|Nisbet, David - Dave|
Submitted to: American Society for Microbiology Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 7/30/2004
Publication Date: 10/6/2004
Citation: Cox, M., Ziprin, R.L., Kubena, L.F., Nisbet, D.J., Ricke, S.C., Kwon, Y.M. 2004. Genome scanning for virulence genes by transposon signature profiling [abstract]. American Society for Microbiology Conference on Functional Genomics and Bioinformatics Approaches to Infectious Disease Research. p. 24. Interpretive Summary:
Technical Abstract: Genome-wide scanning of microorganisms remains a challenging task to simultaneously identify all genes of conditional necessity. Here, we describe a method termed 'transposon signature profiling' (TSP) to determine those genes in pathogenic bacteria that play a role in virulence. Short DNA signatures, representative of a unique transposon insertion in a complex pool of mutants are extracted simultaneously. A profile of the resulting DNA signatures provides a quantitative catalog of all insertions in the library. Next, profiles assembled before and after infection are compared to determine those mutants with a survival defect. We have demonstrated the utility of TSP by screening a pool of ~500 Tn5 mutants of Salmonella enteriditis in a chicken infection model. Among these, we identified 45 signatures that showed at least a 3-fold decrease after infection and located sequence regions where gene disruption occurred. The same studies also revealed 58 signatures that were increased at least 3-fold, indicative of genes conferring hypervirulence when mutated. TSP can be used to study any organism capable of harboring transposons and does not require prior sequence data for signature profiling. The potential for TSP in scanning an entire microbial genome can be a useful tool for identifying all crucial target sequences required in a given condition.