Identification of the genetic elements involved in biofilm formation by Salmonella enterica serovar Tennessee using mini-Tn10 mutagenesis and DNA sequencing.

Biofilms are known to contribute to bacterial tolerance to desiccation and survival in low moisture foods. However, the molecular mechanisms underlying biofilm formation have not been fully elucidated. This study identified some of the genes that are implicated in biofilm formation by Salmonella enterica serovar Tennessee, the "peanut butter outbreak" strain. Mini-Tn10 mutagenesis was used in the study to generate random transposon insertion libraries. The ability of selected mutants in forming biofilms was compared with their wildtype parent using the crystal violet binding assay. Mutants forming significantly less (P ≤ 0.05) biofilm compared to their wildtype parent were selected for whole-genome sequencing. Mini-Tn10 insertion sites on mutant genomes were identified by comparing the acquired sequencing data with those in the Genbank using the BLAST search. In total, 56 mutants were obtained, and five were selected for further analysis according to the result of the biofilm assay. Sequencing analysis revealed that the mini-Tn10 interrupted the S. enterica genes that encode bacterial cell membrane lipoprotein, DNA topoisomerase III, attachment and invasion locus protein, bacteriocin immunity protein, and cell division protein. The information generated from the research should be useful in the control fo S. enterica in low-moisture foods and their production environments.