High-throughput insertion tracking by deep sequencing for the analysis of bacterial pathogens

Area of Science:

• Microbiology
• Genomics
• Infectious Diseases

Background:

• Whole-genome techniques are crucial for identifying microbial genes essential for survival and growth during infection.
• Massively parallel sequencing platforms have accelerated genome-scale analyses, enhancing sensitivity and quantification.

Purpose of the Study:

• To present a genome-scale methodology for identifying genes critical for microbial survival in infection models.
• To demonstrate the application of this method in studying bacterial pathogenesis.

Main Methods:

• High-density transposon mutagenesis using a mariner transposon.
• Deep sequencing to identify genes required for survival.
• Application in experimental models of pathogenesis, specifically a murine pulmonary model for *Haemophilus influenzae*.

Main Results:

• A comprehensive analysis of the relative role of each gene in *Haemophilus influenzae* during a murine pulmonary infection model.
• Identification of genes essential for pathogen survival and growth in vivo.

Conclusions:

• The described genome-scale methodology effectively identifies genes critical for microbial survival during infection.
• This approach is highly adaptable to diverse microbial species amenable to transposon mutagenesis.