Comprehensive identification of essential Staphylococcus aureus genes using Transposon-Mediated Differential Hybridisation (TMDH)
- Equal contributors
1 Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, CB3 0ES, UK
2 Arrow Therapeutics, Britannia House, 7 Trinity Street, London, SE1 1DB, UK
3 Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, UK
4 Current address: Pfizer Australia, Veterinary Medicine Research and Development, 45 Poplar Road, Parkville, 3052 Victoria, Australia
5 Current address: Cancer Research Technology Development Laboratory, Wolfson Institute for Biomedical Research, The Cruciform Building, Gower Street, London, WC1E 6BT, UK
6 Current address: Division of Microbial Diseases, UCL Eastman Dental Institute, 256 Gray's Inn Road, London, WC1X 8LD, UK
7 Current address: Oxford Gene Technology, Begbroke Science Park, Sandy Lane, Yarnton, Oxford, OX5 1PF, UK
8 Current address: Centre for Bioinformatics, Division of Molecular Biosciences, Faculty of Natural Sciences, Imperial College London, London, SW7 2AZ, UK
BMC Genomics 2009, 10:291 doi:10.1186/1471-2164-10-291Published: 1 July 2009
In recent years there has been an increasing problem with Staphylococcus aureus strains that are resistant to treatment with existing antibiotics. An important starting point for the development of new antimicrobial drugs is the identification of "essential" genes that are important for bacterial survival and growth.
We have developed a robust microarray and PCR-based method, Transposon-Mediated Differential Hybridisation (TMDH), that uses novel bioinformatics to identify transposon inserts in genome-wide libraries. Following a microarray-based screen, genes lacking transposon inserts are re-tested using a PCR and sequencing-based approach. We carried out a TMDH analysis of the S. aureus genome using a large random mariner transposon library of around a million mutants, and identified a total of 351 S. aureus genes important for survival and growth in culture. A comparison with the essential gene list experimentally derived for Bacillus subtilis highlighted interesting differences in both pathways and individual genes.
We have determined the first comprehensive list of S. aureus essential genes. This should act as a useful starting point for the identification of potential targets for novel antimicrobial compounds. The TMDH methodology we have developed is generic and could be applied to identify essential genes in other bacterial pathogens.