Paulsen, IT, Cain, AK and Hassan, KA (2017) Physical enrichment of transposon mutants from saturation mutant libraries using the TraDISort approach. Mobile Genetic Elements, 7 (3). pp. 1-7. ISSN 2159-2543
Abstract
Transposon-insertion sequencing methods are finding their way into the molecular toolbox of many fields of microbiology. These methods can identify the genomic locations and density of transposon insertions in saturated transposon mutant libraries and can be used to make inferences on gene function. For example, where no insertions or very few insertions are identified within a gene in a mutant library grown under permissive conditions, the gene may be essential. Furthermore, where mutations are enriched or lost in a gene after passaging the library through a selective process, the gene is likely to be involved in the process. Typically, a fitness based selection such as a stress condition is used in these experiments and the processed sequencing data is used to identify genes required for fitness under the selection. Our research team recently expanded the utility of the transposon directed insertion sequencing (TraDIS) method by applying a physical separation of a transposon mutant library mediated by fluorescence activated cell sorting, rather than a fitness-based selection. This approach, which we have named “TraDISort” is significant because it allows the study of phenotypes that are not linked to cell survival. The TraDISort approach has a broad range of future applications, in drug development, metabolic engineering and in studies of basic bacterial cell physiology.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2017, Taylor & Francis. This is an Accepted Manuscript of an article published by Taylor & Francis in Mobile Genetics Elements on 31 March 2017, available online: http://www.tandfonline.com/10.1080/2159256X.2017.1313805 |
Keywords: | biosensor, cell sorting, FACS, fluorescence, reporter construct, transposon sequencing |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 23 May 2017 10:16 |
Last Modified: | 31 Mar 2018 00:39 |
Status: | Published |
Publisher: | Taylor and Francis |
Identification Number: | 10.1080/2159256X.2017.1313805 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:116756 |