Summary
Human cytomegalovirus (HCMV) infection has a marked effect on promyelocytic leukemia (PML) bodies. Here, we describe a novel real-time monitoring system for HCMV-infected cells in vitro using a newly established cell line that stably expresses GFP-PML protein. Upon infection, HCMV causes specific dispersion of GFP-PML bodies, thereby allowing the infected cells to be monitored by fluorescence microscopy without immunostaining. Quantitative protocols using either an NPB fluorescence assay or a GFP-PML imaging assay are also described. The NPB fluorescence assay is rapid, sensitive, and sufficiently simple for screening of inhibitory reagents, while the GFP-PML imaging assay is highly sensitive and applicable to drug susceptibility testing of low-titer clinical isolates.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Ueno, T., Ogawa-Goto, K. (2009). Use of a GFP-PML-Expressing Cell Line as a Biosensor for Human Cytomegalovirus Infection. In: Hicks, B.W. (eds) Viral Applications of Green Fluorescent Protein. Methods in Molecular Biology™, vol 515. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-559-6_3
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DOI: https://doi.org/10.1007/978-1-59745-559-6_3
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-934115-87-9
Online ISBN: 978-1-59745-559-6
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