P[acman]: A BAC Transgenic Platform for Targeted Insertion of Large DNA Fragments in D. melanogaster
Howard Hughes Medical Institute · Lawrence Berkeley National Laboratory · +1 more institution
Abstract
We describe a transgenesis platform for Drosophila melanogaster that integrates three recently developed technologies: a conditionally amplifiable bacterial artificial chromosome (BAC), recombineering, and bacteriophage PhiC31-mediated transgenesis. The BAC is maintained at low copy number, facilitating plasmid maintenance and recombineering, but is induced to high copy number for plasmid isolation. Recombineering allows gap repair and mutagenesis in bacteria. Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. PhiC31-mediated transgenesis integrates these large DNA fragments at specific sites in the genome, allowing the rescue of lethal mutations in the corresponding…
Citation impact
- FWCI
- 7.50
- Percentile
- 100%
- References
- 25
Authors
4- KJKoen J. T. Venken
Howard Hughes Medical Institute, Lawrence Berkeley National Laboratory, Baylor College of Medicine
- YHYuchun He
Howard Hughes Medical Institute, Lawrence Berkeley National Laboratory, Baylor College of Medicine
- RARoger A. Hoskins
Howard Hughes Medical Institute, Lawrence Berkeley National Laboratory, Baylor College of Medicine
- HJHugo J. BellenCorresponding
Howard Hughes Medical Institute, Lawrence Berkeley National Laboratory, Baylor College of Medicine
Topics & keywords
- Recombineering
- Transgenesis
- Bacterial artificial chromosome
- Biology
- Genetics
- Plasmid
- Mutagenesis
- Drosophila melanogaster