disease virus ,ORF, Open reading frame ,Co-IP, Co-Immunoprecipitation ,DLP, Double-layered particle ,DAPI, 4′, 6-diamidino-2-phenylindole - Abstract
The genome of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) contains 10 double stranded RNA segments (S1–S10). The segment 7 (S7) encodes 50 kDa protein which is considered as a structural protein. The expression pattern and function of p50 in the virus life cycle are still unclear. In this study, the viral structural protein 7 (VP7) polyclonal antibody was prepared with immunized mouse to explore the presence of small VP7 gene-encoded proteins in Bombyx mori cytoplasmic polyhedrosis virus. The expression pattern of vp7 gene was investigated by its overexpression in BmN cells. In addition to VP7, supplementary band was identified with western blotting technique. The virion, BmCPV infected cells and midguts were also examined using western blotting technique. 4, 2 and 5 bands were detected in the corresponding samples, respectively. The replication of BmCPV genome in the cultured cells and midgut of silkworm was decreased by reducing the expression level of vp7 gene using RNA interference. In immunoprecipitation experiments, using a polyclonal antiserum directed against the VP7, one additional shorter band in BmCPV infected midguts was detected, and then the band was analyzed with mass spectrum (MS), the MS results showed thatone candidate interacted protein (VP7 voltage-dependent anion-selective channel-like isoform, VDAC) was identified from silkworm. We concluded that the novel viral product was generated with a leaky scanning mechanism and the VDAC may be an interacted protein with VP7., Highlights • The small VP7 protein was present in BmCPV. • The replication of BmCPV genome was decreased by reducing vp7 gene expression. • This novel product was generated with a leaky scanning mechanism.
- Published
- 2017