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Negative Stain Electron Microscopy (EM) of Baculovirus Cultureassembled Virus Capsids

Published online by Cambridge University Press:  02 July 2020

Charles D. Humphrey
Charles D. Humphrey*
Affiliation:
Infectious Disease Pathology Activity CDC, Atlanta, Ga, 30333
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Abstract

Caliciviruses (CacV) including Norwalk like virus and Sapporolike virus, papillomaviruses (PapV, and parvovirus B19 (B19V) are infectious viruses known to cause a variety of diseases. These viruses are distinctive because of their low titers in body fluids and fastidious nature to culture, making them difficult to detect by EM or serology. The genomes of several CacVs, PapVs, and B19Vs have been characterized facilitating expression of specific proteins via recombinant technologies. Recombinant proteins may be useful for obtaining the antibodies to be used in improved immunoassays and vaccines for preventing diseases associated with these viruses. CDC and Emory University investigators have designed techniques by which capsid proteins for a variety of viruses including CaCV, PapV, and B19V were expressed in Autographies califomicanuclear polyhedrosis virus (AcNPV) infected Spodoptera fruigiperda (Sf9) cells. Those proteins self-assembled into virus-like particles (VLPs)(Figs.l- 4). Culture-expressed capsids were “purified” and concentrated by traditional protein purification techniques including gel filtration, sucrose gradient centrifugation and cesium chloride gradient centrifugation.

Type
Microbiology
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 744 - 745
Copyright
Copyright © Microscopy Society of America 2001

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