PMID:16837071

Citation	McCarthy, SE, Licata, JM and Harty, RN (2006) A luciferase-based budding assay for Ebola virus. J. Virol. Methods 137:115-9
Abstract	The VP40 matrix protein of Ebola virus (EBOV) is capable of budding from mammalian cells as a virus-like particle (VLP) and is the major protein involved in virus egress. A functional budding assay has been developed based upon this characteristic of VP40 to assess the contributions of VP40 sequences as well as host proteins to the budding process. This well-defined assay has been modified for potential use in a high-throughput format in which the detection and quantification of firefly luciferase protein in VLPs represents a direct measure of VP40 budding efficiency. Luciferase was found to be incorporated into budding VP40 VLPs. Furthermore, co-expression of EBOV glycoprotein (GP) enhances release of VLPs containing VP40 and luciferase. In contrast, when luciferase is co-expressed with a budding deficient mutant of VP40, luciferase levels in the VLP fraction decrease significantly. This assay represents a promising high-throughput approach to identify inhibitors of EBOV budding.
Links	PubMed Online version:10.1016/j.jviromet.2006.06.007
Keywords	Cell Line; Ebolavirus/growth & development; Ebolavirus/metabolism; Genes, Reporter; Humans; Luciferases, Firefly/analysis; Luciferases, Firefly/metabolism; Luminescent Measurements/methods; Nucleoproteins/metabolism; Viral Core Proteins/metabolism; Virus Assembly
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Significance

Gene product	Qualifier	GO Term	Evidence Code	Aspect	Notes	Status
EBOZM:VP40	involved_in	GO:0046755: viral budding	ECO:0000315: mutant phenotype evidence used in manual assertion	P	Seeded From UniProt	complete
EBOZM:VP40		GO:0046755: viral budding	ECO:0000315:	P	Fig. 4	complete CACAO 8595

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