Transposon Sequencing in Leptospira interrogans

Our limited understanding of the relationship of genotype to phenotype in the spirochete Leptospira interrogans stems from the inefficiency of the genetic tools available to manipulate the pathogen. The recent development of random transposon mutagenesis in L. interrogans has allowed the creation of large libraries of mutants, permitting the identification of several genes involved in certain functions such as virulence. However, the process of phenotypically screening individual mutants in the library remains time- and labor-intensive. Here, we describe a transposon sequencing technique (Tn-Seq), which combines random transposon mutagenesis with high-throughput sequencing for screening L. interrogans mutants more rapidly with fewer resources than traditional methods.