Microfluorometer assay to measure the expression of β-galactosidase and green fluorescent protein reporter genes in single Drosophila flies

被引:24
作者
Jung, AC [1 ]
Criqui, MC [1 ]
Rutschmann, S [1 ]
Hoffmann, JA [1 ]
Ferrandon, D [1 ]
机构
[1] CNRS, Inst Biol Mol & Cellulaire, UPR 9022, F-67084 Strasbourg, France
关键词
D O I
10.2144/01303rr04
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 whale 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.
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收藏
页码:594 / +
页数:6
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