Biochemical and genetic studies on the arginine-requiring auxotrophs derived from a Serratia marcescens strain were carried out. The arg mutants were classified into seven biochemical groups based on their growth response to five precursors of arginine biosynthesis and enzyme deficiency. Reciprocal transduction tests among these arg mutants divided them into three linkage groups, and the fine mapping in each of the groups by two- or three-point crosses revealed the following arrangement of loci. (1) arg44-thy11-lys1; (2) met1-glt2-argE-(arg19-arg51)-arg120-argG-argH; (3) arg33-pyr4. Five of the seven biochemically distinct arg mutants belonged to the second linkage group, and they constituted an agr-gene cluster. A characteristic feature of the arg-gene cluster of S. marcescens is that it involves argG, which was previously reported only in the Proteus group of Enterobacteriaceae.