期刊
ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY
卷 99, 期 3, 页码 515-522出版社
SPRINGER
DOI: 10.1007/s10482-010-9518-1
关键词
Streptomyces; Cosmid library; Transposon; Insertion
类别
资金
- BBSRC [BB/E019242/1]
- European Commission [LSH-IP 005224]
- BBSRC [BB/E019242/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/E019242/1] Funding Source: researchfish
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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