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Mode of Action

Bacteriocidal. Diffuses through the porinchannels in the outer membrane of gram-negative bacteria. Interacts with at least three ribosomal proteins, inhibiting protein synthesis and increasing translation errors.

Mechanism of Resistance

Kanamycin is inactivated by bacterial aminophosphotransferases (APHs). The APHs inactivate kanamycin by transferring the γ-phosphate of ATP to the hydroxyl group in the 3' position of the pseudosaccharide. The KanR gene codes for kanamycin resistance. Several versions of this gene exist, with varying crossover resistance to other antibiotics such as neomycin or gentamycin.

Working Concentration and Stock Solution

Working concentration is 50 μg/ml, or 25 μg/ml for low-copy plasmids like PACs. Stock solution is 35 mg/ml in water (kanamycin is insoluble in 50% alcohol).


Molecular Cloning: A Laboratory Manual, Vol 1.


Children's Hospital Oakland Research Institute (CHORI) - BAC/PAC resources