01/16/2018
Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria
The Science
Streamlined approaches for testing hundreds to thousands of genetic systems in parallel against a target bacterium.
- Avoids the labor and cost associated with trial-and-error based approaches.
- Opens door for large-scale genetic studies in diverse bacteria.
The Impact
- Uses a “parts-based” synthetic biology cloning strategy.
- Uses random DNA barcode sequences to track the efficacy of many transposon vectors in parallel.
- Successfully adapted the strategy to 5 different genera of bacteria, including 3 from the phylum Bacteroidetes.
Summary
the development of a suitable next-generation sequencing strategy for a given bacterium can be costly and time-consuming. To meet this challenge, researchers describe a part-based strategy for constructing libraries of hundreds of transposon delivery vectors, which they term “magic pools.” Within a magic pool, each transposon vector has a different combination of upstream sequences (promoters and ribosome binding sites) and antibiotic resistance markers as well as a random DNA barcode sequence, which allows the tracking of each vector during mutagenesis experiments. To identify an efficient vector for a given bacterium, researchers mutagenize it with a magic pool and sequence the resulting insertions; they then use this efficient vector to generate a large mutant library. This study used the magic pool strategy to construct transposon mutant libraries in five genera of bacteria, including three genera of the phylum Bacteroidetes.
References
Liu, H., M. N. Price, R. J. Waters, J. Ray, H. K. Carlson, J. S. Lamson, R. Chakraborty, A. P. Arkin, and A. M. Deutschbauer. 2017. “Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria.” mSystems 3(1) e00143-17. DOI:10.1128/mSystems.00143-17.