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.
|Number of pages||8|
|Journal||Antonie van Leeuwenhoek: International Journal of General and Molecular Microbiology|
|Publication status||Published - Mar 2011|
- DNA transposable elements
- mutagenesis, insertional
- streptomyces coelicolor
Fernández-Martínez, L. T., Del Sol, R., Evans, M. C., Fielding, S., Herron, P. R., Chandra, G., & Dyson, P. J. (2011). A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2). Antonie van Leeuwenhoek: International Journal of General and Molecular Microbiology, 99(3), 515-522. https://doi.org/10.1007/s10482-010-9518-1