Microfluorometer assay to measure the expression of beta-galactosidase and green fluorescent protein reporter genes in single Drosophila flies.
beta-galactosidase and green fluorescent protein (GFP) are among the most commonly used reporter genes to monitor gene expression in various organisms including Drosophila melanogaster. Their expression is usually detected in a qualitative way by direct microscopic observations of cells, tissues, or whole animals. To measure in vivo the inducibility of two antimicrobial peptide genes expressed during the Drosophila innate immune response, we have adapted two reporter gene systems based on the beta-galactosidase enzymatic activity and GFP. We have designed a 96-well microplate fluorometric assay sensitive enough to quantify the expression of both reporter genes in single flies. The assay has enabled us to process efficiently and rapidly a large number of individual mutant flies generated during an ethylmethane sulfonate saturation mutagenesis of the Drosophila genome. This method may be used in any screen that requires the quantification of reporter gene activity in individual insects.
Centre National de la Recherche Scientifique, Strasbourg, France.
A C
AC
Jung
M C
MC
Criqui
S
S
Rutschmann
J A
JA
Hoffmann
D
D
Ferrandon
D000818:Animals; D003592:Cytophotometry; D004330:Drosophila; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase