Versatile P[acman] BAC libraries for transgenesis studies in Drosophila melanogaster
Top Cited Papers
Open Access
- 24 May 2009
- journal article
- research article
- Published by Springer Nature in Nature Methods
- Vol. 6 (6), 431-434
- https://doi.org/10.1038/nmeth.1331
Abstract
Two bacterial artificial chromosome (BAC) libraries, spanning almost the entire D. melanogaster genome in insert sizes of 20 and 80 kb, that allow easy integration into the fruit fly genome at defined docking sites provide a rich resource to study gene expression and function. We constructed Drosophila melanogaster bacterial artificial chromosome libraries with 21-kilobase and 83-kilobase inserts in the P[acman] system. We mapped clones representing 12-fold coverage and encompassing more than 95% of annotated genes onto the reference genome. These clones can be integrated into predetermined attP sites in the genome using ΦC31 integrase to rescue mutations. They can be modified through recombineering, for example, to incorporate protein tags and assess expression patterns.Keywords
This publication has 49 references indexed in Scilit:
- Inactivation of clathrin heavy chain inhibits synaptic recycling but allows bulk membrane uptakeThe Journal of cell biology, 2008
- Recombineering-mediated tagging of Drosophila genomic constructs for in vivo localization and acute protein inactivationNucleic Acids Research, 2008
- Tools for neuroanatomy and neurogenetics in DrosophilaProceedings of the National Academy of Sciences, 2008
- BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammalsNature Methods, 2008
- Exploiting position effects and the gypsy retrovirus insulator to engineer precisely expressed transgenesNature Genetics, 2008
- Huntingtin-interacting protein 14, a palmitoyl transferase required for exocytosis and targeting of CSP to synaptic vesiclesThe Journal of cell biology, 2007
- Vector and parameters for targeted transgenic RNA interference in Drosophila melanogasterNature Methods, 2007
- Eps15 and Dap160 control synaptic vesicle membrane retrieval and synapse developmentThe Journal of cell biology, 2007
- An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrasesProceedings of the National Academy of Sciences, 2007
- Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Research, 1997