Open AccessExpression of the firefly luciferase gene in vaccinia virus: a highly sensitive gene marker to follow virus dissemination in tissues of infected animals.
Author(s) -
José F. Rodrígúez,
Dolores Rodrı́guez,
Juan-Ramon Rodriguez,
Eleanor B. McGowan,
Mariano Esteban
Publication year - 1988
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.85.5.1667
Subject(s) - luciferase , vaccinia , luciferin , biology , virus , lampyridae , microbiology and biotechnology , gene , gene expression , virology , orthopoxvirus , poxviridae , cell culture , transfection , firefly protocol , recombinant dna , biochemistry , genetics , zoology
We have introduced the firefly luciferase gene of Photinus pyralis into the vaccinia virus genome. This gene is expressed in a coordinate fashion during virus infection. Luminescence produced by the action of luciferase [Photinus-luciferin:oxygen 4-oxidoreductase(decarboxylating, ATP-hydrolyzing), EC 1.13.12.7] was easily detectable in infected cells in culture as well as in cells of tissues of infected mice. The limits of detection were about one infected cell in a background of a million noninfected cells. The luciferase assay was about 1000-fold more sensitive than that of beta-galactosidase. Our findings show that the luciferase assay can be conveniently used to follow viral gene expression and virus dissemination both in cell cultures and in tissues of infected animals.