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Archives of Virology

, Volume 164, Issue 7, pp 1753–1760 | Cite as

Assembly of tomato blistering mosaic virus-like particles using a baculovirus expression vector system

  • Raquel Medeiros Vasques
  • Roberto Franco Teixeira Correa
  • Leonardo Assis da Silva
  • Rosana Blawid
  • Tatsuya Nagata
  • Bergmann Morais Ribeiro
  • Daniel M. P. Ardisson-AraújoEmail author
Original Article

Abstract

The expression of several structural proteins from a wide variety of viruses in heterologous cell culture systems results in the formation of virus-like particles (VLPs). These VLPs structurally resemble the wild-type virus particles and have been used to study viral assembly process and as antigens for diagnosis and/or vaccine development. Tomato blistering mosaic virus (ToBMV) is a tymovirus that has a 6.3-kb positive-sense ssRNA genome. We have employed the baculovirus expression vector system (BEVS) for the production of tymovirus-like particles (tVLPs) in insect cells. Two recombinant baculoviruses containing the ToBMV wild-type coat protein (CP) gene or a modified short amino-terminal deletion (Δ2-24CP) variant were constructed and used to infect insect cells. Both recombinant viruses were able to express ToBMV CP and Δ2-24CP from infected insect cells that self-assembled into tVLPs. Therefore, the N-terminal residues (2-24) of the native ToBMV CP were shown not to be essential for self-assembly of tVLPs. We also constructed a third recombinant baculovirus containing a small sequence coding for the major epitope of the chikungunya virus (CHIKV) envelope protein 2 (E2) replacing the native CP N-terminal 2-24 amino acids. This recombinant virus also produced tVLPs. In summary, ToBMV VLPs can be produced in a baculovirus/insect cell heterologous expression system, and the N-terminal residues 2-24 of the CP are not essential for this assembly, allowing its potential use as a protein carrier that facilitates antigen purification and might be used for diagnosis.

Notes

Funding

This work was supported by Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF, Grant number 193.001532/2016). Conselho Nacional de Desenvolvimento Científico e Tecnológico (Grant number 428799/2018-3). Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Grant number CAPES/PROEX 23038.005848/2018-31).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Raquel Medeiros Vasques
    • 1
    • 2
  • Roberto Franco Teixeira Correa
    • 2
  • Leonardo Assis da Silva
    • 2
  • Rosana Blawid
    • 3
  • Tatsuya Nagata
    • 1
  • Bergmann Morais Ribeiro
    • 2
  • Daniel M. P. Ardisson-Araújo
    • 4
    Email author
  1. 1.Laboratory of Microscopy and Virology, Cell Biology DepartmentUniversity of BrasiliaBrasíliaBrazil
  2. 2.Laboratory of Baculovirus, Cell Biology DepartmentUniversity of BrasiliaBrasíliaBrazil
  3. 3.Laboratory of Phytovirology, Department of AgronomyRural Federal University of PernambucoRecifeBrazil
  4. 4.Laboratory of Insect Virology, Department of Biochemistry and Molecular BiologyFederal University of Santa MariaSanta MariaBrazil

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