Chromosomal barcoding as a tool for multiplexed phenotypic characterization of laboratory evolved lineages
| Autor: | Daniel Jean Jacques Simon, Christian Munck, Leonie Johanna Jahn, Svetlana Volkova, Morten Otto Alexander Sommer, Andreas Porse |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Prioritization Lineage (evolution) Science 030106 microbiology Computational biology Biology Genome Article Bacterial evolution 03 medical and health sciences Antibiotics Drug Resistance Bacterial Escherichia coli Selection Genetic Increased tolerance Selection (genetic algorithm) Gene Library Experimental evolution Multidisciplinary High-throughput screening Phenotype Bacterial strain Anti-Bacterial Agents 030104 developmental biology Medicine Genetic Fitness Directed Molecular Evolution Genome Bacterial |
| Zdroj: | Jahn, L J, Porse, A, Munck, C, Simon, D, Volkova, S & Sommer, M O A 2018, ' Chromosomal barcoding as a tool for multiplexed phenotypic characterization of laboratory evolved lineages ', Scientific Reports, vol. 8, 6961 . https://doi.org/10.1038/s41598-018-25201-5 Scientific Reports, Vol 8, Iss 1, Pp 1-12 (2018) Scientific Reports |
| DOI: | 10.1038/s41598-018-25201-5 |
| Popis: | Adaptive laboratory evolution is an important tool to evolve organisms to increased tolerance towards different physical and chemical stress. It is applied to study the evolution of antibiotic resistance as well as genetic mechanisms underlying improvements in production strains. Adaptive evolution experiments can be automated in a high-throughput fashion. However, the characterization of the resulting lineages can become a time consuming task, when the performance of each lineage is evaluated individually. Here, we present a novel method for the markerless insertion of randomized genetic barcodes into the genome of Escherichia coli using a novel dual-auxotrophic selection approach. The barcoded E. coli library allows multiplexed phenotyping of evolved strains in pooled competition experiments. We use the barcoded library in an adaptive evolution experiment; evolving resistance towards three common antibiotics. Comparing this multiplexed phenotyping with conventional susceptibility testing and growth-rate measurements we can show a significant positive correlation between the two approaches. Use of barcoded bacterial strain libraries for individual adaptive evolution experiments drastically reduces the workload of characterizing the resulting phenotypes and enables prioritization of lineages for in-depth characterization. In addition, barcoded clones open up new ways to profile community dynamics or to track lineages in vivo or situ. |
| Databáze: | OpenAIRE |
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