Abstract：
:Overexpression screens can be used to explore gene function in Drosophila melanogaster, but to demonstrate their full potential, comprehensive and systematic collections of fly strains are required. Here we provide a protocol for high-throughput cloning of Drosophila open-reading frames (ORFs) that are regulated by upstream activation sequences (UAS sites); the resulting GAL4-inducible UAS-ORF plasmid library is then used to generate Drosophila strains by ΦC31 integrase-mediated site-specific integration. We also provide details for FLP/FRT-mediated in vivo exchange of epitope tags (or regulatory regions) in the ORF library strains, which further extends the potential applications of the library. These transgenic UAS-ORF strains are a useful resource to complement and validate genetic experiments performed with loss-of-function mutants and RNA interference (RNAi) lines. The duration of the complete protocol strongly depends on the number of ORFs required, but embryos can be injected and balanced fly stocks can be established within ∼7-8 weeks for a few genes.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Bischof J,Sheils EM,Björklund M,Basler Kdoi
10.1038/nprot.2014.105subject
Has Abstractpub_date
2014-07-01 00:00:00pages
1607-20issue
7eissn
1754-2189issn
1750-2799pii
nprot.2014.105journal_volume
9pub_type
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