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Antonie van Leeuwenhoek

, Volume 99, Issue 3, pp 515–522 | Cite as

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

  • L. T. Fernández-Martínez
  • R. Del Sol
  • M. C. Evans
  • S. Fielding
  • P. R. Herron
  • G. Chandra
  • P. J. DysonEmail author
Original Paper

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.

Keywords

Streptomyces Cosmid library Transposon Insertion 

Notes

Acknowledgements

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

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • L. T. Fernández-Martínez
    • 1
    • 4
  • R. Del Sol
    • 1
  • M. C. Evans
    • 1
  • S. Fielding
    • 1
  • P. R. Herron
    • 2
  • G. Chandra
    • 3
  • P. J. Dyson
    • 1
    Email author
  1. 1.Institute of Life Science, School of MedicineSwansea UniversitySwanseaUK
  2. 2.Strathclyde Institute of Pharmacy and Biomedical Science, Royal CollegeUniversity of StrathclydeGlasgowUK
  3. 3.Department of Molecular Microbiology, John Innes CentreNorwich Research ParkNorwichUK
  4. 4.Institute of Biotechnology of León (INBIOTEC), AvdaLeónSpain

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