Modular Cloning for Rapid Generation of Recombinant Plasmids for Analyses of Salmonella enterica Pathogenesis

Analyses of virulence functions of Salmonella enterica demand generation of plasmids for complementation of mutated genes, expression of tagged alleles of effector proteins of the type III secretion systems, or fusions to reporter genes for investigation of regulation of expression, or protein localization and fate after translocation into host cells. Here, a simple and versatile approach for generation of plasmids from modular DNA fragments is described. The Modular Cloning approach is based on Gibson assembly and uses standardized overlap sequences that enable various combinations and reuse of fragments. Modular Cloning does not demand the introduction of sites for restriction enzymes or commercial cloning kits and allows rapid generation of plasmids for various applications in cellular and molecular microbiology of S. enterica.