<jats:title>Abstract</jats:title><jats:p>Resistance to streptothricin was studied in bacteria with different resistance mechanisms. The laboratory‐induced streptothricin‐resistant mutant <jats:italic>E. coli</jats:italic> A19 Stc<jats:sup>r</jats:sup> 2/2/1 showed a high level of cross‐resistance to aminoglycosides and other miscoding inducing antibiotics. In contrast, amino‐glycosid‐resistant <jats:italic>E. coli</jats:italic> strains with plasmid‐determined aminoglycoside modifying enzymes were sensitive to streptothricin. Enzymatic inactivation of streptothricin by acetylation was demonstrated for the streptothricin producing <jats:italic>Streptomyces noursei</jats:italic>, strain NG13. This strain showed no cross‐resistance to miscoding inducing aminoglycosides.