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Ultrastructural Characterization of Virus-Like Particles Produced by a Recombinant Baculovirus Expression Vector

Published online by Cambridge University Press:  02 July 2020

A. J. Wasserman ,
H. Wang ,
G. Clark ,
J. R. Megill  and
S.K. Durham
A. J. Wasserman
Affiliation:
Departments of Experimental Pathology, Princeton, NJ08543
H. Wang
Affiliation:
Virology, Wallingford CT, 06492, Bristol-Myers Squibb Pharmaceutical Research Institute. Princeton, NJ08543
G. Clark
Affiliation:
Virology, Wallingford CT, 06492, Bristol-Myers Squibb Pharmaceutical Research Institute. Princeton, NJ08543
J. R. Megill
Affiliation:
Departments of Experimental Pathology, Princeton, NJ08543
S.K. Durham
Affiliation:
Departments of Experimental Pathology, Princeton, NJ08543
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Extract

Analysis of Hepatitis C virus (HCV) assembly and packaging, and the study of virus-cell interactions, have been impeded by the lack of a robust cell culture system and a viable small animal model. Recently, an alternative approach utilizing recombinant gene technology has been implemented. We describe a system in which specific HCV genes are cloned into a baculovirus to form a recombinant baculovirus expression vector that produces HCV proteins that self-assemble into enveloped virus-like particles (VLP). Four HCV structural genes were cloned into the baculovirus expression vector under the control of the polyhedrin promoter, and were then recombined with baculovirus DNA. A baculovirus with no HCV insert served as a negative control (wild-type). Ovary cells (Spodoptera frugiperda) were infected with either the recombinant or wild type baculovirus. Five-days post-infection, a group of cells were harvested, fixed in McDowell-Trumps solution, and routinely prepared for transmission electron microscopy (TEM).

Type
Application of Classical and Novel Microscopy to Tissue Injury and Infectious Disease Pathogenesis
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 1054 - 1055
Copyright
Copyright © Microscopy Society of America

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