In the current study, we generated a pool of Salmonella typhimurium mutants using the Tn10d-cam mini-transposon. This pool of mutants was administered to BALB/c mice through the oral route, and bacteria were recovered from the spleen 3 days post-infection. After three rounds of serial passage, we observed enrichment of two insertion mutants, a yddG insertion and an amyA insertion. These two genes have been implicated in growth on plant products (amyA) and survival in the presence of paraquat (yddG), both of which are natural environments for Salmonella. Thus, while in vivo expression technology has identified S. typhimurium genes that are absolutely necessary for animal infection, other genes involved in vegetative growth also appear to play role in the establishment of pathogenesis.