Journal article
Simultaneous assay of every Salmonella Typhi gene using one million transposon mutants
GC Langridge, MD Phan, DJ Turner, TT Perkins, L Parts, J Haase, I Charles, DJ Maskell, SE Peters, G Dougan, J Wain, J Parkhill, AK Turner
Genome Research | COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT | Published : 2009
Abstract
Very high-throughput sequencing technologies need to be matched by high-throughput functional studies if we are to make full use of the current explosion in genome sequences. We have generated a very large bacterial mutant pool, consisting of an estimated 1.1 million transposon mutants and we have used genomic DNA from this mutant pool, and Illumina nucleotide sequencing to prime from the transposon and sequence into the adjacent target DNA. With this method, which we have called TraDIS (transposon directed insertion-site sequencing), we have been able to map 370,000 unique transposon insertion sites to the Salmonella enterica serovar Typhi chromosome. The unprecedented density and resolutio..
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Awarded by Biotechnology and Biological Sciences Research Council
Funding Acknowledgements
This work was supported by the Wellcome Trust. We thank Elizabeth Huckle, who helped with the design of the sequencing assay.