ARGRII is one of the three regulatory genes controlling arginine metabolism in yeast. From a pool of hybrid plasmids carrying Sau3A fragments representing the entire yeast genome, a DNA fragment containing the regulatory gene ARGRII was cloned by complementation of an argRII- mutation, which prevents growth on ornithine as sole nitrogen source. Cells containing the cloned DNA regained the ability to repress the synthesis of anabolic enzymes and to induce the synthesis of the catabolic ones, when arginine is present. The 6.2 kb cloned DNA fragment encodes five transcripts (2.8 kb, 1.3 kb, 0.75 kb, 0.45 kb, 0.45 kb), which were located by S1 endonuclease mapping. By marker rescue the argRII- mutations were mapped in the DNA region coding for the 2.8 kb transcript, showing its importance in the control mechanism. Subcloning experiments confirm this result. However, at present the role of the 0.75 kb and 1.3 kb transcripts in the ARGR+ phenotype is unclear.