Identification of Genes Required for Swarming Motility in <i>Bacillus subtilis</i> Using Transposon Mutagenesis and High-Throughput Sequencing (TnSeq)
Sandra Sánchez, Elizabeth V. Snider, Xindan Wang, Daniel B. Kearns
Abstract
In TnSeq, transposons are randomly inserted throughout the chromosome at a population level, but insertions that disrupt genes of essential function cause strains that carry them to fall out of the population and appear underrepresented at the sequence level. Here, we apply TnSeq to the nonessential phenotype of motility in B. subtilis and spatially select for cells proficient in swarming. We find that insertions in nearly all genes previously identified as required for swarming are underrepresented in TnSeq analysis, and we identify 36 additional genes that enhance swarming. We demonstrate that TnSeq is a powerful tool for the genetic analysis of motility and likely other nonlethal screens for which enrichment is strong.