Mice were injected with virulent Salmonella typhimurium SR-11 subfractions containing varied amounts of ribonucleic acid (RNA) and protein or with living attenuated S. typhimurium RIA. In these mice, maximal resistance to lethal infection by 1,000 or 5,000 median lethal doses of S. typhimurium SR-11 was seen 2 to 3 weeks after immunization. The S. typhimurium RIA vaccine and a crude ethanol-precipitated RNA fraction (E-RNA) prepared from lysates of S. typhimurium SR-11 were the most efficient immunogens inducing protection against salmonellosis. The contribution of the components present in the E-RNA fractions to host protection against lethal salmonella infection was also examined. RNA-rich fractions (P-RNA) prepared from lysates of the virulent salmonellae contained several bands of protein when examined by disc electrophoresis. P-RNA fractions stimulated protective immunity in mice to infection with S. typhimurium SR-11 but to a much lesser degree than did the E-RNA fractions or strain RIA vaccine. Protein-rich fractions (NP), separated from E-RNA by salt precipitation, exhibited the same number and distribution of protein bands by disc electrophoresis as did the parent E-RNA fractions. Mixtures of either bovine liver soluble RNA or various synthetic polynucleotides and NP were examined, as was NP fraction alone, for the ability to confer protection in mice to challenge infections by the virulent strain of salmonella. Polyadenylic-uridylic acid plus NP conferred significant protective immunity to challenge infections in mice immunized with this mixture, being nearly as effective an immunogen as were the E-RNA fractions of S. typhimurium SR-11 or the attenuated S. typhimurium RIA.