Vectors for efficient and high-throughput construction of fluorescent drosophila reporters using the PhiC31 site-specific integration system

Genesis. 2010 Jul;48(7):452-6. doi: 10.1002/dvg.20637.

Abstract

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.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Binding Sites
  • Cloning, Molecular / methods*
  • Drosophila melanogaster / genetics*
  • Enhancer Elements, Genetic / genetics
  • Gene Expression Regulation / genetics
  • Gene Transfer Techniques
  • Genes, Reporter*
  • Genetic Vectors*
  • High-Throughput Screening Assays / methods*
  • Integrases / genetics*
  • Plasmids
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombination, Genetic

Substances

  • Recombinant Fusion Proteins
  • Integrases