Loci for resistance to several antibiotics in laboratory-derived strains of Neisseria gonorrhoeae were mapped by genetic transformation. Genes for high-level resistance to streptomycin (str) and spectinomycin (spc) and for low-level resistance to tetracycline (tet) and chloramphenicol (chl) were linked. Also, a locus for high-level resistance to rifampin (rif) was linked to str and tet. The apparent order was rif... str... tet... chl... spc. Loci for resistance to other antibiotics (penicillin, erythromycin) were transferred independently of each other and were not linked to the cluster around str. Similar linkage relationships were found with str, tet, chl, and spc loci obtained from naturally occurring (clinical) isolates of N. gonorrhoeae.