Rapid Virulence Annotation (RVA): identification of virulence factors using a bacterial genome library and multiple invertebrate hosts

Nicholas R Waterfield, Maria Sanchez-Contreras, Ioannis Eleftherianos, Andrea Dowling, Guowei Yang, Paul Wilkinson, J Parkhill, N Thomson, Stuart E Reynolds, H Bode, Stephen Dorus, R Ffrench-Constant

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57 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)15967-15972
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume105
Issue number41
Early online date5 Oct 2008
DOIs
Publication statusPublished - 14 Oct 2008

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    Waterfield, N. R., Sanchez-Contreras, M., Eleftherianos, I., Dowling, A., Yang, G., Wilkinson, P., Parkhill, J., Thomson, N., Reynolds, S. E., Bode, H., Dorus, S., & Ffrench-Constant, R. (2008). Rapid Virulence Annotation (RVA): identification of virulence factors using a bacterial genome library and multiple invertebrate hosts. Proceedings of the National Academy of Sciences of the United States of America, 105(41), 15967-15972. https://doi.org/10.1073/pnas.0711114105