The ORFeome of Staphylococcus aureus v 1.1.
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.


CONCLUSIONS
This first ORFeome library for S. aureus provides an essential new tool for investigating the systems biology of this important pathogen.
Brandner
Christina J
CJ
Department of Cell Biology, University of Salzburg, Hellbrunner Strasse 34, A-5020 Salzburg, Austria. christina.brandner@sbg.ac.at
Maier
Richard H
RH
Henderson
Daryl S
DS
Hintner
Helmut
H
Bauer
Johann W
JW
Onder
Kamil
K
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D016208:Databases, Factual; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D016680:Genome, Bacterial; D005982:Glutathione Transferase / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D006801:Humans; D008712:Methicillin / Q000494:pharmacology; D016106:Methicillin Resistance / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D013203:Staphylococcal Infections / Q000175:diagnosis / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification*; D020798:Two-Hybrid System Techniques
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D006639:Histidine; D005982:Glutathione Transferase; D008712:Methicillin