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A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2)

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Abstract

A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83% of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB (http://strepdb.streptomyces.org.uk/). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.

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Acknowledgements

This research was supported by grants from the BBSRC (BB/E019242/1) and European Commission (LSH-IP 005224).

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Authors and Affiliations

  1. Institute of Life Science, School of Medicine, Swansea University, Singleton Park, Swansea, SA2 8PP, UK

    L. T. Fernández-Martínez, R. Del Sol, M. C. Evans, S. Fielding & P. J. Dyson

  2. Strathclyde Institute of Pharmacy and Biomedical Science, Royal College, University of Strathclyde, Glasgow, UK

    P. R. Herron

  3. Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK

    G. Chandra

  4. Institute of Biotechnology of León (INBIOTEC), Avda, Real, no. 1, 24006, León, Spain

    L. T. Fernández-Martínez

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  1. L. T. Fernández-Martínez
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  2. R. Del Sol
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  3. M. C. Evans
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  4. S. Fielding
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  5. P. R. Herron
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  6. G. Chandra
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  7. P. J. Dyson
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Correspondence to P. J. Dyson.

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Fernández-Martínez, L.T., Del Sol, R., Evans, M.C. et al. A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2). Antonie van Leeuwenhoek 99, 515–522 (2011). https://doi.org/10.1007/s10482-010-9518-1

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  • DOI: https://doi.org/10.1007/s10482-010-9518-1

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