beta-lactamase as a marker for gene expression in live zebrafish embryos.
In this report we describe the development of a sensitive assay for gene expression in zebrafish embryos using beta-lactamase as a reporter gene. We show that injection of a green fluorescent substrate for beta-lactamase allows the detection of reporter gene expression in live embryos. The beta-lactamase enzyme catalyzes the hydrolysis of the substrate, thereby disrupting fluorescence resonance energy transfer from the donor to the acceptor dye in the molecule. As a result, a blue fluorescent product is produced and retained specifically in cells within which the enzyme is expressed. beta-Lactamase is therefore suitable for monitoring spatially restricted patterns of gene expression in the early embryo. We suggest that this new reporter system provides a major advancement in sensitivity over the existing methods for monitoring gene expression in vivo during early embryogenesis.
Raz
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Institute for Biology 1, University of Freiburg, Hauptstrasse 1, Freiburg, D-79104, Germany. razerez@ruf.uni-freiburg.de
Zlokarnik
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Tsien
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Driever
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D000818:Animals; D015415:Biomarkers; D005452:Fluoresceins / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D007769:Lactams; D008164:Luminescent Proteins / Q000235:genetics; D015027:Zebrafish / Q000196:embryology*; D001618:beta-Lactamases / Q000235:genetics*; D047090:beta-Lactams / Q000378:metabolism
D015415:Biomarkers; C109764:CCF 2; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007769:Lactams; D008164:Luminescent Proteins; D047090:beta-Lactams; D049452:Green Fluorescent Proteins; D001618:beta-Lactamases