Bacterial Phenotypic Diversity
PI: Dr. Colin ManoilDepartment of Genome Sciences
University of Washington, Seattle, WA
The primary goal of the Bacterial Phenotypic Diversity research project is to identify the Francisella tularensis and Burkholderia seudomallei genes responsible for several intrinsic antibiotic resistance, virulence and stress tolerance traits. The results should provide mechanistic insights into the physiological basis of each and may identify potential antibacterial drug targets. An additional set of experiments will help define of B. pseudomallei species diversity in these and other phenotypes, and may identify diagnostic tests to differentiate natural isolates.
As part of the project, a new-generation sequencing-based technology for assessing the makeup of transposon mutant pools of will be developed. Functions required for intrinsic antibiotic resistance and stress resistance are potential drug targets for combination antibacterial therapy; we anticipate that conserved homeostatic functions will be represented and may represent broad host range targets. The analysis of B. pseudomallei phenotypic species variation may identify diagnostic growth tests for discriminating rapidly between natural (non-laboratory) isolates. Finally, The F. novicida mutant-phenotype database to be constructed should eventually serve as a reference for identifying the mechanisms of action of new antibacterial compounds (“chemogenomic profiling”), including those active against a broad range of organisms.
Enstrom M, Held K, Ramage B, Brittnacher M, Gallagher L, Manoil C (2012) Genotype-phenotype associations in a nonmodel prokaryote. MBio. 2012 Mar 20;3(2). pii: e00001-12. doi: 10.1128/mBio.00001-12. Print 2012. PubMed PMID: 22434848; PubMed Central PMCID: PMC3312209
Gallagher LA, Shendure J, Manoil C (2011) Genome-scale identification of resistance functions in Pseudomonas aeruginosa using Tn-seq. MBio. 2011 Jan 18;2(1):e00315-10. PMCID: PMC3023915
Thongdee, M., L. A. Gallagher, M. Schell, T. Dharakul, S. Songsivilai, and C. Manoil. (2008) Targeted mutagenesis of Burkholderia thailandensis and Burkholderia pseudomallei through natural transformation of PCR fragments. Appl Environ Microbiol 74:2985-9.
Gallagher, L., Turner, C., Ramage, E. and Manoil, C. (2007) Creating recombination-activated genes and sequence-defined mutant libraries using transposons. Methods Enzymol, 421, 126-140.
Gallagher, L.A., Ramage, E., Jacobs, M.A., Kaul, R., Brittnacher, M. and Manoil, C. (2007) A comprehensive transposon mutant library of Francisella novicida, a bioweapon surrogate. Proc Natl Acad Sci U S A, 104, 1009-1014. PMCID: PMC1783355