Rapid Virulence Annotation (RVA): identification of virulence factors using a bacterial genome library and multiple invertebrate hosts.
Current sequence databases now contain numerous whole genome sequences of pathogenic bacteria. However, many of the predicted genes lack any functional annotation. We describe an assumption-free approach, Rapid Virulence Annotation (RVA), for the high-throughput parallel screening of genomic libraries against four different taxa: insects, nematodes, amoeba, and mammalian macrophages. These hosts represent different aspects of both the vertebrate and invertebrate immune system. Here, we apply RVA to the emerging human pathogen Photorhabdus asymbiotica using "gain of toxicity" assays of recombinant Escherichia coli clones. We describe a wealth of potential virulence loci and attribute biological function to several putative genomic islands, which may then be further characterized using conventional molecular techniques. The application of RVA to other pathogen genomes promises to ascribe biological function to otherwise uncharacterized virulence genes.
Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom. bssnw@bath.ac.uk
Nicholas R
NR
Waterfield
Maria
M
Sanchez-Contreras
Ioannis
I
Eleftherianos
Andrea
A
Dowling
Guowei
G
Yang
Paul
P
Wilkinson
Julian
J
Parkhill
Nicholas
N
Thomson
Stuart E
SE
Reynolds
Helge B
HB
Bode
Steven
S
Dorus
Richard H
RH
Ffrench-Constant
D000818:Animals; D019295:Computational Biology / Q000379:methods*; D016680:Genome, Bacterial; D044404:Genomic Islands; D015698:Genomic Library; D007448:Invertebrates / Q000382:microbiology*; D008322:Mammals / Q000382:microbiology; D008969:Molecular Sequence Data; D020637:Photorhabdus / Q000235:genetics / Q000472:pathogenicity; D037521:Virulence Factors / Q000235:genetics*
D037521:Virulence Factors