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Haploid strains of Saccharomyces cerevisiae with mutations in two different rib genes were constructed. These strains were studied by tetrad analysis and by quantitative determination of accumulation products. The genes rib1, rib7, and rib2 are not linked to each other. rib1-rib7 strains and rib1-rib2 strains exhibit the phenotypic properties of rib1 strains. rib7-rib2 strains show the phenotypic properties of rib7 strains. The results support the conclusion that the genes rib1 and rib7 code for the first and second enzyme of the riboflavine pathway, respectively.