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Pathogenomics research overview. Steps. Limitations. Genome sequencing and annotation. (Conserved) hypotheticals and misannotation next-gen sequencing problems with “ mobilome ”. Comprehensive identification of virulence gene candidates by various means.
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Pathogenomics research overview Steps Limitations Genome sequencing and annotation (Conserved) hypotheticals and misannotation next-gen sequencing problems with “mobilome” Comprehensive identification of virulence gene candidates by various means Bioinformatic and experimental screens Bioinformatic prioritization of virulence gene candidates and generation of hypotheses regarding functions of encoded factors Protein structure/function prediction Experimental determination of functions of individual virulence factors “Characterization bottleneck” Integrate knowledge of factors for use in: -eco-evo-systems biology study of pathogen -comparative genomics Ongoing curation
Vivo Andrews-Polymenis et al. 2009
Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa Wolfgang et al. 2003
Gene expression in Pseudomonas aeruginosa biofilms rpoS was downregulated in biofilm Whiteley et al. 2001
Fluorescence-based isolation of bacterial genes expressed within host cells Valdivia and Falkow 1997
A high-throughput, near-saturating screen for type III effector genes from Pseudomonas syringae Chang et al. 2005
Collmer et al. 2002 Lindeberg et al. 2006
An extensive repertoire of type III secretion effectors in Escherichia coli O157 Tobe et al. 2006
Signature-tagged mutagenesis Mazurkiewicz et al. 2006
TraSH = Transposon site hybridization Mazurkiewicz et al. 2006
Comprehensive transposon mutant library of Pseudomonas aeruginosa Jacobs et al. 2003
Comprehensive transposon mutant library of Pseudomonas aeruginosa Jacobs et al. 2003