Construction of Salmonella Transposon Mutant Library.

One of the main challenges in understanding Salmonella biology lies in dissecting the genetic basis of its survival, pathogenicity, and antimicrobial resistance. Here, we describe the construction of a high-density transposon mutant library in Salmonella. Using a mariner-based transposon system delivered through conjugation, we generated a library comprising more than 1 × 105 independent mutants, covering the majority of nonessential genes. This library enables a valuable resource for large-scale functional genomics studies, including the identification of essential genes, virulence determinations, stress adaptation, and potential drug resistance genes, providing a platform for systematic in vitro or in vivo research of Salmonella-host interactions.