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.
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, 99(3), 515-522. https://doi.org/doi:10.1007/s10482-010-9518-1