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. Dyson

Research output: Contribution to journalArticle

30 Citations (Scopus)

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.
LanguageEnglish
Pages515-522
Number of pages8
JournalAntonie van Leeuwenhoek: International Journal of General and Molecular Microbiology
Volume99
Issue number3
DOIs
Publication statusPublished - Mar 2011

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Streptomyces coelicolor
Cosmids
Gene Knockout Techniques
Gene Library
Mutagenesis
Streptomyces
Open Reading Frames
Genes
Libraries
Genome
Databases

Keywords

  • cosmids
  • DNA transposable elements
  • mutagenesis, insertional
  • streptomyces coelicolor

Cite this

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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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A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2). / Fernández-Martínez, L T; Del Sol, R.; Evans, M.C.; Fielding, S.; Herron, P R; Chandra, G.; Dyson, P.J.

In: Antonie van Leeuwenhoek: International Journal of General and Molecular Microbiology, Vol. 99, No. 3, 03.2011, p. 515-522.

Research output: Contribution to journalArticle

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