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Production and Purification of Baculovirus for Gene Therapy Application
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Budded baculovirus particle structure revisited.

Qiushi Wang1, Berend-Jan Bosch2, Just M Vlak3

  • 1Laboratory of Virology, Wageningen University, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands; Virology Division, Department of Infectious Disease and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.

Journal of Invertebrate Pathology
|January 9, 2016
PubMed
Summary

Cryo-electron microscopy reveals the near-native structure of baculoviruses (BV). This study details the elongated, ovoid shape of budded viruses (BVs) and their unique envelope structure, proposing a new model for these insect viruses.

Keywords:
BaculovirusBudded virusCryo-EMSpike structureUltrastructure

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Area of Science:

  • Virology
  • Structural Biology
  • Insect Pathology

Background:

  • Baculoviruses are enveloped, double-stranded DNA insect viruses with distinct budded (BV) and occlusion-derived (ODV) forms.
  • Conventional electron microscopy (EM) methods distort the fragile BV structure, limiting understanding of their native morphology.
  • Budded viruses (BVs) are typically described as rod-shaped with apical protein spikes, but their precise structure remains unclear.

Purpose of the Study:

  • To elucidate the near-native morphology of baculovirus budded virions (BVs) using cryo-electron microscopy (cryo-EM).
  • To analyze the structural differences between Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Spodoptera exigua MNPV (SeMNPV) BVs.
  • To propose a novel structural model for baculovirus BVs based on cryo-EM findings.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) was employed to image baculovirus budded virions (BVs) in a near-native state.
  • Two model baculoviruses, AcMNPV and SeMNPV, were analyzed to compare their BV structures.
  • Structural features, including envelope thickness and protein spike distribution, were examined.

Main Results:

  • Cryo-EM revealed an elongated, ovoid shape for AcMNPV and SeMNPV BVs, with a significant space between the nucleocapsid (NC) and envelope.
  • The viral envelope is approximately 6-7 nm thick, comprising two layers, and is tightly associated with the NC.
  • Protein spikes (GP64 and F) are densely clustered apically, with sparser distribution laterally; AcMNPV and SeMNPV spikes exhibit distinct morphologies.

Conclusions:

  • Baculovirus BVs possess a near-native structure characterized by an ovoid shape and a thick, two-layered envelope.
  • The tight interaction between the nucleocapsid and envelope influences BV morphology and its appearance in conventional EM.
  • A new structural model for baculovirus BVs is proposed, enhancing our understanding of these important insect viruses.