Salmonella typhimurium strains are known to vary greatly in their virulence for mice. A few of the genes affecting their virulence have been described. In this report we have localized at least two genes that affect the ability of S. typhimurium to grow in BALB/cByJ to a 6 unit section of the salmonella chromosome which does not contain any previously described virulence determinants. The genetic mapping was done by interrupted matings using Hfr strains made in a virulent LT2 strain. The Hfr strains were constructed by inserting the plasmid F′(TS)114 lac+ Tn::10 into the LT2 chromosome at specific sites through homologous recombination with chromosomal Tn10s. Short interrupted matings to an avirulent LT2 strain in either direction through the portion of the chromosome from trpD at 34 units to supD at 40 units resulted in transconjugants which were fully virulent. Since we also found several transconjugants with intermediate virulence it appears that more than one virulence gene may exist in this area of the chromosome. The mechanisms of action of these genes are not known. © 1986.