Haemophilus influenzae Database
As part of an NIAID funded proposal and in collaboration with the University of Massachusetts Medical School, we developed a novel, whole-genome scale application of the Illumina sequencing platform to discover genes contributing to bacterial growth and/or persistence under any selective condition. Transposon-insertion-profiling by deep sequencing (HITS) is particularly suitable for in vivo pathogenesis studies where an animal host imposes selective pressure on the infecting bacteria. The basis of the approach is the specific capture, sequencing, and enumeration of all transposon insertion sites present in a population of bacteria that have been subjected to random transposon insertion mutagenesis. Each bacterium in the population harbors a single genomic insertion. As a whole, the population (a bacterial insertion library) can represent disruptions in thousands of genes. Under an experimental selection, such as infectious growth or persistence in a host, a few of the bacteria in the population will harbor gene disruptions that impair (or promote) their ability to grow or persist. HITS will identify the insertions in genes that contribute to growth or persistence and provide a quantitative assessment of the gene?s contribution. Any organism that can be subjected to insertion mutagenesis can be readily analyzed by this procedure; thus, the approach is broadly applicable to almost any bacterial pathogen currently studied.
HITS was used to identify Haemophilus influenzae genes required for persistence in a murine pulmonary model (Gawronski, et al. 2009). As part of this effort we generated draft sequences of H. influenzae strains NT127 and RdAW.
Jeffrey D. Gawronski, Sandy M. S. Wong, Georgia Giannoukos, Doyle V. Ward, and Brian J. Akerley, (2009) Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung. PNAS (in press)