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Hemorrhagic Fever Virus Budding Studies

Part of the Methods in Molecular Biology book series (MIMB,volume 1604)

Abstract

Independent expression of the VP40 or Z matrix proteins of filoviruses (marburgviruses and ebolaviruses) and arenaviruses (Lassa fever and Junín), respectively, gives rise to the production and release of virus-like particles (VLPs) that are morphologically identical to infectious virions. We can detect and quantify VLP production and egress in mammalian cells by transient transfection, SDS-PAGE, Western blotting, and live cell imaging techniques such as total internal reflection fluorescence (TIRF) microscopy. Since the VLP budding assay accurately mimics budding of infectious virus, this BSL-2 assay is safe and useful for the interrogation of both viral and host determinants required for budding and can be used as an initial screen to identify and validate small molecule inhibitors of virus release and spread.

Key words

  • Budding
  • Ebola virus
  • VLPs
  • VP40
  • Matrix protein
  • TIRF

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Acknowledgments

I would like to thank past and present members of the lab who contributed to this work. This work was supported in part by NIH grants to R.N.H.

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Correspondence to Ronald N. Harty .

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Harty, R.N. (2018). Hemorrhagic Fever Virus Budding Studies. In: Salvato, M. (eds) Hemorrhagic Fever Viruses. Methods in Molecular Biology, vol 1604. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6981-4_15

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  • DOI: https://doi.org/10.1007/978-1-4939-6981-4_15

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