The net accumulation of tetracyclines by Escherichia coli as a function of concentration was shown to be biphasic. At concentrations less than the bacteriostatic levels, the mode of uptake was not azide-sensitive and was considered to be physical adsorption on the cell surface. At concentrations above the minimal inhibitory level, a second, azide-sensitive, uptake component was functional in addition to the surface adsorption process. This second energy-requiring mode was judged to represent penetration of the cytoplasmic membrane by tetracycline molecules to their sites of inhibitory action. Each mode for a given tetracycline and culture is expressed algebraically by a characteristic Freundlich equation. Resistance in E. coli is shown to be a result of diminished transport of antibiotic. However, this resistance was due not to a reduction or loss of a transport mechanism but rather to a requirement for higher antibiotic concentrations before the second mode of uptake could become operative.