Current Microbiology

, Volume 70, Issue 4, pp 623–629 | Cite as

Determination of the Proteins Encoded by BmBDV VD1-ORF4 and Their Interacting Proteins in BmBDV-Infected Midguts

  • Guohui Li
  • Qian Zhou
  • Zhaoyang Hu
  • Peng Wang
  • Qi Tang
  • Keping Chen
  • Qin Yao
Article
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Accepted:

Abstract

Bombyx mori bidensovirus (BmBDV) VD1-ORF4 consists of 3,318 nucleotides, which codes for a predicted protein with molecular weight of about 127 kDa. However, the authentic proteins encoded by VD1-ORF4 in silkworm midguts infected with BmBDV and their interacting proteins are still unclear. In this study, Western blot analysis revealed that a 127-kDa protein was confirmed to be translated from the VD1-ORF4 transcript using polyclonal antibodies and monoclonal antibodies against VD1-ORF4 deduced amino acid. Moreover, four smaller proteins with molecular weight of about 70, 60, 53, and 42 kDa were also examined in the infected midguts. Transient expression assay indicated that the expression amount of VD1-ORF4 fused with egfp was at least 30-fold lower than that of egfp gene, and immunofluorescence staining result indicated that these proteins encoded by VD1-ORF4 were located in both the cytoplasm and nucleus. Co-immunoprecipitation result showed that Aminopeptidase and Heat shock protein 90 can be captured by these proteins encoded by VD1-ORF4. In conclusion, multiple proteins were produced from the transcripts of VD1-ORF4 gene by an uncertain expression strategy, which may play important roles in viral replication and assembly.

Keywords

Enhanced Green Fluorescent Protein Viral Life Cycle Mulberry Leave Bimolecular Fluorescence Complementation Baculovirus Expression Vector System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (31000080, 31270192, 31272507 and 31402016), the National basic Research Program of China under Grant No. 2012CB114604, and the Startup Scientific Research Fund from Jiangsu University for Senior Professionals (No. 09JDG057).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Guohui Li
    • 1
  • Qian Zhou
    • 1
  • Zhaoyang Hu
    • 1
  • Peng Wang
    • 1
  • Qi Tang
    • 1
  • Keping Chen
    • 1
  • Qin Yao
    • 1
  1. 1.Institute of Life SciencesJiangsu UniversityZhenjiangChina

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