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A DNA fragment containing the defective virulence gene(s) in the live oral vaccine candidate Salmonella typhimurium SR-11 Fad· was isolated. Genomic DNA fragments from the vaccine candidate, constructed by TnlOd::cam transposon mutagenesis, were probed with the 1.4 Kbp chloramphenicol resistance gene contained in TnlOd::cam and analyzed by Southern hybridization. Two of the fragments, an 8.0 Kbp Hind III fragment and a 4.5 Kbp Pst I :fragment, were isolated, cloned into E. coli HB 101, and mapped. The Pst I fragment, containing approximately 2.9 Kbp of flanking genomic DNA, was subcloned into the sequencing vector pBluescript II SK + (Stratagene, La Jolla, CA) and mapped.

Isolation of the Virulence Gene(s) in the New, Live, Oral Vaccine Candidate Salmonella typhimurium SR-11 Fad