Comprehensive identification of essential Staphylococcus aureus genes using Transposon-Mediated Differential Hybridisation (TMDH).
BACKGROUND
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.


RESULTS
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.


CONCLUSIONS
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.
Chaudhuri
Roy R
RR
Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, CB3 0ES, UK. roy.chaudhuri@gmail.com
Allen
Andrew G
AG
Owen
Paul J
PJ
Shalom
Gil
G
Stone
Karl
K
Harrison
Marcus
M
Burgis
Timothy A
TA
Lockyer
Michael
M
Garcia-Lara
Jorge
J
Foster
Simon J
SJ
Pleasance
Stephen J
SJ
Peters
Sarah E
SE
Maskell
Duncan J
DJ
Charles
Ian G
IG
D019295:Computational Biology; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D020043:Genes, Essential; D016680:Genome, Bacterial; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods*; D012984:Software; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes