Vectors for efficient and high-throughput construction of fluorescent drosophila reporters using the PhiC31 site-specific integration system.
The fruit fly Drosophila is a leading model system for the study of transcriptional control by cis-regulatory elements or enhancers. Here, we present a rapid and highly efficient system for the large-scale analysis of enhancer elements, site-specifically integrated into the Drosophila genome. This system, which is scalable for either small projects or high-throughput approaches, makes use of the Gateway cloning technology and the PhiC31 site-specific integration system, which allows the insertion of constructs at predetermined genomic locations. Thus, this system allows not only a fast and easy analysis of reporter gene expression in live animals, but also the simultaneous analysis of different regulatory outputs on a cellular resolution by recombining in the same animal distinct enhancer elements fused to different fluorescent proteins.
Boy
Aurelia L
AL
CellNetworks-Cluster of Excellence and BIOQUANT Center, University of Heidelberg, D-69120 Heidelberg, Germany.
Zhai
Zongzhao
Z
Habring-Müller
Anette
A
Kussler-Schneider
Yvonne
Y
Kaspar
Petra
P
Lohmann
Ingrid
I
D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D003001:Cloning, Molecular / Q000379:methods*; D004331:Drosophila melanogaster / Q000235:genetics*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D057166:High-Throughput Screening Assays / Q000379:methods*; D019426:Integrases / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic
D011993:Recombinant Fusion Proteins; D019426:Integrases