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Open Access Highly Accessed Research article

The ORFeome of Staphylococcus aureus v 1.1

Christina J Brandner1, Richard H Maier1, Daryl S Henderson2, Helmut Hintner3, Johann W Bauer3 and Kamil Önder13*

Author Affiliations

1 Department of Cell Biology, University of Salzburg, Hellbrunner Strasse 34, A-5020 Salzburg, Austria

2 Department of Pharmacological Sciences, School of Medicine, Stony Brook University, Stony Brook, New York, 11794, USA

3 Division of Molecular Dermatology, Department of Dermatology, Paracelsus Private Medical University Salzburg, Salzburg, Austria

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BMC Genomics 2008, 9:321  doi:10.1186/1471-2164-9-321

Published: 7 July 2008

Abstract

Background

The bacterium Staphylococcus aureus causes significant morbidity and mortality in humans, primarily due to the emergence of strains that are resistant to antibiotics – notably methicillin-resistant S. aureus (MRSA) isolates. Development of effective strategies for the control and treatment of MRSA infections may best be achieved through 'omics' approaches, which first requires cloning the entire set of S. aureus' protein-encoding open reading frames (ORFs), or ORFeome.

Results

The complete genome sequence of S. aureus strain Mu50 has 2697 predicted protein-coding ORFs. Based on the sequence of this strain we designed PCR primers to construct from an S. aureus (non-MRSA) clinical isolate an ORFeome library that contains 2562 unique Gateway® entry clones (95% coverage), each corresponding to a defined ORF. The high quality of the ORFeome library was verified by DNA sequencing and PCR amplification, and its functionality was demonstrated by expressing recombinant proteins and observing protein interactions in a yeast 2-hybrid homodimerization screen.

Conclusion

This first ORFeome library for S. aureus provides an essential new tool for investigating the systems biology of this important pathogen.